Exploring Nutritional Evaluation of Cauliflower (Brassica Oleracea Var Botrytis) Waste on the Thyroid Hormone, Antioxidant Status, Immunity and Expression of Interleukin 6 and HSP 70 Genes in Broilers

Exploring Nutritional Evaluation of Cauliflower (Brassica Oleracea Var Botrytis) Waste on the Thyroid Hormone, Antioxidant Status, Immunity and Expression of Interleukin 6 and HSP 70 Genes in Broilers | 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 Exploring Nutritional Evaluation of Cauliflower (Brassica Oleracea Var Botrytis) Waste on the Thyroid Hormone, Antioxidant Status, Immunity and Expression of Interleukin 6 and HSP 70 Genes in Broilers Mahesh Theerthesh, Avishek Biswas, Gautham Kolluri, Subodh Kumar Saha, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7145387/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 10 Feb, 2026 Read the published version in Waste and Biomass Valorization → Version 1 posted 5 You are reading this latest preprint version Abstract The present study was investigated effect of CLM on thyroid hormone profile, immunity, expression of IL6 & HSP70 gene and antioxidant status of broiler. 240 day-old chicks were distributed into 5 groups having 8 birds in each replicate. The diet was formulated as per BIS (2007) standards for T1, while T2, T3, T4, T5 and T6 were given diets substituted with CLM @ 50, 75, 100, 125, and 150 g/kg, respectively for 42 days. The weight of the spleen, thymus and humoral immune response was significantly (P < 0.05) higher in T6 group. On 21st day, serum IL6 levels was significantly (P < 0.05) lower in T4, T5, and T6. On 42nd day, it was significantly (P < 0.05) lower in T6. Relative expression of the HSP70 gene in liver tissue was significantly (P < 0.05) down regulated in T6. The serum SOD and catalase levels were significantly (P < 0.05) higher in T4, T5, and T6. Serum FRAP and DPPH values were significantly (P < 0.05) higher in T5 and T6. The thyroid hormone profile, weight of bursa, cell-mediated immune response and serum GPx levels were remained comparable. Hence, it is concluded that CLM inclusion in a diet up to 15% improves immunity and antioxidant and alleviates heat stress in chickens. Antioxidant Cauliflower leaf meal Immunity Heat stress Polyphenol Interleukin 6 Heat shock protein 70 Introduction Leaf meal is a protein-rich feed ingredient made from the leaves of plants that can be used in broiler production as a protein source to supplement traditional feed ingredients, such as corn and soybean meal. The use of leaf meal in broiler production can have several benefits considering the shorter production cycle of broilers. It is a highly digestible protein source, which can help improve the growth rate and feed efficiency of broilers, resulting in profitable broiler production [ 68 ]. The leaf meal can be a cost-effective alternative to traditional protein sources, as it is often available at a lower price. The presence of health-promoting substances such as vitamins, carotenoids, phenols, flavonoids, and minerals in leaf meals is responsible for their good health qualities [ 6 , 54 ]. Along with protein, most leaf meals have high ash content, making them a rich source of minerals for poultry [ 68 ]. Cauliflower leaf meal (CLM) is rich also rich in essential amino acids and would offer enough of these nutrients for poultry and animal diets [ 43 ]. Brassicas are known for their antioxidant properties, attributed to the presence of polyphenols and flavonoids [ 19 ]. Efforts have been stepped up to find an alternative to antibiotics for in-feed use. It has been found that using leaf meals improves the health and physiological conditions of broilers on farms. As a result, leaf meal could be used as an alternative to in-feed antibiotics in broiler chickens [ 68 ]. Among the active substances, phenols have been shown to aid in the development of immunological organs and boost the immune system of chicks [ 34 ]. Phenols may also function as antimicrobials and antioxidants, protecting broilers' immune systems from the harmful effects of reactive oxygen species [ 34 ]. One of the most pleiotropic intesrleukins generated in areas of damage or infection is interleukin 6. High levels of IL-6 have been associated with an increased risk of disease and mortality in broiler chickens [ 72 ]. The HSP70 is the most conserved and well-studied member of the HSP family, and its expression is altered by a wide range of substances or stressors. The primary inducer of HSP-related genes is the high ambient temperature [ 77 ]. HSP70 gene expression has been extensively researched and utilised as a marker for heat stress in chickens. With this above brief background, the study was planned to investigate the effect of the inclusion of CLM on the thyroid hormone profile, immunity, heat stress, and antioxidant status of broiler chickens. Materials and Methods Ethics statement The experimental procedures carried out in the present study were approved by the Institutional Animal Ethics Committee (IAEC). The guidelines of the ‘committee for the purpose of control and supervision of experiments on animals (CPCSEA) 2012’ established under the ‘Prevention of Cruelty to Animals Act 1960’ of the Indian Penal Code were followed. Preparation of Cauliflower leaf meal The cauliflower leaves waste was collected from the nearby vegetable market, in Delapeer, Bareilly, India. These wastes were chopped using a chaff cutter. The chopped waste is then sun-dried on a tarpaulin