Novel Limoisilactobacillus reuteri and Helianthus tuberoses synbiotic improved faecal biomarkers, antioxidant status and influenced the fecal microbial antibiotic sensitivity in neonatal dairy calves

Novel Limoisilactobacillus reuteri and Helianthus tuberoses synbiotic improved faecal biomarkers, antioxidant status and influenced the fecal microbial antibiotic sensitivity in neonatal dairy calves | 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 Novel Limoisilactobacillus reuteri and Helianthus tuberoses synbiotic improved faecal biomarkers, antioxidant status and influenced the fecal microbial antibiotic sensitivity in neonatal dairy calves Kasi Sowjanya Lakshmi R, Anju Kala, Neeta Agarwal, Zunjar Baburao Dubal, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7146873/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Antibiotic growth promoters (AGPs) are commonly used in neonatal dairy calves to prevent diarrhea and enhance early growth performance. However, their extensive use contributes to antimicrobial resistance (AMR). This study evaluated the potential of a calf fecal-origin probiotic-based synbiotic as an alternative to AGP tetracycline (TET) in influencing fecal biomarkers, antioxidant status, and the gut resistome in neonatal dairy calves. Eighteen newborn Vrindavani calves’ hand-fed milk, were randomly assigned to three groups (n = 6): no supplementation (CON), TET (1 mg/kg body weight) for 2 weeks, and synbiotic (SYN Limosilactobacillus reuteri RM125 (3–5×10⁸ cfu/mL) and Jerusalem artichoke tuber ( Helianthus tuberoses , JAT) powder (2% of dry matter intake) for 12 weeks. The calves were provided free access to forage and calf starter. Rectal fecal samples collected at 0, 1, 2, 3, 4, 6, 8, 10, and 12 week to assess gut health parameters. Fresh fecal samples were subjected to weekly antibiotic sensitivity tests against six representative antibiotics. Serum samples collected at weeks 0, 6, and 12 were analyzed for biochemical and antioxidant markers. The SYN group showed a significant (P < 0.05) increase in fecal Lactobacillus and Bifidobacteria counts, along with significantly (P < 0.05) reduced pH and ammonia levels, and improved lactic acid and short-chain fatty acids, indicating enhanced gut health. The serum lipid profile significantly (P < 0.05) improved in SYN calves, while liver and kidney function indices and serum mineral status remained comparable across the groups. Antioxidant status improved in the SYN group, with higher malondialdehyde (MDA) and catalase (CAT) enzyme activity. Among the six antibiotics viz, tetracycline, ciprofloxacin, ampicillin, ceftriaxone, cotrimoxazole and chloramphenicol, the fecal microbiome of SYN calves exhibited significantly (P < 0.05) higher zones of inhibition for ciprofloxacin and ceftriaxone, suggesting a favorable shift in the gut resistome. These findings suggest that synbiotic containing calf fecal-origin L. reuteri RM125 and JAT is a promising alternative to AGPs for enhancing fecal biomarkers, antioxidant status, and gut microbial resistance in neonatal dairy calves. Synbiotic Neonatal calves Host-specific probiotic Jerusalem artichoke Gut health Gut resistome Figures Figure 1 Figure 2 Figure 3 1. Introduction The early development of dairy calves is critically influenced by gut health, as a sterile gastrointestinal tract at birth that rapidly becomes colonized by environmental and maternal microbes [ 1 ]. Effective management practices during the neonatal period are crucial, as they influence growth rate, immunity, and overall productivity. Central to this is gut health, which is pivotal for nutrient absorption and immune system development [ 2 ]. In commercial dairy systems, where calves are separated from the dam after colostrum feeding, this rapid microbial colonization, combined with underdeveloped immunity and weaning stress, predisposes the calves to neonatal diarrohea [ 3 ]. Antibiotic growth promoters (AGPs) have historically been employed in neonatal calf feeding programs to prevent diarrhea, promote early growth, and improve feed efficiency [ 4 ]. However, the widespread use of AGPs has contributed to the emergence of antibiotic-resistant pathogens, posing a significant threat to both animal and public health [ 5 ]. This concern has catalyzed a shift towards non-antibiotic alternatives, including functional foods that promote gut health and favorably modulate the gut microbiome. Functional foods such as probiotics, prebiotics, and synbiotics have garnered attention for their ability to enhance gut health and immunity while mitigating the risk of antibiotic resistance [ 6 ]. Probiotics, such as Limosilactobacillus reuteri , inhibit pathogenic colonization, stabilize the gut microbiota, and modulate host immunity [ 7 ]. Prebiotics, including inulin from Jerusalem artichoke tuber (JAT), selectively stimulate beneficial gut bacteria, enhance nutrient absorption, and contribute to the gut's overall health [ 8 ]. Rich in inulin and polyphenolic compounds, JAT has demonstrated prebiotic and antimicrobial properties, making it a promising component of functional foods [ 9 ]. When combined as synbiotic, these functional foods exhibit synergistic effects, creating a favorable gut environment that supports health and growth [ 10 ]. Emerging evidence suggests that functional foods not only enhance growth performance and immunity but also play a critical role in modulating the gut resistome including genes involved in drug resistance and detrimental metabolic pathways [ 11 ]. By promoting the growth of beneficial microbes, synbiotics can suppress antibiotic-resistant pathogen population, potentially reversing antibiotic resistance [ 12 ]. This attribute is particularly significant in combating the global issue of antimicrobial resistance (AMR).Combining Jerusalem artichoke tuber (JAT) with calf fecal-origin L. reuteri RM125, a host-specific probiotic, demonstrated enhanced in-vitro gut health parameters, highlighting its promise as an effective symbiotic [ 13 ]. Host-specific probiotics, provide customized health advantages due to their superior survival and colonization in the host's gut environment [ 14 ], leading to improved growth performance. This research evaluates the effectiveness of the synbiotic, a combination calf gut origin L. reuteri RM125 and JAT as an alternative to the antibiotic growth promoter, tetracycline. In this study, we explored the impact of synbiotic supplementation on fecal biomarkers, antioxidant status, and the fecal microbial antibiotic sensitivity dynamics in neonatal dairy calves. By focusing on gut health and antibiotic resistance modulation, this research aims to provide sustainable strategies for enhancing calf gut health, while addressing AMR concerns. 2. Materials and Methods 1. Ethics statement The experiment was conducted in the calf shed of the dairy cattle unit of the Livestock Production and Management (LPM) Division, ICAR-IVRI, Izatnagar, Uttar Pradesh, India, following standard management protocols. It was carried out over 12 weeks, in accordance with the guidelines set by the Institutional Ethics Committee (Regt. No. 96/GO/ReBiBt-S/ReBi-L/2000/CPCSEA) of ICAR-IVRI, under approval reference no. 26 − 4/2022-23/JDR/IAEC. The experiment took place during the hot summer months (April to August), with temperatures ranging from 28°C to 44°C. 2. Experimental animals and treatment diets Eighteen newborn Vrindavani crossbred dairy calves, with an average body weight of 23.5 ± 0.5 kg, were selected and randomly assigned to three groups of six animals each, following a completely randomized design (Fig. 1 ). The calves were separated from their dams on day third of birth and were hand-fed with milk from the fourth day onward (Fig. 1 ). The groups were Control (CON) that r eceived a basal diet without any supplementation, Antibiotic Group (TET) s upplemented with tetracycline hydrochloride at 1 mg/kg body weight for two weeks, Synbiotic Group (SYN) that were s upplemented with Limisilactobacillusreuteri (3–5 × 10⁸ cfu/mL) and Jerusalem artichoke tuber (JAT) powder at 2% of dry matter intake for 12 weeks (5). The synbiotic combination ( L. reuteri RM125, NCBI accession No. MT903467) and JAT powder was selected based on in-vitro hindgut fermentation results [ 13 ]. Tetracycline hydrochloride was mixed with sterile distilled water and administered orally [ 16 ] at a dose of 1 mg/kg body weight. All calves were pail-fed with milk according to standard milk feeding schedule followed by weaning at 72 days. Green forage and calf starter were provided ad libitum . The calf starter comprised of maize (50%), soybean meal (37%), wheat bran (10%), mineral mixture (2%), and salt (1%), prepared every four weeks by hand-mixing the ingredients. Standard practices such as deworming, debudding, and deticking were uniformly followed for all calves throughout the study. Calves were observed for weekly weight changes, DMI intake from calf starter, green fodder and milk to evaluate the intake and growth performance. 3. Fecal sample collection and processing Fecal samples (4–5 g) were collected directly from the rectum at 0, 1, 2, 3, 4, 6, 8, 10, and 12 weeks to assess fecal Lactobacillus, Bifidobacteria , and coliform counts, as well as biochemical indices including pH, lactic acid, ammonia, and short-chain fatty acids (SCFA). Bacterial loads were estimated and converted to log10 cfu/g of fresh feces [ 17 ]. Fecal pH was measured immediately using a digital pH meter (Hanna Instruments, H198127, Mumbai). Fecal ammonia was determined by the Berthelot reaction [ 18 ], lactic acid was estimated by colorimetric method [ 19 ] and SCFAs like acetate, propionate, butyrate, total volatile fatty acids (TVFA), and acetate-to-propionate ratio were quantified using gas-liquid chromatography (GLC, Neucon II, AshcoNiulab Equipment Company, Mumbai) [ 20 ]. The stationary phase consisted of a liquid adsorbed on an inert solid matrix, and nitrogen was used as the mobile phase. The gas flow for nitrogen, hydrogen and air were 30, 30 and 300 ml/min, respectively. Temperature of injector oven, column oven and detector were 270ºC, 172ºC and 270ºC, respectively. 4. Serum biochemicals and antioxidant markers Blood samples were collected from all calves at 0, 6, and 12 weeks in clot activator tubes for serum separation. Liver and renal profiles, along with calcium and phosphorus levels, were analyzed using commercial serum kits (Coral Clinical Systems, Tulip Diagnostics Pvt Ltd.) following the manufacturer’s protocols. Serum lipid profiles were determined using quantitative enzymatic colorimetric methods with commercial test kits from the same manufacturer. The antioxidant status was evaluated by measuring superoxide dismutase (SOD), glutathione peroxidase (GPx), and malondialdehyde (MDA) levels using sandwich ELISA kits with pre-coated plates containing specific bovine serum antibodies (BT LAB Bioassay Technology Laboratories, New Delhi). Catalase (CAT) activity was measured using a CAT assay kit (ImmunoTag, Immuneel Therapeutics Pvt Ltd). 5. Fecal Microbial Antibiotic Sensitivity Test (ABST) Sterile fecal samples were collected at weekly intervals (0–12 weeks) to study the fecal microbial sensitivity to selected antibiotics by antibiotic disc sensitivity test (ABST) [ 21 ]. The rectal swabs of calves were transferred to tubes containing sterile normal saline, vortexed 90 sec to release the fecal material and 10 µl of fecal suspension was transferred to sterile BHI (brain heart infusion) broth tubes and incubated overnight at 37 0 C. Microbial pellet was prepared by taking 100 µl of incubated BHI broth and adjusted with sterile NS to standardize with 0.5 Mcfarland`s standard. These adjusted cultures were spread over sterile MHA (Muller Hinton Agar) plates using sterile cotton swabs and antibiotic Hexadisc (hexa-G minus21 Combi 90 - Himedia, cat. # OD291R-1PK) with ampicillin (10 mcg), chloramphenicol (30 mcg), ceftriaxone (20 mcg), cotrimoxazole (25 mcg), ciprofloxacin (5 mcg) and tetracycline (30 mcg) was placed on the culture spread plates. These plates were incubated at 37 0 C for 24 h to measure the zones of inhibition (ZOI, mm). After 24 h incubation the plates were read with the help of ZOI scale to record the resistance pattern against each antibiotic disc. The influence of synbiotic supplementation on the fecal microbiome's antibiotic sensitivity was evaluated by conducting weekly ABST and measuring ZOI against each antibiotic disc. Weekly ZOI values for treatment groups were represented as heat maps (Fig. 3 ). 6. Statistical analysis: The data generated in the current study were analyzed statistically following the methods of Snedecor and Cochran (1985) using SPSS version 26.0 (Chicago, USA). Fecal microbial counts, biochemical indices, serum biochemistry, antioxidant profile, and fecal antibiotic sensitivity tests were analyzed using two-way analysis of variance (ANOVA), considering treatment and period as the two factors. Heat maps depicting the dynamics of zones of inhibition (ZOI) against the tested antibiotic discs were constructed using GraphPad Prism version 9.0 (released in 2020). Statistical significance was determined at 5% level of significance (P < 0.05) using Tukey. 3. Results 1. Growth performance The growth performance of calves under different treatments was evaluated considering the body weight changes, DMI intake from calf starter, forage and milk for a period of 12 weeks. The performance of calves under current study showed that sybiotic supplementation significantly resulted in higher body weight gain (15% higher than control) and DMI intake in SYN calves over TET and CON calves (Fig. 2). 