Impact of feeding Murraya koenigii and Aegle marmelos leaves on metabolic and reproductive performance in crossbred cows

Impact of feeding Murraya koenigii and Aegle marmelos leaves on metabolic and reproductive performance in crossbred cows | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Impact of feeding Murraya koenigii and Aegle marmelos leaves on metabolic and reproductive performance in crossbred cows Juli Chakma, N. Dutta, S. E. Jadhav, S.K. Singh, D. R. Choravada, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4557558/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 03 Mar, 2025 Read the published version in Tropical Animal Health and Production → Version 1 posted 4 You are reading this latest preprint version Abstract The study was carried out to ascertain the effect of herbal formulation on metabolic profile and reproductive performance of crossbred cows. Crossbred cows (24) were randomly assigned to four treatment groups. Cows in CON (control) group were fed a diet comprised of green fodder, concentrate and wheat straw, while, the cows in HP (herbal powder), HPL (herbal pellets) and HEX (herbal extract) groups were offered CON diet having herbal powder (200gd − 1 ), herbal pellets (200gd − 1 ) and methanolic herbal extract (equivalent to 200g herbal powder d − 1 ), respectively. The cows were fed on treatment diets for 7 d and continued with CON diet up to one month. Hb levels were analogous among the treatments; however, haematocrit values were significantly higher (P < 0.05) in the HEX group relative to CON. Total protein, albumin, globulin, A:G ratio, serum glucose, total cholesterol, AST and ALT were comparable across the dietary treatments. Serum NEFA, urea and creatinine were lower (P < 0.01) in treatment groups relative to CON. Serum minerals viz. i-P, Cu, Zn, Mn and iron were significantly (P < 0.01) higher in all the treatments than CON. The conception at first insemination was considerably higher in HPL, HEX and HP groups relative to CON. Spinnbarkeit and pH values of cervical mucus were noticeably higher in HP, HPL and HEX groups. The typical fern pattern of cervical mucus exhibited higher conception rate in treatment groups. It may be concluded that the herbal formulation positively influenced metabolic parameters and reproductive performance in post-partum crossbred cows. Aegle marmelos Cattle Cervical mucus Murraya koenigii Reproduction Figures Figure 1 Figure 2 Figure 3 Figure 4 1. Introduction Natural products derived from plants have garnered significant attention within the scientific community and among the general public due to their potential role in treating, preventing, and enhancing health. The accessibility and affordability of these medicinal herbs have contributed to their popularity as a reasonably priced source of therapeutic compounds (Abeysinghe et al. 2021 ). In the context of livestock management, reproductive efficiency is a pivotal determinant of a sound and profitable enterprise. Reproductive diseases in dairy animals pose a substantial challenge, leading to poor reproductive efficiency and subsequent reductions in productivity (Kuralkar and Kuralkar, 2021 ). Achieving a phase of transition free of diseases, increased artificial insemination (AI), and higher pregnancy rates per service becomes imperative to augment reproductive efficiency in bovines. Despite advancements in breeding, feeding, and management practices, the reproductive efficiency of dairy animals has witnessed a decline over the past few decades, contrasting with increased production levels. The use of therapeutic agents, both hormonal and non-hormonal, has been extensive in attempts to enhance reproductive efficiency in dairy animals, albeit with varying degrees of success (Jadhav and Bhutani, 2005 ; Khade et al. 2011 ; Lone et al. 2012 ; Purkayastha et al. 2015 ; Dutta et al. 2019 ; Kumar et al. 2019 ; Verma et al. 2019 ). However, hormonal treatments have raised concerns about potential adverse effects on neuroendocrine and physiological functions, along with implications for public health due to residues in food animals and their products (Dutt et al. 2018 ). The need for alternative, affordable methods is highlighted by the disadvantages of utilizing synthetic hormones and medications for animal reproductive management, such as their high costs, variable outcomes, and difficulty in obtaining commercial formulations. Approaches to reduce disease incidence and enhance reproductive function are being developed in order to minimize financial losses in dairy animals (Kumar et al. 2014 ). India, known as the "Botanical Garden of the World," boasts a diverse range of medicinal plants, positioning it as the leading producer of medicinal herbs globally. These herbs have significant implications for both human and animal health. Recent attention has been directed towards several herbs, including garlic, fenugreek, raspberry leaves, shatavari, thyme, black pepper, ashwagandha, and clove, which have demonstrated extraordinary therapeutic potential in combating various reproductive diseases and disorders in dairy animals (Mishra et al. 2002 ; Kabir et al. 2001 ; Das et al. 2002 ; Kumar et al. 2008 ; Rajkumar et al. 2008 ; Kumar et al. 2018 ; Chakma et al., 2020 ; Shafi et al. 2020). Commercially available herbal remedies have shown variable success rates in restoring cyclicity in dairy animals, stimulating ovarian function and improving reproductive health. Notably, the past two decades have seen systematic studies on the use of medicinal plants, such as pipal ( Ficus religiosa ), curry ( Murraya koenigii ), and bel ( Aegle marmelos ), to enhance fertility in dairy animals (Jondhale, 2007 ; Kumar, 2008 ; Jondhale et al. 2009a ; b ). Researches have demonstrated that extracts and powders of A. marmelos and M. koenigii, either alone or in combination, can lower inflammation and uterine infections while concurrently improving lipid profiles, insulin, glucose, and antioxidant enzyme activity (Sabu and Kuttan, 2004 ; Upadhya et al. 2004 ; Xie et al. 2006 ; Sharma et al. 2007 ; Abdallah et al. 2017 ). Recent research has demonstrated encouraging outcomes in enhancing reproductive parameters, antioxidant responses, and immunological functions in rats through the supplementation of herbal extracts from M. koenigii , M. oleifera , and A. marmelos (Dhivagar, 2022 ). The abundance of phytochemicals in bel and curry leaves, such as marmelosin, tannins, limonene, aegelin, marmesinin, marmelin, koenigine, mukonicine, murrayacinine, isomahanine, and more, underscores their therapeutic potential (Bansal and Bansal, 2011 ; Naczk and Shahidi, 2004 ; Reddy, 2008 ; Jain et al. 2012 ; Snehlata et al. 2018; Ganesan et al. 2013 ; Chakma et al., 2024 ). These phytochemicals have been associated with various health benefits, including antioxidant, antimicrobial, antidiabetic, anticholesterol, anticancer, and hepatoprotective properties. Among the studied herbs, curry ( M. koenigii ) and bel ( A. marmelos ) stand out as significant plants examined for ovarian dynamic investigations in both laboratory and farm animals. Their effects have been evaluated in different reproductive disorders including anestrus, repeat breeding, and endometritis, showing promising results as therapeutics (Hegde et al. 2002 ; Mehrotra et al. 2003 ; Satheshkumar and Punniamurthy, 2009 ; Nandini et al. 2010 ; Dutt et al. 2011 ). Supplementation with powder or extract of M. koenigii and A. marmelos for 5 to 9 days has been observed to improve genital health, increase estrus expression, and enhance conception rates in dairy animals facing reproductive challenges (Baitule et al. 2016 ; Kumawat et al. 2016 ; Tiwari et al. 2021 ; Dhami et al. 2022 ). In light of these findings, it may be possible to turn powdered herbs into extracts and pellets with the goal of lowering dosages, increasing utilization, and creating formulations based on active ingredients. Therefore, the present study hypothesizes using curry and bel leaves as a safe, potent, and cost-effective alternative strategy to improve the reproductive efficiency of crossbred cows during the postpartum period. The research will specifically address metabolic profiles, antioxidant indices, immune responses, and reproductive performances in the pursuit of sustainable and effective strategies for livestock management. 2. Material and methods The research was conducted at the cattle and buffalo farm of the ICAR-Indian Veterinary Research Institute (IVRI) in Izatnagar, Uttar Pradesh, India, situated at the coordinates 28.4059ºN and 79.4332ºE. In this region, the integration of crops and livestock, known as mixed farming, has been a long-standing tradition. Despite the importance of livestock, small mixed farms often suffer from low productivity due to the prevalence of non-descript breeds and inadequate nutrition. The main crops cultivated in this area are rice, wheat, and sugarcane, with cereal straws and stovers constituting the primary diet for ruminants. All experimental protocols received approval from the Institutional Animal Ethics Committee and the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), India. 2.1. Preparation of the plant material 2.1.1. Collection, drying and storage of plant material The M. koenigii and A. marmelos leaves were collected in a single batch from the ICAR-IVRI, Izatnagar in September 2022. After being air-dried at a room temperature of 27ºC, the leaves were ground using an electric grinder. The dried and ground leaves were then combined in equal parts, creating a 1:1 ratio herbal formulation. 2.1.2. Preparation of herbal pellets The herbal powder was further used to prepare pellets, which underwent processing in a feed pelleting unit. Within the pellet-making machine, this mixture was passed through the die which was of 10 mm size, the mixture was churned and pressed, transforming it into pellet form. The resulting pellets averaged 4–5 cm in length and had a diameter of 10 mm. 2.1.3. Methanolic extract preparation In a Stoppard conical flask, 100 g of dried herbal powder underwent extraction with 1000 mL of methanol-water solution (70:30 ratio) at room temperature. The mixture was agitated for 12 h using an electric shaker. Subsequently, the extracts were filtered using a cotton wool plug and filter paper. The filtrate was then dried at 39°C for future use. The resulting dried powder was crushed and stored in bottles at 4°C, as described by Begashaw et al. ( 2017 ). 2.2. Animals and treatments Twenty-four crossbred cows (60 d post-partum) of 2nd to 4th parity were distributed into four treatment groups consisting of six animals in each in a CRD. The cows in all the treatment groups were provided iso-nitrogenous and iso-caloric diets. The experimental cows were randomly divided into 4 treatment groups (CON, HPL, HEX and HP). CON (control) group animals were fed a diet comprised of green fodder, concentrate mixture and wheat straw, while, the experimental cows in HP (herbal powder), HPL (herbal pellets) and HEX (herbal extract) groups were offered CON diet having herbal powder (200gd − 1 ), herbal pellets (200gd − 1 ) and methanolic herbal extract (equivalent to 200g herbal powder d − 1 ), respectively. The experimental cows were fed on treatment diets for 7 d and continued with CON diet up to one month, further after one month of treatment the non-responsive cows (not exhibited oestrus) were given treatment again in different groups and monitored for another one month to assess the response in respect of oestrus induction and pregnancy establishment, metabolic profile, health, production and reproduction performance. The pregnancy diagnosis was done per rectal after 60 d of AI. The total duration of the experiment was of 120 d. 2.3. Collection of blood and separation of serum The blood samples were collected from jugular venipuncture on 0, 3, 7, 14 and 21 day of feeding by following standard protocol with with clean, sterilized needles, early in the morning before feeding and watering. About 10 mL blood was collected, out of which 2 mL was placed into vacutainers containing anticoagulant (EDTA) for estimation of Hb and haematocrit. Remaining 8.0 mL blood was dispensed into serum vacutainers and allowed to clot at ambient temperature for 45 minutes by keeping at slanting position. The tubes were spun at 2,000 xg for 10 minutes using a refrigerated centrifuge. The collected sera were transferred into Eppendorf tube with Pasteur pipette and stored in deep freeze at -20 0 C for further serum-biochemical analysis. Haemoglobin (Hb) and haematocrit were measured in whole blood immediately after blood collection. Haemoglobin (Hb) was measured by using the cyanomethaemoglobin technique (Dacie and Lewis, 1969 ) and hematocrit was measured using Wintrobe’s tube. The serum glucose, NEFA, total proteins, albumin, urea, creatinine, total cholesterol, aspartate transaminase (AST) and alanine aminotransferase (ALT) were analyzed spectrophotometrically (MultiskanTM FC Microplate Photometer with incubator, Thermo Scientific Ltd.) by using diagnostic kits manufactured by Coral Clinical Systems (Goa, India) following the standard protocol provided with the kits. The major minerals (Ca and P) were analyzed by using the method of Talapatra et al. ( 1940 ) and trace minerals such as Cu, Zn, Mn and iron were analyzed by Atomic Absorption Spectroscopy (AAS) method 2.4. Chemical analysis Samples of the concentrate mixture, wheat straw, green fodder, herbal pellets, herbal extracts and herbal powder were milled to pass through a 1.0 mm sieve and then examined for proximate principles (AOAC, 2012 ). The neutral detergent fibre (NDF) and acid detergent fibre (ADF) were measured following the methods of Van Soest et al. ( 1991 ). 2.5. Gene expression study For gene expression study, already published primer of progesterone (PGR) TAATCTGTGGGGATGAAGCA-F and TAATCTGTGGGGATGAAGCA-R was standardized and used (Driver et al. 2009 ). The PCR conditions were optimized by amplifying the cDNA with specific primers at various annealing temperatures. The specificity of each primer was verified before performing the qRT-PCR analysis. The relative change in gene expression was analyzed using the 2-ΔΔCT method, as detailed by Livak and Schnittger (2001). Relative fold change = 2 −ΔΔCT , where, ΔΔCT= (C T , target gene- C T , reference gene) test – (C T , target gene- C T , reference gene) housekeeping gene. 2.6. Reproductive performance Oestrus detection was done for all the lactating crossbred cows twice a day throughout the study period with the help of a trained teaser bull. All the postpartum crossbred cows were inseminated artificially with frozen semen which were exhibited in oestrus. All the non-return cows were diagnosed for pregnancy after 60 d of insemination by trans-rectal palpation. The different postpartum reproductive parameters like onset of oestrus, services per conception and conception rate was recorded. All the experimental cows were closely monitored every day for any types of disease or disorders such as mastitis, metritis etc. 2.6.1. Cervical mucus characteristics Cervical mucus was obtained prior to artificial insemination using mid-cervical aspiration utilizing sterile blue sheaths and a universal artificial insemination gun via the recto-vaginal method. The blue sheath was attached to the universal artificial insemination gun to act as an extended syringe. The insemination gun's piston functioned as a plunger, facilitating the extraction of the cervical mucus. Following the collection, 5 mL of the cervical mucus was transferred immediately to the laboratory in a sterile petri dish for analysis of the specified parameters: 2.6.1.1. Spinnbarkeit value Few drops (2–3 drops) of cervical mucus were placed on a grease-free slide, with another clean slide positioned on top. The slides were then gently pulled apart to stretch the mucus until it broke. The distance between the two slides at the point of breakage was measured using a centimeter scale mounted on a wall. (Panigrahi, 1964 ). 2.6.1.2. Arborization pattern and pH To analyze the arborization pattern, 2–3 drops of cervical mucus were placed on a grease-free glass slide and spread evenly. The sample was air-dried and examined under a low-power objective (10X) of a microscope. Fern patterns were classified into typical, atypical, and absent categories. pH was determined by using a pH indicator stick (full range, pH 1.0 to 14.0) - Qualigens (#Q38141, Thermo Fisher Scientific) which was saturated with a drop of cervical mucus placed on a clean slide. At least 2 sticks with overlapping pH ranges (5.4 to 7.0 and 6.4 to 8.0) were used to test the same sample. 2.7. Statistical analysis The results obtained were statistically analyzed using IBM, SPSS (20.0) and through Duncan’s Multiple Range test, means were categorized accordingly. The parameters from periodic collections were scrutinized by implementing repeated measures protocol through GLM of SPSS; which encompassed between-subjects principal effect of treatments, within-subjects principal effect of period and interaction amid the treatment and period. The P < 0.05 was used to declare the means to be significant. All the statistical procedures were used as per Snedecor and Cochran (1994). 3. Results 3.6. Chemical composition of feedstuffs offered (% DM basis) The chemical composition and fibre fractions of wheat straw, green, concentrate, pellets, extract and powder supplements are mentioned in Table 1 . The chemical composition of herbal pellets and powder was found to be comparable, however, herbal extract showed lower values of EE, TA, NDF and ADF as compared to herbal pellets and powder. Table 1 Chemical composition of feeds offered (% DM basis) Attributes (%) Wheat straw Green fodder Concentrate mixture Herbal pellets Herbal extract Herbal powder DM 91.82 10.60 86.82 92.94 90.16 94.11 OM 93.17 87.44 92.68 87.33 85.42 88.15 CP 2.72 10.82 17.86 11.25 13.41 12.14 EE 1.06 1.33 2.48 2.81 3.50 1.21 TA 6.83 12.57 7.32 12.67 14.58 11.85 NDF 85.42 75.61 43.53 53.68 31.56 51.16 ADF 51.21 41.24 16.85 32.38 23.45 29.36 [DM: Dry matter; OM: Organic matter; CP: Crude protein; EE: Ether extract; TA: Total ash; NDF: Neutral detergent fibre; ADF: Acid detergent fibre] 3.7. Metabolic profile 3.2.1. Hb and haematocrit The haematocrit (%) and Hb (gdL − 1 ) levels are presented in Table 2 . The levels of Hb (gdL − 1 ) were analogous (P > 0.05) among the treatment groups. Mean values of haematocrit were significantly higher (P < 0.05) in the HEX group relative to CON, while HPL and HP groups had intermediate response between CON and HEX. 3.7.1. Serum biochemical parameters The results related with serum glucose, NEFA, total protein and its fractions, serum urea, total cholesterol, creatinine, AST and ALT are presented in Table 2 . The serum glucose (mgdL − 1 ) levels were comparable (P > 0.05) amongst the dietary treatments, though serum NEFA (mmolL − 1 ) values were significantly (P < 0.01) lower in HPL group followed by HEX, HP and CON groups. Serum NEFA was significantly reduced (P 0.05) amongst the dietary treatments. The serum urea level was significantly (P < 0.01) lower in HPL and HEX groups relative to CON, however, HP group had an intermediate position among HPL, HEX and CON. The serum creatinine (mg %) concentration was significantly (P < 0.01) lower in HPL, HEX and HP as compared to CON. Activities of serum enzymes viz. AST and ALT were similar among the treatment groups. Table 2 Effect of herbal formulations supplementation on metabolic profile of crossbred cows Attributes Treatments SEM P-value CON HPL HP HP Hb, gdl − 1 11.57 12.18 12.27 11.67 0.22 0.213 Haematocrit, % 34.73 a 36.56 ab 36.82 b 35.03 ab 0.32 0.049 Glucose, mgdl − 1 47.27 48.15 48.02 48.25 0.34 0.740 NEFA, mmolL − 1 0.43 c 0.37 a 0.39 b 0.39 b 0.00 < 0.001 Total protein, gdl − 1 7.75 7.78 8.08 7.87 0.05 0.137 Albumin, gdl − 1 3.30 3.30 3.41 3.38 0.02 0.260 Globulin, gdl − 1 4.45 4.47 4.67 4.49 0.05 0.400 A: G ratio 0.75 0.74 0.74 0.79 Total cholesterol, mgdl − 1 142.47 140.84 141.4 141.55 0.96 0.964 Urea, mgdl − 1 35.96 b 32.48 a 31.79 a 33.55 ab 0.55 0.02 Creatinine, mg% 1.23 b 1.10 a 1.11 a 1.11 a 0.01 0.00 ALT, UL − 1 32.08 31.80 32.47 32.17 0.31 0.903 AST, UL − 1 90.65 92.14 88.62 89.91 0.53 0.135 abc and xyz means bearing different superscript in a row and column differ significantly 3.7.2. Serum mineral profile Serum mineral such as Ca, i-P, Cu, Zn, Mn and iron are presented in Table 3 . The mean values for serum Ca (mgdL − 1 ) were parallel (P > 0.05) among the treatment groups, however, serum i-P (mgdL − 1 ) levels were significantly (P < 0.01) higher in HPL, HEX and HP in relative to CON. The mean for serum Ca were significantly (P < 0.05) higher at 3rd d, however, i-P values were increased 3rd d onwards. The mean values of Cu, Zn, Mn and iron were significantly (P < 0.01) higher in treatment groups as compared to CON. Serum Cu and Zn levels were noticeably (P < 0.01) higher in HPL followed by HEX and HP and CON. Serum Mn values were significantly (P < 0.01) higher in HPL followed by HEX, HP and CON group. The levels of serum iron were significantly (P < 0.01) higher in HPL and HP groups relative to CON, while HEX had an intermediate position between CON and HP groups. The values of all the trace minerals were increased significantly (P 0.01). Table 3 Serum mineral profile of crossbred cow supplemented with herbal formulations Treatments Period (s) Treatment mean SEM P-value 0 3 7 14 21 T P TxP Ca, mgdl − 1 CON 11.83 12.45 10.02 10.30 10.81 11.08 0.22 0.213 0.015 0.828 HPL 11.66 12.53 10.55 11.13 13.85 11.94 HEX 11.93 12.77 12.02 11.03 12.58 12.07 HP 10.65 13.45 10.03 10.36 10.62 11.02 Period mean 11.52 xy 12.80 y 10.66 x 10.71 x 11.96 xy i-P, mgdl − 1 CON 5.86 7.03 7.22 6.97 7.17 6.85 a 0.08 0.025 0.003 0.752 HPL 6.94 7.65 7.51 7.08 7.77 7.39 b HEX 6.75 7.73 7.81 7.20 8.07 7.51 b HP 6.95 7.58 7.25 7.73 7.10 7.32 b Period mean 6.63 x 7.50 y 7.45 y 7.24 y 7.53 y Cu, µgml − 1 CON 0.74 0.75 0.78 0.79 0.74 0.76 a 0.01 0.000 0.000 0.002 HPL 0.80 1.27 1.23 1.20 1.21 1.14 c HEX 0.74 1.03 1.13 1.10 1.09 1.02 b HP 0.70 1.07 1.09 1.11 0.99 0.99 b Period mean 0.75 x 1.03 y 1.06 y 1.05 y 1.01 y Zn, µgml − 1 CON 0.70 0.72 0.79 0.82 0.83 0.77 a 0.01 0.000 0.000 0.053 HPL 0.74 1.04 1.29 1.24 1.31 1.12 c HEX 0.74 1.02 1.17 1.07 1.10 1.02 b HP 0.74 1.00 1.15 1.14 1.19 1.05 b Period mean 0.73 x 0.95 y 1.10 z 1.07 z 1.11 z Mn, µgml − 1 CON 0.25 0.25 0.26 0.27 0.30 0.27 a 0.00 0.000 0.000 0.018 HPL 0.26 0.35 0.38 0.39 0.39 0.35 d HEX 0.25 0.35 0.36 0.37 0.37 0.34 c HP 0.25 0.30 0.33 0.35 0.36 0.32 b Period mean 0.25 w 0.31 x 0.33 xy 0.34 yz 0.35 z Iron, µgml − 1 CON 3.48 3.37 3.54 4.00 3.67 3.61 a 0.03 0.000 0.000 0.000 HPL 3.44 4.07 4.57 4.80 5.14 4.40 c HEX 3.50 4.05 4.53 4.68 4.58 4.27 bc HP 3.50 4.05 4.24 4.44 4.58 4.16 b Period mean 3.48 w 3.88 x 4.22 y 4.48 z 4.49 z abc and xyz means bearing different superscript in a row and column differ significantly 3.8. Reproductive performance Supplementation of herbal formulation in different forms showed considerable improvement on confirmed pregnancy rate in crossbred cows. The conception at first insemination was considerably higher in HPL, HEX and HP groups relative to CON, while, conception at subsequent insemination was found higher in HPL, HEX and CON than HP group. The cows in treatment groups came into heat (first oestrus) earlier (P < 0.01) as compared to CON. However, 66.7, 16.67, 16.7 and 33.3% of cows in CON, HPL, HEX and HP did not show any sign of oestrus and were non-responsive. The number of AIs per conception were higher (P < 0.01) in CON relative to treatment groups (Fig. 1). 3.3.1. Physical characteristic of cervical mucus The pH and spinnbarkeit values (cm) of cervical mucus were significantly higher (P < 0.05) in the HP, HPL, and HEX groups compared to the CON (Figs. 2 and 3). The arborization pattern (fern pattern) of cervical mucus with typical fern pattern exhibited higher conception rate in all the treatment (HP, HPL and HEX) groups in relation to atypical fern pattern in CON group is depicted in Table 7 . Table 7 Conception rate in relation to fern pattern of cervical mucus in crossbred cows fed herbal formulation Treatments Arborization pattern (fern pattern) Conception rate (%) CON Typical 16.66 Atypical 16.66 HPL Typical 66.66 Atypical 16.66 HEX Typical 83.33 Atypical 0.00 HP Typical 50.00 Atypical 16.66 3.9. Relative mRNA expression of PGR genes The data pertaining to gene expression of progesterone (PGR) is depicted in Fig. 4. The fold expression of PGR gene was significantly higher (P < 0.01) in HPL, HEX and HP groups compared to CON. 4. Discussion 4.6. Chemical composition of feeds The chemical composition of wheat straw and green fodder offered to experimental cows was within the normal range as reported by previous workers (Negi et al. 2015 ; Kumar et al. 2019 ; Wadhwa and Bakshi, 2023 ; Yatto et al. 2023). The DM, OM, CP, EE, TA, NDF and ADF of the concentrate mixture was also observed to be consistent with the results obtained by earlier workers (Choudhary et al. 2022 ; Patir et al. 2023; Singh et al. 2023). The composition of herbal powder and pellets was similar, while herbal extract had lower EE, TA, NDF and ADF values. 