followed by milling using an attrition mill and stored in an airtight container. Proximate analysis and determination of the chemical composition of cauliflower leaf meal The proximate analysis was carried out as per the method suggested by AOAC, (1990). The Crude Protein (CP) was estimated by the Kjeldahl method by taking a pooled sample, ether extract using soxhlet ether extraction apparatus, and crude fibre was estimated by acid (2.04N H2SO4) digestion followed by alkali (2.5 N NaOH) digestion, expressed as a difference in oven-dry weight and weight after ashing. The total ash (TA) was determined by ashing the samples in a muffle furnace at 550˚C after decarbonisation. Total phosphorus (TP) was estimated by the calorimetric method using phospho molybdo-vanadate reagent [ 9 ] and Calcium as per the methods of [ 71 ]. Glucosinolates and the total phenolic content were estimated by the method suggested by Abaza et al. [ 1 ] and Makkar et al. [ 47 ], respectively. Gross energy (kcal/kg) was estimated by using Gallenkamp adiabatic bomb calorimeter. The metabolisable energy was calculated by multiplying the gross energy value by a factor of 0.82 [ 50 ]. Experimental birds The experiment was conducted for a total duration of 42 days. 240 straight run day old CARI BRO VISHAL chicks were allotted to six groups in a completely randomized design, fourty in each group. Each group had five replicates and each replicate had eight birds. These birds were fed increasing level of cauliflower leaf meal (CLM) at 5, 7.5, 10, 12.5 and 15% from the day old. Feeds and feeding The ingredients and nutrient composition of the basal experimental diet are given in the Table 1 . The BIS (2007) feeding standards were followed to fulfill the nutrient requirements of broilers. As per BIS standard, the birds were fed three types of diet during the 42-day trial i.e. , pre-starter (0–7 days), starter (8–21 days), and finisher (22–42 days). The T1 group of birds was fed a control diet and T2, T3, T4, T5 & T6 groups were fed CLM included diet by partial replacement of maize and soya bean meal in the experimental diet at the level of 50, 75, 100, 125, and 150 g/kg, respectively. The diet of all the groups were made iso-caloric and iso-nitrogenous. Table 1 Ingredient and chemical composition of the control experimental diets Ingredients (g/kg) Broiler pre-starter (0–7 days) Broiler starter (8–21 days) Broiler finisher (22–42 days) Maize 534.1 579.9 656.4 SBM 410 364.1 279 Oil 18 28 36 Limestone 9 12.5 11 DCP 17 4.2 6 Methionine 1.7 0.3 0.6 Lysine HCL 0 1 1 Salt 5 5 5 Mineral mix* 1 1.5 1.5 Vitamin premix** 2.2 1.5 1.5 Coccidiostat 1 1 1 Toxin binder 1 1 1 Analysed (% DM) CP 22.8 22.2 20.3 CF 2.61 3.8 3.7 EE 3.56 3.7 3.9 AIA 0.28 0.78 0.58 Calculated ME (kcal/kg) 3017.88 3106.4 3198.8 E:P ratio 131.2 140.8 160.2 AP (g/kg) 4.5 4.5 4.5 Lys (g/kg) 13 12 10.8 Met (g/kg) 5 5 4.5 SBM soya bean meal; DCP digestible crude protein; CP crude protein; CF crude fibre; EE ether extract; AIA acid insoluble ash; AP available phosphorus; Lys lysine; Met methionine *Mineral mix supplied mg/kg diet: Mg 300, Mn 55, I 0.4, Fe 56, Zn 30, Cu 4.2. **Vitamin premix supplied per kg diet: Vit. A 8250 IU, Vit. D 3 2400 IU, Vit. E 15 mg, Vit. K 1 mg, Vit. B 1 6 mg, Vit. B 12 8 mg, Niacin 12 mg, Pyridoxin 1.6 mg, Choline chloride 500 mg. Housing and management The chicks of different experimental groups were reared in cages with a wire mesh bottom. The electrically heated battery brooders were used to brood the chicks. 23 hrs lighting program was practiced. The experimental birds were fed in the morning every day. Ad-libitum feed and water were made available throughout the day. All the birds were reared under uniform and standard management practices. Thyroid hormone profile Thyroid hormone profile namely, T3, T4 and TSH was estimated by Quanti Microlisa® (J. Mitra and Co. Pvt. Ltd) based on the principle of competitive ELISA. Microwells are coated with respective anti-antibodies. Sample is added to microwell followed by addition of enzyme conjugate. Incubation is followed by a washing step to remove unbound compounds. The colour reaction is started by addition of substrate and stopped after a definite time. The colour intensity is proportional to the concentration of hormone in the sample. Weight of immune organs At the end of forty two days of experimental period, 10 birds from each dietary group (2 birds per replicate) were randomly selected and sacrificed after 17 hrs of fasting. Spleen, bursa, and thymus were collected and weighed. The weight of immune organs was expressed as % live weight (Pre-slaughter live weight) as shown below. $$\:Spleen\:\left(\%\right)=\frac{Weight\:of\:spleen\:\left(g\right)}{\text{P}\text{r}\text{e}\:\text{s}\text{l}\text{a}\text{u}\text{g}\text{h}\text{t}\text{e}\text{r}\:\text{l}\text{i}\text{v}\text{e}\:\text{w}\text{e}\text{i}\text{g}\text{h}\text{t}\:\left(\text{g}\right)}\times\:100$$ $$\:Bursa\:\left(\%\right)=\frac{Weight\:of\:bursa\:\left(g\right)}{\text{P}\text{r}\text{e}\:\text{s}\text{l}\text{a}\text{u}\text{g}\text{h}\text{t}\text{e}\text{r}\:\text{l}\text{i}\text{v}\text{e}\:\text{w}\text{e}\text{i}\text{g}\text{h}\text{t}\:\left(\text{g}\right)}\times\:100$$ $$\:Thymus\:\left(\%\right)=\frac{Weight\:of\:thymus\:\left(g\right)}{\text{P}\text{r}\text{e}\:\text{s}\text{l}\text{a}\text{u}\text{g}\text{h}\text{t}\text{e}\text{r}\:\text{l}\text{i}\text{v}\text{e}\:\text{w}\text{e}\text{i}\text{g}\text{h}\text{t}\:\left(\text{g}\right)}\times\:100$$ Cell mediated immunity Using a method recommended by Cheng and Lamont (1988), the CMI response to the Phytohemagglutinin-P (PHA-P) mitogen was evaluated on day 27 of the experiment. PHA-P (1.0 mg/ml of PBS, 100microgram /bird) was administered intradermally to right foot web of ten birds in each treatment using a tuberculin syringe. Foot web of selected birds was cleaned using the cotton before injection. The left foot web of same bird was given 0.1 ml of freshly prepared autoclaved PBS serving as a control. The mean skin thickness of the right and left foot webs of the injected birds of each treatment was measured using a micrometre at 0 hours and 24 hours after PHA-P mitogen injection. The difference in thickness of the control foot web at 0 and 24 hours was subtracted from the difference in thickness of the mitogen-injected foot web to calculate the foot web index (FWI). FWI (mm) = (R2 - R1) - (L2 - L1) Where, R 1 = Thickness (millimetre) before injection of PHA-P injection R 2 = Thickness (millimetre) after 24 hours of PHA-P injection L 1 = Thickness (millimetre) before injection of PBS solution L 2 = Thickness (millimetre) after 24 hours of PBS injection Humoral immune response As a measure of antibody titre against 1% SRBC suspension, humoral immunity was assessed. 10 birds in each treatment which were not involved in cell mediated immunity test were injected 1% SRBC via intravenous route on 29th day of the experiment. After five days post-inoculation, about 2.0 ml of blood was collected in serum vacutainers from immunized birds. The separated serum was used for microtiter haemagglutination (HA) test following a method reported by Siegel and Gross [ 64 ]. Estimation of serum IL6 and HSP70 To evaluate the serum IL6 and HSP70, blood was collected on 21st day and 42nd day from 10 birds/treatment in serum vacutainers. The separated serum was transferred to 2ml Eppendorf tubes and stored at -20 o C until analysed. Amount of IL6 in serum was quantified using Fine Test® Chicken Interleukin-6 ELISA kit (Catalogue no ECH0046). Likewise, serum HSP70 using Fine Test® Chicken HSP70 ELISA kit (Catalogue no ECH0072). Gene expression Gene expression study of IL6 gene in spleen and HSP 70 gene in liver was done at 21st and 42nd day of experiment, respectively. Four birds per treatment were randomly chosen and sacrificed to collect 100 mg of spleen and liver samples (following standard protocol) from each bird. Collected samples were immediately transferred to nuclease-free, 2.0 ml Eppendorf tubes that contained 1.0 ml of RNAprotect® reagent. The tissues were homogenised using an automated homogenizer (Polytron) on a high setting for 30 to 60 seconds. The RNA was isolated from the homogenised samples using the TRIzol™ reagent. Isolated RNA samples were checked for purity in a Nano drop UV spectrophotometer (Nano Drop 1000®, Thermo-Scientific, Singapore).The concentration of each RNA sample was calculated after obtaining the O.D. at 260/280. Revert Aid™ first-strand cDNA synthesis kit (K 1622, Thermo Fisher Scientific, U.S.) was used to create the first-strand cDNA from the appropriate amount of RNA using random hexamer primers. Already reported primers (Table 2 ) were used to quantify IL6 and HSP70 genes. mRNA expression folds of target genes were quantified relative to GAPDH and 28s RNA as housekeeping genes respectively. The 2- ΔΔ Ct method was used to analyse the real-time PCR data. Relative expression of genes IL-6 and HSP-70 in spleen and liver tissues respectively were quantified by a real-time PCR detection system (QIAquant 96® thermocycler) using 2X SYBR green master mix (RR420A, Takara TB Green™ Premix Ex Taq™). All molecular reactions were performed by using a nuclease-free 8 tube-strips containing optically clear flat caps (Axygen Scientific®, Inc. USA). Table 2 Primers (forward and reverse) used for real-time assay S. No Gene Sequence (5’-3’) Annealing Temperature (˚C) Length (bp) Accession no 1 IL6 F-AAATCCCTCCTCGCCAATCT R-CCCTCACGGTCTTCTCCATAAA 61 106 NM204628.1 2 HSP70 F-ACGACCAGGGGAACCGCACCAC R-GGCTTGCCACCCTCGTTCACCAC 56 74 HM587997 3 GAPDH F-CCGTCCTCTCTGGCAAAGTCC R-AGCCCCAGCCTTCTCCATG 58 266 NM204305 4 28S RNA F-CAGGTGCAGATCTTGGTGGTAGTA R- GCTCCCGCTGGCTTCTCC 58 273 JN639848 IL6 Interleukin 6; HSP70 Heat shock protein 70; GAPDH Glyceraldehyde 3-phosphate dehydrogenase; RNA Ribonucleic acid; F Forward; R Reverse Temperature humidity index (THI) THI was calculated using the formula suggested by Habeeb et al . [ 29 ]Z. THI = db° F–[(0.55–0.55 x RH) (db° F–58)].Where, db° F = dry bulb temperature (in Fahrenheit) and RH = relative humidity percentage (RH%)/100.THI is classified as: <82 = absence of heat stress, 82 to < 84 = moderate heat stress, 84 to < 86 = severe heat stress, and 86 and more = very severe heat stress. Antioxidant status Serum superoxide dismutase was quantified using Amplite™ colorimetric superoxide dismutase assay kit by AAT Bioquest® (Catalogue no 11305).Serum catalase enzyme using SIGMA-ALDRICH® catalase assay kit (Catalogue no CAT100).Serum glutathione peroxidase enzyme using Cayman® glutathione peroxidase assay kit (Item no. 703102).Total antioxidant capacity of serum is evaluated by ferric reducing ability of plasma (FRAP) as suggested by Benzie et al. , 1999 and 2,2-diphenyl-1-picrylhydrazyl (DPPH) by Brand et al. , [ 14 ]. Statistical Analysis The experimental data were analysed using the SPSS computer package (IBMSPSS software version 26.0, SPSS Inc., Chicago, USA) adopting