2. Fecal microbial load, metabolites and fermentative indices The average fecal Lactobacillus, Bifidobacterial and coliforms count for 0–12 week period expressed as log cfu/g of fresh feces (Table 1 ). The total viable fecal microbial load as affected by treatments and time post supplementation of TET or SYN is presented as supplementary data (Table S1 ). The fecal coliform counts varied non-significantly (P > 0.05) among CON, TET and SYN groups. There was significant reduction in fecal coliform load with advancement of age (8.70 at week 0 to 6.07 at week 12) and the interaction of treatment and time also affected the coliform count. The fecal Lactobacillus, Bifidobacterial loads improved significantly (P < 0.05) with the supplementation of either synbiotic (SYN) or antibiotic tetracycline (TET) as compared to CON calves. No specific pattern was observed fecal Lactobacillus (8.9 at 0-week vs 6.6 at week 12) and Bifidobacterial loads with the advancement of calf age. Hind gut health indicators (lower fecal pH, ammonia, higher lactic acid and SCFAs) improved significantly (P < 0.05) in SYN calves over CON calves (Table 1 ). The fecal lactic acid of TET calves was found comparable with CON and SYN groups. The fecal SCFA fractions Viz. , acetate, propionate, butyrate and TVFA were improved significantly (P < 0.05) in SYN group over CON and TET group calves. Temporal changes were observed in the fecal SCFA fractions (Table S3) and TVFA but the interactive effect of time and treatment was found negligible. Similar to fecal microbial loads, significant (P < 0.05) variations in fecal pH, metabolites (lactic acid, ammonia) were observed with increasing age of calves (Table S2). Table 1 Effect of tetracycline or synbiotic supplementation on fecal microbial load, metabolites and fermentative indices Fecal Attribute CON TET SYN SEM P value Fecal microbial load Lactobacillus 7.97 b 8.40 a 8.50 a 0.033 < 0.001 Bifidobacterial 7.73 b 8.13 a 8.24 a 0.072 < 0.001 Coliforms 6.67 6.79 6.60 0.168 0.321 Hindgut metabolites pH 6.65 b 6.81 a 6.58 c 0.276 0.040 Ammonia 19.97 a 19.84 a 16.66 b 1.301 0.018 Lactic acid 1.61 b 1.89 ab 2.05 a 0.145 0.038 Hind gut fermentative indices Acetate 3.56 b 3.99 b 4.48 a 0.683 0.029 Propionate 1.58 b 1.60 b 1.81 a 0.310 0.004 Butyrate 0.49 b 0.49 b 0.62 a 0.021 0.025 TVFA 5.63 b 6.07 ab 7.02 a 1.054 0.041 Acetate: Propionate 2.32 2.64 2.38 0.423 0.996 abc Means bearing different superscripts in a row differ significantly (P < 0.05)CON, milk + calf starter-forage based diets; TET tetracycline 1 mg/kg body weight; ,SYN, L.reuteri (3–5 × 10 9 cfu/mL) and JAT powder at 2% of DM 3. Serum biochemical profile and antioxidant status of calves Serum biochemical profile of calves with respect to liver and kidney function indices in different treatment groups (Table 2) were comparable at all tested stages (0, and 12 weeks). Interestingly, improved lipid profile was observed in SYN calves observed with reduced total cholesterol, LDL cholesterol and improved Triglycerides: Total Cholesterol and HDL: LDL ratio. Temporal variations were found significant (P<0.05) for triglycerides, VLDL cholesterol that decreased significantly with age of calves (Table S4). The serum calcium and phosphorus levels were comparable among three treatments, whereas the ratio of Ca:P increased significantly (P<0.05) in SYN over CON group. Serum antioxidants MDA (ng/Dl) and CAT were significantly (P<0.05)lower and higher in the treated groups (SYN and TET) as compared to CON group (Table 3) meanwhile, significant (P<0.05) temporal variations, treatment time interactive effect was observed (Table S5) in serum SOD and GPx levels (ng/Dl). Table 2 Effect of tetracycline or synbiotic supplementation on serum biochemical indices of calves Serum attribute CON TET SYN SEM P value Liver function indices Total protein(g/dL) 7.64 7.59 7.37 0.423 0.867 Albumin (g/dL) 3.16 3.13 2.74 0.192 0.257 Globulin (g/dL) 4.48 4.54 4.63 0.065 0.959 A:G ratio 0.93 0.83 0.63 0.090 0.067 Kidney function indices Urea (mg/dL) 35.44 35.01 27.07 3.546 0.182 Creatinine (mg/dL) 1.89 1.61 1.74 0.261 0.745 Blood urea nitrogen (mg/dL) 16.56 16.36 12.65 4.443 0.443 Lipid profile Total cholesterol (mg/dL) 75.06 a 72.19 ab 59.92 b 4.844 0.034 Triglycerides(mg/dL) 40.91 57.02 46.43 5.828 0.151 HDL cholesterol (mg/dL) 24.85 22.37 22.56 0.749 0.807 LDL Cholesterol (mg/dL) 42.03 a 38.42 ab 28.07 b 5.09 0.040 VLDL cholesterol (mg/dL) 8.18 11.40 9.28 1.17 0.151 Triglycerides :Total Cholesterol 0.549 b 0.827 a 0.799 a 0.115 0.044 HDL:LDL 0.623 c 0.732 b 0.958 a 0.122 0.012 Serum Mineral status Calcium 8.30 8.74 8.90 0.114 0.215 Phosphorus 8.16 8.11 7.85 0.154 0.332 Calcium: Phosphorus 1.01 a 1.07 ab 1.13 b 0.036 0.047 abc Means bearing different superscripts in a row differ significantly (P<0.05) CON, milk + calf starter-forage based diets; TET tetracycline 1 mg/kg body weight; ,SYN, L.reuteri (3-5 × 10 9 cfu/mL) and JAT powder at 2% of DM Table 3 Effect of tetracycline or synbiotic supplementation on serum antioxidant status of calves Serum enzyme CON TET SYN SEM P value Malondialdehyde (ng/dL) 9.24 a 7.46 b 8.68 ab 0.249 0.021 Reduced Superoxide dismutase (ng/dL) 7.49 6.86 7.04 0.999 0.726 Reduced Glutathione (ng/dL) 13.0 11.2 12.1 1.940 0.447 Catalase (IU/dL) 2.89 c 4.77 a 3.97 b 0.678 0.014 ab Means bearing different superscripts in a row differ significantly (P<0.05) CON, milk + calf starter-forage based diets; TET tetracycline 1 mg/kg body weight; ,SYN, L.reuteri (3-5 × 10 9 cfu/mL) and JAT powder at 2% of DM 4. Fecal microbial ABST The weekly ZOI values of fecal microbiota against each antibiotic tested are represented as heat maps (Fig. 3 ). Though there was no definite trend, however the fecal microbiota of all the three groups was highly resistant to ampicillin and cotrimoxazole compared to other antibiotics. Comparison of mean ZOI values revealed no significant difference among the three groups against ampicillin, chloramphenicol, cotrimoxazole and tetracycline. Significantly, higher mean ZOI values were recorded in SYN group for ceftriaxone as compared to TET and CON, For ciprofloxacin, the ZOI values were significantly higher in both TET and SYN group over CON (Table 4 ). The ZOI values against chloramphenicol, ceftriaxone, ciprofloxacin and tetracycline were significantly influenced by age and interactive effect of treatment and age was evident (Table 4 ). Table 4 Effect of antibiotic and synbiotic supplementation on temporal changes in fecal microbial sensitivity against different antibiotics Attribute Treatment groups* Period average SEM P CON AB SYN Ampicillin Week 0 6.00 6.00 6.00 6.00 0.196 T = 0.484 P = 0.319 T*P = 0.484 Week 1 6.00 6.00 6.00 6.00 Week 2 6.00 6.00 6.00 6.00 Week 3 6.00 6.00 6.00 6.00 Week 4 6.00 6.00 6.67 6.22 Week 6 6.00 6.00 7.17 6.39 Week 8 6.00 6.00 6.00 6.00 Week 7 6.00 6.00 6.00 6.00 Week 8 6.00 6.00 6.00 6.00 Week 9 6.00 6.00 6.00 6.00 Week 10 6.00 6.00 6.00 6.00 Week 11 6.00 6.67 6.00 6.22 Week 12 6.00 6.00 6.00 6.00 Treatment means 6.00 6.05 6.14 6.06 Chloramphenicol Week 0 13.83 8.33 11.50 11.22 D 2.356 T = 0.088 P < 0.001 T*P < 0.001 Week 1 16.17 8.33 8.17 10.89 D Week 2 15.67 9.83 18.00 14.50 BCD Week 3 23.50 21.67 27.17 24.11 A Week 4 16.00 21.83 20.00 19.28 ABC Week 5 10.33 24.50 25.17 20.00 AB Week 6 19.00 16.00 23.17 19.39 ABC Week 7 12.17 15.50 12.33 13.33 CD Week 8 15.17 14.20 14.00 14.22 BCD Week 9 10.83 17.50 17.83 15.39 BCD Week 10 10.00 13.67 12.50 12.06 D Week 11 21.00 20.67 16.17 19.16 ABC Week 12 10.17 16.00 14.00 13.39 CD Treatment means 14.87 15.95 16.92 15.91 Ceftriaxone Week 0 13.00 10.33 15.83 12.28 BC 1.243 T < 0.001 P < 0.001 T*P = 0.001 Week 1 12.50 14.83 17.17 14.83 AB Week 2 16.83 14.83 17.33 16.33 A Week 3 14.33 15.00 15.33 14.89 AB Week 4 12.83 15.33 13.00 13.72 ABC Week 5 14.00 14.50 15.33 14.61 AB Week 6 13.00 12.00 15.00 13.33 ABC Week 7 8.83 15.17 12.67 12.22 BC Week 8 14.00 10.50 14.50 13.00 ABC Week 9 8.83 13.67 13.33 11.94 BC Week 10 8.00 12.33 12.33 10.89 C Week 11 11.20 13.50 13.83 12.84 BC Week 12 11.83 13.33 13.00 12.72 BC Treatment means 12.25 Y 13.49 Y 14.51 X 13.36 Cotrimaxazole Week 0 6.00 6.00 6.00 6.00 0.684 T = 0.989 P = .355 T*P = .297 Week 1 6.00 6.00 6.00 6.00 Week 2 8.50 6.00 6.00 6.83 Week 3 6.83 6.00 8.33 7.06 Week 4 6.00 6.00 6.00 6.00 Week 5 6.00 6.00 6.67 6.22 Week 6 6.00 8.83 6.00 6.94 Week 7 6.00 6.00 6.00 6.00 Week 8 6.00 6.00 6.00 6.00 Week 9 6.00 6.00 6.00 6.00 Week 10 6.00 6.00 6.00 6.00 Week 11 6.00 6.00 6.00 6.00 Week 12 6.00 6.00 6.00 6.00 Treatment means 6.26 6.22 6.23 6.24 Ciprofloxacin Week 0 7.33 6.00 6.00 6.44 E 1.925 T < 0.001 P < 0.001 T*P < 0.001 Week 1 7.33 6.00 6.00 6.44 E Week 2 12.00 7.33 14.17 11.17 ABCDE Week 3 13.17 12.50 17.17 14.28 AB Week 4 8.33 13.83 15.50 12.56 ABC Week 5 7.83 13.67 14.33 11.94 ABCD Week 6 12.00 18.00 18.00 16.00 A Week 7 8.50 14.83 14.83 12.72 ABC Week 8 6.00 9.33 17.33 10.89 ABCDE Week 9 6.00 15.83 8.67 10.17 BCDE Week 10 6.00 8.50 7.33 7.28 DE Week 11 14.20 12.83 10.50 12.51 ABCD Week 12 11.17 14.50 12.67 12.78 ABC Treatment means 9.55 y 11.78 x 12.50 x 11.28 Tetracycline Week 0 12.17 11.00 11.33 11.50 B 1.070 T = 0.074 P = 0.002 T*P = 0.017 Week 1 14.00 12.00 13.67 13.22 AB Week 2 15.33 10.83 13.17 13.11 AB Week 3 12.67 10.83 12.67 12.06 AB Week 4 11.17 14.67 13.33 13.06 AB Week 5 11.33 10.33 12.00 11.22 B Week 6 12.50 14.83 16.67 14.67 A Week 7 11.17 13.83 13.83 12.94 AB Week 8 13.00 13.33 15.67 14.00 AB Week 9 12.17 12.00 14.67 12.94 AB Week 10 12.67 11.50 13.50 12.56 AB Week 11 15.33 14.67 13.50 14.50 A Week 12 14.67 12.83 11.17 12.89 AB Treatment means 12.97 12.51 13.47 12.99 4. Discussion In this study, supplementation with a synbiotic containing host-specific probiotic Lactobacillus reuteri RM125 and JAT significantly influenced the fecal microbial populations of select microbes, hindgut health indicators, serum biochemicals, antioxidant status, and antimicrobial resistance patterns in neonatal calves compared to the antibiotic growth promoter, tetracycline (TET) and the control group (CON). The synbiotic supplementation (SYN) resulted in a substantial increase in beneficial gut bacteria, particularly Lactobacillus and Bifidobacteria , compared to the control group, consistent with previous findings that synbiotics promote beneficial gut microbiota [ 22 , 23 ]. Notably, fecal coliform counts decreased with advancing age across all groups, reflecting the natural maturation of the gut microbiota. However, this reduction was more pronounced in SYN and TET groups, with SYN calves maintaining a better balance of beneficial and potentially pathogenic bacteria. These results suggest that while both synbiotic and tetracycline supplementation improved gut microbial composition, synbiotics were more effective in promoting beneficial bacteria while suppressing pathogenic ones, supporting the competitive exclusion principle of lactic acid bacteria (LAB) as reported in earlier studies [ 24 ]. The favorable shift in gut microbial populations in SYN was further corroborated by improvements in hindgut health indicators. These enhancements in biochemical indices are consistent with our earlier in vitro findings of synbiotic testing using calf fecal fermentation medium[ 13 ]. The SYN group exhibited a significant reduction in fecal pH and ammonia levels, accompanied by increased concentrations of lactic acid and short-chain fatty acids (SCFAs), particularly acetate, propionate, and butyrate, compared to CON and TET groups. These changes indicate enhanced saccharolytic fermentation, which is advantageous for gut health, as lower ammonia levels reduce the risk of intestinal toxicity and inflammation[ 25 ]. Additionally, increased SCFA levels contribute to gut epithelial health, improved nutrient absorption, and modulation of lipid metabolism. This was reflected in the improved serum lipid profiles observed in SYN calves, characterized by lower total cholesterol, LDL cholesterol, and improved HDL:LDL ratios. These changes suggest a systemic impact of the altered gut microbiota and its metabolites, likely mediated by SCFA-driven modulation of hepatic cholesterol synthesis and bile acid metabolism, aligning with earlier findings on the role of gut microbiota in lipid metabolism [ 26 , 27 ]. The improved lipid profile was accompanied by enhanced antioxidant status in SYN calves, as evidenced by lower malondialdehyde (MDA) levels and increased catalase (CAT) activity, indicating reduced oxidative stress. The significant temporal variations in superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels further highlight the dynamic interplay between gut microbiota and host antioxidant mechanisms with advancing age. These observations are consistent with previous studies reporting the antioxidative properties of synbiotics through enhanced gut microbial metabolism and SCFA production[ 23 , 24 ]. Furthermore, no significant changes were observed in serum calcium and phosphorus levels across all groups, but the improved Ca:P ratio in SYN calves indicates better mineral utilization, possibly facilitated by enhanced SCFA production and superior gut health. This supports earlier findings linking synbiotic supplementation with improved mineral absorption due to SCFA-mediated pH reduction and increased mineral solubility [ 23 ]. These metabolic and antioxidant benefits were mirrored by improved growth performance and health status, where calves