4.7. Hb and haematocrit The present investigation demonstrates a noteworthy increase (P < 0.05) in haematocrit levels in the HEX group following administration of herbal formulation, while levels of Hb remained consistent across all treatment cohorts. These findings align with the established normal range for cattle, as noted by Kaneko et al. ( 2008 ). This elevation in haematocrit may be attributed to the potential enhancement of blood circulation facilitated by the herbal mixtures, ensuring efficient transportation of red blood cells throughout the body. The observed rise in haematocrit values could also be associated with the presence of flavonoids and quercetin, known for their hematopoietic properties (Raja et al. 2011 ). Furthermore, it's noteworthy that the herbal formulation contains volatile compounds (Mendoza et al. 2019 ; Sánchez et al. 2021 ). Our findings corroborate those reported by Nanda et al. ( 2013 ), who documented a significant increase (P < 0.05) in haematocrit concentration in goats following supplementation with a polyherbal formulation. Similarly, Barkakati and Kalita ( 2020 ) observed a significant elevation in Hb levels with polyherbal formulation supplementation compared to the CON group in cattle. Choudhury and Sinha ( 2015 ) observed a significant increase (P < 0.05) in haematocrit concentration in rats with the addition of M. koenigii leaves extract at doses of 250–500 mg/kg BW compared to the CON group. Lee-Rangel et al. ( 2022 ) also reported a significant increase in haematocrit concentration in calves treated with a polyherbal phytogenic source. 4.8. Serum glucose and NEFA The synthesis, breakdown, and transport of lipids in the body have all been demonstrated to be influenced by herbal formulations containing polyphenols and flavonoids, which may decrease the release of NEFA from adipose tissue and increase lipid clearance from the bloodstream, thus lowering circulating NEFA levels (Hashemzadeh- Cigari et al. 2014; Baghbadorani et al. 2022 ). Furthermore, the decreased NEFA concentration could be due to the presence of bioactive compounds in herbal formulations ( M. koenigii and A. marmelos ) that enhanced energy metabolism and nutrient absorption in cows (Lopreiato et al. 2020 ). The improved energy utilization could lead to a reduction in circulating NEFA levels, as the cow’s body is better able to use energy sources from the diet. Nevertheless, these herbs are known to have hepatoprotective properties, supporting the liver’s function and metabolism. A healthier liver might contribute to better lipid metabolism, reducing the release of NEFAs into the bloodstream (Adewuyi et al. 2005 ). The present results are in concurrence with Barjibhe et al. ( 2019 ), who reported that polyherbal supplementation to cows significantly reduced the NEFA concentration in serum. In addition, cows supplemented with herbal extract in the treatment group had a lower (P < 0.01) concentration of serum NEFA (Baghbadorani et al. 2022 ). Razo Ortiz et al. ( 2020 ) reported that supplementation of polyherbal mixture (Ashwagandha, tulsi, gooseberry and giloy) to lambs did not have any significant effect to glucose level. Lozano-Sánchez et al. ( 2021 ) also observed that there was no significant effect on serum glucose with supplementation of polyherbal supplement in finishing lambs. Hashemzadeh-Cigari et al. ( 2015 ) observed that glucose level was not affected, but serum NEFA reduced in cows supplemented with herbal mixture as compared to the CON group, despite their analogous energy balance. Although the precise mechanisms responsible for the lower NEFA levels in cows supplemented with the herbal mixture remain unclear, it is proposed that the herbal mixture may inhibit lipolysis, thereby improving lipid metabolism and enhancing metabolic status in periparturient cows by increasing insulin sensitivity. 4.9. Total protein, albumin, globulin and A: G ratio Blood metabolites such as total protein, albumin, globulin, and A:G ratio were assessed as indicators of the general health and vitality of the animals. The results revealed that these blood metabolites did not show significant differences (P > 0.05) among the various dietary treatments. These observation align with previous research conducted by Birudu et al. ( 2020 ), who similarly found no significant changes in A/G ratio and albumin levels in rats treated with methanolic leaf extracts of A. marmelos compared to control groups. Adebajo et al. ( 2006 ) also observed no significant impact on serum total protein, albumin, and globulin levels in Swiss Albino rats when supplemented with methanolic extracts of M. koenigii at different dosage rates. However, contrary results were reported by Gandhi et al. ( 2012 ), who noted a significant reduction in total protein levels in rats treated with A. marmelos . Furthermore, Dhami et al. ( 2022 ) reported varying plasma protein concentrations among different animal categories upon bi-herbal therapy with M. koenigii and A. marmelos , with lowered levels observed in buffaloes and infertile heifers compared to cows. Rivero et al. ( 2012 ) observed an increase in plasma protein levels after adding an extract rich in phenols to lamb diets, consistent with the findings of Lee-Rangel et al. ( 2022 ), who observed increased serum protein concentration in calves treated with a polyherbal phytogenic source. Furthermore, Dorantes-Iturbide et al. ( 2022 ) found that lambs fed a dietary supplement with a polyherbal additive containing hydrolyzable tannins, flavonoids, and essential oils exhibited a significant increase in serum albumin concentration. The analogous values of serum proteins observed in our study may be attributed to the relatively short duration of the treatment (7 days) compared to previous experiments, which were conducted for varying durations. 4.10. Total cholesterol The feeding of the herbal formulation to dairy cows did not show a significant effect (P > 0.05) among the treatments. In this study, supplementation with the herbal formulation had no effect on serum cholesterol levels, suggesting that the bioactive metabolites did not impact intestinal absorption or lipid synthesis in the cells of cows (Dorantes-Iturbide et al. 2022 ). However, Razo Ortiz et al. ( 2020 ), who also reported that supplementation of polyherbal mixture ( Ashwagandha, tulsi, gooseberry and giloy ) to lambs do not have any significant effect to serum cholesterol level. In contrast to our findings, Satheesh and Pari, ( 2008 ) reported that saponins extracted from A. marmelos reduce blood cholesterol levels by competing with cholesterol for binding sites or by interfering with cholesterol biosynthesis in the liver. Priyadarshinee et al. ( 2021 ) observed that M. koenigii and A. marmelos leaves supplementation in cows significantly reduced the total cholesterol in treatment groups. The comparable cholesterol concentration in present study may be attributed to the short duration (7 d) of herbal treatment. 4.11. Serum urea, creatinine and serum enzymes A significant (P < 0.01) reduction was observed in serum urea concentration in the HEX and HPL groups, indicating a decrease in rumen protein breakdown. This suggests enhanced EAA absorption in HEX, HPL, and HP groups (Pathak et al. 2017 ; Wankhede et al. 2022 ). Moreover, the decrease in serum urea indicates minimal protein catabolism and normal kidney function (Roseler et al. 1993 ; Hosten, 1990 ; Shavit et al. 2012 ). The serum creatinine value was also significantly (P < 0.01) lower in treatment groups as compared to CON. This beneficial effect may be due to the presence of polyphenols, tannins, and flavonoids in the herbal formulations. These compounds exhibit antioxidant activity and anti-inflammatory effects, and provide cardiovascular health benefits, potentially aiding kidney health by reducing oxidative stress and inflammation in renal tissues. Previous studies have associated these compounds with potential protection against kidney damage (Arulselvan et al. 2006 ). The findings are consistent with the observations of Arulselvan et al. ( 2006 ), who found that blood urea and plasma creatinine decreased in treatment groups when experimental rats were treated with M. koenigii , which provides addition al evidence to prove the anti-diabetogenic property of M. koenigii leaf extract. The primary serum markers for assessing liver changes include AST, ALT, total protein, and total albumin were unaffected by the treatment involving herbal formulation mixture, indicating no liver damage (Díaz Galván et al. 2021 ). Furthermore, its polyphenols might provide an antioxidant benefit (Biswas and Giri, 2015 ), along with other advantageous effects on liver function due to this nutrient (Mehedint and Zeisel, 2013 ). 4.12. Serum mineral profile Macro and micro elements play a crucial role in maintaining immune function and overall health. The high magnesium content in curry leaves suggests that they could serve as an excellent source of magnesium, which is known to activate various enzyme systems (Olusanya et al. 2008). Ca and P are also critical, as they influence an animal's ability to utilize other micronutrients and can modulate reproductive hormone sensitivity through various enzyme systems (Dutta et al. 2001 ). This could explain why the supplementation of curry leaves might lead to higher serum Ca and P levels and, in turn, support ovarian activity in anestrous heifers. Additionally, both M. koenigii and A. marmelos leaves are rich in essential minerals such as C, P, iron, zinc, manganese, and copper (Shantala and Prakash, 2005 ; Igara et al. 2016 ; Vijayalakshmi and Venkatalakshmi, 2017 ; Kujur et al. 2022 ). Our results align with Orzuna-Orzuna et al. ( 2021 ), who found that serum Ca levels among treatment groups were within the normal range for ovine, indicating that herbal mixtures do not disrupt mineral balance or nutritional status in lambs. In a similar vein, Sathesh Kumar and Punniamurthy (2009) observed that daily supplementation with 100 gd − 1 of M. koenigii leaves for 30 days significantly improved serum inorganic i-P levels and induced estrus in anestrous heifers. Similarly, Razo Ortiz et al. ( 2020 ) found that serum calcium levels were not affected by polyphenol and flavonoid doses, but serum phosphorus concentrations rose with increased doses of these compounds. Dhami et al. ( 2019 ) reported that supplementing cows with a mixture of M. koenigii and A. marmelos led to significant increases in plasma Ca, i-P, and other micro-minerals such as Zn, iron, and Cu. It is obvious that the mineral composition of the tree foliage is superior to that of tropical grasses. Goodchild and McMeniman ( 1994 ) reported that inclusion of tree foliage in ruminants’ diet increased the supply of minerals to the animals and rumen microbes and ultimately improved their performance. However, in contrary to our findings, Baitule et al. ( 2016 ) noted significant increases in serum Ca levels on day 9, the day of estrus, and on day 8 after estrus onset in treated groups, possibly due to supplementation with A. marmelos and M. koenigii . Likewise, Kumawat et al. ( 2014 ) found significantly higher serum Ca levels on days 4, 6, 8, and 10 after treating delayed pubertal heifers with dried leaf powder from A. marmelos and M. koenigii , compared to non-treated groups. This study also observed that the effect of these leaves on serum P levels was significant, with marked variations on days 2, 4, and 8 after treatment compared to non-treated heifers. 4.13. Reproduction Supplementation of herbal formulation in different forms showed considerable improvement on confirmed pregnancy rate in crossbred cows (Patton et al. 2007 ; Ambrose, 2021 ). The beneficial effects of herbal formulations ( A. marmelos and M. koenigii ) supplementation could be due to the presence of phytoestrogens, minerals, and antioxidants. These components might stimulate gonadotropin hormones from the anterior pituitary, potentially contributing to the restoration of cyclicity in anestrus animals (Dutt et al., 2010 ; 2011 ). The antioxidants present may have enhanced folliculogenesis by preventing ROS-induced apoptosis of oocytes (Vijayalakshmi and Venkatalakshmi, 2017 ). The phytoconstituents might have stimulated steroidogenic activity in the hypothalamus, promoted follicular growth, and facilitated steroid production, leading to the initiation of cyclicity (Jondhale et al. 2009a ; Nandini et al. 2010 ). Additionally, the presence of bioactive compounds in the herbal formulation may have contributed to follicular development, particularly during the dominance phase leading up to ovulation, resulting in the resumption of estrus. In the present study, though the conception rate was significantly higher in all the treatment groups relative to CON, however, HPL and HEX groups showed better conception rate and established pregnancy than HP (herbal powder) group, which could be probably due to the less wastage and complete consumption of herbal powder in pellets forms as compared to powder (Wanapat et al. 2013 ). The findings of the current study align with those of Patel and Gupta ( 2020 ), who reported the highest conception rate (71.42%) in delayed pubertal heifers supplemented with M. koenigii , A. marmelos , and a mineral mixture. Additionally, M. koenigii , whether used alone or in combination with a mineral supplement and a low dose of GnRH injection, was found to be equally effective in inducing fertility in anestrus buffaloes (Umashanker et al. 2006 ). Delayed pubertal heifers have shown induced estrus behavior after being supplemented with A. marmelos combined with M. koenigii (Kumar et al. 2016 ; Kumawat et al. 2016 ). Baitule et al. ( 2016 ) found 75% oestrus induction after supplementing A. marmelos and M. koenigii for 9 days in anestrous buffaloes. Satheshkumar and Punniamurthy ( 2009 ) also studied that supplemental effect of M. koenigii leaves for a period of 30 days and found induced oestrus in 60% of the anestrus heifers. Mehrotra ( 2002 ) have also documented that M. koenigii has been found to enhance ovarian function in rats and improve the therapeutic response in acyclic goats and heifers, particularly regarding oestrus and conception. Tiwari et al. ( 2021 ) reported that the oestrus response was 83.33% in postpartum buffaloes when supplemented with a mixture of A. marmelos and M. koenigii . However, Das et al. ( 2012 ) observed 92.90% oestrus induction in delayed pubertal buffalo heifers after feeding of A. marmelos and M. koenigii herbs for 9 days. Das et al. ( 2016 ) and Kumawat et al. ( 2016 ) reported that 90.9% and 92.3% of delayed pubertal heifers, respectively, began cycling after being fed a herbal mixture. Additionally, they observed a pregnancy rate of 54.1% in the group treated with the herbal mixture, compared to 16.7% in the control group. Therefore, it may be speculated that favourable effects of herbal formulations (mixture of M. koenigii and A. marmelos ) in terms of estrus induction, ovulation, established pregnancy and shorter time period taken in induction of estrus are exhibited as compared to individual plant treatment which could be a reflection of synergistic actions of herbs active principles (Nandini et al. 2010 ; Hedge et al. 2002; Mehrotra et al. 2005 ). The active principles seem to operate either through mimicking gonadotrophins activity or stimulating the central mechanism for endogenous release of gonadotrophins along with possibility of local action (Dutt et al. 2018 ). 4.14. Physical characteristic of cervical mucus It is believed that the herbal formulations (mixture of M. koenigii and A. marmelos leaves) are having hormonal regulatory properties that might have supported the oestrous cycle and overall reproductive health cows. Since, this herbal formulation having antioxidant and anti-inflammatory properties which might have helped in reducing inflammation in the reproductive tract, promoting better cervical mucus production and a favourable environment for sperm transport (Dhankhar et al. 2011 ; Gahlawat et al. 2014 ; Nigam and Nambiar, 2015 ; Rautela et al. 2018 ). Our findings align with those of Dhami et al. ( 2022 ), who observed that administering a bi-herbal treatment ( M. koenigii and A. marmelos ) @ 200 g daily for 5 days resulted in improved mucus quality at subsequent next oestrus in infertile animals. On contrary, Rahi et al. ( 2013 ) reported that on feeding of herbal extract containing polyphenols have reduced the pH which may be due to decline in bacterial load and inflammatory process in uterus after treatment. The fertility is associated with an arborization pattern, as in our study we found that typical fern pattern was having higher conception rate in treatment groups thereby it was observed higher conception rate with typical fern pattern in all the treatment groups. The enhanced conception rate in crossbred cows with typical fern pattern indicates higher sperm penetration (Rangnekar et al. 2002 ; Layek et al. 2013 ). Alkaline pH of cervical mucus is favourable for higher sperm motility. The increase in pH in the treatment groups had higher fertility rate and also suitable for uterine environment with maximum sperm penetration in alkaline pH (Pattabiraman et al. 1967 ; Rangnekar et al. 2002 ). Similarly, the treatment groups exhibited a higher conception rate, correlated with an increased spinnbarkeit value. Additionally, the conception rate was influenced by the arborization pattern, as a typical arborization pattern is indicative of a proper hormonal balance, which supports higher conception rates (Alena et al. 2008 ).4.10. Relative mRNA expression of PGR genes The progesterone gene’s expression was associated with the progesterone hormone, which was likewise shown to be significantly greater in the treatment groups. The fold expression of PGR gene was significantly higher (P < 0.01) in HPL, HEX and HP groups as compared to CON. Progesterone is a crucial ovarian steroid that regulates the estrous cycle, maternal recognition of pregnancy, and gestation in farm animals (Spencer et al., 2004). Beyond these roles, progesterone is also vital for various reproductive functions, including the behavioral expression of estrus, ovulation, maintenance of uterine quiescence, embryo survival and normal parturition. Conclusions In conclusion, the herbal formulation exhibited a positive influence on both metabolic parameters and reproductive performance in postpartum crossbred cows. The observed improvements in blood parameters, and early onset of estrus suggest promising prospects for integrating herbal supplements in dairy cattle management. These findings underscore the potential of herbal formulations as a supportive measure for enhancing the overall health and fertility of postpartum cows. Declarations Ethical approval Before conducting the experimental trails, the animals (crossbred cattle) studies were approved by institutional ethics committee IAEC, ICAR-IVRI, Izatnagar-243122, Bareilly, Uttar Pradesh (Regd No: IAEC/07.07.2022/B6), and was conducted in accordance with the ethics standards as applicable international, national and/or institutional guidelines. Conflict of interest The authors do not have any conflict of interest regarding this research. Funding This study was financially supported by funds provided by Indian Council of Agriculture Research (AICRP project), New Delhi, India. Authors contributions J. C: conducted the research as a part of her Ph.D. research work. She has conducted the experimental trial deligently along with handling and management of the cows and laboratory tests including laboratory and molecular works. She also wrote the main manuscript text. Acknowledgements The authors would like to thank ICAR-IVRI, Izatnagar, UP-243122 for providing the facilities for conducting the research. Availability of data and materials Data provided within the manuscript is complete and can be provided on request to the authors. References Abdallah, I.Z., Salem, I., El-Salam, A. and Nayrouz, A.S., 2017. Evaluation of antidiabetic and antioxidant activity of A. marmelos L. Correa fruit extract in diabetic rats. Egyptian Journal of Hospital Medicine 67, 731–741. Abeysinghe, D.T., Kumara, K.A.H., Kaushalya, K.A.D., Chandrika, U.G. and Alwis, D.D.D.H., 2021. Phytochemical screening, total polyphenol, flavonoid content, in vitro antioxidant and antibacterial activities of Sri Lankan varieties of Murraya koenigii and Micromelum minutum leaves. Heliyon 7(7), 07449. https://doi.org/10.1016/j.heliyon.2021.e07449 Adebajo, A.C., Ayoola, F.O., Iwalewa, E.O., Akindahunsi, A.A., Omisore, N.O.A., Adewunmi, C.O. and Adenowo, T.K., 2006. Anti-trichomonal, biochemical and toxicological activities of methanolic extract and some carbazole alkaloids isolated from the leaves of Murraya koenigii growing in Nigeria. Phytomedicine 13(4), 246–254. https://doi.org/10.1016/j.phymed.2004.12.002 Adewuyi, A.A., Gruys, E. and Van Eerdenburg, F.J.C.M., 2005. Non esterified fatty acids (NEFA) in dairy cattle. A review. Veterinary Quarterly 27(3), 117–126. https://doi.org/10.1080/01652176.2005.9695192 Alena, J., Ludek, S., Mojmir, V. and Franstisek, L., 2008. Factors affecting the cervical mucus crystallization, the sperm survival in cervical mucus and pregnancy rate in Holstein cows. Journal of Central European Agriculture 9(2), 377–384. AOAC, 2012. Official method of analysis of AOAC International. 19th ed. Association of Analytical Communities International, Virginia, USA. Ambrose, D.J., 2021. Postpartum anestrus and its management in dairy cattle. Bovine Reproduction, 408–430. https://doi.org/10.1002/9781119602484.ch34 Arulselvan, P., Senthilkumar, G.P., Sathish Kumar, D. and Subramanian, S., 2006. Anti-diabetic effect of A. marmelos leaves on streptozotocin induced diabetic rats. Die Pharmazie-International Journal of Pharmaceutical Sciences Research 61(10), 874–877. Baghbadorani, M.K., Mehrzad, J., Vodjgani, M., Khosravi, A. and Akbarinejad, V., 2022. Evaluation of biochemical and hematological parameters in postpartum Holstein dairy cows following supplementation of Immunofin® herbal extract. Iranian Journal of Veterinary Medicine 16(3), 274–287. https://dx.doi.org/10.22059/IJVM.2021.315021.1005144 Baitule, M.M., Gawande, A.P., Kumar, U., Sahatpure, S.K., Patil, M.S. and Baitule, M.M., 2016. Effect of A. marmelos and A. marmelos in treatment of delayed pubertal buffalo heifers. Veterinary World 9(12), 1375. https://doi.org/10.14202%2Fvetworld.2016.1375-1380 Bansal, Y. and Bansal, G., 2011. Analytical methods for standardization of A. marmelos: a review. Journal of Pharmaceutical Education and Research 2, 37–44. Barjibhe, S., Oberoi, P.S., Patel, B. and Patel, P.K., 2019. Improving reproductive efficiency through the supplementation of mustard oil, poly-herbal mixture and butyric acid during the periparturient period in Sahiwal cows. Journal of Entomology and Zoology Studies 7(2), 668–673. Barkakati, J. and Kalita, D.J., 2020. Alteration in hematological profile of pregnant crossbred cattle of Assam after feeding some medicinal leaves/herbs and polyherbal preparation (Restobal) during their transition period. The Pharma Innovation 9(9), 197–201. https://doi.org/10.22271/tpi.2020.v9.i9c.5110 Begashaw, B., Mishra, B., Tsegaw, A. and Shewamene, Z., 2017. Methanol leaves extract Hibiscus micranthus Linn exhibited antibacterial and wound healing activities. BMC Complementary and Alternative Medicine 17(1), 1–11. https://doi.org/10.1186/s12906-017-1841-x Birudu, R.B., Pamulapati, P. and Manoharan, S.K., 2020. Evaluation of biochemical changes in diabetic rats treated with Aegle marmelos (L.) methanolic leaf extract. Pharmacognosy Research 12(2), 127–130. http://dx.doi.org/10.4103/pr.pr_53_19 Biswas, S. and Giri, S., 2015. Importance of choline as essential nutrient and its role in prevention of various toxicities. Prague Medical Report 116, 5–15. doi: 10.14712/23362936.2015.40 Chakma, J., Samanta, A., Dutta, T. and Arya, R.S., 2020. Effect of supplementing Moringa oleifera leaf extract and clove bud oil to the diet on microflora population and intestinal morphology of broiler birds. Indian Journal of Animal Health 59, 222–227. doi: 10.36062/ijah.59.2SPL.2020.222-227 Chakma, J., Kaur, N., Namdeo, S., Patir, M., Dutta, N., Jadhav, S.E., Singh, G. and Singh, S.K., 2024. Effect of Murraya koenigii and Aegle marmelos leaves supplementation on gas production kinetics and feed degradation with cattle inoculum. Animal Nutrition and Feed Technology 24(1), 137–148. doi: 10.5958/0974-181X.2024.00010.6 Choudhary, S., Chaudhary, L.C., Kala, A., Agarwal, N. and Rahman, H., 2022. Effect of dietary sulphur supplementation at graded levels on rumen feed fermentation and microbial profile in buffaloes. Animal Nutrition and Feed Technology 22(3), 543–552. doi: 10.5958/0974-181X.2022.00043.9 Choudhury, S. and Sinha, M.P., 2015. Effect of aqueous extract of Murraya koenigii on haematological, hormonal and lipid profile of Albino rats. Journal of Coastal Life Medicine 3, 901–905. doi: 10.12980/jclm.3.2015j5-63 Dacie, Z. and Lewis, S., 1969. Practical Haematology. 