standard statistical procedures of Snedecor and Cochran, [ 66 ]. The parameters were analysed by one-way ANOVA using the general linear model and Tukey post-hoc analysis. Significance was declared at P < 0.05 unless otherwise stated. Results Chemical composition of cauliflower leaf meal The chemical composition of cauliflower leaf meal (CLM) was given in the Table 3 . The dry matter (DM), crude protein (CP), ether extract (EE), crude fibre (CF), nitrogen-free extract (NFE), total ash (TA), and acid insoluble ash (AIA)content of the fresh cauliflower waste were 13.6, 22.3, 6.93, 9.47, 49.76, 11.54 and 1.37 percent, respectively. Similarly, the calcium (Ca), phosphorus (P), total phenolics (TP), and glucosinolates were 3.60, 0.6, 7.68, and 0.042 percent respectively. The gross energy (GE) and metabolisable energy (ME) content of CLM was 2560 kcal/kg and 2099 kcal/kg respectively. Table 3 Chemical composition (%) of the cauliflower leaf meal Particulars % CP 22.3 EE 6.93 CF 9.47 NFE 49.76 TA 11.54 AIA 1.37 Ca 0.61 P 0.28 TPC 7.68 Glucosinolates 0.042 Gross Energy (kcal/kg) 2560 Metabolisable energy (kcal/kg) 2099 CP crude protein; EE ether extract; CF crude fibre; NFE nitrogen free extract; TA total ash; AIA acid insoluble ash; Ca calcium; P phosphorus; TPC total phenolic content Thyroid hormone profile The thyroid hormone profiles (T3, T4, and TSH) of experimental group of birds fed CLM in the present investigation were comparable with the control group of birds on the 21st and 42nd day of the experiment (Table 4 ). Table 4 Effect of feeding cauliflower leaf meal on the thyroid hormone profile Treatment 21st day 42nd day T3 (ng/ml) T4 (µg/ml) TSH (µIU/ml) T3 (ng/ml) T4 (µg/ml) TSH (µIU/ml) T1 1.12 19.61 0.86 1.99 21.79 1.25 T2 1.21 17.70 0.91 2.08 19.55 1.52 T3 1.33 17.75 0.87 2.16 18.80 1.28 T4 1.82 17.77 0.94 1.78 19.07 1.19 T5 1.25 18.08 0.98 2.05 18.88 1.20 T6 1.45 17.85 0.91 2.22 19.04 1.18 SEM 0.452 0.283 0.025 0.259 0.452 0.058 P-value 0.851 0.242 0.779 0.745 0.289 0.618 T1 Control diet; T2 (50 mg/kg CLM); T3 (75 mg/kg CLM); T4 (100 mg/kg CLM); T5 (125 mg/kg CLM); T6 (150 mg/kg CLM); CLM Cauliflower; T3 Triiodothyronine; T4 Thyroxine; TSH Thyroid stimulating hormone; SEM standard error mean. Weight of immune organs The weight of different immune organs in the current investigation was presented in the Table 5 .The weight of the spleen was significantly (P < 0.05) higher in T6 group as compared to the control group. The weight of the thymus was significantly (P < 0.05) higher in the T6 group of birds as compared to the control group. The weight of bursa of experimental group of birds fed CLM was comparable with the control group of birds. Table 5 Effect of feeding cauliflower leaf meal on the weight of immune organs Live weight (%) Treatment Spleen Bursa Thymus T1 0.245 a 0.146 0.282 a T2 0.250 a 0.140 0.300 a T3 0.248 a 0.148 0.313 a T4 0.258 a 0.144 0.343 ab T5 0.271 a 0.158 0.354 ab T6 0.320 b 0.151 0.440 b SEM 0.041 0.010 0.017 P-value 0.038 0.517 0.029 T1 Control diet; T2 (50 mg/kg CLM); T3 (75 mg/kg CLM); T4 (100 mg/kg CLM); T5 (125 mg/kg CLM); T6 (150 mg/kg CLM); CLM Cauliflower leaf meal; SEM standard error mean abc Mean with superscript in a column differ significantly (P < 0.05) Cell-mediated and Humoral immune response The cell-mediated immune (CMI) response of the experimental group of birds fed CLM in the current investigation was comparable with the control group (Table 6 ). The humoral immune response of experimental group of birds against sheep RBCs (SRBCs) was significantly (P < 0.05) higher in T6 group when compared with the control group of birds (Table 6 ). Table 6 Effect of feeding cauliflower leaf meal on cell-mediated immunity, humoral immunity, serum IL6 levels, and relative expression of IL6 gene Treatment FWI (mm) HA titre (log2) IL6 ELISA (pg/ml) IL6 Fold change 21st day 42nd day T1 0.283 18.98 a 171.71 b 287.81 b 1 a T2 0.410 19.29 a 144.57 ab 284.00 b 0.96 a T3 0.355 19.06 a 120.57 ab 278.29 ab 1.10 a T4 0.321 25.33 ab 76.00 a 276.86 ab 1.18 ab T5 0.372 26.57 ab 68.76 a 259.72 ab 1.27 ab T6 0.390 31.22 b 70.42 a 193.52 a 1.77 b SEM 0.015 1.055 9.340 10.08 0.076 P-value 0.182 0.003 < 0.001 0.049 0.012 T1 Control diet; T2 (50 mg/kg CLM); T3 (75 mg/kg CLM); T4 (100 mg/kg CLM); T5 (125 mg/kg CLM); T6 (150 mg/kg CLM); CLM Cauliflower leaf meal; FWI Foot web index; HA Haemagglutination; IL6 Interleukin 6; SEM standard error mean abc Mean with superscript in a column differ significantly (P < 0.05). Serum IL-6 levels The serum interleukin-6 (IL6) levels on 21st & 42nd day of the experiment were presented in the Table 6 . In the current investigation, the serum IL-6 level (pg/ml) on the 21st day was significantly (P < 0.05) lower in T4, T5 and T6 group of birds. On the 42nd day, the serum IL 6 was significantly (P < 0.05) lower in T6 group and the serum IL6 levels in the other experimental group of birds fed CLM were comparable with the control group. IL-6 gene expression study The results of relative expression of IL6 gene were presented in the Table 6 . The relative expression of IL6 gene was significantly (P < 0.05) higher in T6 group of birds as compared to the control group. Temperature humidity index The weekly temperature humidity index (THI) during the experimental were 80.69, 82.01, 83.02, 85.20, 85.61, and 89.32 during the 1st, 2nd, 3rd, 4th, 5 th, and 6th week of the experiment, respectively. The weekly temperature humidity index (THI) was significantly (P < 0.05) higher