supplemented with the synbiotic showed enhanced body weight gains, better dry matter intake, improved feed conversion efficiency (Fig. 2), and overall better health and immune status. These findings suggest that positive modulation of the gut microbiome directly influenced overall performance and health of the animals, highlighting the systemic benefits of synbiotic supplementation. The neonatal gut microbiome is initially dominated by environmental microbes, many of which are antibiotic-resistant, leading to a high resistome load in young calves compared to adults[ 28 ]. As calves mature, their gastrointestinal microbiota becomes more calf-specific, and antimicrobial resistance tends to decline [ 1 , 29 ]. In our study, during the first week of life, calves showed lower zones of inhibition (ZOI) against most antibiotics, reflecting the presence of environmentally acquired resistant strains. These microbes are gradually replaced by calf-specific communities, influencing the gut resistome dynamics in response to dietary transitions[ 30 ] Calves experiencing diarrhea exhibited a higher prevalence of antibiotic resistance genes (ARGs), compared to healthy calves, with a. Notable, 100% prevalence of the quinolone resistance gene gyrA in diarrheic calves [ 31 ]. In contrast, synbiotic (SYN) supplementation enhanced microbial sensitivity to ciprofloxacin (quinolone) and ceftriaxone, indicating its potential to reduce antimicrobial resistance. This aligns with earlier findings that early-life dietary interventions can influence resistome dynamics [ 30 ]. Additionally, ZOI values against chloramphenicol, ceftriaxone, ciprofloxacin, and tetracycline were significantly influenced by age and the interactive effects of treatment and age, highlighting the role of age in shaping the fecal microbiome and its antibiotic response. Higher levels of beta lactamase resistant (blaTEM) and chloramphenicol resistant (floR and catA) genes in neonatal calf feces [ 31 ]. The non-significant changes observed for ampicillin, chloramphenicol, cotrimoxazole or tetracycline in the present study might be due to presence of resistance genes for the respective antibiotics. Overall, present findings underscore the influence of age and dietary interventions such as synbiotic supplementation, on the fecal microbiome antibiotic sensitivity dynamics of neonatal calves and paves a way for future research on resistance gene load as affected by different factors to combat the problem of AMR in dairy industry. 5. Conclusion This study demonstrates that early-life supplementation with a synbiotic containing L. reuteri RM125 and JAT improves gut health, lipid metabolism, antioxidant status, immune responses, and decelerate the gut antimicrobial resistance in neonatal calves. These benefits highlight its potential as an alternative to antibiotic growth promoters, enhancing calf health, growth, and disease resilience while reducing antimicrobial resistance, supporting sustainable animal nutrition. Declarations Conflict of interest: 6. The authors declare that they have no conflict of interest. Author Contribution Anju Kala: Supervised the research work, performed statistical analysis, and critically reviewed the manuscript draft.Kasi Sowjanya Lakshmi R: Executed the field work and was the primary contributor to manuscript writing.Neeta Agarwal: Contributed to the critical revision and refinement of the manuscript.Zunjar Baburao Dubal: Provided monitoring support and participated in manuscript review.Ashok Kumar Verma: Offered overall supervision of the research and provided technical guidance throughout the study.Lal Chandra Chaudhary: Facilitated laboratory infrastructure and provided scientific oversight.Akhilesh Kumar: Conducted serum sample analysis and supervised blood collection procedures.Bharat Shindhe: Assisted in conducting the animal trial and contributed to data collection.Mohammad Younus Ali: Supported statistical analysis and participated in manuscript review. Acknowledgement This research was carried out as part of the SAARC Agriculture PhD Fellowship Programme-2020, undertaken by the first author, with the generous support of the Director of the SAARC Agriculture Centre (SAC), Dhaka, Bangladesh. The authors sincerely appreciate the research facilities made available by the Director of ICAR-Indian Veterinary Research Institute, Izatnagar. Financial support for this study was provided by CAAST-NAHEP, ICAR New Delhi, under the ICAR-World Bank-funded National Agricultural Higher Education Project (NAHEP). The authors confirm that there are no conflicts of interest associated with this work. References Malmuthuge N, Guan LL. Understanding the gut microbiome of dairy calves (2017) Opportunities to improve early-life gut health. J Dairy Sci.;100(7):5996–6005. Alawneh JI, Barreto MO, Moore RJ, Soust M, Al-harbi H, James AS, et al. (2020) Systematic review of an intervention: the use of probiotics to improve health and productivity of calves. Prev Vet Med.183:105147. https://www.sciencedirect.com/science/article/abs/pii/S0167587720306681 Berge ACB, Lindeque P, Moore DA, Sischo WM. A (2005) Clinical Trial Evaluating Prophylactic and Therapeutic Antibiotic Use on Health and Performance of Preweaned Calves. J Dairy Sci 88(6):2166–77. https://www.sciencedirect.com/science/article/pii/S0022030205728927 Morrill JL, Dayton AD, Mickelsen R.(1977) Cultured Milk and Antibiotics for Young Calves. J Dairy Sci. 60(7):1105–9. https://www.sciencedirect.com/science/article/pii/S0022030277839957 Rochegüe T, Haenni M, Mondot S, Astruc C, Cazeau G, Ferry T, Madec JY, Lupo A. (2021) Impact of antibiotic therapies on resistance genes dynamic and composition of the animal gut microbiota. Animals;11(11):3280. https://doi.org/10.3390/ani11113280 Swanson KS, Gibson GR, Hutkins R, Reimer RA, Reid G, Verbeke K, Scott KP, Holscher HD, Azad MB, Delzenne NM, Sanders ME. (2020) The International Scientific Association for Probiotics and Prebiotics (ISAPP) consensus statement on the definition and scope of synbiotics. Nature reviews Gastroenterology & hepatology. 17(11):687-701. https://doi.org/10.1038/s41575-020-0344-2 Elshaghabee FM, Rokana N. . (2022) Mitigation of antibiotic resistance using probiotics, prebiotics and synbiotics A review. Environmental Chemistry Letters. 20(2):1295-308. https://doi.org/10.1007/s10311-021-01382-w Samal L, Chaturvedi VB, Saikumar G, Somvanshi R, Pattanaik AK. (2015) Prebiotic potential of Jerusalem artichoke (Helianthus tuberosus L.) in Wistar rats: effects of levels of supplementation on hindgut fermentation, intestinal morphology, blood metabolites and immune response. Journal of the Science of Food and Agriculture. 95(8):1689-96. https://doi.org/10.1002/jsfa.6873 Ashwin K, Pattanaik AK, Howarth GS (2021) Polyphenolic bioactives as an emerging group of nutraceuticals for promotion of gut health: A review. Food Bioscience. 1;44:101376. https://doi.org/10.1016/j.fbio.2021.101376 Singh M, Varada VV, Kumar S. Synbiotic supplementation influence select antioxidants markers and immune response of Murrah buffalo calves. Emerging Animal Species. 2023 May 1;7:100026. https://doi.org/10.1016/j.eas.2023.100026 Zhang W, An Y, Qin X, Wu X, Wang X, Hou H, Song X, Liu T, Wang B, Huang X, Cao H. (2021) Gut microbiota-derived metabolites in colorectal cancer: the bad and the challenges. Frontiers in oncology. 18;11:739648. https://doi.org/10.3389/fonc.2021.739648 Huang L, Wu C, Gao H, Xu C, Dai M, Huang L, Hao H, Wang X, Cheng G. (2022) Bacterial multidrug efflux pumps at the frontline of antimicrobial resistance: an overview. Antibiotics 13;11(4):520. https://doi.org/10.3390/antibiotics11040520 Lakshmi KS, Kala A, Verma AK, Chaudhary LC, Agarwal N, Ali MY, Shindhe B. (2024) Calf fecal origin Limosilactobacillus reuteri and Helianthus tuberosus synbiotic improved gut health with calf fecal inoculum using In vitro hind gut model. Animal Nutrition and Feed Technology. 2024;24(3):477-93. DOI: 10.5958/0974-181X.2024.00036.1 Martín R, Langella P (2019)Emerging health concepts in the probiotics field: streamlining the definitions. Frontiers in microbiology.21;10:1047. https://doi.org/10.3389/fmicb.2019.01047 Issac YM, Pattanaik AK, Singh A, Gupta M, Jadhav SE, Gaur GK, Kala A. (2024) Influence of Jerusalem artichoke tuber as a prebiotic on the nutrient utilization, weaning stress indicators and immunocompetence of crossbred calves during the pre-and post-weaning phases. Bioactive Carbohydrates and Dietary Fibre. 31:100423. https://doi.org/10.1016/j.bcdf.2024.100423 Thames CH, Pruden A, James RE, Ray PP, Knowlton KF. (2012) Excretion of antibiotic resistance genes by dairy calves fed milk replacers with varying doses of antibiotics. Frontiers in microbi10;3:139. https://doi.org/10.3389/fmicb.2012.00139 Kumar M, Kala A, Chaudhary LC, Agarwal N, Kochewad SA. (2022) Microencapsulated and lyophilized Lactobacillus acidophilus improved gut health and immune status of preruminant calves. Probiotics and Antimicrobial Proteins. 14(3):523-34. https://doi.org/10.1007/s12602-021-09821-4 Patton CJ, Crouch SR. (1977) Spectrophotometric and kinetics investigation of the Berthelot reaction for the determination of ammonia. Analytical chemistry. 49(3):464-9. Barker SB, W, Summerson WH (1941)The colorimetric determination of lactic acid in biological material. Journal of Biological Chemistry. 138:535–54. Cottyn BG, Boucque CV (1968) Rapid method for the gas-chromatographic determination of volatile fatty acids in rumen fluid. Journal of Agricultural and Food Chemistry. 16(1):105-7. Langford FM, Weary DM, Fisher L. (2003) Antibiotic resistance in gut bacteria from dairy calves: a dose response to the level of antibiotics fed in milk. Journal of Dairy Science. 86(12):3963-6. https://doi.org/10.3168/jds.S0022-0302(03)74006-5 Varada VV, Kumar S, Chhotaray S, Tyagi AK (2022) Host-specific probiotics feeding influence growth, gut microbiota, and fecal biomarkers in buffalo calves. AMB Express. 12(1):118. https://doi.org/10.1186/s13568-022-01460-4 Sharma AN, Chaudhary P, Grover CR, Kumar S, Mondal G. (2024) Impact of synbiotics on growth performance and gut health in Murrah buffalo calves. Veterinary Research Communications. 48(1):179-90. https://doi.org/10.1007/s11259-023-10194-y Fan J, Cui H, Mu Z, Yao C, Yang M, Jin Y, Ning C, Zhang H.(2024) Non-targeted metabolomics analysis of fermented traditional Chinese medicine and its impact on growth performance, serum biochemistry, and intestinal microbiome of weaned lambs. Scientific Reports. 14(1):20385. https://doi.org/10.1038/s41598-024-71516-x Jayswal K, Kala A, Chaudhary LC, Kumar A, Tomar AK. (2023) Autochthonous buffalo-gut origin Pediococcus pentosaceus RM119 decreased diarrhea and enhanced gut health, immunity in neonatal Murrah calves. Animal Biotechnology. 34(8):3306-14. https://doi.org/10.1080/10495398.2022.2138415 Hashmi A, Naeem N, Farooq Z, Masood S, Iqbal S, Naseer R. (2016) Effect of prebiotic galacto-oligosaccharides on serum lipid profile of hypercholesterolemics. Probiotics and antimicrobial proteins. 8(1):19-30. https://doi.org/10.1007/s12602-016-9206-1 Kellogg TF (1974) Steroid balance and tissue cholesterol accumulation in germfree and conventional rats fed diets containing saturated and polyunsaturated fats. Journal of Lipid Research. 15(6):574-9. DOI: 10.1016/S0022-2275(20)36758-4 Liu J, Taft DH, Maldonado-Gomez MX, Johnson D, Treiber ML, Lemay DG, DePeters EJ, Mills DA. (2019) The fecal resistome of dairy cattle is associated with diet during nursing. Nature communications. 10(1):4406. https://doi.org/10.1038/s41467-019-12111-x Oikonomou G, Teixeira AG, Foditsch C, Bicalho ML, Machado VS, Bicalho RC (2013) Fecal microbial diversity in pre-weaned dairy calves as described by pyrosequencing of metagenomic 16S rDNA. Associations of Faecalibacterium species with health and growth. PloS one. 8(4):e63157. https://doi.org/10.1371/journal.pone.0063157 Liu J, Taft DH, Maldonado-Gomez MX, Johnson D, Treiber ML, Lemay DG, DePeters EJ, Mills DA (2019) The fecal resistome of dairy cattle is associated with diet during nursing. Nature communications. 