4th ed. J and A., Churchill, London. Das, G.K., Mehrotra, S., Kumar, B., Jan, M.H., Pande, M., Singh, S.K., Kumar, H., Khan, T.A., Agarwal, S.K. and Shankar, U., 2012. Pattern and estrus induction response in delayed pubertal buffalo heifers treated with a combined treatment of Murraya koenigii and Aegle marmelos. In: National symposium of XXVIII Annual Convention of the Indian Society for Study of Animal Reproduction. Khanapara, Guwahati, India. pp. 50. Das, P., Das, S.K., Arya, H.P.S., Subba Reddy, G. and Mishra, A., 2002. Veterinary Science and Animal Husbandry. Inventory of indigenous technical knowledge in agriculture, Document-1, ICAR, New Delhi, pp.185–285. Das, G.K., Mehrotra, S., Narayanan, K., Kumawat, B.L., De, U.K., Khan, T.A., 2016. Estrus induction response and fertility performance in delayed pubertal heifers treated with Aegle marmelos and Murraya koenigii. Journal of Animal Research 6(1), 151–156. Dhami, A.J., Parmar, S.C., Patel, J.A., Hadiya, K.K., Joshi, R.S. and Sarvaiya, N.P., 2022. Current status of reproductive performance and problems in dairy animals of Gujarat and their amelioration including study of fertility gene markers. Markers 18(2), 12–20. Dhami, A.J., Patel, J.A., Hadiya, K.K., Parmar, S.C. and Chaudhari, D.V., 2019. Nutritional infertility and ameliorative measures in dairy animals of middle Gujarat. Indian Journal of Veterinary Science and Biotechnology 14(3), 5–9. Dhankhar, S., Ruhil, S., Balhara, M., Dhankhar, S. and Chhillar, A.K., 2011. Aegle marmelos (Linn.) Correa: A potential source of phytomedicine. Journal of Medicinal Plants Research 5(9), 1497–1507. Dhivagar, M., 2022. Effect of herbal extracts supplementation on performance of wistar rats. M.V.Sc. Thesis, Indian Veterinary Research Institute, Izatnagar (Deemed University), Bareilly, India. Díaz Galván, C., Méndez Olvera, E.T., Martínez Gómez, D., Gloria Trujillo, A., Hernández García, P.A., Espinosa Ayala, E., Palacios Martínez, M., Lara Bueno, A., Mendoza Martínez, G.D. and Velázquez Cruz, L.A., 2021. Influence of a polyherbal mixture in dairy calves: growth performance and gene expression. Frontiers in Veterinary Science 7, 623710. https://doi.org/10.3389/fvets.2020.623710 Dorantes-Iturbide, G., Orzuna-Orzuna, J.F., Lara-Bueno, A., Miranda-Romero, L.A. and Mendoza-Martínez, G.D., 2022. Effects of a polyherbal dietary additive on performance, dietary energetics, carcass traits and blood metabolites of finishing lambs. Metabolites 12(5), 413. https://doi.org/10.3390/metabo12050413 Driver, A.M., Huang, W., Gajic, S., Monson, R.L., Rosa, G.J.M. and Khatib, H., 2009. Effects of the progesterone receptor variants on fertility traits in cattle. Journal of Dairy Science 92(8), 4082–4085. https://doi.org/10.3168/jds.2009-2013 Dutt, R., Dalal, J., Singh, G., Gahalot, S.C. and Chandolia, R.K., 2018. Medicinal uses of Murraya koenigii and Aegle marmelos for fertility augmentation in animals: a review. International Journal of Current Microbiology and Applied Sciences 7(9), 645–657. https://doi.org/10.20546/ijcmas.2018.709.077 Dutt, R., Mehrotra, S., Hoque, M., Shanker, U., Singh, G., Agarwal, S.K., Das, G.K. and Singh, S.K., 2010. Effect of Murraya koenigii and Aegle marmelos combination on resumption of fertility in anestrous goats. Journal of Applied Animal Research 38(2), 275–278. https://doi.org/10.1080/09712119.2010.10539520 Dutt, R., Mehrotra, S., Shanker, U. and Singh, G., 2011. Effect of Murraya koenigii and Aegle marmelos feeding on anestrus buffaloes. Indian Journal of Animal Reproduction 32(1), 47–49. Dutta, A., Baruah, B., Sarmah, B.C., Baruah, K.K. and Goswami, R.N., 2001. Macromineral levels in cyclic, postpartum anestrus and repeat breeding local cows in lower Brahmaputra valley of Assam. Indian Journal of Animal Reproduction 22(1), 41–44. Dutta, L.J., Nath, K.C., Deka, B.C., Bhuyan, D., Borah, P., Saikia, G.K., Bora, D.P., Deka, R., Acharya, C., Bharali, D., 2019. Therapeutic management of true anestrous crossbred cows through nutritional and hormonal intervention. Journal of Entomology and Zoology Studies 7(3), 09–12. Gahlawat, D.K., Jakhar, S. and Dahiya, P., 2014. Murraya koenigii (L.) Spreng: An ethnobotanical, phytochemical and pharmacological review. Journal of Pharmacognosy and Phytochemistry 3(3), 109–119. Gandhi, G.R., Ignacimuthu, S. and Paulraj, M.G., 2012. Hypoglycemic and β-cells regenerative effects of Aegle marmelos (L.) Corr. bark extract in streptozotocin-induced diabetic rats. Food and Chemical Toxicology 50(5), 1667–1674. Ganesan, P., Phaiphan, A., Murugan, Y. and Baharin, B.S., 2013. Comparative study of bioactive compounds in curry and coriander leaves: An update. Journal of Chemical and Pharmaceutical Research 5(11), 590–594. Goodchild, A.V. and McMeniman, N.P., 1994. Intake and digestibility of low quality roughages when supplemented with leguminous browse. Journal of Agricultural Science 122(1), 151–160. doi: 10.1017/S0021859600065904 Hashemzadeh-Cigari, F., Ghorbani, G.R., Khorvash, M., Riasi, A., Taghizadeh, A. and Zebeli, Q., 2015. Supplementation of herbal plants differently modulated metabolic profile, insulin sensitivity and oxidative stress in transition dairy cows fed various extruded oil seeds. Preventive Veterinary Medicine 118(1), 45–55. https://doi.org/10.1016/j.prevetmed.2014.10.013 Hegde, G.V., Nimbalkar, S.D., Hegde, G.S. and Naik, K., 2002. Use of herbal plants for repeat breeding problem in dairy animals. Indian Veterinary Journal 79(8), 861–862. Hosten, A.O., 1990. BUN and creatinine. In Clinical Methods: The History, Physical, and Laboratory Examinations. 3rd edition. Butterworths. Igara, C.E., Omoboyowa, D.A., Ahuchaogu, A.A., Orji, N.U. and Ndukwe, M.K., 2016. Phytochemical and nutritional profile of Murraya koenigii (Linn) Spreng leaf. Journal of Pharmacognosy and Phytochemistry 5(5), 7. Jadhav, A.N. and Bhutani, K.K., 2005. Ayurveda and gynaecological disorders. Journal of Ethnopharmacology 97, 151–159. https://doi.org/10.1016/j.jep.2004.10.020 Jain, V., Momin, M. and Laddha, K., 2012. Murraya koenigii: an updated review. International Journal of Ayurvedic and Herbal Medicine 2(4), 607–627. Jondhale, N.S., 2007. Effect of Aegle marmelos and Ficus religiosa on the ovarian function in rats and follicular dynamics in goat. MVSc Thesis, Indian Veterinary Research Institute, Izatnagar (Deemed University), Bareilly, India. Jondhale, N.S., Mehrotra, S., Agarwal, S.K., Singh, S.K. and Deori, S., 2009a. Effect of Aegle marmelos and Ficus religiosa on the onset of puberty in immature female rats. Indian Veterinary Journal 86(4), 353–355. Jondhale, N.S., Uma, S., Mehrotra, S., Agarwal, S.K., Singh, S.K., Das, G.K. and Deori, S., 2009b. Effect of Aegle marmelos and Ficus religiosa leaves extracts on the ovarian function in rats. Indian Veterinary Journal 86(11), 1141–1144. Kabir, K.K., Rawal, C.V.S., Ansari, M.R., Varshney, J.P. and Srivastava, R.S., 2001. Comparative efficacy of herbal preparations in the management of anoestrus in non-descript rural buffaloes. Indian Journal of Animal Reproduction 22(2), 143–145. Kaneko, J.J., Harvey, J.W. and Bruss, M.L., 2008. Clinical biochemistry of domestic animals. 6th ed. Academic Press, San Diego, California, USA. Khade, N.B., Patel, D.M., Naikoo, M., Dhami, A.J., Sarvaiya, N.P. and Gohel, M.M., 2011. Estrus induction in pubertal anestrus Gir heifers using different hormone protocols. Indian Journal of Field Veterinarians 7(1), 4–8. Kujur, A., Srivastava, N., Jasrotia, N. and Arunpandian, J., 2022. The role of medicinal plants in domestic animals and estrus induction with particular reference to Aegle marmelos and Murraya koenigii. Animal Reproduction Update 2(1), 56–62. https://doi.org/10.48165/aru.2022.2102 Kumar, P.R., Singh, S.K., Kharche, S.D., Govindaraju, C.S., Behera, B.K., Shukla, S.N., Kumar, H. and Agarwal, S.K., 2014. Anestrus in cattle and buffalo: Indian perspective. Advances in Animal and Veterinary Sciences 2(3), 124–138. http://dx.doi.org/10.14737/journal.aavs/2014/2.3.124.138 Kumar, A., Katiyar, R., Ahmad, S.F., Balamurugan, B., Deepak, D. and Prasad, J.K., 2018. Current treatment aspects of bovine reproductive disorders. Theriogenology Insight 8(3), 101–109. http://dx.doi.org/10.30954/2277-3371.03.2018.2 Kumar, R., 2008. Effect of Aegle marmelos (Bel) and Ficus religiosa (Pipal) on the reproductive performance in anoestrus goat and buffalo. MVSc Thesis, Indian Veterinary Research Institute, Izatnagar, India. Kumar, S., Mehla, R.K. and Dang, A.K., 2008. Use of shatavari (Asparagus racemosus) as a galactopoietic and therapeutic herb: a review. Agricultural Reviews 29(2), 132–138. Kumar, S., Bodla, R.B. and Bansal, H., 2016. Antioxidant activity of leaf extract of Aegle marmelos Correa ex Roxb. Pharmacognosy Journal 8(5), 447–450. http://dx.doi.org/10.5530/pj.2016.5.6 Kumar, S., Kumar, G. and Gahalot, S.C., 2019. Anestrous in buffaloes and different treatment regimens: a mini review. International Journal of Current Microbiology and Applied Sciences 8, 1162–1179. https://doi.org/10.20546/ijcmas.2019.803.138 Kumawat, B., Das, G., Narayanan, K., Mehrotra, S., Prasad, J. and Ghosh, S., 2016. Effect of Aegle marmelos and Murraya koenigii on estrus induction, conception and calving rate in delayed pubertal heifers under field condition. Indian Journal of Animal Reproduction 37(2), 17–19. Kumawat, B.L., Das, G.K., Narayanan, K., Mehrotra, S., Choadhary, R.K., Yogi, R.K., Dey, S. and Kumar, H., 2014. Effect of herbs supplementation on certain serum mineral levels in delayed pubertal heifers. National Symposium and XXX Annual Convention of ISSAR, Mathura, (U.P.) India, November 20–22, 2014. Compendium and Souvenir, F.I. 20, 25. Kuralkar, P. and Kuralkar, S.V., 2021. Role of herbal products in animal production: an updated review. Journal of Ethnopharmacology 278, 114246. https://doi.org/10.1016/j.jep.2021.114246 Layek, S.S., Mohanty, T.K., Kumaresan, A., Behera, K. and Chand, S., 2013. Cervical mucus characteristics and periestrual hormone concentration in relation to ovulation time in Zebu (Sahiwal) cattle. Livestock Science 152(2–3), 273–281. https://doi.org/10.1016/j.livsci.2012.12.023 . Lee-Rangel, H.A., Mendoza-Martinez, G.D., Martínez-García, J.A., Espinosa-Ayala, E., Hernández-García, P.A., Cifuentes-López, R.O., Vazquez-Valladolid, A., García-López, J.C., Lara-Bueno, A. and Roque-Jiménez, J.A., 2022. An Indian polyherbal phytogenic source improved blood serum biochemistry and immune response of dairy calves. Food and Agricultural Immunology 33(1), 97–112. https://doi.org/10.1080/09540105.2021.2024150 . Livak, K.J. and Schmittgen, T.D., 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2 – ∆∆CT method. Methods 25(4), 402–408. https://doi.org/10.1006/meth.2001.1262 . Lone, S.A., Ganie, A.A., Quadri, M.A., Haq, M.R. and Reshi, I.M., 2012. Effect of exogenous insulin on fertility response and serum hormonal profile in delayed pubertal crossbred heifers. Veterinary Practitioner 13(2), 268–269. Lopreiato, V., Mezzetti, M., Cattaneo, L., Ferronato, G., Minuti, A. and Trevisi, E., 2020. Role of nutraceuticals during the transition period of dairy cows: A review. Journal of Animal Science and Biotechnology 11, 1–18. https://doi.org/10.1186/s40104-020-00501-x . Lozano-Sánchez, M., Mendoza-Martínez, G.D., Martínez-García, J.A., la Torre-Hernández, M.E.D., Chamorro-Ramírez, F.H., Martínez-Aispuro, J.A., Cordero-Mora, J.L., Sánchez-Torres, M.T., Hernández-García, P.A. and Jones, R., 2021. Evaluation of polyherbal with vitamin C activity on lamb performance and meat characteristics. Revista Brasileira de Zootecnia 50, e20200166. https://doi.org/10.37496/rbz5020200166 . Mehedint, M.G. and Zeisel, S.H., 2013. Choline's role in maintaining liver function: new evidence for epigenetic mechanisms. Current Opinion in Clinical Nutrition and Metabolic Care 16, 339–345. doi: 10.1097/MCO.0b013e3283600d46 . Mehrotra, S., 2002. Studies on ovarian function using certain medicinal plants in rats, goats and cattle. Doctoral dissertation, Indian Veterinary Research Institute; Bareilly, India. Mehrotra, S., Singh, S.K., Shanker, U.S. and Agarwal, S.K., 2005. Effect of certain indigenous medicinal plants on ovarian hormone profile and reproductive performance in anestrus cattle. Indian Journal of Animal Reproduction 26(1), 20–23. Mehrotra, S., Umashanker, J., Majumdar, A.C. and Agarwal, S.K., 2003. Effect of indigenous medicinal plants on onset of puberty in immature female rats. Indian Journal of Animal Reproduction 24(2), 131–133. Mendoza, G.D., Oviedo, M.F., Pinos, J.M., Lee-Rangel, H.A., Vázquez, A., Flores, R. and Pérez, F., 2019. Milk production in dairy cows supplemented with herbal choline and methionine. Revista de la Facultad de Ciencias Agrarias 52(1), 1–12. Mishra, B.P., Nayak, N. and Mishra, D., 2002. Indigenous methods for animal health care. International seminar on traditional knowledge, health and environment. Bubaneshwar, Abs, pp. 124. Naczk, M. and Shahidi, F., 2004. Extraction and analysis of phenolics in food. Journal of Chromatography A 1054(1–2), 95–111. https://doi.org/10.1016/j.chroma.2004.08.059 . Nanda, S., Tiwari, S.P. and Rathore, R., 2013. Effect of polyherbal formulations on blood haematological constituents and immunity in non-descript goats. Iranian Journal of Applied Animal Science 3(2), 327–331. Nandini, M.S., Veena, T. and Swamy, M.N., 2010. Effect of extracts of Murraya koenigii spreng. and Morus alba linn. on the age of attainment of puberty and ovarian folliculogenesis in rats. Journal of Basic and Clinical Pharmacy 1(4), 203. Negi, D., Mondal, B.C. and Tiwari, D.P., 2015. Effect of phase-feeding on milk production, milk composition, nutrient utilization and haematobiochemical constituents in lactating crossbred cattle. Animal Nutrition and Feed Technology 15(2), 235–244. http://dx.doi.org/10.5958/0974-181X.2015.00026.8 . Nigam, V. and Nambiar, V.S., 2015. Therapeutic potential of Aegle marmelos (L.) Correa leaves as an antioxidant and anti-diabetic agent: A review. International Journal of Pharmaceutical Sciences and Research 6(3), 611–621. Olusanya, J.O., 2008. The food nutrients. In: Essentials of Food and Nutrition, Apex Book Limited Lagos, xxi. Orzuna-Orzuna, J.F., Dorantes-Iturbide, G., Lara-Bueno, A., Mendoza-Martínez, G.D., Miranda-Romero, L.A. and Hernández-García, P.A., 2021. Growth performance, carcass characteristics and blood metabolites of lambs supplemented with a polyherbal mixture. Animals 11(4), 955. https://doi.org/10.3390/ani11040955 . Panigrahi, B., 1964. Spinnbarkeit test of bovine cervical mucus—an index to optimal fertility time. Indian Veterinary Journal 41(6), 410–412. Patel, R.K. and Gupta, D.K., 2020. An in vivo assessment of a non-antibiotic therapy for specific subclinical mastitis in dairy cows. Journal of Entomology and Zoology Studies 8(5), 721–725. Pathak, A.K., Dutta, N., Pattanaik, A.K., Chaturvedi, V.B. and Sharma, K., 2017. Effect of condensed tannins from Ficus infectoria and Psidium guajava leaf meal mixture on nutrient metabolism, methane emission and performance of lambs. Asian-Australasian Journal of Animal Sciences 30(12), 1702. https://doi.org/10.5713/ajas.17.0060 . Patir, M., 2022. Effect of Moringa oleifera foliage supplementation on the performance of crossbred cattle. Thesis. PhD. Deemed University, Indian Veterinary Research Institute, Izatnagar, India. Pattabiraman, S.R., Venkataswamy, V. and Thangraj, T.M., 1967. Physicochemical properties of oestrual mucus of cows. Indian Veterinary Journal 10, 413–417. Patton, J., Kenny, D.A., McNamara, S., Mee, J.F., O’mara, F.P., Diskin, M.G. and Murphy, J.J., 2007. Relationships among milk production, energy balance, plasma analytes and reproduction in Holstein-Friesian cows. Journal of Dairy Science 90(2), 649–658. https://doi.org/10.3168/jds.S0022-0302(07)71547-3 . Priyadarshinee, P., Sethy, K., Swain, R.K., Mishra, S.K., Behera, K., Pradhan, S.K. and Tripathy, S., 2021. Combined effect of Bel and curry leaves supplementation on the performance of anestrous cattle. Indian Journal of Animal Research 38(3), 240–245. https://doi.org/10.5958/2231-6744.2021.00035.9 . Purkayastha, R.D., Shukla, S.N., Shrivastava, O.P. and Kumar, P.R., 2015. A comparative therapeutic management of anoestrus in buffaloes using insulin and GnRH. Veterinary World 8(6), 804–807. https://doi.org/10.14202/vetworld.2015.804-807 . Rahi, S., Gupta, H.P., Prasad, S. and Baithalu, R.K., 2013. Phytotherapy for endometritis and subsequent conception rate in repeat breeding crossbred cows. Indian Journal of Animal Reproduction 34(1), 9–12. Raja, S.B., Murali, M.R., Kumar, N.K. and Devaraj, S.N., 2011. Isolation and partial characterization of a novel lectin from Aegle marmelos fruit and its effect on adherence and invasion of Shigellae to HT29 cells. PLoS One 6(1), e16231. https://doi.org/10.1371/journal.pone.0016231 . Rajkumar, R., Srivastava, S.K., Varshney, V.P. and Mahmood, S., 2008. Effect of medicinal plants Saraca asoca and Trigonella foenum-graecum in anoestrus cows. Indian Veterinary Journal 85(12), 1281–1283. Rangnekar, M.N., Dhoble, R.L., Gacche, M.G., Ingawale, M.V., Sawale, A.G. and Jadhav, J.M., 2002. Physical properties of oestrual cervical mucus in repeat breeding crossbred (Holstein Friesian) cows with reference to fertility. Indian Journal of Animal Sciences 72(12), 1122–1124. Rautela, R., Das, G.K., Khan, F.A., Prasad, S., Kumar, A., Prasad, J.K., Ghosh, S.K., Dhanze, H., Katiyar, R. and Srivastava, S.K., 2018. Antibacterial, anti-inflammatory and antioxidant effects of Aegle marmelos and Murraya koenigii in dairy cows with endometritis. Livestock Science 214, 142–148. https://doi.org/10.1016/j.livsci.2018.05.015 . Razo Ortiz, P.B., Mendoza Martinez, G.D., Silva, G.V., Osorio Teran, A.I., Gonzalez Sanchez, J.F., Hernandez Garcia, P.A., de la Torre Hernández, M.E., Espinosa Ayala, E., 2020. Polyherbal feed additive for lambs: effects on performance, blood biochemistry and biometry. Journal of Applied Animal Research 48(1), 419–424. https://doi.org/10.1080/09712119.2020.1814786 . Reddy, K.N., 2008. Ethnobotany of Andhra Pradesh: A Review. Ethnobotanical Leaflets 12, 305–310. Rivero, N., Salem, A.Z.M., Gado, H.M., Gonzalez-Ronquillo, M., Pliego, A.B., Peñuelas, C.G. and Odongo, N.E., 2012. Effect of exogenous enzymes and Salix babylonica extract or their combination on haematological parameters in growing lambs. Journal of Animal Feed Science 21, 577–586. Roseler, D.K., Ferguson, J.D., Sniffen, C.J. and Herrema, J., 1993. Dietary protein degradability effects on plasma and milk urea nitrogen and milk nonprotein nitrogen in Holstein cows. Journal of Dairy Science 76(2), 525–534. https://doi.org/10.3168/jds.S0022-0302(93)77372-5 . Sabu, M.C. and Kuttan, R., 2004. Antidiabetic activity of Aegle marmelos and its relationship with its antioxidant properties. Indian Journal of Physiology and Pharmacology 48(1), 81–88. Sánchez, N., Lee Rangel, H.A., Martínez-Cortés, I., Mendoza, G.D., Hernández, P.A., Espinoza, E., Vazquez-Valladolid, A., Flores-Ramírez, R., Roque-Jiménez, A., Campillo-Navarro, M. and Relling, A.E., 2021. A polyherbal phytogenic additive improved growth performance, health, and immune response in dairy calves. Food and Agricultural Immunology 32(1), 482–498. https://doi.org/10.1080/09540105.2021.1967296 . Shantala, M. and Prakash, J., 2005. Acceptability of curry leaf (Murraya koenigii) incorporated products and attitude toward consumption. Journal of Food Processing and Preservation 29, 33–44. https://doi.org/10.1111/j.1745-4549.2005.00011.x . Satheesh, M.A. and Pari, L., 2008. Effect of pterostilbene on lipids and lipid profiles in streptozotocin-nicotinamide induced type 2 diabetes mellitus. Journal of Applied Biomedicine 6(1), 31–37. Satheshkumar, S. and Punniamurthy, N., 2009. Estrus induction by supplementation of Murraya koenigii in anestrus heifers. Indian Journal of Animal Reproduction 30, 66–67. Shafi, T.A., Bansal, B.K. and Gupta, D.K., 2018. In vitro antibacterial activity and minimum inhibitory concentration of Ocimum sanctum leaves against common bovine mastitis pathogens. Journal of Dairy, Veterinary and Animal Research 7(6), 322–324. Sharma, P.C., Bhatia, N. and Sharama, A., 2007. A Review on Bael tree. Natural Product Radiance 6(2), 171–178. Shavit, L., Lifschitz, M. and Galperin, I., 2012. Influence of enteric nutrition on blood urea nitrogen (BUN) in very old patients with chronic kidney disease (CKD). Archives of Gerontology and Geriatrics 54(1), 228–231. https://doi.org/10.1016/j Talapatra, S.K., Ray, S.C. and Sen, K.C., 1940. The analysis of mineral constituents in biological materials. Estimation of phosphorus, chlorine, calcium, magnesium, sodium and potassium in food-stuffs. Indian Journal of Veterinary Science 10, 243–258. Tiwari, U.S., Shukla, S.N., Bajaj, N.K., Jain, A.K., Mishra, P., Kumar, J. and Tiwari, A., 2021. Fertility response using timed insemination protocols in postpartum buffaloes. Journal of Pharmacy Innovation 10(10), 1136–1138. Umashanker, A., Mehrotra, S.K., Singh, S.K. and Hoque, M., 2006. Restoration of fertility in anoestrus buffaloes using M. koenigii (Curry leaf plant). In: XXII Annual Convention and National Symposium on Innovative Technologies for Fertility Enhancement in Livestock, MHOW, Madhya Pradesh, India, pp. 283. Upadhya, S., Shanbhag, K.K., Suneetha, G. and Naidu, M.B., 2004. A study of hypoglycemic and antioxidant activity of Aegle marmelos in alloxan induced diabetic rat. Indian Journal of Physiology and Pharmacology 48, 476–480. Van Soest, P.V., Robertson, J.B. and Lewis, B.A., 1991. Methods for dietary fiber, neutral detergent fiber and non-starch polysaccharides in relation to animal nutrition. Journal of Dairy Science 74(10), 3583–3597. https://doi.org/10.3168/jds.S0022-0302(91)78551-2 . Verma, A., Kumar, P., Pandey, A.K., Kumari, S. and Kumar, S., 2019. Anestrus is a major infertility issue in perspective of buffalo reproduction in India: A review. Pharmacy Innovation Journal 8(2), 208–211. Vijayalakshmi, S. and Venkatalakshmi, P., 2017. An updated review on the phytopharmacological significance of Aegle marmelos (L.). World Journal of Pharmaceutical Research 6(16), 1390–1422. Wadhwa, M. and Bakshi, M.P.S., 2023. Effect of feeding detoxified waste bread in total mixed ration on nutrient utilization and rumen fermentation pattern in adult buffaloes. Animal Nutrition and Feed Technology 23(2), 221–231. http://dx.doi.org/10.5958/0974-181X.2023.00020.3 . Wanapat, M., Kang, S. and Polyorach, S., 2013. Development of feeding systems and strategies of supplementation to enhance rumen fermentation and ruminant production in the tropics. Journal of Animal Science and Biotechnology 4(32), 1–11. https://doi.org/10.1186/2049-1891-4-32 . Wankhede, S.D., Tambe, M.B., Dutta, N., Kaur, N., Singh, P., Jadhav, S.E. and Pattanaik, A., 2022. Effect of moringa foliage supplementation on in vitro ruminal gas production kinetics and substrate degradation in cattle. Indian Journal of Animal Science 92, 871–875. https://doi.org/10.56093/ijans.v92i7.114850 . Xie, J.T., Chang, W.T., Wang, C.Z., Mehendale, S.R., Li, J., Ambihaipahar, R., Ambihaipahar, U., Fong, H.H. and Yuan, C.S., 2006. Curry leaf (Murraya koenigii Spreng.) reduces blood cholesterol and glucose levels in ob/ob mice. American Journal of Chinese Medicine 34(02), 279–284. https://doi.org/10.1142/S0192415X06003825 . Yattoo, M.A., Chaudhary, L.C., Kala, A., Agarwal, N. and Kamra, D.N., 2023. Effect of blend of essential oils as feed additive on rumen metabolites and rumen microbial status of buffaloes. Animal Nutrition and Feed Technology 