during 4th, 5th, and 6th week of the experiment. HSP 70 gene expression study The results of relative expression of HSP70 gene was presented in the Table 7 . The relative expression of HSP70 gene was significantly (P < 0.05) lower in T6 group of birds as compared to the control group. Table 7 Effect of feeding cauliflower leaf meal on relative expression HSP70 gene and serum HSP70 levels Serum HSP 70 (ng/ml) Treatment HSP70 Fold change 21st day 42nd day T1 1 b 1.187 1.227 b T2 0.93 ab 1.1617 1.193 ab T3 0.79 ab 1.1718 1.172 ab T4 0.81 ab 1.163 1.155 ab T5 0.68 ab 1.1383 1.093 a T6 0.52 a 1.137 1.077 a SEM 0.048 0.009 0.014 P value 0.025 0.557 0.005 T1 Control diet; T2 (50 mg/kg CLM); T3 (75 mg/kg CLM); T4 (100 mg/kg CLM); T5 (125 mg/kg CLM); T6 (150 mg/kg CLM); CLM Cauliflower leaf meal; HSP70 Heat shock protein 70; SEM Standard error mean abc Mean with superscript in a column differ significantly (P < 0.05). Serum HSP 70 levels On the 21st day, the serum HSP70 levels of the experimental group of birds fed CLM were comparable with the control group however, on the 42nd day of the experiment, the mean serum levels of HSP 70 were significantly (P < 0.05) lower in T5 and T6 groups than T2, T3, T4, and the control group (Table 7 ). Antioxidant status The results pertaining to the concentration of serum antioxidant enzymes and total antioxidant capacity of serum was presented in the Table 8 . The serum SOD levels were significantly (P < 0.05) higher in the T6 group of birds. Catalase level was significantly (P < 0.05) higher in the T4, T5 and T6 group of birds. The serum GPx levels in the experimental group of birds were comparable with the control group. The serum FRAP values were significantly (P < 0.05) higher in T5 and T6 group as compared to the control group of birds (Table 8 ). The serum DPPH values were significantly (P < 0.05)higher in the T6 group of birds and the DDPH values of the other experimental group of birds fed CLM were comparable with the control group (Table 8 ). Table 8 Effect of feeding cauliflower leaf meal on the antioxidant status Treatment SOD (U/ml) Catalase (µmol/min/ml) GPx (nmol/min/ml) FRAP (mM/ml) DPPH (% Antioxidant activity) T1 66.16 a 11.24 a 6.64 2.822 a 14.82 a T2 67.43 ab 11.51 ab 5.09 3.157 ab 15.03 a T3 69.05 ab 11.67 b 5.10 3.177 ab 16.07 a T4 68.64 ab 11.72 c 4.97 3.147 ab 15.38 a T5 70.34 bc 12.29 c 4.25 3.388 b 17.19 ab T6 73.88 c 12.54 c 5.82 3.444 b 18.66 b SEM 0.625 0.113 0.662 0.045 0.637 P-value < 0.001 0.003 0.169 < 0.001 0.042 T1 Control diet; T2 (50 mg/kg CLM); T3 (75 mg/kg CLM); T4 (100 mg/kg CLM); T5 (125 mg/kg CLM); T6 (150 mg/kg CLM); CLM Cauliflower leaf meal; SOD Super oxide dismutase; GPx Glutathione peroxidase; FRAP Ferric reducing ability of plasma; DPPH2,2-diphenylpicrylhydrazyl; SEM Standard error mean abc Mean with superscript in a column differ significantly (P < 0.05). Discussion Chemical composition of cauliflower leaf meal The dry matter (DM) content of cauliflower leaves in the present experiment was similar to the DM content reported by Wadhwa and Bakshi [ 73 ] and (2016) however, it was found to be higher than values reported by Sharma and Prasad [ 62 ] and Raza et al . [ 58 ].The crude protein (CP) content of CLM in the current investigation, was very close to the value reported by Mahgoub et al. [ 45 ] and Gupta and Wagle [ 28 ]. However, the CP content was lower than the value reported by Wani et al . [ 75 ] and higher than the value reported by Mogra et al . [ 48 ] and Bakshi et al . [ 11 ]. Similarly, the ether extract (EE) content of CLM was higher than the EE content reported earlier [28, 48 and 11). The crude fibre (CF) content of CLM (9.47%) was comparable to the values reported by Gupta and Wagle [ 28 ] and lower than the value reported by Mogra et al . [ 48 ]. The nitrogen-free extract (NFE) of CLM was comparable to the value reported by Wani et al . [ 75 ] and Ahmed et al . [ 5 ]. The total ash (TA) content of CLM was consistent with the findings of Ahmed et al . [ 5 ] & Gupta and Wagle [ 28 ] and lower than the value reported by Mogra et al . [ 48 ]. The acid insoluble ash (AIA) of the CLM was higher than the reports of Gupta and Wagle [ 28 ]and it may be due to different ways of collection and drying. The calcium (Ca) content of CLM was comparable to the value reported by Revathi et al . [ 59 ] and higher than the value reported by Ahmed et al . [ 5 ]. The phosphorus (P) content of CLM was also comparable to the value reported by Revathi et al . [ 59 ] and higher than the value reported by Ahmed et al . (2013).The total phenolic content (TPC) of CLM in the present experiment was higher than the values reported by Abul-Fadl [ 3 ] and Wu et al . [ 76 ]. However, the TPC values in the current study were lower than those reported by Collado-Alvarado et al . [ 17 ]. The glucosinolates content of CLM in the present investigation was comparable to the value reported by Gupta and Wagle [ 28 ] & Drabińska et al . [ 20 ] and slightly lower than the levels reported by Abul-Fadl [ 3 ] & Chandra [ 15 ]. Thyroid hormone profile The thyroid hormone profile of birds in the current investigation were consistent with the findings of Rabie et al . [ 56 ] who reported that the feeding canola meal ( Brassica napus ) upto 15% in broilers did not affect the plasma thyroid hormone profile. Further, Abdel-Moneim et al . [ 2 ] reported that the serum T3 concentration was unaffected in the Japanese quail fed a diet containing full-fat canola seeds at 