10(1):4406. https://doi.org/10.1038/s41467-019-12111-x Zhang L, Bai J, Guo Q, Li L, Jia Y, Qiu X, Zhou D, Zhang Z, Niu H (2024) Gut Microbial Composition and Antibiotic Resistance Profiles in Dairy Calves with Diarrhea. Life. 15(1):10. https://doi.org/10.3390/life15010010 Additional Declarations No competing interests reported. Supplementary Files SupplementarytablesforCALFARTICLE2.docx Cite Share Download PDF Status: Posted Version 1 posted 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-7146873","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":508087667,"identity":"e031f37a-9878-4166-98c1-664ebb4967df","order_by":0,"name":"Kasi Sowjanya Lakshmi R","email":"","orcid":"","institution":"ICAR-Indian Veterinary Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Kasi","middleName":"Sowjanya Lakshmi","lastName":"R","suffix":""},{"id":508087668,"identity":"093199cd-99ca-4b04-8230-f317d2ed66e2","order_by":1,"name":"Anju 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Centre","correspondingAuthor":false,"prefix":"","firstName":"Mohammad","middleName":"Younus","lastName":"Ali","suffix":""}],"badges":[],"createdAt":"2025-07-17 08:53:23","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7146873/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7146873/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":90316231,"identity":"11c61452-b9ea-4a6b-b990-21564b7f18ed","added_by":"auto","created_at":"2025-09-01 10:20:09","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":136899,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ePictorial representation of effect of complimentary symbiotic on neonatal calf performance\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7146873/v1/d2b37abae7d2ade3aa77950e.jpg"},{"id":90315265,"identity":"121291bc-d80d-427e-b508-45a27aabf168","added_by":"auto","created_at":"2025-09-01 10:12:09","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":88021,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of synbiotic and tetracycline supplementation on performance of neonatal calves\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003csup\u003eabc\u003c/sup\u003e bars bearing different superscript differ significantly for a parameter\u003c/p\u003e\n\u003cp\u003eCON, milk + calf starter-forage based diets;\u0026nbsp; TET tetracycline 1 mg/kg body weight; ,SYN,\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003cem\u003eL.reuteri\u003c/em\u003e (3-5 ×10\u003csup\u003e9\u003c/sup\u003ecfu/mL) and JAT powder at 2% of DM\u0026nbsp;\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7146873/v1/4e6a64d22f4590e969c73b57.jpg"},{"id":90315267,"identity":"7cbc8b74-1cdf-4fdf-84ac-01c74ca67a73","added_by":"auto","created_at":"2025-09-01 10:12:09","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":167549,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eHeat maps of zone of inhibition against different antibiotic discs (mm)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ea) Tetracycline, b) Ciprofloxacin, c) Ampicillin, d) Ceftriaxone, e) Cotrimoxazole , f) Chloramphenicol\u003c/p\u003e\n\u003cp\u003eCON, milk + calf starter-forage based diets;\u0026nbsp; TET tetracycline 1 mg/kg body weight; ,SYN,\u0026nbsp; \u003cem\u003eL.reuteri\u003c/em\u003e (3-5 × 10\u003csup\u003e9\u003c/sup\u003ecfu/mL) and JAT powder at 2% of DM\u0026nbsp;\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7146873/v1/27cfb22f8a47672d1d0f9d31.jpg"},{"id":92360315,"identity":"1670a9a9-f515-4d87-a92f-04d6f2510697","added_by":"auto","created_at":"2025-09-28 16:31:56","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2047871,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7146873/v1/c2e891cc-3d38-41d1-b34f-53899d7dd74c.pdf"},{"id":90315266,"identity":"0fc832ed-0eaf-4548-9de8-13cd1311924e","added_by":"auto","created_at":"2025-09-01 10:12:09","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":58111,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementarytablesforCALFARTICLE2.docx","url":"https://assets-eu.researchsquare.com/files/rs-7146873/v1/87e04d0bc09ac44faa6ed762.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Novel Limoisilactobacillus reuteri and Helianthus tuberoses synbiotic improved faecal biomarkers, antioxidant status and influenced the fecal microbial antibiotic sensitivity in neonatal dairy calves","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eThe early development of dairy calves is critically influenced by gut health, as a sterile gastrointestinal tract at birth that rapidly becomes colonized by environmental and maternal microbes [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Effective management practices during the neonatal period are crucial, as they influence growth rate, immunity, and overall productivity. Central to this is gut health, which is pivotal for nutrient absorption and immune system development [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. In commercial dairy systems, where calves are separated from the dam after colostrum feeding, this rapid microbial colonization, combined with underdeveloped immunity and weaning stress, predisposes the calves to neonatal diarrohea [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eAntibiotic growth promoters (AGPs) have historically been employed in neonatal calf feeding programs to prevent diarrhea, promote early growth, and improve feed efficiency [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. However, the widespread use of AGPs has contributed to the emergence of antibiotic-resistant pathogens, posing a significant threat to both animal and public health [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. This concern has catalyzed a shift towards non-antibiotic alternatives, including functional foods that promote gut health and favorably modulate the gut microbiome. Functional foods such as probiotics, prebiotics, and synbiotics have garnered attention for their ability to enhance gut health and immunity while mitigating the risk of antibiotic resistance [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Probiotics, such as \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e, inhibit pathogenic colonization, stabilize the gut microbiota, and modulate host immunity [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Prebiotics, including inulin from Jerusalem artichoke tuber (JAT), selectively stimulate beneficial gut bacteria, enhance nutrient absorption, and contribute to the gut's overall health [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Rich in inulin and polyphenolic compounds, JAT has demonstrated prebiotic and antimicrobial properties, making it a promising component of functional foods [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. When combined as synbiotic, these functional foods exhibit synergistic effects, creating a favorable gut environment that supports health and growth [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eEmerging evidence suggests that functional foods not only enhance growth performance and immunity but also play a critical role in modulating the gut resistome including genes involved in drug resistance and detrimental metabolic pathways [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. By promoting the growth of beneficial microbes, synbiotics can suppress antibiotic-resistant pathogen population, potentially reversing antibiotic resistance [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. This attribute is particularly significant in combating the global issue of antimicrobial resistance (AMR).Combining Jerusalem artichoke tuber (JAT) with calf fecal-origin \u003cem\u003eL. reuteri\u003c/em\u003e RM125, a host-specific probiotic, demonstrated enhanced \u003cem\u003ein-vitro\u003c/em\u003e gut health parameters, highlighting its promise as an effective symbiotic [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Host-specific probiotics, provide customized health advantages due to their superior survival and colonization in the host's gut environment [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], leading to improved growth performance.\u003c/p\u003e\u003cp\u003eThis research evaluates the effectiveness of the synbiotic, a combination calf gut origin \u003cem\u003eL. reuteri\u003c/em\u003e RM125 and JAT as an alternative to the antibiotic growth promoter, tetracycline. In this study, we explored the impact of synbiotic supplementation on fecal biomarkers, antioxidant status, and the fecal microbial antibiotic sensitivity dynamics in neonatal dairy calves. By focusing on gut health and antibiotic resistance modulation, this research aims to provide sustainable strategies for enhancing calf gut health, while addressing AMR concerns.\u003c/p\u003e"},{"header":"2. Materials and Methods","content":"\u003cp\u003e\u003cb\u003e1. Ethics statement\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe experiment was conducted in the calf shed of the dairy cattle unit of the Livestock Production and Management (LPM) Division, ICAR-IVRI, Izatnagar, Uttar Pradesh, India, following standard management protocols. It was carried out over 12 weeks, in accordance with the guidelines set by the Institutional Ethics Committee (Regt. No. 96/GO/ReBiBt-S/ReBi-L/2000/CPCSEA) of ICAR-IVRI, under approval reference no. 26\u0026thinsp;\u0026minus;\u0026thinsp;4/2022-23/JDR/IAEC. The experiment took place during the hot summer months (April to August), with temperatures ranging from 28\u0026deg;C to 44\u0026deg;C.\u003c/p\u003e\n\u003ch3\u003e2. Experimental animals and treatment diets\u003c/h3\u003e\n\u003cp\u003eEighteen newborn Vrindavani crossbred dairy calves, with an average body weight of 23.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5 kg, were selected and randomly assigned to three groups of six animals each, following a completely randomized design (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The calves were separated from their dams on day third of birth and were hand-fed with milk from the fourth day onward (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The groups were Control (CON) that \u003cb\u003er\u003c/b\u003eeceived a basal diet without any supplementation, Antibiotic Group (TET) \u003cb\u003es\u003c/b\u003eupplemented with tetracycline hydrochloride at 1 mg/kg body weight for two weeks, Synbiotic Group (SYN) that were \u003cb\u003es\u003c/b\u003eupplemented with \u003cem\u003eLimisilactobacillusreuteri\u003c/em\u003e (3\u0026ndash;5 \u0026times; 10⁸ cfu/mL) and Jerusalem artichoke tuber (JAT) powder at 2% of dry matter intake for 12 weeks (5). The synbiotic combination (\u003cem\u003eL. reuteri\u003c/em\u003e RM125, NCBI accession No. MT903467) and JAT powder was selected based on \u003cem\u003ein-vitro\u003c/em\u003e hindgut fermentation results [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Tetracycline hydrochloride was mixed with sterile distilled water and administered orally [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e] at a dose of 1 mg/kg body weight.\u003c/p\u003e\u003cp\u003eAll calves were pail-fed with milk according to standard milk feeding schedule followed by weaning at 72 days. Green forage and calf starter were provided \u003cem\u003ead libitum\u003c/em\u003e. The calf starter comprised of maize (50%), soybean meal (37%), wheat bran (10%), mineral mixture (2%), and salt (1%), prepared every four weeks by hand-mixing the ingredients. Standard practices such as deworming, debudding, and deticking were uniformly followed for all calves throughout the study. Calves were observed for weekly weight changes, DMI intake from calf starter, green fodder and milk to evaluate the intake and growth performance.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\n\u003ch3\u003e3. Fecal sample collection and processing\u003c/h3\u003e\n\u003cp\u003eFecal samples (4\u0026ndash;5 g) were collected directly from the rectum at 0, 1, 2, 3, 4, 6, 8, 10, and 12 weeks to assess fecal \u003cem\u003eLactobacillus, Bifidobacteria\u003c/em\u003e, and coliform counts, as well as biochemical indices including pH, lactic acid, ammonia, and short-chain fatty acids (SCFA). Bacterial loads were estimated and converted to log10 cfu/g of fresh feces [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Fecal pH was measured immediately using a digital pH meter (Hanna Instruments, H198127, Mumbai). Fecal ammonia was determined by the Berthelot reaction [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e], lactic acid was estimated by colorimetric method [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e] and SCFAs like acetate, propionate, butyrate, total volatile fatty acids (TVFA), and acetate-to-propionate ratio were quantified using gas-liquid chromatography (GLC, Neucon II, AshcoNiulab Equipment Company, Mumbai) [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. The stationary phase consisted of a liquid adsorbed on an inert solid matrix, and nitrogen was used as the mobile phase. The gas flow for nitrogen, hydrogen and air were 30, 30 and 300 ml/min, respectively. Temperature of injector oven, column oven and detector were 270\u0026ordm;C, 172\u0026ordm;C and 270\u0026ordm;C, respectively.\u003c/p\u003e\n\u003ch3\u003e4. Serum biochemicals and antioxidant markers\u003c/h3\u003e\n\u003cp\u003eBlood samples were collected from all calves at 0, 6, and 12 weeks in clot activator tubes for serum separation. Liver and renal profiles, along with calcium and phosphorus levels, were analyzed using commercial serum kits (Coral Clinical Systems, Tulip Diagnostics Pvt Ltd.) following the manufacturer\u0026rsquo;s protocols. Serum lipid profiles were determined using quantitative enzymatic colorimetric methods with commercial test kits from the same manufacturer.\u003c/p\u003e\u003cp\u003eThe antioxidant status was evaluated by measuring superoxide dismutase (SOD), glutathione peroxidase (GPx), and malondialdehyde (MDA) levels using sandwich ELISA kits with pre-coated plates containing specific bovine serum antibodies (BT LAB Bioassay Technology Laboratories, New Delhi). Catalase (CAT) activity was measured using a CAT assay kit (ImmunoTag, Immuneel Therapeutics Pvt Ltd).