23(2), 373–380. http://dx.doi.org/10.5958/0974-181X.2023.00031.8 . Cite Share Download PDF Status: Published Journal Publication published 03 Mar, 2025 Read the published version in Tropical Animal Health and Production → Version 1 posted Reviewers agreed at journal 01 Aug, 2024 Reviewers invited by journal 27 Jun, 2024 Editor assigned by journal 21 Jun, 2024 First submitted to journal 10 Jun, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4557558","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":319945636,"identity":"7ffa9d83-3ad6-4359-b09a-15f9d256716a","order_by":0,"name":"Juli Chakma","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABA0lEQVRIie2PsWrDMBRFJQSd4i/I0C8oxATeJJwP6SIhSKbS1dBA5EVe0t1Df6Fr6PiCoFlEvRqyxEunDt7abLFdugSsdCxUZxJX73C5hAQCfxCqmUaqyYQQtj00Ke+yDP0K/VGuVFy4eZ95le6AfCsjGEfG9plXYYXM8PiSTG9ynIxHrkyec9u2LPntYEch9fbRKQAnRPyU7tXGyVZ5nd9pj2KpYRxQoPhwewUou3X2krLiUNYaI/Om+scvFAtQKZpFBhOoLrWs63aL2U2heme0cEpA1bYIz5Y4V7Y5mod4U95/fjVpMoNyUR+aJR9Wzj9kH4iB847r82DmOQ4EAoF/ygkeI3MmPrRy7gAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0009-0006-1303-9010","institution":"ICAR-Indian Veterinary Research Institute: Indian Veterinary Research Institute","correspondingAuthor":true,"prefix":"","firstName":"Juli","middleName":"","lastName":"Chakma","suffix":""},{"id":319945637,"identity":"8357990f-dcac-41ab-9c59-eddebd4bc6f9","order_by":1,"name":"N. Dutta","email":"","orcid":"","institution":"Indian Veterinary Research Institute","correspondingAuthor":false,"prefix":"","firstName":"N.","middleName":"","lastName":"Dutta","suffix":""},{"id":319945638,"identity":"52c19e28-1d6a-48f0-9af7-d804ece2122a","order_by":2,"name":"S. E. Jadhav","email":"","orcid":"","institution":"Indian Veterinary Research Institute","correspondingAuthor":false,"prefix":"","firstName":"S.","middleName":"E.","lastName":"Jadhav","suffix":""},{"id":319945639,"identity":"164eb265-263e-4f74-8c38-af8e7ceecede","order_by":3,"name":"S.K. Singh","email":"","orcid":"","institution":"Indian Veterinary Research Institute","correspondingAuthor":false,"prefix":"","firstName":"S.K.","middleName":"","lastName":"Singh","suffix":""},{"id":319945640,"identity":"f8760805-d049-4a95-92ea-8ebd79f60cbb","order_by":4,"name":"D. R. Choravada","email":"","orcid":"","institution":"Indian Veterinary Research Institute","correspondingAuthor":false,"prefix":"","firstName":"D.","middleName":"R.","lastName":"Choravada","suffix":""},{"id":319945641,"identity":"f66d07c9-05ca-4918-bbc2-ff0f595f8423","order_by":5,"name":"A. Champati","email":"","orcid":"","institution":"Central Avian Research Institute","correspondingAuthor":false,"prefix":"","firstName":"A.","middleName":"","lastName":"Champati","suffix":""},{"id":319945642,"identity":"6e60d857-15e0-45ed-9a55-8b9839a3d87c","order_by":6,"name":"S. Namdeo","email":"","orcid":"","institution":"Indian Veterinary Research Institute","correspondingAuthor":false,"prefix":"","firstName":"S.","middleName":"","lastName":"Namdeo","suffix":""},{"id":319945643,"identity":"ce481454-900b-429d-9dde-6bca51db2f29","order_by":7,"name":"N. Kaur","email":"","orcid":"","institution":"Indian Veterinary Research Institute","correspondingAuthor":false,"prefix":"","firstName":"N.","middleName":"","lastName":"Kaur","suffix":""}],"badges":[],"createdAt":"2024-06-10 10:57:17","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4557558/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4557558/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s11250-025-04351-5","type":"published","date":"2025-03-03T15:58:23+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":60699722,"identity":"69c287a7-e953-460f-b0a0-c891c250c549","added_by":"auto","created_at":"2024-07-19 17:35:56","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":20549,"visible":true,"origin":"","legend":"\u003cp\u003eReproductive performance of crossbred cows supplemented with herbal formulations (a) conception rate (b) first oestrus after start of feeding (days) and (c) number of inseminations per conception\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4557558/v1/a62594009b8a37c14f11b2d0.png"},{"id":60699725,"identity":"d6770b3f-b8c6-45b8-b1f1-947038221019","added_by":"auto","created_at":"2024-07-19 17:35:57","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":451842,"visible":true,"origin":"","legend":"\u003cp\u003eFern pattern of the cervical mucus\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-4557558/v1/0cddea37cca8614c23d75e85.png"},{"id":60699724,"identity":"dbff9e9f-889c-4090-97f7-1ec3c84bae18","added_by":"auto","created_at":"2024-07-19 17:35:56","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":11434,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of herbal formulations on pH (a) and spinnbarkeit values (b) of cervical mucus in crossbred cows\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-4557558/v1/9a980e846ed00fca7e43abbf.png"},{"id":60700085,"identity":"4fc61b1c-bee6-44db-9d3c-4baa68ab3a10","added_by":"auto","created_at":"2024-07-19 17:43:57","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":7368,"visible":true,"origin":"","legend":"\u003cp\u003emRNA expression of progesterone gene\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-4557558/v1/2380921af58c26ef90a45eee.png"},{"id":78190476,"identity":"fc8ec3a3-cd51-4d79-98bb-8e59c69d447e","added_by":"auto","created_at":"2025-03-10 19:49:21","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2083848,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4557558/v1/898e3ade-e885-498d-8626-6565bfd5446d.pdf"}],"financialInterests":"","formattedTitle":"Impact of feeding Murraya koenigii and Aegle marmelos leaves on metabolic and reproductive performance in crossbred cows","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eNatural products derived from plants have garnered significant attention within the scientific community and among the general public due to their potential role in treating, preventing, and enhancing health. The accessibility and affordability of these medicinal herbs have contributed to their popularity as a reasonably priced source of therapeutic compounds (Abeysinghe et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). In the context of livestock management, reproductive efficiency is a pivotal determinant of a sound and profitable enterprise. Reproductive diseases in dairy animals pose a substantial challenge, leading to poor reproductive efficiency and subsequent reductions in productivity (Kuralkar and Kuralkar, \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Achieving a phase of transition free of diseases, increased artificial insemination (AI), and higher pregnancy rates per service becomes imperative to augment reproductive efficiency in bovines. Despite advancements in breeding, feeding, and management practices, the reproductive efficiency of dairy animals has witnessed a decline over the past few decades, contrasting with increased production levels. The use of therapeutic agents, both hormonal and non-hormonal, has been extensive in attempts to enhance reproductive efficiency in dairy animals, albeit with varying degrees of success (Jadhav and Bhutani, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Khade et al. \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Lone et al. \u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Purkayastha et al. \u003cspan citationid=\"CR89\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Dutta et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Kumar et al. \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Verma et al. \u003cspan citationid=\"CR112\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). However, hormonal treatments have raised concerns about potential adverse effects on neuroendocrine and physiological functions, along with implications for public health due to residues in food animals and their products (Dutt et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe need for alternative, affordable methods is highlighted by the disadvantages of utilizing synthetic hormones and medications for animal reproductive management, such as their high costs, variable outcomes, and difficulty in obtaining commercial formulations. Approaches to reduce disease incidence and enhance reproductive function are being developed in order to minimize financial losses in dairy animals (Kumar et al. \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). India, known as the \"Botanical Garden of the World,\" boasts a diverse range of medicinal plants, positioning it as the leading producer of medicinal herbs globally. These herbs have significant implications for both human and animal health. Recent attention has been directed towards several herbs, including garlic, fenugreek, raspberry leaves, shatavari, thyme, black pepper, ashwagandha, and clove, which have demonstrated extraordinary therapeutic potential in combating various reproductive diseases and disorders in dairy animals (Mishra et al. \u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Kabir et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Das et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Kumar et al. \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Rajkumar et al. \u003cspan citationid=\"CR92\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Kumar et al. \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Chakma et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Shafi et al. 2020). Commercially available herbal remedies have shown variable success rates in restoring cyclicity in dairy animals, stimulating ovarian function and improving reproductive health. Notably, the past two decades have seen systematic studies on the use of medicinal plants, such as pipal (\u003cem\u003eFicus religiosa\u003c/em\u003e), curry (\u003cem\u003eMurraya koenigii\u003c/em\u003e), and bel (\u003cem\u003eAegle marmelos\u003c/em\u003e), to enhance fertility in dairy animals (Jondhale, \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Kumar, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Jondhale et al. \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2009a\u003c/span\u003e; \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003eb\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eResearches have demonstrated that extracts and powders of A. marmelos and M. koenigii, either alone or in combination, can lower inflammation and uterine infections while concurrently improving lipid profiles, insulin, glucose, and antioxidant enzyme activity (Sabu and Kuttan, \u003cspan citationid=\"CR99\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Upadhya et al. \u003cspan citationid=\"CR110\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Xie et al. \u003cspan citationid=\"CR117\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Sharma et al. \u003cspan citationid=\"CR105\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Abdallah et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Recent research has demonstrated encouraging outcomes in enhancing reproductive parameters, antioxidant responses, and immunological functions in rats through the supplementation of herbal extracts from \u003cem\u003eM. koenigii\u003c/em\u003e, \u003cem\u003eM. oleifera\u003c/em\u003e, and \u003cem\u003eA. marmelos\u003c/em\u003e (Dhivagar, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The abundance of phytochemicals in bel and curry leaves, such as marmelosin, tannins, limonene, aegelin, marmesinin, marmelin, koenigine, mukonicine, murrayacinine, isomahanine, and more, underscores their therapeutic potential (Bansal and Bansal, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Naczk and Shahidi, \u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Reddy, \u003cspan citationid=\"CR96\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Jain et al. \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Snehlata et al. 2018; Ganesan et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Chakma et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). These phytochemicals have been associated with various health benefits, including antioxidant, antimicrobial, antidiabetic, anticholesterol, anticancer, and hepatoprotective properties. Among the studied herbs, curry (\u003cem\u003eM. koenigii\u003c/em\u003e) and bel (\u003cem\u003eA. marmelos\u003c/em\u003e) stand out as significant plants examined for ovarian dynamic investigations in both laboratory and farm animals. Their effects have been evaluated in different reproductive disorders including anestrus, repeat breeding, and endometritis, showing promising results as therapeutics (Hegde et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Mehrotra et al. \u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Satheshkumar and Punniamurthy, \u003cspan citationid=\"CR103\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Nandini et al. \u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Dutt et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). Supplementation with powder or extract of \u003cem\u003eM. koenigii\u003c/em\u003e and \u003cem\u003eA. marmelos\u003c/em\u003e for 5 to 9 days has been observed to improve genital health, increase estrus expression, and enhance conception rates in dairy animals facing reproductive challenges (Baitule et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Kumawat et al. \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Tiwari et al. \u003cspan citationid=\"CR108\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Dhami et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn light of these findings, it may be possible to turn powdered herbs into extracts and pellets with the goal of lowering dosages, increasing utilization, and creating formulations based on active ingredients. Therefore, the present study hypothesizes using curry and bel leaves as a safe, potent, and cost-effective alternative strategy to improve the reproductive efficiency of crossbred cows during the postpartum period. The research will specifically address metabolic profiles, antioxidant indices, immune responses, and reproductive performances in the pursuit of sustainable and effective strategies for livestock management.\u003c/p\u003e"},{"header":"2. Material and methods","content":"\u003cp\u003eThe research was conducted at the cattle and buffalo farm of the ICAR-Indian Veterinary Research Institute (IVRI) in Izatnagar, Uttar Pradesh, India, situated at the coordinates 28.4059\u0026ordm;N and 79.4332\u0026ordm;E. In this region, the integration of crops and livestock, known as mixed farming, has been a long-standing tradition. Despite the importance of livestock, small mixed farms often suffer from low productivity due to the prevalence of non-descript breeds and inadequate nutrition. The main crops cultivated in this area are rice, wheat, and sugarcane, with cereal straws and stovers constituting the primary diet for ruminants. All experimental protocols received approval from the Institutional Animal Ethics Committee and the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), India.\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. \u003cem\u003ePreparation of the plant material\u003c/em\u003e\u003c/h2\u003e \u003cdiv id=\"Sec4\" class=\"Section3\"\u003e \u003ch2\u003e2.1.1. \u003cem\u003eCollection, drying and storage of plant material\u003c/em\u003e\u003c/h2\u003e \u003cp\u003eThe \u003cem\u003eM. koenigii\u003c/em\u003e and \u003cem\u003eA. marmelos\u003c/em\u003e leaves were collected in a single batch from the ICAR-IVRI, Izatnagar in September 2022. After being air-dried at a room temperature of 27\u0026ordm;C, the leaves were ground using an electric grinder. The dried and ground leaves were then combined in equal parts, creating a 1:1 ratio herbal formulation.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section3\"\u003e \u003ch2\u003e2.1.2. Preparation of herbal pellets\u003c/h2\u003e \u003cp\u003eThe herbal powder was further used to prepare pellets, which underwent processing in a feed pelleting unit. Within the pellet-making machine, this mixture was passed through the die which was of 10 mm size, the mixture was churned and pressed, transforming it into pellet form. The resulting pellets averaged 4\u0026ndash;5 cm in length and had a diameter of 10 mm.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section3\"\u003e \u003ch2\u003e2.1.3. Methanolic extract preparation\u003c/h2\u003e \u003cp\u003eIn a Stoppard conical flask, 100 g of dried herbal powder underwent extraction with 1000 mL of methanol-water solution (70:30 ratio) at room temperature. The mixture was agitated for 12 h using an electric shaker. Subsequently, the extracts were filtered using a cotton wool plug and filter paper. The filtrate was then dried at 39\u0026deg;C for future use. The resulting dried powder was crushed and stored in bottles at 4\u0026deg;C, as described by Begashaw et al. (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Animals and treatments\u003c/h2\u003e \u003cp\u003eTwenty-four crossbred cows (60 d post-partum) of 2nd to 4th parity were distributed into four treatment groups consisting of six animals in each in a CRD. The cows in all the treatment groups were provided iso-nitrogenous and iso-caloric diets. The experimental cows were randomly divided into 4 treatment groups (CON, HPL, HEX and HP). CON (control) group animals were fed a diet comprised of green fodder, concentrate mixture and wheat straw, while, the experimental cows in HP (herbal powder), HPL (herbal pellets) and HEX (herbal extract) groups were offered CON diet having herbal powder (200gd\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), herbal pellets (200gd\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) and methanolic herbal extract (equivalent to 200g herbal powder d\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), respectively. The experimental cows were fed on treatment diets for 7 d and continued with CON diet up to one month, further after one month of treatment the non-responsive cows (not exhibited oestrus) were given treatment again in different groups and monitored for another one month to assess the response in respect of oestrus induction and pregnancy establishment, metabolic profile, health, production and reproduction performance. The pregnancy diagnosis was done per rectal after 60 d of AI. The total duration of the experiment was of 120 d.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.3. Collection of blood and separation of serum\u003c/h2\u003e \u003cp\u003eThe blood samples were collected from jugular venipuncture on 0, 3, 7, 14 and 21 day of feeding by following standard protocol with with clean, sterilized needles, early in the morning before feeding and watering. About 10 mL blood was collected, out of which 2 mL was placed into vacutainers containing anticoagulant (EDTA) for estimation of Hb and haematocrit. Remaining 8.0 mL blood was dispensed into serum vacutainers and allowed to clot at ambient temperature for 45 minutes by keeping at slanting position. The tubes were spun at 2,000 xg for 10 minutes using a refrigerated centrifuge. The collected sera were transferred into Eppendorf tube with Pasteur pipette and stored in deep freeze at -20\u003csup\u003e0\u003c/sup\u003eC for further serum-biochemical analysis. Haemoglobin (Hb) and haematocrit were measured in whole blood immediately after blood collection. Haemoglobin (Hb) was measured by using the cyanomethaemoglobin technique (Dacie and Lewis, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e1969\u003c/span\u003e) and hematocrit was measured using Wintrobe\u0026rsquo;s tube. The serum glucose, NEFA, total proteins, albumin, urea, creatinine, total cholesterol, aspartate transaminase (AST) and alanine aminotransferase (ALT) were analyzed spectrophotometrically (MultiskanTM FC Microplate Photometer with incubator, Thermo Scientific Ltd.) by using diagnostic kits manufactured by Coral Clinical Systems (Goa, India) following the standard protocol provided with the kits. The major minerals (Ca and P) were analyzed by using the method of Talapatra et al. (\u003cspan citationid=\"CR107\" class=\"CitationRef\"\u003e1940\u003c/span\u003e) and trace minerals such as Cu, Zn, Mn and iron were analyzed by Atomic Absorption Spectroscopy (AAS) method\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e2.4. Chemical analysis\u003c/h2\u003e \u003cp\u003eSamples of the concentrate mixture, wheat straw, green fodder, herbal pellets, herbal extracts and herbal powder were milled to pass through a 1.0 mm sieve and then examined for proximate principles (AOAC, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). The neutral detergent fibre (NDF) and acid detergent fibre (ADF) were measured following the methods of Van Soest et al. (\u003cspan citationid=\"CR111\" class=\"CitationRef\"\u003e1991\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e2.5. Gene expression study\u003c/h2\u003e \u003cp\u003eFor gene expression study, already published primer of progesterone (PGR) TAATCTGTGGGGATGAAGCA-F and TAATCTGTGGGGATGAAGCA-R was standardized and used (Driver et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). The PCR conditions were optimized by amplifying the cDNA with specific primers at various annealing temperatures. The specificity of each primer was verified before performing the qRT-PCR analysis. The relative change in gene expression was analyzed using the 2-ΔΔCT method, as detailed by Livak and Schnittger (2001).\u003c/p\u003e \u003cp\u003eRelative fold change\u0026thinsp;=\u0026thinsp;2\u003csup\u003e\u0026minus;ΔΔCT\u003c/sup\u003e, where, ΔΔCT= (C\u003csub\u003eT\u003c/sub\u003e, target gene- C\u003csub\u003eT\u003c/sub\u003e, reference gene) test \u0026ndash; (C\u003csub\u003eT\u003c/sub\u003e, target gene- C\u003csub\u003eT\u003c/sub\u003e, reference gene) housekeeping gene.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e2.6. Reproductive performance\u003c/h2\u003e \u003cp\u003eOestrus detection was done for all the lactating crossbred cows twice a day throughout the study period with the help of a trained teaser bull. All the postpartum crossbred cows were inseminated artificially with frozen semen which were exhibited in oestrus. All the non-return cows were diagnosed for pregnancy after 60 d of insemination by trans-rectal palpation. The different postpartum reproductive parameters like onset of oestrus, services per conception and conception rate was recorded. All the experimental cows were closely monitored every day for any types of disease or disorders such as mastitis, metritis etc.\u003c/p\u003e \u003cdiv id=\"Sec12\" class=\"Section3\"\u003e \u003ch2\u003e2.6.1. Cervical mucus characteristics\u003c/h2\u003e \u003cp\u003eCervical mucus was obtained prior to artificial insemination using mid-cervical aspiration utilizing sterile blue sheaths and a universal artificial insemination gun via the recto-vaginal method. The blue sheath was attached to the universal artificial insemination gun to act as an extended syringe. The insemination gun's piston functioned as a plunger, facilitating the extraction of the cervical mucus. Following the collection, 5 mL of the cervical mucus was transferred immediately to the laboratory in a sterile petri dish for analysis of the specified parameters:\u003c/p\u003e \u003cdiv id=\"Sec13\" class=\"Section4\"\u003e \u003ch2\u003e2.6.1.1. Spinnbarkeit value\u003c/h2\u003e \u003cp\u003eFew drops (2\u0026ndash;3 drops) of cervical mucus were placed on a grease-free slide, with another clean slide positioned on top. The slides were then gently pulled apart to stretch the mucus until it broke. The distance between the two slides at the point of breakage was measured using a centimeter scale mounted on a wall. (Panigrahi, \u003cspan citationid=\"CR82\" class=\"CitationRef\"\u003e1964\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section4\"\u003e \u003ch2\u003e2.6.1.2. Arborization pattern and pH\u003c/h2\u003e \u003cp\u003eTo analyze the arborization pattern, 2\u0026ndash;3 drops of cervical mucus were placed on a grease-free glass slide and spread evenly. The sample was air-dried and examined under a low-power objective (10X) of a microscope. Fern patterns were classified into typical, atypical, and absent categories. pH was determined by using a pH indicator stick (full range, pH 1.0 to 14.0) - Qualigens (#Q38141, Thermo Fisher Scientific) which was saturated with a drop of cervical mucus placed on a clean slide. At least 2 sticks with overlapping pH ranges (5.4 to 7.0 and 6.4 to 8.0) were used to test the same sample.