150g/kg. Kermanshahi and Pour [ 36 ] reported no significant difference in thyroid hormone profile of broilers fed rapeseed meal upto 20% level. Similarly, Pena [ 53 ] reported that there was no significant difference in serum T3, and T4 hormone levels of laying hens fed Broccoli Stems and Leaves Meal upto 15% level. Weight of immune organs The higher weight of immune organs in the 15% CLM fed group of birds in the current experiment corroborated with that of Ao and Kim [ 7 ] who reported higher relative weight of the spleen in broilers fed grape seed extract rich in polyphenols. The similar findings were also reported by Omar et al . [ 52 ] who reported a higher weight of thymus in broilers fed onion extract rich in polyphenols. The improved humoral immune response against sheep red blood cells (SRBC) seen in this experiment may be co-related with the increased weight of immune organs. Similar co-relation between greater spleen, thymus, and other immune organ weight and higher antibody titre against SRBCs was also reported by Kabir et al . [ 33 ]. Increased weight of immune organs may be attributed due to the proliferation of lymphocytes. When challenged with an antigen, lymphocytes respond by creating lymphokines, proliferating their antigen-specific lymphocyte clones, and enhancing immune responses [ 65 ]. Yang et al . [ 78 ] reported an increased splenic lymphocyte proliferation response and spleen weight in broilers supplemented with quercetin (phenolic compound). Cell-mediated and Humoral immune response The cell mediated immune response of experimental group of birds recorded in the current investigation was consistent with the findings of Kumar et al . [ 39 ] who reported that there was no significant difference in CMI response of broilers fed pomegranate peel extract rich in polyphenols during the summer. Similar findings were reported by Rao et al . [ 57 ] where he reported that there was no significant difference in cell mediated immune response to PHA-P in broilers fed Moringa olefera leaf meal and pomegranate peal meal at 1000mg/kg & 500mg/kg of basal diet respectively. In contrast, to the present findings, Gopi et al . [ 27 ] reported that pigeons fed polyphenols isolated from pomegranate peels had better cell mediated immune response against intradermal inoculation of PHA-P. Dumaup and Ampode [ 23 ] also reported significantly (P < 0.05) higher CMI in broiler chickens fed diets containing water hyacinth meal. The humoral immune response of experimental group of birds recorded in the current investigation were consistent with the findings of Pourhossein et al . [ 55 ], who reported the enhanced immune response against SRBCs in birds fed sweet orange peel extract rich in polyphenols. Emadi and Kermanshahi [ 25 ] reported that the broiler chicks fed turmeric rhizome powder rich in polyphenols upto 0.75% level had a higher antibody titre against SRBCs. In further support of these findings, Gholamrezaie et al . [ 26 ] reported that broiler chickens supplemented with Artemsia annua extract and leaf powder (contains flavonoids and quercetin) at 1.5% level increased total antibody titre against SRBC in the experimental groups when compared with the control group. Increased antibody titre against SRBC in the current study is suggestive of increase in humoral immunity of experimental birds when compared with the control group. Serum IL-6 levels IL6 is a pleiotropic cytokine, and its concentration is used as an indicator to study the inflammatory status of animals [ 60 ]. In the present investigation, the results were in line with the reports of Sun et al . [ 70 ], who reported that the supplementation of a polyphenol-rich Yucca schidigera extract at 300mg/kg of basal diet in broiler chickens, resulted in lower serum IL-6 concentrations when compared with the control group. Adeyemi et al . [ 4 ] also reported lower serum IL6 levels in broiler chickens fed diets supplemented with Anacardium occidentale leaf powder upto 4g/kg. In further support of these findings, Liu et al . [ 42 ] reported that the grape seed extract rich in polyphenols reduced inflammation in mice fed a high-fat diet by significantly (P < 0.05) lowering serum pro-inflammatory cytokine (IL6) concentrations. Similarly, Kilany et al . [ 38 ] reported that the there was a significant decrease in serum Il6 levels of broilers fed a diet containing 5% Moringa olefera leaf meal and 0.5% sweet basil. The findings of the present investigation were consistent with the results of Nootash et al . [ 49 ], who reported that the Interleukin-6 gene expression in the spleen of rainbow trout was upregulated on feeding green tea extract ( Camellia sinensis ) rich in polyphenol at a rate of 500 mg/kg. Further, Shekarabi et al . [ 63 ] reported that the transcription levels of interleukin -6 genes were up-regulated in the fish fed dandelion flower extract (DFE) at 3 and 4 g/kg of basal diet. In further support of this study, Awad et al . [ 10 ] reported that the IL-6 gene was up-regulated in the spleen of broiler chickens fed rutin a bioactive chemical found in apple and tea leaves at a rate of 1g/kg diet. Similarly, Khalid et al . [ 37 ] reported significantly increased IL6 gene expression in rabbits fed Moringa olefera leaf powder at 200 mg/kg body weight. Temperature humidity index During the last 3 weeks of the experimental trial the THI was in severe to very severe heat stress. The effects of heat stress on laying hens can be compared between various