\u003c/p\u003e\n\u003ch3\u003e5. Fecal Microbial Antibiotic Sensitivity Test (ABST)\u003c/h3\u003e\n\u003cp\u003eSterile fecal samples were collected at weekly intervals (0\u0026ndash;12 weeks) to study the fecal microbial sensitivity to selected antibiotics by antibiotic disc sensitivity test (ABST) [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. The rectal swabs of calves were transferred to tubes containing sterile normal saline, vortexed 90 sec to release the fecal material and 10 \u0026micro;l of fecal suspension was transferred to sterile BHI (brain heart infusion) broth tubes and incubated overnight at 37\u003csup\u003e0\u003c/sup\u003eC. Microbial pellet was prepared by taking 100 \u0026micro;l of incubated BHI broth and adjusted with sterile NS to standardize with 0.5 Mcfarland`s standard. These adjusted cultures were spread over sterile MHA (Muller Hinton Agar) plates using sterile cotton swabs and antibiotic Hexadisc (hexa-G minus21 Combi 90 - Himedia, cat. # OD291R-1PK) with ampicillin (10 mcg), chloramphenicol (30 mcg), ceftriaxone (20 mcg), cotrimoxazole (25 mcg), ciprofloxacin (5 mcg) and tetracycline (30 mcg) was placed on the culture spread plates. These plates were incubated at 37 \u003csup\u003e0\u003c/sup\u003eC for 24 h to measure the zones of inhibition (ZOI, mm). After 24 h incubation the plates were read with the help of ZOI scale to record the resistance pattern against each antibiotic disc. The influence of synbiotic supplementation on the fecal microbiome's antibiotic sensitivity was evaluated by conducting weekly ABST and measuring ZOI against each antibiotic disc. Weekly ZOI values for treatment groups were represented as heat maps (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\n\u003ch3\u003e6. Statistical analysis:\u003c/h3\u003e\n\u003cp\u003eThe data generated in the current study were analyzed statistically following the methods of Snedecor and Cochran (1985) using SPSS version 26.0 (Chicago, USA). Fecal microbial counts, biochemical indices, serum biochemistry, antioxidant profile, and fecal antibiotic sensitivity tests were analyzed using two-way analysis of variance (ANOVA), considering treatment and period as the two factors. Heat maps depicting the dynamics of zones of inhibition (ZOI) against the tested antibiotic discs were constructed using GraphPad Prism version 9.0 (released in 2020). Statistical significance was determined at 5% level of significance (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) using Tukey.\u003c/p\u003e"},{"header":"3. Results","content":"\u003cp\u003e\u003cb\u003e1. Growth performance\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe growth performance of calves under different treatments was evaluated considering the body weight changes, DMI intake from calf starter, forage and milk for a period of 12 weeks. The performance of calves under current study showed that sybiotic supplementation significantly resulted in higher body weight gain (15% higher than control) and DMI intake in SYN calves over TET and CON calves (Fig.\u0026nbsp;2).\u003c/p\u003e\n\u003ch3\u003e2. Fecal microbial load, metabolites and fermentative indices\u003c/h3\u003e\n\u003cp\u003eThe average fecal \u003cem\u003eLactobacillus, Bifidobacterial\u003c/em\u003e and coliforms count for 0\u0026ndash;12 week period expressed as log cfu/g of fresh feces (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The total viable fecal microbial load as affected by treatments and time post supplementation of TET or SYN is presented as supplementary data (Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). The fecal coliform counts varied non-significantly (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05) among CON, TET and SYN groups. There was significant reduction in fecal coliform load with advancement of age (8.70 at week 0 to 6.07 at week 12) and the interaction of treatment and time also affected the coliform count. The fecal \u003cem\u003eLactobacillus, Bifidobacterial\u003c/em\u003e loads improved significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) with the supplementation of either synbiotic (SYN) or antibiotic tetracycline (TET) as compared to CON calves. No specific pattern was observed fecal \u003cem\u003eLactobacillus\u003c/em\u003e (8.9 at 0-week vs 6.6 at week 12) and \u003cem\u003eBifidobacterial\u003c/em\u003e loads with the advancement of calf age.\u003c/p\u003e\u003cp\u003eHind gut health indicators (lower fecal pH, ammonia, higher lactic acid and SCFAs) improved significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) in SYN calves over CON calves (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The fecal lactic acid of TET calves was found comparable with CON and SYN groups. The fecal SCFA fractions \u003cem\u003eViz.\u003c/em\u003e, acetate, propionate, butyrate and TVFA were improved significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) in SYN group over CON and TET group calves. Temporal changes were observed in the fecal SCFA fractions (Table S3) and TVFA but the interactive effect of time and treatment was found negligible. Similar to fecal microbial loads, significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) variations in fecal pH, metabolites (lactic acid, ammonia) were observed with increasing age of calves (Table S2).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eEffect of tetracycline or synbiotic supplementation on fecal microbial load, metabolites and fermentative indices\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"6\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eFecal Attribute\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCON\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eTET\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eSYN\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eSEM\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eP value\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colspan=\"6\" nameend=\"c6\" namest=\"c1\"\u003e\u003cp\u003e\u003cem\u003eFecal microbial load\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eLactobacillus\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e7.97\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e8.40\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e8.50\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.033\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eBifidobacterial\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e7.73\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e8.13\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e8.24\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.072\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eColiforms\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.79\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.60\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.168\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.321\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"6\" nameend=\"c6\" namest=\"c1\"\u003e\u003cp\u003e\u003cb\u003eHindgut metabolites\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003epH\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.65\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.81\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.58\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.276\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.040\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAmmonia\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e19.97\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e19.84\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e16.66\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1.301\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.018\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLactic acid\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.61\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.89\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e2.05\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.145\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.038\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"6\" nameend=\"c6\" namest=\"c1\"\u003e\u003cp\u003e\u003cb\u003eHind gut fermentative indices\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAcetate\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.56\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e3.99\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e4.48\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.683\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.029\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePropionate\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.58\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.60\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.81\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.310\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.004\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eButyrate\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.49\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.49\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.62\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.021\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.025\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTVFA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e5.63\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.07\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e7.02\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1.054\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.041\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAcetate: Propionate\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2.32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e2.64\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e2.38\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.423\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.996\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003csup\u003eabc\u003c/sup\u003eMeans bearing different superscripts in a row differ significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05)CON, milk\u0026thinsp;+\u0026thinsp;calf starter-forage based diets; TET tetracycline 1 mg/kg body weight; ,SYN, \u003cem\u003eL.reuteri\u003c/em\u003e (3\u0026ndash;5 \u0026times; 10\u003csup\u003e9\u003c/sup\u003ecfu/mL) and JAT powder at 2% of DM\u003c/p\u003e\u003cp\u003e\u003cstrong\u003e\u003cem\u003e3. Serum biochemical profile and antioxidant status of calves\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSerum biochemical profile of calves with respect to \u0026nbsp; liver and kidney function indices in different treatment groups (Table 2)\u0026nbsp; were comparable at all tested stages (0, and 12 weeks). Interestingly, improved lipid profile was observed in SYN calves observed with reduced total cholesterol, LDL cholesterol and improved Triglycerides: Total Cholesterol and HDL: LDL ratio. Temporal variations were found significant (P\u0026lt;0.05) for triglycerides, VLDL cholesterol that decreased significantly with age of calves\u0026nbsp;(Table S4). The serum calcium and phosphorus levels were comparable among three treatments, whereas the ratio of Ca:P increased significantly (P\u0026lt;0.05) in SYN over CON group. \u0026nbsp;Serum antioxidants MDA (ng/Dl) and CAT were significantly (P\u0026lt;0.05)lower and higher in the treated groups (SYN and TET) as compared to CON group (Table 3)\u0026nbsp;meanwhile, significant (P\u0026lt;0.05) temporal variations, treatment time interactive effect was observed (Table S5)\u0026nbsp;in serum SOD and GPx levels (ng/Dl).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2 \u0026nbsp;Effect of tetracycline or synbiotic supplementation on serum biochemical indices of calves\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"652\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eSerum attribute\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eCON\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eTET\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eSYN\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eSEM\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eP value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003e\n \u003cp\u003e\u003cstrong\u003eLiver function indices\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eTotal protein(g/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e7.64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e7.59\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e7.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.423\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.867\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eAlbumin (g/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3.16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.192\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.257\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eGlobulin (g/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.065\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.959\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eA:G ratio\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.090\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.067\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003e\n \u003cp\u003e\u003cstrong\u003eKidney function indices\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eUrea (mg/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e35.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e35.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e27.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3.546\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.182\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCreatinine (mg/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1.61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.261\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.745\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eBlood urea nitrogen (mg/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e16.56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e16.