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003e2.7. Statistical analysis\u003c/h2\u003e \u003cp\u003eThe results obtained were statistically analyzed using IBM, SPSS (20.0) and through Duncan\u0026rsquo;s Multiple Range test, means were categorized accordingly. The parameters from periodic collections were scrutinized by implementing repeated measures protocol through GLM of SPSS; which encompassed between-subjects principal effect of treatments, within-subjects principal effect of period and interaction amid the treatment and period. The P\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was used to declare the means to be significant. All the statistical procedures were used as per Snedecor and Cochran (1994).\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\n \u003ch2\u003e3.6. Chemical composition of feedstuffs offered (% DM basis)\u003c/h2\u003e\n \u003cp\u003eThe chemical composition and fibre fractions of wheat straw, green, concentrate, pellets, extract and powder supplements are mentioned in Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e. The chemical composition of herbal pellets and powder was found to be comparable, however, herbal extract showed lower values of EE, TA, NDF and ADF as compared to herbal pellets and powder.\u0026nbsp;\u003c/p\u003e\n \u003ctable id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eChemical composition of feeds offered (% DM basis)\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eAttributes (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eWheat straw\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eGreen fodder\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eConcentrate mixture\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHerbal pellets\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHerbal extract\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHerbal powder\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eDM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e91.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10.60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e86.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e92.94\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e90.16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e94.11\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e93.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e87.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e92.68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e87.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e85.42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e88.15\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e17.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e13.41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.14\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eEE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.21\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e7.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11.85\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNDF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e85.42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e75.61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e43.53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e53.68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e31.56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e51.16\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eADF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e51.21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e41.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e16.85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e32.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e23.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e29.36\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"7\"\u003e[DM: Dry matter; OM: Organic matter; CP: Crude protein; EE: Ether extract; TA: Total ash; NDF: Neutral detergent fibre; ADF: Acid detergent fibre]\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\n \u003ch2\u003e3.7. Metabolic profile\u003c/h2\u003e\n \u003cdiv id=\"Sec19\" class=\"Section3\"\u003e\n \u003ch2\u003e3.2.1. Hb and haematocrit\u003c/h2\u003e\n \u003cp\u003eThe haematocrit (%) and Hb (gdL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) levels are presented in Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. The levels of Hb (gdL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) were analogous (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05) among the treatment groups. Mean values of haematocrit were significantly higher (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) in the HEX group relative to CON, while HPL and HP groups had intermediate response between CON and HEX.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec20\" class=\"Section3\"\u003e\n \u003ch2\u003e3.7.1. Serum biochemical parameters\u003c/h2\u003e\n \u003cp\u003eThe results related with serum glucose, NEFA, total protein and its fractions, serum urea, total cholesterol, creatinine, AST and ALT are presented in Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. The serum glucose (mgdL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) levels were comparable (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05) amongst the dietary treatments, though serum NEFA (mmolL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) values were significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) lower in HPL group followed by HEX, HP and CON groups. Serum NEFA was significantly reduced (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) from 3rd d onwards. The mean values of total protein, albumin, globulin (gdL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), A: G ratio and total cholesterol (mgdL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) were parallel (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05) amongst the dietary treatments. The serum urea level was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) lower in HPL and HEX groups relative to CON, however, HP group had an intermediate position among HPL, HEX and CON. The serum creatinine (mg %) concentration was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) lower in HPL, HEX and HP as compared to CON. Activities of serum enzymes viz. AST and ALT were similar among the treatment groups.\u0026nbsp;\u003c/p\u003e\n \u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eEffect of herbal formulations supplementation on metabolic profile of crossbred cows\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eAttributes\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003eTreatments\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eSEM\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eP-value\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eCON\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHPL\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHP\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHP\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHb, gdl\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.213\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHaematocrit, %\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e34.73\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e36.56\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e36.82\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e35.03\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.049\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGlucose, mgdl\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e47.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e48.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e48.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e48.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.740\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNEFA, mmolL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.43\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.37\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.39\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.39\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTotal protein, gdl\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.87\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.137\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAlbumin, gdl\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.260\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGlobulin, gdl\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.400\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA: G ratio\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTotal cholesterol, mgdl\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e142.47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e140.84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e141.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e141.55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.964\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUrea, mgdl\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e35.96\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e32.48\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e31.79\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e33.55\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.02\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCreatinine, mg%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.23\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.10\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.11\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.11\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eALT, UL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e32.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e31.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e32.47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e32.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.903\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAST, UL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e90.65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e92.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e88.62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e89.91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.135\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"7\"\u003e\u003csup\u003eabc\u003c/sup\u003e and \u003csup\u003exyz\u003c/sup\u003e means bearing different superscript in a row and column differ significantly\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec21\" class=\"Section3\"\u003e\n \u003ch2\u003e3.7.2. Serum mineral profile\u003c/h2\u003e\n \u003cp\u003eSerum mineral such as Ca, i-P, Cu, Zn, Mn and iron are presented in Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e. The mean values for serum Ca (mgdL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) were parallel (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05) among the treatment groups, however, serum i-P (mgdL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) levels were significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) higher in HPL, HEX and HP in relative to CON. The mean for serum Ca were significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher at 3rd d, however, i-P values were increased 3rd d onwards. The mean values of Cu, Zn, Mn and iron were significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) higher in treatment groups as compared to CON. Serum Cu and Zn levels were noticeably (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) higher in HPL followed by HEX and HP and CON. Serum Mn values were significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) higher in HPL followed by HEX, HP and CON group. The levels of serum iron were significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) higher in HPL and HP groups relative to CON, while HEX had an intermediate position between CON and HP groups. The values of all the trace minerals were increased significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) from 3rd d onwards. The treatments and periods interactions were also significantly (P\u0026thinsp;\u0026gt;\u0026thinsp;0.01).\u0026nbsp;\u003c/p\u003e\n \u003ctable id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eSerum mineral profile of crossbred cow supplemented with herbal formulations\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eTreatments\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"5\"\u003e\n \u003cp\u003ePeriod (s)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eTreatment mean\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eSEM\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003eP-value\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0\u003c/strong\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e3\u003c/strong\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e7\u003c/strong\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e14\u003c/strong\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e21\u003c/strong\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eT\u003c/strong\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eP\u003c/strong\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eTxP\u003c/strong\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"12\"\u003e\n \u003cp\u003eCa, mgdl\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCON\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.213\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" rowspan=\"5\"\u003e\n \u003cp\u003e0.015\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.828\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHPL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13.85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.94\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHEX\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePeriod mean\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.52\u003csup\u003exy\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.80\u003csup\u003ey\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.66\u003csup\u003ex\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.71\u003csup\u003ex\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.96\u003csup\u003exy\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"12\"\u003e\n \u003cp\u003ei-P, mgdl\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCON\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.85\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.025\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" rowspan=\"5\"\u003e\n \u003cp\u003e0.003\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.752\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHPL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.94\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.39\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHEX\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.51\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.95\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.32\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePeriod mean\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.63\u003csup\u003ex\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.50\u003csup\u003ey\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.45\u003csup\u003ey\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.24\u003csup\u003ey\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.53\u003csup\u003ey\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"12\"\u003e\n \u003cp\u003eCu, \u0026micro;gml\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCON\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.76\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" rowspan=\"5\"\u003e\n \u003cp\u003e0.000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.002\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHPL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.14\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHEX\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.02\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.99\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePeriod mean\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.75\u003csup\u003ex\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.03\u003csup\u003ey\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.06\u003csup\u003ey\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.05\u003csup\u003ey\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.01\u003csup\u003ey\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"12\"\u003e\n \u003cp\u003eZn, \u0026micro;gml\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCON\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.77\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" rowspan=\"5\"\u003e\n \u003cp\u003e0.000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.053\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHPL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.12\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHEX\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.02\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.05\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePeriod mean\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.73\u003csup\u003ex\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.95\u003csup\u003ey\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.10\u003csup\u003ez\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.07\u003csup\u003ez\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.11\u003csup\u003ez\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"12\"\u003e\n \u003cp\u003eMn, \u0026micro;gml\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCON\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.27\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" rowspan=\"5\"\u003e\n \u003cp\u003e0.000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.018\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHPL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.35\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHEX\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.34\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.32\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePeriod mean\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.25\u003csup\u003ew\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.31\u003csup\u003ex\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.33\u003csup\u003exy\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.34\u003csup\u003eyz\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.35\u003csup\u003ez\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"12\"\u003e\n \u003cp\u003eIron, \u0026micro;gml\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCON\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.61\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" rowspan=\"5\"\u003e\n \u003cp\u003e0.000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e0.000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHPL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.40\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHEX\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.27\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.16\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePeriod mean\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.48\u003csup\u003ew\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.88\u003csup\u003ex\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.22\u003csup\u003ey\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.48\u003csup\u003ez\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.49\u003csup\u003ez\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"14\"\u003e\u003csup\u003eabc\u003c/sup\u003e and \u003csup\u003exyz\u003c/sup\u003e means bearing different superscript in a row and column differ significantly\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec22\" class=\"Section2\"\u003e\n \u003ch2\u003e3.8. Reproductive performance\u003c/h2\u003e\n \u003cp\u003eSupplementation of herbal formulation in different forms showed considerable improvement on confirmed pregnancy rate in crossbred cows. The conception at first insemination was considerably higher in HPL, HEX and HP groups relative to CON, while, conception at subsequent insemination was found higher in HPL, HEX and CON than HP group. The cows in treatment groups came into heat (first oestrus) earlier (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) as compared to CON. However, 66.7, 16.67, 16.7 and 33.3% of cows in CON, HPL, HEX and HP did not show any sign of oestrus and were non-responsive. The number of AIs per conception were higher (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) in CON relative to treatment groups (Fig. 1).\u003c/p\u003e\n \u003cdiv id=\"Sec23\" class=\"Section3\"\u003e\n \u003ch2\u003e3.3.1. Physical characteristic of cervical mucus\u003c/h2\u003e\n \u003cp\u003eThe pH and spinnbarkeit values (cm) of cervical mucus were significantly higher (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) in the HP, HPL, and HEX groups compared to the CON (Figs. 2 and 3). The arborization pattern (fern pattern) of cervical mucus with typical fern pattern exhibited higher conception rate in all the treatment (HP, HPL and HEX) groups in relation to atypical fern pattern in CON group is depicted in Table \u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003e.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\n \u003cdiv align=\"left\" class=\"colspec\"\u003eTable 7 Conception rate in relation to fern pattern of cervical mucus in crossbred cows fed herbal formulation\u003c/div\u003e\u0026nbsp;\u003ctable id=\"Tabb\" border=\"1\"\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTreatments\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eArborization pattern (fern pattern)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eConception rate (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eCON\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTypical\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.66\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAtypical\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.66\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eHPL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTypical\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e66.66\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAtypical\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.66\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eHEX\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTypical\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e83.33\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAtypical\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eHP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTypical\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAtypical\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.66\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec24\" class=\"Section2\"\u003e\n \u003ch2\u003e3.9. Relative mRNA expression of PGR genes\u003c/h2\u003e\n \u003cp\u003eThe data pertaining to gene expression of progesterone (PGR) is depicted in Fig. 4. The fold expression of PGR gene was significantly higher (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) in HPL, HEX and HP groups compared to CON.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cdiv id=\"Sec26\" class=\"Section2\"\u003e \u003ch2\u003e4.6. Chemical composition of feeds\u003c/h2\u003e \u003cp\u003eThe chemical composition of wheat straw and green fodder offered to experimental cows was within the normal range as reported by previous workers (Negi et al. \u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Kumar et al. \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Wadhwa and Bakshi, \u003cspan citationid=\"CR114\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Yatto et al. 2023). The DM, OM, CP, EE, TA, NDF and ADF of the concentrate mixture was also observed to be consistent with the results obtained by earlier workers (Choudhary et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Patir et al. 2023; Singh et al. 2023). The composition of herbal powder and pellets was similar, while herbal extract had lower EE, TA, NDF and ADF values.