study conditions by utilising THI, which takes into account both ambient temperature and humidity [ 35 ]. El-Tarabany, [ 24 ] reported that, Japanese quail exposed to heat stress showed decreased immunological and production characteristics as well as negative welfare consequences. HSP 70 gene expression study The heat shock transcription factor activity or concentration can be increased at high temperatures to cause the synthesis of HSP70 mRNA [ 18 ]. The HSP70 is the most conserved and well-studied members of the HSP family, and their expression is altered by a wide range of substances or stressors. It is reported that the primary inducer of HSP-related gene expression is high ambient temperature [ 77 ]. The results of the present investigation were consistent with the findings of Hajati et al . [ 29 ], who reported that the broilers supplemented with grape seed extract rich in polyphenols and vitamin C during chronic heat stress conditions resulted in a down-regulation of HSP-70 gene expression in the liver and heart when compared with the control group. Further, Sugito et al . [ 69 ] reported the downregulation of HSP-70 gene in the kidney of broilers fed with salix extract under heat stress conditions. Wan et al . [ 74 ] reported that the mRNA expression levels of HSP70 was decreased in breast muscle of broilers supplemented with Artemisia annua L. when compared with the control group. Similarly, Song et al . [ 67 ] reported significantly (P < 0.05) decreased HSP70 mRNA expression in the intestine of broilers supplemented with Artemisia annua at 1g/kg of diet. Serum HSP 70 levels When analysing the broiler's response to heat stress, it is crucial to take oxidative stress into account [ 40 ]. Bird's maintaining heat homeostasis under stressful conditions results in higher reactive oxygen species (ROS) concentrations. It is reported that the body begins producing heat shock proteins to defend itself from ROS' damaging effects on the cells [ 22 ]. The results of the present findings were consistent with the findings of Hassan and Asim [ 30 ], who reported that the broilers supplemented with vitamin C and acetylsalicylic acid had lower serum HSP70 levels than controls when raised in heat stress conditions (40°C for 4 hours per day). Hu et al . [ 32 ] reported that the serum HSP70 levels were significantly (P < 0.05) reduced in broilers fed the diet supplemented with phloretin (a type of natural phenol that is found in apple tree leaves) at 200 mg/kg. In further support of these findings, Zhang et al . [ 79 ] and Liu et al . [ 41 ] reported the significant (P < 0.05) decrease in HSP70 levels in broilers fed a diet supplemented with the plant antioxidants like curcumin and resveratrol. The reduced serum HSP70 levels in treatment groups may also be attributed to the fact that the polyphenols present in the CLM will scavenge the ROS which is also one of the factors responsible for HSP production as suggested by Mahmoud et al . [ 46 ]. Antioxidant status Broilers require higher antioxidant capacity since low antioxidant capacity has been linked to cellular damage, immune system degeneration, and metabolic abnormalities [ 68 ]. The results of present investigation were similar with the Drazbo et al . [ 21 ] who reported that there was a significant (P < 0.05) increase in serum SOD levels of turkey fed rape seed meal at a 15% level. Wan et al . [ 74 ] reported that the broilers fed Artemisia annua L. leaves rich in polyphenols showed greater blood levels of SOD and catalase. Further, Hu et al . [ 31 ] reported that the serum catalase and glutathione peroxidase concentration were significantly (P < 0.05) increased in broilers fed the diet supplemented with phloretin (natural phenol in apple tree leaves) at 200 mg/kg diet. These results corroborated with the findings of Drazbo et al . [ 21 ], who reported significantly (P < 0.05) higher FRAP in the turkey fed a rape seed meal at a 15% level. Similar results were reported by Ognik et al . [ 51 ] in turkey hens fed a diet supplemented with the quercetin (phenolic compound). Llorach et al . [ 44 ], reported that the testing of a cauliflower by-product extract using FRAP method revealed a significant antioxidant potential. He also reported that there is a linear correlation between FRAP readings and the overall phenolic content of cauliflower leaves. These results are consistent with the findings of Wan et al . [ 74 ] who reported the broilers fed Artemisia annua L. leaves rich in polyphenols showed significantly (P < 0.05) higher serum DPPH levels when compared with the control. Llorach et al . [ 44 ] reported that an extract of cauliflower by-products effectively scavenges DPPH free radicals. Similarly, Seong et al . [ 61 ] reported the significantly higher DPPH free radical scavenging capacity of Chinese cabbage extract. Conclusions Therefore, it is concluded that cauliflower leaf meal (CLM) inclusion in the diet upto 15% improves immunity, which is evident by increased immune organ weight, higher haemagglutination titre against sheep RBCs, and upregulation of IL6 gene in the spleen. CLM feeding also increases the antioxidant status and alleviates heat stress in broiler chickens. Declarations Competing Interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Funding The work has been done under the institute project entitled “Studies on the utilization of fruit and vegetable waste as an alternative feed’’ and funded by Indian Veterinary Research Institute, Bareilly, India. Author Contributions TM is involved in conducting experiment, analysis, and writing the manuscript. SS and AB planned and executed of both experiments, edited manuscript, coordinated studies. GK and AB are involved in conducting experimental trial and analysis part. RM edited manuscript. Data Availability Corresponding author should be contacted to get data available on request. References A.O.A.C. Methods of analysis. Association of Official Analytical Chemists. (15th Edn.) Washington, USA : 20–44. (1990) A.O.A.C. Official Methods of analysis. AOAC international. 