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e12.65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4.443\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.443\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eLipid profile\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eTotal cholesterol (mg/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e75.06\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e72.19\u003csup\u003e\u0026nbsp;ab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e59.92\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4.844\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.034\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eTriglycerides(mg/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e40.91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e57.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e46.43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e5.828\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.151\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHDL cholesterol (mg/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e24.85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e22.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e22.56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.749\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.807\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eLDL Cholesterol (mg/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e42.03\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e38.42\u003csup\u003e\u0026nbsp;ab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e28.07\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e5.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.040\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eVLDL cholesterol (mg/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e8.18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e11.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e9.28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.151\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eTriglycerides :Total Cholesterol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.549\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.827\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.799\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.115\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.044\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHDL:LDL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.623\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.732\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.958\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.122\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.012\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eSerum Mineral status\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCalcium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e8.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e8.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e8.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.114\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.215\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003ePhosphorus\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e8.16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e8.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e7.85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.154\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.332\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCalcium: Phosphorus\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1.01\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1.07\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1.13\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.036\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.047\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003csup\u003eabc\u003c/sup\u003eMeans bearing different superscripts in a row \u0026nbsp;differ significantly (P\u0026lt;0.05)\u003c/p\u003e\n\u003cp\u003eCON, milk + calf starter-forage based diets; \u0026nbsp;TET tetracycline 1 mg/kg body weight; ,SYN, \u0026nbsp;\u003cem\u003eL.reuteri\u003c/em\u003e (3-5 \u0026times; 10\u003csup\u003e9\u003c/sup\u003ecfu/mL) and JAT powder at 2% of DM \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3 Effect of tetracycline or synbiotic supplementation on serum antioxidant status of calves\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"634\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eSerum enzyme\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eCON\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eTET\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eSYN\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eSEM\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eP value\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eMalondialdehyde (ng/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e9.24\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e7.46\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e8.68\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.249\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.021\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eReduced Superoxide dismutase (ng/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e7.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e6.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e7.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.999\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.726\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eReduced Glutathione (ng/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e13.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e11.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e12.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1.940\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.447\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCatalase (IU/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2.89\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4.77\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3.97\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.678\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.014\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003csup\u003eab\u0026nbsp;\u003c/sup\u003eMeans bearing different superscripts in a row \u0026nbsp;differ significantly (P\u0026lt;0.05)\u003c/p\u003e\n\u003cp\u003eCON, milk + calf starter-forage based diets; \u0026nbsp;TET tetracycline 1 mg/kg body weight; ,SYN, \u0026nbsp;\u003cem\u003eL.reuteri\u003c/em\u003e (3-5 \u0026times; 10\u003csup\u003e9\u003c/sup\u003ecfu/mL) and JAT powder at 2% of DM \u0026nbsp;\u003c/p\u003e\n\u003ch3\u003e4. Fecal microbial ABST\u003c/h3\u003e\n\u003cp\u003eThe weekly ZOI values of fecal microbiota against each antibiotic tested are represented as heat maps (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Though there was no definite trend, however the fecal microbiota of all the three groups was highly resistant to ampicillin and cotrimoxazole compared to other antibiotics. Comparison of mean ZOI values revealed no significant difference among the three groups against ampicillin, chloramphenicol, cotrimoxazole and tetracycline. Significantly, higher mean ZOI values were recorded in SYN group for ceftriaxone as compared to TET and CON, For ciprofloxacin, the ZOI values were significantly higher in both TET and SYN group over CON (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). The ZOI values against chloramphenicol, ceftriaxone, ciprofloxacin and tetracycline were significantly influenced by age and interactive effect of treatment and age was evident (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eEffect of antibiotic and synbiotic supplementation on temporal changes in fecal microbial sensitivity against different antibiotics\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"7\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eAttribute\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"3\" nameend=\"c4\" namest=\"c2\"\u003e\u003cp\u003eTreatment groups*\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003ePeriod average\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eSEM\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eP\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCON\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAB\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eSYN\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colspan=\"7\" nameend=\"c7\" namest=\"c1\"\u003e\u003cp\u003e\u003cem\u003eAmpicillin\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\" morerows=\"13\" rowspan=\"14\"\u003e\u003cp\u003e0.196\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\" morerows=\"13\" rowspan=\"14\"\u003e\u003cp\u003eT\u0026thinsp;=\u0026thinsp;0.484\u003c/p\u003e\u003cp\u003eP\u0026thinsp;=\u0026thinsp;0.319\u003c/p\u003e\u003cp\u003eT*P\u0026thinsp;=\u0026thinsp;0.484\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.22\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e7.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.39\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.22\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTreatment means\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e6.00\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e6.05\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e6.14\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.06\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"7\" nameend=\"c7\" namest=\"c1\"\u003e\u003cp\u003e\u003cb\u003eChloramphenicol\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e13.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e8.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e11.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e11.22\u003csup\u003eD\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\" morerows=\"13\" rowspan=\"14\"\u003e\u003cp\u003e2.356\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\" morerows=\"13\" rowspan=\"14\"\u003e\u003cp\u003eT\u0026thinsp;=\u0026thinsp;0.088\u003c/p\u003e\u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003cp\u003eT*P\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e16.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e8.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e8.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e10.89\u003csup\u003eD\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e15.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e9.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e18.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e14.50\u003csup\u003eBCD\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e23.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e21.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e27.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e24.11\u003csup\u003eA\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e16.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e21.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e20.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e19.28\u003csup\u003eABC\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e10.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e24.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e25.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e20.00\u003csup\u003eAB\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e19.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e16.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e23.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e19.39\u003csup\u003eABC\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e12.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e15.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e12.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e13.33\u003csup\u003eCD\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e15.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e14.20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e14.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e14.22\u003csup\u003eBCD\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e10.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e17.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e17.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e15.39\u003csup\u003eBCD\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e10.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e13.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e12.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.06\u003csup\u003eD\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e21.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e20.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e16.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e19.16\u003csup\u003eABC\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e10.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e16.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e14.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e13.39\u003csup\u003eCD\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTreatment means\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e14.87\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e15.95\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e16.92\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e15.91\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"7\" nameend=\"c7\" namest=\"c1\"\u003e\u003cp\u003e\u003cb\u003eCeftriaxone\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e13.