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec27\" class=\"Section2\"\u003e \u003ch2\u003e4.7. Hb and haematocrit\u003c/h2\u003e \u003cp\u003eThe present investigation demonstrates a noteworthy increase (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) in haematocrit levels in the HEX group following administration of herbal formulation, while levels of Hb remained consistent across all treatment cohorts. These findings align with the established normal range for cattle, as noted by Kaneko et al. (\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). This elevation in haematocrit may be attributed to the potential enhancement of blood circulation facilitated by the herbal mixtures, ensuring efficient transportation of red blood cells throughout the body. The observed rise in haematocrit values could also be associated with the presence of flavonoids and quercetin, known for their hematopoietic properties (Raja et al. \u003cspan citationid=\"CR91\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). Furthermore, it's noteworthy that the herbal formulation contains volatile compounds (Mendoza et al. \u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; S\u0026aacute;nchez et al. \u003cspan citationid=\"CR100\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Our findings corroborate those reported by Nanda et al. (\u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e2013\u003c/span\u003e), who documented a significant increase (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) in haematocrit concentration in goats following supplementation with a polyherbal formulation. Similarly, Barkakati and Kalita (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) observed a significant elevation in Hb levels with polyherbal formulation supplementation compared to the CON group in cattle. Choudhury and Sinha (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) observed a significant increase (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) in haematocrit concentration in rats with the addition of \u003cem\u003eM. koenigii\u003c/em\u003e leaves extract at doses of 250\u0026ndash;500 mg/kg BW compared to the CON group. Lee-Rangel et al. (\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) also reported a significant increase in haematocrit concentration in calves treated with a polyherbal phytogenic source.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec28\" class=\"Section2\"\u003e \u003ch2\u003e4.8. Serum glucose and NEFA\u003c/h2\u003e \u003cp\u003eThe synthesis, breakdown, and transport of lipids in the body have all been demonstrated to be influenced by herbal formulations containing polyphenols and flavonoids, which may decrease the release of NEFA from adipose tissue and increase lipid clearance from the bloodstream, thus lowering circulating NEFA levels (Hashemzadeh- Cigari et al. 2014; Baghbadorani et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Furthermore, the decreased NEFA concentration could be due to the presence of bioactive compounds in herbal formulations (\u003cem\u003eM. koenigii\u003c/em\u003e and \u003cem\u003eA. marmelos\u003c/em\u003e) that enhanced energy metabolism and nutrient absorption in cows (Lopreiato et al. \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). The improved energy utilization could lead to a reduction in circulating NEFA levels, as the cow\u0026rsquo;s body is better able to use energy sources from the diet. Nevertheless, these herbs are known to have hepatoprotective properties, supporting the liver\u0026rsquo;s function and metabolism. A healthier liver might contribute to better lipid metabolism, reducing the release of NEFAs into the bloodstream (Adewuyi et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). The present results are in concurrence with Barjibhe et al. (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), who reported that polyherbal supplementation to cows significantly reduced the NEFA concentration in serum. In addition, cows supplemented with herbal extract in the treatment group had a lower (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) concentration of serum NEFA (Baghbadorani et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Razo Ortiz et al. (\u003cspan citationid=\"CR95\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) reported that supplementation of polyherbal mixture (Ashwagandha, tulsi, gooseberry and giloy) to lambs did not have any significant effect to glucose level. Lozano-S\u0026aacute;nchez et al. (\u003cspan citationid=\"CR100\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) also observed that there was no significant effect on serum glucose with supplementation of polyherbal supplement in finishing lambs. Hashemzadeh-Cigari et al. (\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) observed that glucose level was not affected, but serum NEFA reduced in cows supplemented with herbal mixture as compared to the CON group, despite their analogous energy balance. Although the precise mechanisms responsible for the lower NEFA levels in cows supplemented with the herbal mixture remain unclear, it is proposed that the herbal mixture may inhibit lipolysis, thereby improving lipid metabolism and enhancing metabolic status in periparturient cows by increasing insulin sensitivity.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec29\" class=\"Section2\"\u003e \u003ch2\u003e4.9. Total protein, albumin, globulin and A: G ratio\u003c/h2\u003e \u003cp\u003eBlood metabolites such as total protein, albumin, globulin, and A:G ratio were assessed as indicators of the general health and vitality of the animals. The results revealed that these blood metabolites did not show significant differences (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05) among the various dietary treatments. These observation align with previous research conducted by Birudu et al. (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), who similarly found no significant changes in A/G ratio and albumin levels in rats treated with methanolic leaf extracts of \u003cem\u003eA. marmelos\u003c/em\u003e compared to control groups. Adebajo et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2006\u003c/span\u003e) also observed no significant impact on serum total protein, albumin, and globulin levels in Swiss Albino rats when supplemented with methanolic extracts of \u003cem\u003eM. koenigii\u003c/em\u003e at different dosage rates. However, contrary results were reported by Gandhi et al. (\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2012\u003c/span\u003e), who noted a significant reduction in total protein levels in rats treated with \u003cem\u003eA. marmelos\u003c/em\u003e. Furthermore, Dhami et al. (\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) reported varying plasma protein concentrations among different animal categories upon bi-herbal therapy with \u003cem\u003eM. koenigii\u003c/em\u003e and \u003cem\u003eA. marmelos\u003c/em\u003e, with lowered levels observed in buffaloes and infertile heifers compared to cows. Rivero et al. (\u003cspan citationid=\"CR97\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) observed an increase in plasma protein levels after adding an extract rich in phenols to lamb diets, consistent with the findings of Lee-Rangel et al. (\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), who observed increased serum protein concentration in calves treated with a polyherbal phytogenic source. Furthermore, Dorantes-Iturbide et al. (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) found that lambs fed a dietary supplement with a polyherbal additive containing hydrolyzable tannins, flavonoids, and essential oils exhibited a significant increase in serum albumin concentration. The analogous values of serum proteins observed in our study may be attributed to the relatively short duration of the treatment (7 days) compared to previous experiments, which were conducted for varying durations.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec30\" class=\"Section2\"\u003e \u003ch2\u003e4.10. Total cholesterol\u003c/h2\u003e \u003cp\u003eThe feeding of the herbal formulation to dairy cows did not show a significant effect (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05) among the treatments. In this study, supplementation with the herbal formulation had no effect on serum cholesterol levels, suggesting that the bioactive metabolites did not impact intestinal absorption or lipid synthesis in the cells of cows (Dorantes-Iturbide et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). However, Razo Ortiz et al. (\u003cspan citationid=\"CR95\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), who also reported that supplementation of polyherbal mixture (\u003cem\u003eAshwagandha, tulsi, gooseberry and giloy\u003c/em\u003e) to lambs do not have any significant effect to serum cholesterol level. In contrast to our findings, Satheesh and Pari, (\u003cspan citationid=\"CR102\" class=\"CitationRef\"\u003e2008\u003c/span\u003e) reported that saponins extracted from A. marmelos reduce blood cholesterol levels by competing with cholesterol for binding sites or by interfering with cholesterol biosynthesis in the liver. Priyadarshinee et al. (\u003cspan citationid=\"CR88\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) observed that \u003cem\u003eM. koenigii\u003c/em\u003e and \u003cem\u003eA. marmelos\u003c/em\u003e leaves supplementation in cows significantly reduced the total cholesterol in treatment groups. The comparable cholesterol concentration in present study may be attributed to the short duration (7 d) of herbal treatment.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec31\" class=\"Section2\"\u003e \u003ch2\u003e4.11. Serum urea, creatinine and serum enzymes\u003c/h2\u003e \u003cp\u003eA significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) reduction was observed in serum urea concentration in the HEX and HPL groups, indicating a decrease in rumen protein breakdown. This suggests enhanced EAA absorption in HEX, HPL, and HP groups (Pathak et al. \u003cspan citationid=\"CR84\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Wankhede et al. \u003cspan citationid=\"CR116\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Moreover, the decrease in serum urea indicates minimal protein catabolism and normal kidney function (Roseler et al. \u003cspan citationid=\"CR98\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Hosten, \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e1990\u003c/span\u003e; Shavit et al. \u003cspan citationid=\"CR106\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). The serum creatinine value was also significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) lower in treatment groups as compared to CON. This beneficial effect may be due to the presence of polyphenols, tannins, and flavonoids in the herbal formulations. These compounds exhibit antioxidant activity and anti-inflammatory effects, and provide cardiovascular health benefits, potentially aiding kidney health by reducing oxidative stress and inflammation in renal tissues. Previous studies have associated these compounds with potential protection against kidney damage (Arulselvan et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). The findings are consistent with the observations of Arulselvan et al. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2006\u003c/span\u003e), who found that blood urea and plasma creatinine decreased in treatment groups when experimental rats were treated with \u003cem\u003eM. koenigii\u003c/em\u003e, which provides addition al evidence to prove the anti-diabetogenic property of \u003cem\u003eM. koenigii\u003c/em\u003e leaf extract.\u003c/p\u003e \u003cp\u003eThe primary serum markers for assessing liver changes include AST, ALT, total protein, and total albumin were unaffected by the treatment involving herbal formulation mixture, indicating no liver damage (D\u0026iacute;az Galv\u0026aacute;n et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Furthermore, its polyphenols might provide an antioxidant benefit (Biswas and Giri, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), along with other advantageous effects on liver function due to this nutrient (Mehedint and Zeisel, \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e2013\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec32\" class=\"Section2\"\u003e \u003ch2\u003e4.12. Serum mineral profile\u003c/h2\u003e \u003cp\u003eMacro and micro elements play a crucial role in maintaining immune function and overall health. The high magnesium content in curry leaves suggests that they could serve as an excellent source of magnesium, which is known to activate various enzyme systems (Olusanya et al. 2008). Ca and P are also critical, as they influence an animal's ability to utilize other micronutrients and can modulate reproductive hormone sensitivity through various enzyme systems (Dutta et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). This could explain why the supplementation of curry leaves might lead to higher serum Ca and P levels and, in turn, support ovarian activity in anestrous heifers. Additionally, both \u003cem\u003eM. koenigii\u003c/em\u003e and \u003cem\u003eA. marmelos\u003c/em\u003e leaves are rich in essential minerals such as C, P, iron, zinc, manganese, and copper (Shantala and Prakash, \u003cspan citationid=\"CR101\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Igara et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Vijayalakshmi and Venkatalakshmi, \u003cspan citationid=\"CR113\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Kujur et al. \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Our results align with Orzuna-Orzuna et al. (\u003cspan citationid=\"CR81\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), who found that serum Ca levels among treatment groups were within the normal range for ovine, indicating that herbal mixtures do not disrupt mineral balance or nutritional status in lambs. In a similar vein, Sathesh Kumar and Punniamurthy (2009) observed that daily supplementation with 100 gd\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of \u003cem\u003eM. koenigii\u003c/em\u003e leaves for 30 days significantly improved serum inorganic i-P levels and induced estrus in anestrous heifers. Similarly, Razo Ortiz et al. (\u003cspan citationid=\"CR95\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) found that serum calcium levels were not affected by polyphenol and flavonoid doses, but serum phosphorus concentrations rose with increased doses of these compounds. Dhami et al. (\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) reported that supplementing cows with a mixture of \u003cem\u003eM. koenigii\u003c/em\u003e and \u003cem\u003eA. marmelos\u003c/em\u003e led to significant increases in plasma Ca, i-P, and other micro-minerals such as Zn, iron, and Cu. It is obvious that the mineral composition of the tree foliage is superior to that of tropical grasses. Goodchild and McMeniman (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e1994\u003c/span\u003e) reported that inclusion of tree foliage in ruminants\u0026rsquo; diet increased the supply of minerals to the animals and rumen microbes and ultimately improved their performance. However, in contrary to our findings, Baitule et al. (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) noted significant increases in serum Ca levels on day 9, the day of estrus, and on day 8 after estrus onset in treated groups, possibly due to supplementation with \u003cem\u003eA. marmelos\u003c/em\u003e and \u003cem\u003eM. koenigii\u003c/em\u003e. Likewise, Kumawat et al. (\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) found significantly higher serum Ca levels on days 4, 6, 8, and 10 after treating delayed pubertal heifers with dried leaf powder from \u003cem\u003eA. marmelos\u003c/em\u003e and \u003cem\u003eM. koenigii\u003c/em\u003e, compared to non-treated groups. This study also observed that the effect of these leaves on serum P levels was significant, with marked variations on days 2, 4, and 8 after treatment compared to non-treated heifers.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec33\" class=\"Section2\"\u003e \u003ch2\u003e4.13. Reproduction\u003c/h2\u003e \u003cp\u003eSupplementation of herbal formulation in different forms showed considerable improvement on confirmed pregnancy rate in crossbred cows (Patton et al. \u003cspan citationid=\"CR87\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Ambrose, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The beneficial effects of herbal formulations (\u003cem\u003eA. marmelos\u003c/em\u003e and \u003cem\u003eM. koenigii\u003c/em\u003e) supplementation could be due to the presence of phytoestrogens, minerals, and antioxidants. These components might stimulate gonadotropin hormones from the anterior pituitary, potentially contributing to the restoration of cyclicity in anestrus animals (Dutt et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). The antioxidants present may have enhanced folliculogenesis by preventing ROS-induced apoptosis of oocytes (Vijayalakshmi and Venkatalakshmi, \u003cspan citationid=\"CR113\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The phytoconstituents might have stimulated steroidogenic activity in the hypothalamus, promoted follicular growth, and facilitated steroid production, leading to the initiation of cyclicity (Jondhale et al. \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2009a\u003c/span\u003e; Nandini et al. \u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Additionally, the presence of bioactive compounds in the herbal formulation may have contributed to follicular development, particularly during the dominance phase leading up to ovulation, resulting in the resumption of estrus. In the present study, though the conception rate was significantly higher in all the treatment groups relative to CON, however, HPL and HEX groups showed better conception rate and established pregnancy than HP (herbal powder) group, which could be probably due to the less wastage and complete consumption of herbal powder in pellets forms as compared to powder (Wanapat et al. \u003cspan citationid=\"CR115\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). The findings of the current study align with those of Patel and Gupta (\u003cspan citationid=\"CR83\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), who reported the highest conception rate (71.42%) in delayed pubertal heifers supplemented with \u003cem\u003eM. koenigii\u003c/em\u003e, \u003cem\u003eA. marmelos\u003c/em\u003e, and a mineral mixture. Additionally, \u003cem\u003eM. koenigii\u003c/em\u003e, whether used alone or in combination with a mineral supplement and a low dose of GnRH injection, was found to be equally effective in inducing fertility in anestrus buffaloes (Umashanker et al. \u003cspan citationid=\"CR109\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). Delayed pubertal heifers have shown induced estrus behavior after being supplemented with \u003cem\u003eA. marmelos\u003c/em\u003e combined with \u003cem\u003eM. koenigii\u003c/em\u003e (Kumar et al. \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Kumawat et al. \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Baitule et al. (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) found 75% oestrus induction after supplementing \u003cem\u003eA. marmelos\u003c/em\u003e and \u003cem\u003eM. koenigii\u003c/em\u003e for 9 days in anestrous buffaloes. Satheshkumar and Punniamurthy (\u003cspan citationid=\"CR103\" class=\"CitationRef\"\u003e2009\u003c/span\u003e) also studied that supplemental effect of \u003cem\u003eM. koenigii\u003c/em\u003e leaves for a period of 30 days and found induced oestrus in 60% of the anestrus heifers. Mehrotra (\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e2002\u003c/span\u003e) have also documented that \u003cem\u003eM. koenigii\u003c/em\u003e has been found to enhance ovarian function in rats and improve the therapeutic response in acyclic goats and heifers, particularly regarding oestrus and conception. Tiwari et al. (\u003cspan citationid=\"CR108\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) reported that the oestrus response was 83.33% in postpartum buffaloes when supplemented with a mixture of \u003cem\u003eA. marmelos\u003c/em\u003e and \u003cem\u003eM. koenigii\u003c/em\u003e. However, Das et al. (\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) observed 92.90% oestrus induction in delayed pubertal buffalo heifers after feeding of \u003cem\u003eA. marmelos\u003c/em\u003e and \u003cem\u003eM. koenigii\u003c/em\u003e herbs for 9 days. Das et al. (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) and Kumawat et al. (\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) reported that 90.9% and 92.3% of delayed pubertal heifers, respectively, began cycling after being fed a herbal mixture. Additionally, they observed a pregnancy rate of 54.1% in the group treated with the herbal mixture, compared to 16.7% in the control group. Therefore, it may be speculated that favourable effects of herbal formulations (mixture of \u003cem\u003eM. koenigii\u003c/em\u003e and \u003cem\u003eA. marmelos\u003c/em\u003e) in terms of estrus induction, ovulation, established pregnancy and shorter time period taken in induction of estrus are exhibited as compared to individual plant treatment which could be a reflection of synergistic actions of herbs active principles (Nandini et al. \u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Hedge et al. 2002; Mehrotra et al. \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). The active principles seem to operate either through mimicking gonadotrophins activity or stimulating the central mechanism for endogenous release of gonadotrophins along with possibility of local action (Dutt et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec34\" class=\"Section2\"\u003e \u003ch2\u003e4.14. Physical characteristic of cervical mucus\u003c/h2\u003e \u003cp\u003eIt is believed that the herbal formulations (mixture of \u003cem\u003eM. koenigii\u003c/em\u003e and \u003cem\u003eA. marmelos\u003c/em\u003e leaves) are having hormonal regulatory properties that might have supported the oestrous cycle and overall reproductive health cows. Since, this herbal formulation having antioxidant and anti-inflammatory properties which might have helped in reducing inflammation in the reproductive tract, promoting better cervical mucus production and a favourable environment for sperm transport (Dhankhar et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Gahlawat et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Nigam and Nambiar, \u003cspan citationid=\"CR79\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Rautela et al. \u003cspan citationid=\"CR94\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Our findings align with those of Dhami et al. (\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), who observed that administering a bi-herbal treatment (\u003cem\u003eM. koenigii\u003c/em\u003e and \u003cem\u003eA. marmelos\u003c/em\u003e) @ 200 g daily for 5 days resulted in improved mucus quality at subsequent next oestrus in infertile animals. On contrary, Rahi et al. (\u003cspan citationid=\"CR90\" class=\"CitationRef\"\u003e2013\u003c/span\u003e) reported that on feeding of herbal extract containing polyphenols have reduced the pH which may be due to decline in bacterial load and inflammatory process in uterus after treatment.\u003c/p\u003e \u003cp\u003eThe fertility is associated with an arborization pattern, as in our study we found that typical fern pattern was having higher conception rate in treatment groups thereby it was observed higher conception rate with typical fern pattern in all the treatment groups. The enhanced conception rate in crossbred cows with typical fern pattern indicates higher sperm penetration (Rangnekar et al. \u003cspan citationid=\"CR93\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Layek et al. \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Alkaline pH of cervical mucus is favourable for higher sperm motility. The increase in pH in the treatment groups had higher fertility rate and also suitable for uterine environment with maximum sperm penetration in alkaline pH (Pattabiraman et al. \u003cspan citationid=\"CR86\" class=\"CitationRef\"\u003e1967\u003c/span\u003e; Rangnekar et al. \u003cspan citationid=\"CR93\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). Similarly, the treatment groups exhibited a higher conception rate, correlated with an increased spinnbarkeit value. Additionally, the conception rate was influenced by the arborization pattern, as a typical arborization pattern is indicative of a proper hormonal balance, which supports higher conception rates (Alena et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2008\u003c/span\u003e).4.10. \u003cem\u003eRelative mRNA expression of PGR genes\u003c/em\u003e\u003c/p\u003e \u003cp\u003eThe progesterone gene\u0026rsquo;s expression was associated with the progesterone hormone, which was likewise shown to be significantly greater in the treatment groups. The fold expression of PGR gene was significantly higher (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) in HPL, HEX and HP groups as compared to CON. Progesterone is a crucial ovarian steroid that regulates the estrous cycle, maternal recognition of pregnancy, and gestation in farm animals (Spencer et al., 2004). Beyond these roles, progesterone is also vital for various reproductive functions, including the behavioral expression of estrus, ovulation, maintenance of uterine quiescence, embryo survival and normal parturition.\u003c/p\u003e \u003cp\u003e \u003cb\u003eConclusions\u003c/b\u003e \u003c/p\u003e \u003cp\u003eIn conclusion, the herbal formulation exhibited a positive influence on both metabolic parameters and reproductive performance in postpartum crossbred cows. The observed improvements in blood parameters, and early onset of estrus suggest promising prospects for integrating herbal supplements in dairy cattle management. These findings underscore the potential of herbal formulations as a supportive measure for enhancing the overall health and fertility of postpartum cows.\u003c/p\u003e \u003c/div\u003e"},{"header":"Declarations","content":" \u003ch2\u003eEthical approval\u003c/h2\u003e \u003cp\u003e Before conducting the experimental trails, the animals (crossbred cattle) studies were approved by institutional ethics committee IAEC, ICAR-IVRI, Izatnagar-243122, Bareilly, Uttar Pradesh (Regd No: IAEC/07.07.2022/B6), and was conducted in accordance with the ethics standards as applicable international, national and/or institutional guidelines.\u003c/p\u003e \u003ch2\u003eConflict of interest\u003c/h2\u003e \u003cp\u003eThe authors do not have any conflict of interest regarding this research.\u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eThis study was financially supported by funds provided by Indian Council of Agriculture Research (AICRP project), New Delhi, India.\u003c/p\u003e\u003ch2\u003eAuthors contributions\u003c/h2\u003e \u003cp\u003eJ. C: conducted the research as a part of her Ph.D. research work. She has conducted the experimental trial deligently along with handling and management of the cows and laboratory tests including laboratory and molecular works. She also wrote the main manuscript text.\u003c/p\u003e\u003ch2\u003eAcknowledgements\u003c/h2\u003e \u003cp\u003eThe authors would like to thank ICAR-IVRI, Izatnagar, UP-243122 for providing the facilities for conducting the research.\u003c/p\u003e\u003ch2\u003eAvailability of data and materials\u003c/h2\u003e \u003cp\u003eData provided within the manuscript is complete and can be provided on request to the authors.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbdallah, I.Z., Salem, I., El-Salam, A. and Nayrouz, A.S., 2017. Evaluation of antidiabetic and antioxidant activity of A. marmelos L. Correa fruit extract in diabetic rats. Egyptian Journal of Hospital Medicine 67, 731\u0026ndash;741.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAbeysinghe, D.T., Kumara, K.A.H., Kaushalya, K.A.D., Chandrika, U.G. and Alwis, D.D.D.H., 2021. Phytochemical screening, total polyphenol, flavonoid content, in vitro antioxidant and antibacterial activities of Sri Lankan varieties of Murraya koenigii and Micromelum minutum leaves. Heliyon 7(7), 07449. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.heliyon.2021.e07449\u003c/span\u003e\u003cspan address=\"10.1016/j.heliyon.2021.e07449\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAdebajo, A.C., Ayoola, F.O., Iwalewa, E.O., Akindahunsi, A.A., Omisore, N.O.A., Adewunmi, C.O. and Adenowo, T.K., 2006. Anti-trichomonal, biochemical and toxicological activities of methanolic extract and some carbazole alkaloids isolated from the leaves of Murraya koenigii growing in Nigeria. Phytomedicine 13(4), 246\u0026ndash;254. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.phymed.2004.12.002\u003c/span\u003e\u003cspan address=\"10.1016/j.phymed.2004.12.002\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAdewuyi, A.A., Gruys, E. and Van Eerdenburg, F.J.C.M., 2005. Non esterified fatty acids (NEFA) in dairy cattle. A review. Veterinary Quarterly 27(3), 117\u0026ndash;126. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/01652176.2005.9695192\u003c/span\u003e\u003cspan address=\"10.1080/01652176.2005.9695192\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAlena, J., Ludek, S., Mojmir, V. and Franstisek, L., 2008. Factors affecting the cervical mucus crystallization, the sperm survival in cervical mucus and pregnancy rate in Holstein cows. Journal of Central European Agriculture 9(2), 377\u0026ndash;384.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAOAC, 2012. Official method of analysis of AOAC International. 19th ed. Association of Analytical Communities International, Virginia, USA.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAmbrose, D.J., 2021. Postpartum anestrus and its management in dairy cattle. Bovine Reproduction, 408\u0026ndash;430. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/9781119602484.ch34\u003c/span\u003e\u003cspan address=\"10.1002/9781119602484.ch34\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eArulselvan, P., Senthilkumar, G.P., Sathish Kumar, D. and Subramanian, S., 2006. Anti-diabetic effect of A. marmelos leaves on streptozotocin induced diabetic rats. Die Pharmazie-International Journal of Pharmaceutical Sciences Research 61(10), 874\u0026ndash;877.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBaghbadorani, M.K., Mehrzad, J., Vodjgani, M., Khosravi, A. and Akbarinejad, V., 2022. Evaluation of biochemical and hematological parameters in postpartum Holstein dairy cows following supplementation of Immunofin\u0026reg; herbal extract. Iranian Journal of Veterinary Medicine 16(3), 274\u0026ndash;287. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://dx.doi.org/10.22059/IJVM.2021.315021.1005144\u003c/span\u003e\u003cspan address=\"10.22059/IJVM.2021.315021.1005144\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBaitule, M.M., Gawande, A.P., Kumar, U., Sahatpure, S.K., Patil, M.S. and Baitule, M.M., 2016. Effect of A. marmelos and A. marmelos in treatment of delayed pubertal buffalo heifers. Veterinary World 9(12), 1375. https://doi.org/10.14202%2Fvetworld.2016.1375-1380\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBansal, Y. and Bansal, G., 2011. Analytical methods for standardization of A. marmelos: a review. Journal of Pharmaceutical Education and Research 2, 37\u0026ndash;44.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBarjibhe, S., Oberoi, P.S., Patel, B. and Patel, P.K., 2019. Improving reproductive efficiency through the supplementation of mustard oil, poly-herbal mixture and butyric acid during the periparturient period in Sahiwal cows. Journal of Entomology and Zoology Studies 7(2), 668\u0026ndash;673.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBarkakati, J. and Kalita, D.J., 2020. Alteration in hematological profile of pregnant crossbred cattle of Assam after feeding some medicinal leaves/herbs and polyherbal preparation (Restobal) during their transition period. The Pharma Innovation 9(9), 197\u0026ndash;201. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.22271/tpi.2020.v9.i9c.5110\u003c/span\u003e\u003cspan address=\"10.22271/tpi.2020.v9.i9c.5110\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBegashaw, B., Mishra, B., Tsegaw, A. and Shewamene, Z., 2017. Methanol leaves extract Hibiscus micranthus Linn exhibited antibacterial and wound healing activities. BMC Complementary and Alternative Medicine 17(1), 1\u0026ndash;11. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s12906-017-1841-x\u003c/span\u003e\u003cspan address=\"10.1186/s12906-017-1841-x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBirudu, R.B., Pamulapati, P. and Manoharan, S.K., 2020. Evaluation of biochemical changes in diabetic rats treated with Aegle marmelos (L.) methanolic leaf extract. Pharmacognosy Research 12(2), 127\u0026ndash;130. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.4103/pr.pr_53_19\u003c/span\u003e\u003cspan address=\"10.4103/pr.pr_53_19\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBiswas, S. and Giri, S., 2015. Importance of choline as essential nutrient and its role in prevention of various toxicities. Prague Medical Report 116, 5\u0026ndash;15. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.14712/23362936.2015.40\u003c/span\u003e\u003cspan address=\"10.14712/23362936.2015.40\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChakma, J., Samanta, A., Dutta, T. and Arya, R.S., 2020. Effect of supplementing Moringa oleifera leaf extract and clove bud oil to the diet on microflora population and intestinal morphology of broiler birds. Indian Journal of Animal Health 59, 222\u0026ndash;227. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.36062/ijah.59.2SPL.2020.222-227\u003c/span\u003e\u003cspan address=\"10.36062/ijah.59.2SPL.2020.222-227\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChakma, J., Kaur, N., Namdeo, S., Patir, M., Dutta, N., Jadhav, S.E., Singh, G. and Singh, S.K., 2024. Effect of Murraya koenigii and Aegle marmelos leaves supplementation on gas production kinetics and feed degradation with cattle inoculum. Animal Nutrition and Feed Technology 24(1), 137\u0026ndash;148. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.5958/0974-181X.2024.00010.6\u003c/span\u003e\u003cspan address=\"10.5958/0974-181X.2024.00010.6\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChoudhary, S., Chaudhary, L.C., Kala, A., Agarwal, N. and Rahman, H., 2022. Effect of dietary sulphur supplementation at graded levels on rumen feed fermentation and microbial profile in buffaloes. Animal Nutrition and Feed Technology 22(3), 543\u0026ndash;552. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.5958/0974-181X.2022.00043.9\u003c/span\u003e\u003cspan address=\"10.5958/0974-181X.2022.00043.9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChoudhury, S. and Sinha, M.P., 2015. Effect of aqueous extract of Murraya koenigii on haematological, hormonal and lipid profile of Albino rats. Journal of Coastal Life Medicine 3, 901\u0026ndash;905. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.12980/jclm.3.2015j5-63\u003c/span\u003e\u003cspan address=\"10.12980/jclm.3.2015j5-63\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDacie, Z. and Lewis, S., 1969. Practical Haematology. 4th ed. J and A., Churchill, London.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDas, G.K., Mehrotra, S., Kumar, B., Jan, M.H., Pande, M., Singh, S.K., Kumar, H., Khan, T.A., Agarwal, S.K. and Shankar, U., 2012. Pattern and estrus induction response in delayed pubertal buffalo heifers treated with a combined treatment of Murraya koenigii and Aegle marmelos. In: National symposium of XXVIII Annual Convention of the Indian Society for Study of Animal Reproduction. Khanapara, Guwahati, India. pp. 50.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDas, P., Das, S.K., Arya, H.P.S., Subba Reddy, G. and Mishra, A., 2002. Veterinary Science and Animal Husbandry. Inventory of indigenous technical knowledge in agriculture, Document-1, ICAR, New Delhi, pp.185\u0026ndash;285.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDas, G.K., Mehrotra, S., Narayanan, K., Kumawat, B.L., De, U.K., Khan, T.A., 2016. Estrus induction response and fertility performance in delayed pubertal heifers treated with Aegle marmelos and Murraya koenigii. Journal of Animal Research 6(1), 151\u0026ndash;156.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDhami, A.J., Parmar, S.C., Patel, J.A., Hadiya, K.K., Joshi, R.S. and Sarvaiya, N.P., 2022. Current status of reproductive performance and problems in dairy animals of Gujarat and their amelioration including study of fertility gene markers. Markers 18(2), 12\u0026ndash;20.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDhami, A.J., Patel, J.A., Hadiya, K.K., Parmar, S.C. and Chaudhari, D.V., 2019. Nutritional infertility and ameliorative measures in dairy animals of middle Gujarat. Indian Journal of Veterinary Science and Biotechnology 14(3), 5\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDhankhar, S., Ruhil, S., Balhara, M., Dhankhar, S. and Chhillar, A.K., 2011. Aegle marmelos (Linn.) Correa: A potential source of phytomedicine. Journal of Medicinal Plants Research 5(9), 1497\u0026ndash;1507.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDhivagar, M., 2022. Effect of herbal extracts supplementation on performance of wistar rats. M.V.Sc. Thesis, Indian Veterinary Research Institute, Izatnagar (Deemed University), Bareilly, India.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eD\u0026iacute;az Galv\u0026aacute;n, C., M\u0026eacute;ndez Olvera, E.T., Mart\u0026iacute;nez G\u0026oacute;mez, D., Gloria Trujillo, A., Hern\u0026aacute;ndez Garc\u0026iacute;a, P.A., Espinosa Ayala, E., Palacios Mart\u0026iacute;nez, M., Lara Bueno, A., Mendoza Mart\u0026iacute;nez, G.D. and Vel\u0026aacute;zquez Cruz, L.A., 2021. Influence of a polyherbal mixture in dairy calves: growth performance and gene expression. Frontiers in Veterinary Science 7, 623710. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fvets.2020.623710\u003c/span\u003e\u003cspan address=\"10.3389/fvets.2020.623710\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDorantes-Iturbide, G., Orzuna-Orzuna, J.F., Lara-Bueno, A., Miranda-Romero, L.A. and Mendoza-Mart\u0026iacute;nez, G.D., 2022. Effects of a polyherbal dietary additive on performance, dietary energetics, carcass traits and blood metabolites of finishing lambs. Metabolites 12(5), 413. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/metabo12050413\u003c/span\u003e\u003cspan address=\"10.3390/metabo12050413\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDriver, A.M., Huang, W., Gajic, S., Monson, R.L., Rosa, G.J.M. and Khatib, H., 2009. Effects of the progesterone receptor variants on fertility traits in cattle. Journal of Dairy Science 92(8), 4082\u0026ndash;4085. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3168/jds.2009-2013\u003c/span\u003e\u003cspan address=\"10.3168/jds.2009-2013\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDutt, R., Dalal, J., Singh, G., Gahalot, S.C. and Chandolia, R.K., 2018. Medicinal uses of Murraya koenigii and Aegle marmelos for fertility augmentation in animals: a review. International Journal of Current Microbiology and Applied Sciences 7(9), 645\u0026ndash;657. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.20546/ijcmas.2018.709.077\u003c/span\u003e\u003cspan address=\"10.20546/ijcmas.2018.709.077\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDutt, R., Mehrotra, S., Hoque, M., Shanker, U., Singh, G., Agarwal, S.K., Das, G.K. and Singh, S.K., 2010. Effect of Murraya koenigii and Aegle marmelos combination on resumption of fertility in anestrous goats. Journal of Applied Animal Research 38(2), 275\u0026ndash;278. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/09712119.2010.10539520\u003c/span\u003e\u003cspan address=\"10.1080/09712119.2010.10539520\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDutt, R., Mehrotra, S., Shanker, U. and Singh, G., 2011. Effect of Murraya koenigii and Aegle marmelos feeding on anestrus buffaloes. Indian Journal of Animal Reproduction 32(1), 47\u0026ndash;49.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDutta, A., Baruah, B., Sarmah, B.C., Baruah, K.K. and Goswami, R.N., 2001. Macromineral levels in cyclic, postpartum anestrus and repeat breeding local cows in lower Brahmaputra valley of Assam. Indian Journal of Animal Reproduction 22(1), 41\u0026ndash;44.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDutta, L.J., Nath, K.C., Deka, B.C., Bhuyan, D., Borah, P., Saikia, G.K., Bora, D.P., Deka, R., Acharya, C., Bharali, D., 2019. Therapeutic management of true anestrous crossbred cows through nutritional and hormonal intervention. Journal of Entomology and Zoology Studies 7(3), 09\u0026ndash;12.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGahlawat, D.K., Jakhar, S. and Dahiya, P., 2014. Murraya koenigii (L.) Spreng: An ethnobotanical, phytochemical and pharmacological review. Journal of Pharmacognosy and Phytochemistry 3(3), 109\u0026ndash;119.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGandhi, G.R., Ignacimuthu, S. and Paulraj, M.G., 2012. Hypoglycemic and β-cells regenerative effects of Aegle marmelos (L.) Corr. bark extract in streptozotocin-induced diabetic rats. Food and Chemical Toxicology 50(5), 1667\u0026ndash;1674.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGanesan, P., Phaiphan, A., Murugan, Y. and Baharin, B.S., 2013. Comparative study of bioactive compounds in curry and coriander leaves: An update. Journal of Chemical and Pharmaceutical Research 5(11), 590\u0026ndash;594.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGoodchild, A.V. and McMeniman, N.P., 1994. Intake and digestibility of low quality roughages when supplemented with leguminous browse. Journal of Agricultural Science 122(1), 151\u0026ndash;160. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1017/S0021859600065904\u003c/span\u003e\u003cspan address=\"10.1017/S0021859600065904\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHashemzadeh-Cigari, F., Ghorbani, G.R., Khorvash, M., Riasi, A., Taghizadeh, A. and Zebeli, Q., 2015. Supplementation of herbal plants differently modulated metabolic profile, insulin sensitivity and oxidative stress in transition dairy cows fed various extruded oil seeds. Preventive Veterinary Medicine 118(1), 45\u0026ndash;55. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.prevetmed.2014.10.013\u003c/span\u003e\u003cspan address=\"10.1016/j.prevetmed.2014.10.013\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHegde, G.V., Nimbalkar, S.D., Hegde, G.S. and Naik, K., 2002. Use of herbal plants for repeat breeding problem in dairy animals. Indian Veterinary Journal 79(8), 861\u0026ndash;862.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHosten, A.O., 1990. BUN and creatinine. In Clinical Methods: The History, Physical, and Laboratory Examinations. 3rd edition. Butterworths.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIgara, C.E., Omoboyowa, D.A., Ahuchaogu, A.A., Orji, N.U. and Ndukwe, M.K., 2016. Phytochemical and nutritional profile of Murraya koenigii (Linn) Spreng leaf. Journal of Pharmacognosy and Phytochemistry 5(5), 7.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJadhav, A.N. and Bhutani, K.K., 2005. Ayurveda and gynaecological disorders. Journal of Ethnopharmacology 97, 151\u0026ndash;159. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jep.2004.10.020\u003c/span\u003e\u003cspan address=\"10.1016/j.jep.2004.10.020\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJain, V., Momin, M. and Laddha, K., 2012. Murraya koenigii: an updated review. International Journal of Ayurvedic and Herbal Medicine 2(4), 607\u0026ndash;627.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJondhale, N.S., 2007. Effect of Aegle marmelos and Ficus religiosa on the ovarian function in rats and follicular dynamics in goat. MVSc Thesis, Indian Veterinary Research Institute, Izatnagar (Deemed University), Bareilly, India.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJondhale, N.S., Mehrotra, S., Agarwal, S.K., Singh, S.K. and Deori, S., 2009a. Effect of Aegle marmelos and Ficus religiosa on the onset of puberty in immature female rats. Indian Veterinary Journal 86(4), 353\u0026ndash;355.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJondhale, N.S., Uma, S., Mehrotra, S., Agarwal, S.K., Singh, S.K., Das, G.K. and Deori, S., 2009b. Effect of Aegle marmelos and Ficus religiosa leaves extracts on the ovarian function in rats. Indian Veterinary Journal 86(11), 1141\u0026ndash;1144.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKabir, K.K., Rawal, C.V.S., Ansari, M.R., Varshney, J.P. and Srivastava, R.S., 2001. Comparative efficacy of herbal preparations in the management of anoestrus in non-descript rural buffaloes. Indian Journal of Animal Reproduction 22(2), 143\u0026ndash;145.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKaneko, J.J., Harvey, J.W. and Bruss, M.L., 2008. Clinical biochemistry of domestic animals. 6th ed. Academic Press, San Diego, California, USA.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKhade, N.B., Patel, D.M., Naikoo, M., Dhami, A.J., Sarvaiya, N.P. and Gohel, M.M., 2011. Estrus induction in pubertal anestrus Gir heifers using different hormone protocols. Indian Journal of Field Veterinarians 7(1), 4\u0026ndash;8.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKujur, A., Srivastava, N., Jasrotia, N. and Arunpandian, J., 2022. The role of medicinal plants in domestic animals and estrus induction with particular reference to Aegle marmelos and Murraya koenigii. Animal Reproduction Update 2(1), 56\u0026ndash;62. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.48165/aru.2022.2102\u003c/span\u003e\u003cspan address=\"10.48165/aru.2022.2102\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar, P.R., Singh, S.K., Kharche, S.D., Govindaraju, C.S., Behera, B.K., Shukla, S.N., Kumar, H. and Agarwal, S.K., 2014. Anestrus in cattle and buffalo: Indian perspective. Advances in Animal and Veterinary Sciences 2(3), 124\u0026ndash;138. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.14737/journal.aavs/2014/2.3.124.138\u003c/span\u003e\u003cspan address=\"10.14737/journal.aavs/2014/2.3.124.138\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar, A., Katiyar, R., Ahmad, S.F., Balamurugan, B., Deepak, D. and Prasad, J.K., 2018. Current treatment aspects of bovine reproductive disorders. Theriogenology Insight 8(3), 101\u0026ndash;109. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.30954/2277-3371.03.2018.2\u003c/span\u003e\u003cspan address=\"10.30954/2277-3371.03.2018.2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar, R., 2008. Effect of Aegle marmelos (Bel) and Ficus religiosa (Pipal) on the reproductive performance in anoestrus goat and buffalo. MVSc Thesis, Indian Veterinary Research Institute, Izatnagar, India.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar, S., Mehla, R.K. and Dang, A.K., 2008. Use of shatavari (Asparagus racemosus) as a galactopoietic and therapeutic herb: a review. Agricultural Reviews 29(2), 132\u0026ndash;138.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar, S., Bodla, R.B. and Bansal, H., 2016. Antioxidant activity of leaf extract of Aegle marmelos Correa ex Roxb. Pharmacognosy Journal 8(5), 447\u0026ndash;450. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.5530/pj.2016.5.6\u003c/span\u003e\u003cspan address=\"10.5530/pj.2016.5.6\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar, S., Kumar, G. and Gahalot, S.C., 2019. Anestrous in buffaloes and different treatment regimens: a mini review. International Journal of Current Microbiology and Applied Sciences 8, 1162\u0026ndash;1179. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.20546/ijcmas.2019.803.138\u003c/span\u003e\u003cspan address=\"10.20546/ijcmas.2019.803.138\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumawat, B., Das, G., Narayanan, K., Mehrotra, S., Prasad, J. and Ghosh, S., 2016. Effect of Aegle marmelos and Murraya koenigii on estrus induction, conception and calving rate in delayed pubertal heifers under field condition. Indian Journal of Animal Reproduction 37(2), 17\u0026ndash;19.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumawat, B.L., Das, G.K., Narayanan, K., Mehrotra, S., Choadhary, R.K., Yogi, R.K., Dey, S. and Kumar, H., 2014. Effect of herbs supplementation on certain serum mineral levels in delayed pubertal heifers. National Symposium and XXX Annual Convention of ISSAR, Mathura, (U.P.) India, November 20\u0026ndash;22, 2014. Compendium and Souvenir, F.I. 20, 25.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKuralkar, P. and Kuralkar, S.V., 2021. Role of herbal products in animal production: an updated review. Journal of Ethnopharmacology 278, 114246. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jep.2021.114246\u003c/span\u003e\u003cspan address=\"10.1016/j.jep.2021.114246\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLayek, S.S., Mohanty, T.K., Kumaresan, A., Behera, K. and Chand, S., 2013. Cervical mucus characteristics and periestrual hormone concentration in relation to ovulation time in Zebu (Sahiwal) cattle. Livestock Science 152(2\u0026ndash;3), 273\u0026ndash;281. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.livsci.2012.12.023\u003c/span\u003e\u003cspan address=\"10.1016/j.livsci.2012.12.023\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLee-Rangel, H.A., Mendoza-Martinez, G.D., Mart\u0026iacute;nez-Garc\u0026iacute;a, J.A., Espinosa-Ayala, E., Hern\u0026aacute;ndez-Garc\u0026iacute;a, P.A., Cifuentes-L\u0026oacute;pez, R.O., Vazquez-Valladolid, A., Garc\u0026iacute;a-L\u0026oacute;pez, J.C., Lara-Bueno, A. and Roque-Jim\u0026eacute;nez, J.A., 2022. An Indian polyherbal phytogenic source improved blood serum biochemistry and immune response of dairy calves. Food and Agricultural Immunology 33(1), 97\u0026ndash;112. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/09540105.2021.2024150\u003c/span\u003e\u003cspan address=\"10.1080/09540105.2021.2024150\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLivak, K.J. and Schmittgen, T.D., 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2\u0026thinsp;\u0026ndash; ∆∆CT method. Methods 25(4), 402\u0026ndash;408. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1006/meth.2001.1262\u003c/span\u003e\u003cspan address=\"10.1006/meth.2001.1262\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLone, S.A., Ganie, A.A., Quadri, M.A., Haq, M.R. and Reshi, I.M., 2012. Effect of exogenous insulin on fertility response and serum hormonal profile in delayed pubertal crossbred heifers. Veterinary Practitioner 13(2), 268\u0026ndash;269.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLopreiato, V., Mezzetti, M., Cattaneo, L., Ferronato, G., Minuti, A. and Trevisi, E., 2020. Role of nutraceuticals during the transition period of dairy cows: A review. Journal of Animal Science and Biotechnology 11, 1\u0026ndash;18. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s40104-020-00501-x\u003c/span\u003e\u003cspan address=\"10.1186/s40104-020-00501-x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLozano-S\u0026aacute;nchez, M., Mendoza-Mart\u0026iacute;nez, G.D., Mart\u0026iacute;nez-Garc\u0026iacute;a, J.A., la Torre-Hern\u0026aacute;ndez, M.E.D., Chamorro-Ram\u0026iacute;rez, F.H., Mart\u0026iacute;nez-Aispuro, J.A., Cordero-Mora, J.L., S\u0026aacute;nchez-Torres, M.T., Hern\u0026aacute;ndez-Garc\u0026iacute;a, P.A. and Jones, R., 2021. Evaluation of polyherbal with vitamin C activity on lamb performance and meat characteristics. Revista Brasileira de Zootecnia 50, e20200166. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.37496/rbz5020200166\u003c/span\u003e\u003cspan address=\"10.37496/rbz5020200166\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMehedint, M.G. and Zeisel, S.H., 2013. Choline's role in maintaining liver function: new evidence for epigenetic mechanisms. Current Opinion in Clinical Nutrition and Metabolic Care 16, 339\u0026ndash;345. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1097/MCO.0b013e3283600d46\u003c/span\u003e\u003cspan address=\"10.1097/MCO.0b013e3283600d46\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMehrotra, S., 2002. Studies on ovarian function using certain medicinal plants in rats, goats and cattle. Doctoral dissertation, Indian Veterinary Research Institute; Bareilly, India.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMehrotra, S., Singh, S.K., Shanker, U.S. and Agarwal, S.K., 2005. Effect of certain indigenous medicinal plants on ovarian hormone profile and reproductive performance in anestrus cattle. Indian Journal of Animal Reproduction 26(1), 20\u0026ndash;23.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMehrotra, S., Umashanker, J., Majumdar, A.C. and Agarwal, S.K., 2003. Effect of indigenous medicinal plants on onset of puberty in immature female rats. Indian Journal of Animal Reproduction 24(2), 131\u0026ndash;133.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMendoza, G.D., Oviedo, M.F., Pinos, J.M., Lee-Rangel, H.A., V\u0026aacute;zquez, A., Flores, R. and P\u0026eacute;rez, F., 2019. Milk production in dairy cows supplemented with herbal choline and methionine. Revista de la Facultad de Ciencias Agrarias 52(1), 1\u0026ndash;12.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMishra, B.P., Nayak, N. and Mishra, D., 2002. Indigenous methods for animal health care. International seminar on traditional knowledge, health and environment. Bubaneshwar, Abs, pp. 124.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNaczk, M. and Shahidi, F., 2004. Extraction and analysis of phenolics in food. Journal of Chromatography A 1054(1\u0026ndash;2), 95\u0026ndash;111. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.chroma.2004.08.059\u003c/span\u003e\u003cspan address=\"10.1016/j.chroma.2004.08.059\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNanda, S., Tiwari, S.P. and Rathore, R., 2013. Effect of polyherbal formulations on blood haematological constituents and immunity in non-descript goats. Iranian Journal of Applied Animal Science 3(2), 327\u0026ndash;331.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNandini, M.S., Veena, T. and Swamy, M.N., 2010. Effect of extracts of Murraya koenigii spreng. and Morus alba linn. on the age of attainment of puberty and ovarian folliculogenesis in rats. Journal of Basic and Clinical Pharmacy 1(4), 203.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNegi, D., Mondal, B.C. and Tiwari, D.P., 2015. Effect of phase-feeding on milk production, milk composition, nutrient utilization and haematobiochemical constituents in lactating crossbred cattle. Animal Nutrition and Feed Technology 15(2), 235\u0026ndash;244. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.5958/0974-181X.2015.00026.8\u003c/span\u003e\u003cspan address=\"10.5958/0974-181X.2015.00026.8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNigam, V. and Nambiar, V.S., 2015. Therapeutic potential of Aegle marmelos (L.) Correa leaves as an antioxidant and anti-diabetic agent: A review. International Journal of Pharmaceutical Sciences and Research 6(3), 611\u0026ndash;621.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOlusanya, J.O., 2008. The food nutrients. In: Essentials of Food and Nutrition, Apex Book Limited Lagos, xxi.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOrzuna-Orzuna, J.F., Dorantes-Iturbide, G., Lara-Bueno, A., Mendoza-Mart\u0026iacute;nez, G.D., Miranda-Romero, L.A. and Hern\u0026aacute;ndez-Garc\u0026iacute;a, P.A., 2021. Growth performance, carcass characteristics and blood metabolites of lambs supplemented with a polyherbal mixture. Animals 11(4), 955. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/ani11040955\u003c/span\u003e\u003cspan address=\"10.3390/ani11040955\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePanigrahi, B., 1964. Spinnbarkeit test of bovine cervical mucus\u0026mdash;an index to optimal fertility time. Indian Veterinary Journal 41(6), 410\u0026ndash;412.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePatel, R.K. and Gupta, D.K., 2020. An in vivo assessment of a non-antibiotic therapy for specific subclinical mastitis in dairy cows. Journal of Entomology and Zoology Studies 8(5), 721\u0026ndash;725.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePathak, A.K., Dutta, N., Pattanaik, A.K., Chaturvedi, V.B. and Sharma, K., 2017. Effect of condensed tannins from Ficus infectoria and Psidium guajava leaf meal mixture on nutrient metabolism, methane emission and performance of lambs. Asian-Australasian Journal of Animal Sciences 30(12), 1702. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.5713/ajas.17.0060\u003c/span\u003e\u003cspan address=\"10.5713/ajas.17.0060\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePatir, M., 2022. Effect of Moringa oleifera foliage supplementation on the performance of crossbred cattle. Thesis. PhD. Deemed University, Indian Veterinary Research Institute, Izatnagar, India.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePattabiraman, S.R., Venkataswamy, V. and Thangraj, T.M., 1967. Physicochemical properties of oestrual mucus of cows. Indian Veterinary Journal 10, 413\u0026ndash;417.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePatton, J., Kenny, D.A., McNamara, S., Mee, J.F., O\u0026rsquo;mara, F.P., Diskin, M.G. and Murphy, J.J., 2007. Relationships among milk production, energy balance, plasma analytes and reproduction in Holstein-Friesian cows. Journal of Dairy Science 90(2), 649\u0026ndash;658. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3168/jds.S0022-0302(07)71547-3\u003c/span\u003e\u003cspan address=\"10.3168/jds.S0022-0302(07)71547-3\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePriyadarshinee, P., Sethy, K., Swain, R.K., Mishra, S.K., Behera, K., Pradhan, S.K. and Tripathy, S., 2021. Combined effect of Bel and curry leaves supplementation on the performance of anestrous cattle. Indian Journal of Animal Research 38(3), 240\u0026ndash;245. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.5958/2231-6744.2021.00035.9\u003c/span\u003e\u003cspan address=\"10.5958/2231-6744.2021.00035.9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePurkayastha, R.D., Shukla, S.N., Shrivastava, O.P. and Kumar, P.R., 2015. A comparative therapeutic management of anoestrus in buffaloes using insulin and GnRH. Veterinary World 8(6), 804\u0026ndash;807. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.14202/vetworld.2015.804-807\u003c/span\u003e\u003cspan address=\"10.14202/vetworld.2015.804-807\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRahi, S., Gupta, H.P., Prasad, S. and Baithalu, R.K., 2013. Phytotherapy for endometritis and subsequent conception rate in repeat breeding crossbred cows. Indian Journal of Animal Reproduction 34(1), 9\u0026ndash;12.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRaja, S.B., Murali, M.R., Kumar, N.K. and Devaraj, S.N., 2011. Isolation and partial characterization of a novel lectin from Aegle marmelos fruit and its effect on adherence and invasion of Shigellae to HT29 cells. PLoS One 6(1), e16231. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1371/journal.pone.0016231\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0016231\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRajkumar, R., Srivastava, S.K., Varshney, V.P. and Mahmood, S., 2008. Effect of medicinal plants Saraca asoca and Trigonella foenum-graecum in anoestrus cows. Indian Veterinary Journal 85(12), 1281\u0026ndash;1283.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRangnekar, M.N., Dhoble, R.L., Gacche, M.G., Ingawale, M.V., Sawale, A.G. and Jadhav, J.M., 2002. Physical properties of oestrual cervical mucus in repeat breeding crossbred (Holstein Friesian) cows with reference to fertility. Indian Journal of Animal Sciences 72(12), 1122\u0026ndash;1124.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRautela, R., Das, G.K., Khan, F.A., Prasad, S., Kumar, A., Prasad, J.K., Ghosh, S.K., Dhanze, H., Katiyar, R. and Srivastava, S.K., 2018. Antibacterial, anti-inflammatory and antioxidant effects of Aegle marmelos and Murraya koenigii in dairy cows with endometritis. Livestock Science 214, 142\u0026ndash;148. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.livsci.2018.05.015\u003c/span\u003e\u003cspan address=\"10.1016/j.livsci.2018.05.015\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRazo Ortiz, P.B., Mendoza Martinez, G.D., Silva, G.V., Osorio Teran, A.I., Gonzalez Sanchez, J.F., Hernandez Garcia, P.A., de la Torre Hern\u0026aacute;ndez, M.E., Espinosa Ayala, E., 2020. Polyherbal feed additive for lambs: effects on performance, blood biochemistry and biometry. Journal of Applied Animal Research 48(1), 419\u0026ndash;424. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/09712119.2020.1814786\u003c/span\u003e\u003cspan address=\"10.1080/09712119.2020.1814786\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eReddy, K.N., 2008. Ethnobotany of Andhra Pradesh: A Review. Ethnobotanical Leaflets 12, 305\u0026ndash;310.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRivero, N., Salem, A.Z.M., Gado, H.M., Gonzalez-Ronquillo, M., Pliego, A.B., Pe\u0026ntilde;uelas, C.G. and Odongo, N.E., 2012. Effect of exogenous enzymes and Salix babylonica extract or their combination on haematological parameters in growing lambs. Journal of Animal Feed Science 21, 577\u0026ndash;586.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRoseler, D.K., Ferguson, J.D., Sniffen, C.J. and Herrema, J., 1993. Dietary protein degradability effects on plasma and milk urea nitrogen and milk nonprotein nitrogen in Holstein cows. Journal of Dairy Science 76(2), 525\u0026ndash;534. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3168/jds.S0022-0302(93)77372-5\u003c/span\u003e\u003cspan address=\"10.3168/jds.S0022-0302(93)77372-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSabu, M.C. and Kuttan, R., 2004. Antidiabetic activity of Aegle marmelos and its relationship with its antioxidant properties. Indian Journal of Physiology and Pharmacology 48(1), 81\u0026ndash;88.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eS\u0026aacute;nchez, N., Lee Rangel, H.A., Mart\u0026iacute;nez-Cort\u0026eacute;s, I., Mendoza, G.D., Hern\u0026aacute;ndez, P.A., Espinoza, E., Vazquez-Valladolid, A., Flores-Ram\u0026iacute;rez, R., Roque-Jim\u0026eacute;nez, A., Campillo-Navarro, M. and Relling, A.E., 2021. A polyherbal phytogenic additive improved growth performance, health, and immune response in dairy calves. Food and Agricultural Immunology 32(1), 482\u0026ndash;498. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/09540105.2021.1967296\u003c/span\u003e\u003cspan address=\"10.1080/09540105.2021.1967296\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShantala, M. and Prakash, J., 2005. Acceptability of curry leaf (Murraya koenigii) incorporated products and attitude toward consumption. Journal of Food Processing and Preservation 29, 33\u0026ndash;44. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.1745-4549.2005.00011.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1745-4549.2005.00011.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSatheesh, M.A. and Pari, L., 2008. Effect of pterostilbene on lipids and lipid profiles in streptozotocin-nicotinamide induced type 2 diabetes mellitus. Journal of Applied Biomedicine 6(1), 31\u0026ndash;37.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSatheshkumar, S. and Punniamurthy, N., 2009. Estrus induction by supplementation of Murraya koenigii in anestrus heifers. Indian Journal of Animal Reproduction 30, 66\u0026ndash;67.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShafi, T.A., Bansal, B.K. and Gupta, D.K., 2018. In vitro antibacterial activity and minimum inhibitory concentration of Ocimum sanctum leaves against common bovine mastitis pathogens. Journal of Dairy, Veterinary and Animal Research 7(6), 322\u0026ndash;324.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSharma, P.C., Bhatia, N. and Sharama, A., 2007. A Review on Bael tree. Natural Product Radiance 6(2), 171\u0026ndash;178.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShavit, L., Lifschitz, M. and Galperin, I., 2012. Influence of enteric nutrition on blood urea nitrogen (BUN) in very old patients with chronic kidney disease (CKD). Archives of Gerontology and Geriatrics 54(1), 228\u0026ndash;231. \u003cdiv class=\"ExternalRefDOI\"\u003ehttps://doi.org/10.1016/j\u003c/div\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTalapatra, S.K., Ray, S.C. and Sen, K.C., 1940. The analysis of mineral constituents in biological materials. Estimation of phosphorus, chlorine, calcium, magnesium, sodium and potassium in food-stuffs. Indian Journal of Veterinary Science 10, 243\u0026ndash;258.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTiwari, U.S., Shukla, S.N., Bajaj, N.K., Jain, A.K., Mishra, P., Kumar, J. and Tiwari, A., 2021. Fertility response using timed insemination protocols in postpartum buffaloes. Journal of Pharmacy Innovation 10(10), 1136\u0026ndash;1138.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eUmashanker, A., Mehrotra, S.K., Singh, S.K. and Hoque, M., 2006. Restoration of fertility in anoestrus buffaloes using M. koenigii (Curry leaf plant). In: XXII Annual Convention and National Symposium on Innovative Technologies for Fertility Enhancement in Livestock, MHOW, Madhya Pradesh, India, pp. 283.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eUpadhya, S., Shanbhag, K.K., Suneetha, G. and Naidu, M.B., 2004. A study of hypoglycemic and antioxidant activity of Aegle marmelos in alloxan induced diabetic rat. Indian Journal of Physiology and Pharmacology 48, 476\u0026ndash;480.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVan Soest, P.V., Robertson, J.B. and Lewis, B.A., 1991. Methods for dietary fiber, neutral detergent fiber and non-starch polysaccharides in relation to animal nutrition. Journal of Dairy Science 74(10), 3583\u0026ndash;3597. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3168/jds.S0022-0302(91)78551-2\u003c/span\u003e\u003cspan address=\"10.3168/jds.S0022-0302(91)78551-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVerma, A., Kumar, P., Pandey, A.K., Kumari, S. and Kumar, S., 2019. Anestrus is a major infertility issue in perspective of buffalo reproduction in India: A review. Pharmacy Innovation Journal 8(2), 208\u0026ndash;211.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVijayalakshmi, S. and Venkatalakshmi, P., 2017. An updated review on the phytopharmacological significance of Aegle marmelos (L.). World Journal of Pharmaceutical Research 6(16), 1390\u0026ndash;1422.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWadhwa, M. and Bakshi, M.P.S., 2023. Effect of feeding detoxified waste bread in total mixed ration on nutrient utilization and rumen fermentation pattern in adult buffaloes. Animal Nutrition and Feed Technology 23(2), 221\u0026ndash;231. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.5958/0974-181X.2023.00020.3\u003c/span\u003e\u003cspan address=\"10.5958/0974-181X.2023.00020.3\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWanapat, M., Kang, S. and Polyorach, S., 2013. Development of feeding systems and strategies of supplementation to enhance rumen fermentation and ruminant production in the tropics. Journal of Animal Science and Biotechnology 4(32), 1\u0026ndash;11. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/2049-1891-4-32\u003c/span\u003e\u003cspan address=\"10.1186/2049-1891-4-32\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWankhede, S.D., Tambe, M.B., Dutta, N., Kaur, N., Singh, P., Jadhav, S.E. and Pattanaik, A., 2022. Effect of moringa foliage supplementation on in vitro ruminal gas production kinetics and substrate degradation in cattle. Indian Journal of Animal Science 92, 871\u0026ndash;875. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.56093/ijans.v92i7.114850\u003c/span\u003e\u003cspan address=\"10.56093/ijans.v92i7.114850\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eXie, J.T., Chang, W.T., Wang, C.Z., Mehendale, S.R., Li, J., Ambihaipahar, R., Ambihaipahar, U., Fong, H.H. and Yuan, C.S., 2006. Curry leaf (Murraya koenigii Spreng.) reduces blood cholesterol and glucose levels in ob/ob mice. American Journal of Chinese Medicine 34(02), 279\u0026ndash;284. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1142/S0192415X06003825\u003c/span\u003e\u003cspan address=\"10.1142/S0192415X06003825\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYattoo, M.A., Chaudhary, L.C., Kala, A., Agarwal, N. and Kamra, D.N., 2023. Effect of blend of essential oils as feed additive on rumen metabolites and rumen microbial status of buffaloes. Animal Nutrition and Feed Technology 23(2), 373\u0026ndash;380. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.5958/0974-181X.2023.00031.8\u003c/span\u003e\u003cspan address=\"10.5958/0974-181X.2023.00031.8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"tropical-animal-health-and-production","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"trop","sideBox":"Learn more about [Tropical Animal Health and Production](https://www.springer.com/journal/11250)","snPcode":"11250","submissionUrl":"https://submission.nature.com/new-submission/11250/3","title":"Tropical Animal Health and Production","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Aegle marmelos, Cattle, Cervical mucus, Murraya koenigii, Reproduction","lastPublishedDoi":"10.21203/rs.3.rs-4557558/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4557558/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe study was carried out to ascertain the effect of herbal formulation on metabolic profile and reproductive performance of crossbred cows. Crossbred cows (24) were randomly assigned to four treatment groups. Cows in CON (control) group were fed a diet comprised of green fodder, concentrate and wheat straw, while, the cows in HP (herbal powder), HPL (herbal pellets) and HEX (herbal extract) groups were offered CON diet having herbal powder (200gd\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), herbal pellets (200gd\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) and methanolic herbal extract (equivalent to 200g herbal powder d\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), respectively. The cows were fed on treatment diets for 7 d and continued with CON diet up to one month. Hb levels were analogous among the treatments; however, haematocrit values were significantly higher (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) in the HEX group relative to CON. Total protein, albumin, globulin, A:G ratio, serum glucose, total cholesterol, AST and ALT were comparable across the dietary treatments. Serum NEFA, urea and creatinine were lower (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) in treatment groups relative to CON. Serum minerals viz. i-P, Cu, Zn, Mn and iron were significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) higher in all the treatments than CON. The conception at first insemination was considerably higher in HPL, HEX and HP groups relative to CON. Spinnbarkeit and pH values of cervical mucus were noticeably higher in HP, HPL and HEX groups. The typical fern pattern of cervical mucus exhibited higher conception rate in treatment groups. It may be concluded that the herbal formulation positively influenced metabolic parameters and reproductive performance in post-partum crossbred cows.\u003c/p\u003e","manuscriptTitle":"Impact of feeding Murraya koenigii and Aegle marmelos leaves on metabolic and reproductive performance in crossbred cows","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-07-19 17:35:52","doi":"10.21203/rs.3.rs-4557558/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"","date":"2024-08-01T19:02:41+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-06-27T20:21:13+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-06-21T09:20:19+00:00","index":"","fulltext":""},{"type":"submitted","content":"Tropical Animal Health and Production","date":"2024-06-10T06:56:54+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"tropical-animal-health-and-production","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"trop","sideBox":"Learn more about [Tropical Animal Health and Production](https://www.springer.com/journal/11250)","snPcode":"11250","submissionUrl":"https://submission.nature.com/new-submission/11250/3","title":"Tropical Animal Health and Production","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"204c850a-495d-48f9-b341-e3c631d046c7","owner":[],"postedDate":"July 19th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-03-10T19:45:53+00:00","versionOfRecord":{"articleIdentity":"rs-4557558","link":"https://doi.org/10.1007/s11250-025-04351-5","journal":{"identity":"tropical-animal-health-and-production","isVorOnly":false,"title":"Tropical Animal Health and Production"},"publishedOn":"2025-03-03 15:58:23","publishedOnDateReadable":"March 3rd, 2025"},"versionCreatedAt":"2024-07-19 17:35:52","video":"","vorDoi":"10.1007/s11250-025-04351-5","vorDoiUrl":"https://doi.org/10.1007/s11250-025-04351-5","workflowStages":[]},"version":"v1","identity":"rs-4557558","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4557558","identity":"rs-4557558","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}