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e10.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e15.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.28 \u003csup\u003eBC\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\" morerows=\"13\" rowspan=\"14\"\u003e\u003cp\u003e1.243\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\" morerows=\"13\" rowspan=\"14\"\u003e\u003cp\u003eT\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003cp\u003eT*P\u0026thinsp;=\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e12.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e14.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e17.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e14.83\u003csup\u003eAB\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e16.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e14.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e17.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e16.33\u003csup\u003eA\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e14.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e15.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e15.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e14.89\u003csup\u003eAB\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e12.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e15.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e13.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e13.72\u003csup\u003eABC\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e14.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e14.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e15.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e14.61\u003csup\u003eAB\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e13.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e15.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e13.33\u003csup\u003eABC\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e8.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e15.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e12.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.22 \u003csup\u003eBC\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e14.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e10.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e14.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e13.00\u003csup\u003eABC\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e8.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e13.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e13.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e11.94 \u003csup\u003eBC\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e8.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e12.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e10.89 \u003csup\u003eC\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e11.20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e13.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e13.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.84\u003csup\u003eBC\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e11.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e13.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e13.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.72 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namest=\"c1\"\u003e\u003cp\u003e\u003cb\u003eCotrimaxazole\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\" morerows=\"13\" rowspan=\"14\"\u003e\u003cp\u003e0.684\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\" morerows=\"13\" rowspan=\"14\"\u003e\u003cp\u003eT\u0026thinsp;=\u0026thinsp;0.989\u003c/p\u003e\u003cp\u003eP\u0026thinsp;=\u0026thinsp;.355\u003c/p\u003e\u003cp\u003eT*P\u0026thinsp;=\u0026thinsp;.297\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e8.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.83\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e8.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e7.06\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.22\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e8.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.94\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTreatment means\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e6.26\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e6.22\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e6.23\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e6.24\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"7\" nameend=\"c7\" namest=\"c1\"\u003e\u003cp\u003e\u003cb\u003eCiprofloxacin\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e7.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.44 \u003csup\u003eE\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\" morerows=\"13\" rowspan=\"14\"\u003e\u003cp\u003e1.925\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\" morerows=\"13\" rowspan=\"14\"\u003e\u003cp\u003eT\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003cp\u003eT*P\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e7.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.44\u003csup\u003eE\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e12.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e7.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e14.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e11.17\u003csup\u003eABCDE\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e13.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e17.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e14.28\u003csup\u003eAB\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e8.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e13.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e15.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.56\u003csup\u003eABC\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e7.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e13.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e14.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e11.94\u003csup\u003eABCD\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e12.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e18.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e18.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e16.00\u003csup\u003eA\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e8.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e14.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e14.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.72\u003csup\u003eABC\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e9.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e17.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e10.89 \u003csup\u003eABCDE\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e15.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e8.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e10.17 \u003csup\u003eBCDE\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e8.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e7.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e7.28 \u003csup\u003eDE\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e14.20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e10.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.51\u003csup\u003eABCD\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e11.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e14.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e12.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.78\u003csup\u003eABC\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTreatment means\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e9.55\u003c/b\u003e\u003csup\u003e\u003cb\u003ey\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e11.78\u003c/b\u003e\u003csup\u003e\u003cb\u003ex\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e12.50\u003c/b\u003e\u003csup\u003e\u003cb\u003ex\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e11.28\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"7\" nameend=\"c7\" namest=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTetracycline\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e12.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e11.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e11.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e11.50\u003csup\u003eB\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\" morerows=\"13\" rowspan=\"14\"\u003e\u003cp\u003e1.070\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\" morerows=\"13\" rowspan=\"14\"\u003e\u003cp\u003eT\u0026thinsp;=\u0026thinsp;0.074\u003c/p\u003e\u003cp\u003eP\u0026thinsp;=\u0026thinsp;0.002\u003c/p\u003e\u003cp\u003eT*P\u0026thinsp;=\u0026thinsp;0.017\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e14.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e13.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e13.22\u003csup\u003eAB\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e15.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e10.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e13.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e13.11\u003csup\u003eAB\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e12.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e10.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e12.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.06 \u003csup\u003eAB\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e11.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e14.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e13.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e13.06 \u003csup\u003eAB\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e11.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e10.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e12.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e11.22\u003csup\u003eB\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e12.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e14.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e16.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e14.67\u003csup\u003eA\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e11.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e13.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e13.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.94 \u003csup\u003eAB\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e13.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e13.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e15.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e14.00\u003csup\u003eAB\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e12.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e14.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.94 \u003csup\u003eAB\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e12.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e11.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e13.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.56 \u003csup\u003eAB\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e15.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e14.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e13.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e14.50\u003csup\u003eA\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeek 12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e14.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e11.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.89 \u003csup\u003eAB\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTreatment means\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e12.97\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e12.51\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e13.47\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e12.99\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eIn this study, supplementation with a synbiotic containing host-specific probiotic \u003cem\u003eLactobacillus reuteri\u003c/em\u003e RM125 and JAT significantly influenced the fecal microbial populations of select microbes, hindgut health indicators, serum biochemicals, antioxidant status, and antimicrobial resistance patterns in neonatal calves compared to the antibiotic growth promoter, tetracycline (TET) and the control group (CON). The synbiotic supplementation (SYN) resulted in a substantial increase in beneficial gut bacteria, particularly \u003cem\u003eLactobacillus\u003c/em\u003e and \u003cem\u003eBifidobacteria\u003c/em\u003e, compared to the control group, consistent with previous findings that synbiotics promote beneficial gut microbiota [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. Notably, fecal coliform counts decreased with advancing age across all groups, reflecting the natural maturation of the gut microbiota. However, this reduction was more pronounced in SYN and TET groups, with SYN calves maintaining a better balance of beneficial and potentially pathogenic bacteria. These results suggest that while both synbiotic and tetracycline supplementation improved gut microbial composition, synbiotics were more effective in promoting beneficial bacteria while suppressing pathogenic ones, supporting the competitive exclusion principle of lactic acid bacteria (LAB) as reported in earlier studies [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe favorable shift in gut microbial populations in SYN was further corroborated by improvements in hindgut health indicators. These enhancements in biochemical indices are consistent with our earlier \u003cem\u003ein vitro\u003c/em\u003e findings of synbiotic testing using calf fecal fermentation medium[\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. The SYN group exhibited a significant reduction in fecal pH and ammonia levels, accompanied by increased concentrations of lactic acid and short-chain fatty acids (SCFAs), particularly acetate, propionate, and butyrate, compared to CON and TET groups. These changes indicate enhanced saccharolytic fermentation, which is advantageous for gut health, as lower ammonia levels reduce the risk of intestinal toxicity and inflammation[\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Additionally, increased SCFA levels contribute to gut epithelial health, improved nutrient absorption, and modulation of lipid metabolism. This was reflected in the improved serum lipid profiles observed in SYN calves, characterized by lower total cholesterol, LDL cholesterol, and improved HDL:LDL ratios. These changes suggest a systemic impact of the altered gut microbiota and its metabolites, likely mediated by SCFA-driven modulation of hepatic cholesterol synthesis and bile acid metabolism, aligning with earlier findings on the role of gut microbiota in lipid metabolism [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe improved lipid profile was accompanied by enhanced antioxidant status in SYN calves, as evidenced by lower malondialdehyde (MDA) levels and increased catalase (CAT) activity, indicating reduced oxidative stress. The significant temporal variations in superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels further highlight the dynamic interplay between gut microbiota and host antioxidant mechanisms with advancing age. These observations are consistent with previous studies reporting the antioxidative properties of synbiotics through enhanced gut microbial metabolism and SCFA production[\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. Furthermore, no significant changes were observed in serum calcium and phosphorus levels across all groups, but the improved Ca:P ratio in SYN calves indicates better mineral utilization, possibly facilitated by enhanced SCFA production and superior gut health. This supports earlier findings linking synbiotic supplementation with improved mineral absorption due to SCFA-mediated pH reduction and increased mineral solubility [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThese metabolic and antioxidant benefits were mirrored by improved growth performance and health status, where calves supplemented with the synbiotic showed enhanced body weight gains, better dry matter intake, improved feed conversion efficiency (Fig.\u0026nbsp;2), and overall better health and immune status. These findings suggest that positive modulation of the gut microbiome directly influenced overall performance and health of the animals, highlighting the systemic benefits of synbiotic supplementation.\u003c/p\u003e\u003cp\u003eThe neonatal gut microbiome is initially dominated by environmental microbes, many of which are antibiotic-resistant, leading to a high resistome load in young calves compared to adults[\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. As calves mature, their gastrointestinal microbiota becomes more calf-specific, and antimicrobial resistance tends to decline [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. In our study, during the first week of life, calves showed lower zones of inhibition (ZOI) against most antibiotics, reflecting the presence of environmentally acquired resistant strains. These microbes are gradually replaced by calf-specific communities, influencing the gut resistome dynamics in response to dietary transitions[\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e] Calves experiencing diarrhea exhibited a higher prevalence of antibiotic resistance genes (ARGs), compared to healthy calves, with a. Notable, 100% prevalence of the quinolone resistance gene gyrA in diarrheic calves [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eIn contrast, synbiotic (SYN) supplementation enhanced microbial sensitivity to ciprofloxacin (quinolone) and ceftriaxone, indicating its potential to reduce antimicrobial resistance. This aligns with earlier findings that early-life dietary interventions can influence resistome dynamics [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. Additionally, ZOI values against chloramphenicol, ceftriaxone, ciprofloxacin, and tetracycline were significantly influenced by age and the interactive effects of treatment and age, highlighting the role of age in shaping the fecal microbiome and its antibiotic response.\u003c/p\u003e\u003cp\u003eHigher levels of beta lactamase resistant (blaTEM) and chloramphenicol resistant (floR and catA) genes in neonatal calf feces [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. The non-significant changes observed for ampicillin, chloramphenicol, cotrimoxazole or tetracycline in the present study might be due to presence of resistance genes for the respective antibiotics. Overall, present findings underscore the influence of age and dietary interventions such as synbiotic supplementation, on the fecal microbiome antibiotic sensitivity dynamics of neonatal calves and paves a way for future research on resistance gene load as affected by different factors to combat the problem of AMR in dairy industry.\u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eThis study demonstrates that early-life supplementation with a synbiotic containing \u003cem\u003eL. reuteri\u003c/em\u003e RM125 and JAT improves gut health, lipid metabolism, antioxidant status, immune responses, and decelerate the gut antimicrobial resistance in neonatal calves. These benefits highlight its potential as an alternative to antibiotic growth promoters, enhancing calf health, growth, and disease resilience while reducing antimicrobial resistance, supporting sustainable animal nutrition.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003ch2\u003eConflict of interest:\u003c/h2\u003e\u003cp\u003e6. The authors declare that they have no conflict of interest.\u003c/p\u003e\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eAnju Kala: Supervised the research work, performed statistical analysis, and critically reviewed the manuscript draft.Kasi Sowjanya Lakshmi R: Executed the field work and was the primary contributor to manuscript writing.Neeta Agarwal: Contributed to the critical revision and refinement of the manuscript.Zunjar Baburao Dubal: Provided monitoring support and participated in manuscript review.Ashok Kumar Verma: Offered overall supervision of the research and provided technical guidance throughout the study.Lal Chandra Chaudhary: Facilitated laboratory infrastructure and provided scientific oversight.Akhilesh Kumar: Conducted serum sample analysis and supervised blood collection procedures.Bharat Shindhe: Assisted in conducting the animal trial and contributed to data collection.Mohammad Younus Ali: Supported statistical analysis and participated in manuscript review.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eThis research was carried out as part of the SAARC Agriculture PhD Fellowship Programme-2020, undertaken by the first author, with the generous support of the Director of the SAARC Agriculture Centre (SAC), Dhaka, Bangladesh. The authors sincerely appreciate the research facilities made available by the Director of ICAR-Indian Veterinary Research Institute, Izatnagar. Financial support for this study was provided by CAAST-NAHEP, ICAR New Delhi, under the ICAR-World Bank-funded National Agricultural Higher Education Project (NAHEP). The authors confirm that there are no conflicts of interest associated with this work.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eMalmuthuge N, Guan LL. Understanding the gut microbiome of dairy calves (2017) Opportunities to improve early-life gut health. J Dairy Sci.;100(7):5996\u0026ndash;6005. \u003c/li\u003e\n\u003cli\u003eAlawneh JI, Barreto MO, Moore RJ, Soust M, Al-harbi H, James AS, et al. (2020) Systematic review of an intervention: the use of probiotics to improve health and productivity of calves. 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DOI: 10.1016/S0022-2275(20)36758-4 \u003c/li\u003e\n\u003cli\u003eLiu J, Taft DH, Maldonado-Gomez MX, Johnson D, Treiber ML, Lemay DG, DePeters EJ, Mills DA. (2019) The fecal resistome of dairy cattle is associated with diet during nursing. Nature communications. 10(1):4406. https://doi.org/10.1038/s41467-019-12111-x\u003c/li\u003e\n\u003cli\u003eOikonomou G, Teixeira AG, Foditsch C, Bicalho ML, Machado VS, Bicalho RC (2013) Fecal microbial diversity in pre-weaned dairy calves as described by pyrosequencing of metagenomic 16S rDNA. Associations of Faecalibacterium species with health and growth. PloS one. 8(4):e63157. https://doi.org/10.1371/journal.pone.0063157\u003c/li\u003e\n\u003cli\u003eLiu J, Taft DH, Maldonado-Gomez MX, Johnson D, Treiber ML, Lemay DG, DePeters EJ, Mills DA (2019) The fecal resistome of dairy cattle is associated with diet during nursing. Nature communications. 10(1):4406. https://doi.org/10.1038/s41467-019-12111-x\u003c/li\u003e\n\u003cli\u003eZhang L, Bai J, Guo Q, Li L, Jia Y, Qiu X, Zhou D, Zhang Z, Niu H (2024) Gut Microbial Composition and Antibiotic Resistance Profiles in Dairy Calves with Diarrhea. Life. 15(1):10. \u003cstrong\u003ehttps://doi.org/10.3390/life15010010\u003c/strong\u003e\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Synbiotic, Neonatal calves, Host-specific probiotic, Jerusalem artichoke, Gut health, Gut resistome","lastPublishedDoi":"10.21203/rs.3.rs-7146873/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7146873/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eAntibiotic growth promoters (AGPs) are commonly used in neonatal dairy calves to prevent diarrhea and enhance early growth performance. However, their extensive use contributes to antimicrobial resistance (AMR). This study evaluated the potential of a calf fecal-origin probiotic-based synbiotic as an alternative to AGP tetracycline (TET) in influencing fecal biomarkers, antioxidant status, and the gut resistome in neonatal dairy calves. Eighteen newborn Vrindavani calves\u0026rsquo; hand-fed milk, were randomly assigned to three groups (n\u0026thinsp;=\u0026thinsp;6): no supplementation (CON), TET (1 mg/kg body weight) for 2 weeks, and synbiotic (SYN \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003eRM125 (3\u0026ndash;5\u0026times;10⁸ cfu/mL) and Jerusalem artichoke tuber (\u003cem\u003eHelianthus tuberoses\u003c/em\u003e, JAT) powder (2% of dry matter intake) for 12 weeks. The calves were provided free access to forage and calf starter. Rectal fecal samples collected at 0, 1, 2, 3, 4, 6, 8, 10, and 12 week to assess gut health parameters. Fresh fecal samples were subjected to weekly antibiotic sensitivity tests against six representative antibiotics. Serum samples collected at weeks 0, 6, and 12 were analyzed for biochemical and antioxidant markers. The SYN group showed a significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) increase in fecal \u003cem\u003eLactobacillus\u003c/em\u003e and \u003cem\u003eBifidobacteria\u003c/em\u003e counts, along with significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) reduced pH and ammonia levels, and improved lactic acid and short-chain fatty acids, indicating enhanced gut health. The serum lipid profile significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) improved in SYN calves, while liver and kidney function indices and serum mineral status remained comparable across the groups. Antioxidant status improved in the SYN group, with higher malondialdehyde (MDA) and catalase (CAT) enzyme activity. Among the six antibiotics viz, tetracycline, ciprofloxacin, ampicillin, ceftriaxone, cotrimoxazole and chloramphenicol, the fecal microbiome of SYN calves exhibited significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher zones of inhibition for ciprofloxacin and ceftriaxone, suggesting a favorable shift in the gut resistome. These findings suggest that synbiotic containing calf fecal-origin \u003cem\u003eL. reuteri\u003c/em\u003e RM125 and JAT is a promising alternative to AGPs for enhancing fecal biomarkers, antioxidant status, and gut microbial resistance in neonatal dairy calves.\u003c/p\u003e","manuscriptTitle":"Novel Limoisilactobacillus reuteri and Helianthus tuberoses synbiotic improved faecal biomarkers, antioxidant status and influenced the fecal microbial antibiotic sensitivity in neonatal dairy calves","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-01 10:12:04","doi":"10.21203/rs.3.rs-7146873/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"5d179dd2-6528-4596-8158-a8c2021ec170","owner":[],"postedDate":"September 1st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-09-28T16:23:31+00:00","versionOfRecord":[],"versionCreatedAt":"2025-09-01 10:12:04","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7146873","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7146873","identity":"rs-7146873","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}