18th edn., Virginia, USA (2000) Abaza, R.H., Blake, J.T.: Fisher. Oxalate determination: Analytical problems encountered with certain plant species. J. Association Official Anal. 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PloS one. 9 (7), e102204 (2014). https://doi.org/10.1371/journal.pone.0102204 Yang, J.X., Maria, T.C., Zhou, B., Xiao, F.L., Wang, M., Mao, Y.J., Li, Y.: Quercetin improves immune function in Arbor Acre broilers through activation of NF-κB signaling pathway. Poult. sci. 99 (2), 906–913 (2020). https://doi.org/10.1016/j.psj.2019.12.021 Zhang, G.F., Yang, Z.B., Wang, Y., Yang, W.R., Jiang, S.Z.: Gai. Effects of ginger root (Zingiber officinale) processed to different particle sizes on growth performance, antioxidant status, and serum metabolites of broiler chickens. Poult. sci. 88 (10), 2159–2166 (2009). https://doi.org/10.3382/ps.2009-00165 Cite Share Download PDF Status: Published Journal Publication published 10 Feb, 2026 Read the published version in Waste and Biomass Valorization → Version 1 posted Reviewers agreed at journal 01 Dec, 2025 Reviewers invited by journal 14 Aug, 2025 Editor invited by journal 09 Aug, 2025 Editor assigned by journal 17 Jul, 2025 First submitted to journal 16 Jul, 2025 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. 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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-7145387","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":500317578,"identity":"d393a1a7-6527-4703-a90b-afb8bdc8c14c","order_by":0,"name":"Mahesh Theerthesh","email":"","orcid":"","institution":"ICAR-Indian Veterinary Research Institute: Indian Veterinary Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Mahesh","middleName":"","lastName":"Theerthesh","suffix":""},{"id":500317579,"identity":"616256c8-8662-4cf3-b9f1-01dec0a0f6d9","order_by":1,"name":"Avishek Biswas","email":"","orcid":"","institution":"Central Avian Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Avishek","middleName":"","lastName":"Biswas","suffix":""},{"id":500317580,"identity":"81c76981-f496-41eb-a5ee-6d220b704c2c","order_by":2,"name":"Gautham Kolluri","email":"","orcid":"","institution":"Central Avian Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Gautham","middleName":"","lastName":"Kolluri","suffix":""},{"id":500317581,"identity":"8a9605c0-97c5-425b-9194-da349def58f1","order_by":3,"name":"Subodh Kumar Saha","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA3ElEQVRIiWNgGAWjYFACHgYG3oYEBgYJBsYHDAwHSNPCbECyFjYJorTIz8g9+ODtjjR7/tnNz6p5au7I8TMwP3x0A48Wgxt5yYZzz+QkzrhzzOw2z7FnxpINbMbGOfi0SOSYSfO2VSQw3EgAamE7nLjhAA+bND4t8jMgWuzlb6R/K+b5R4QWhhtgLTmMG4AMZt42IrQYnHkH9EtbWuLGGznFknP7DhtLNhPwi3w7KMTaku3lbqRv/PDm22E5fvbmh4/xOgwZMPGASGZilYMA4w9SVI+CUTAKRsGIAQB5WVBqiMPh5wAAAABJRU5ErkJggg==","orcid":"","institution":"ICAR-Indian Veterinary Research Institute: Indian Veterinary Research Institute","correspondingAuthor":true,"prefix":"","firstName":"Subodh","middleName":"Kumar","lastName":"Saha","suffix":""},{"id":500317582,"identity":"52bd15b0-e85c-4a22-88a9-a5b7726013c9","order_by":4,"name":"Ashvini Pundalik Bansod","email":"","orcid":"","institution":"Indian Veterinary Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Ashvini","middleName":"Pundalik","lastName":"Bansod","suffix":""},{"id":500317583,"identity":"212f0713-f92b-47d1-8bcd-5279d1931fc5","order_by":5,"name":"Rajeshwar Khandare","email":"","orcid":"","institution":"Indian Veterinary Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Rajeshwar","middleName":"","lastName":"Khandare","suffix":""}],"badges":[],"createdAt":"2025-07-17 06:19:55","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7145387/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7145387/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s12649-026-03499-x","type":"published","date":"2026-02-10T15:56:56+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":102785117,"identity":"9cf8f2b7-1ade-4496-81c2-4bf58fc162cd","added_by":"auto","created_at":"2026-02-16 15:59:51","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1771355,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7145387/v1/ed382cd4-2efd-483a-b46f-1470e07bc4e5.pdf"}],"financialInterests":"","formattedTitle":"Exploring Nutritional Evaluation of Cauliflower (Brassica Oleracea Var Botrytis) Waste on the Thyroid Hormone, Antioxidant Status, Immunity and Expression of Interleukin 6 and HSP 70 Genes in Broilers","fulltext":[{"header":"Introduction","content":"\u003cp\u003eLeaf meal is a protein-rich feed ingredient made from the leaves of plants that can be used in broiler production as a protein source to supplement traditional feed ingredients, such as corn and soybean meal. The use of leaf meal in broiler production can have several benefits considering the shorter production cycle of broilers. It is a highly digestible protein source, which can help improve the growth rate and feed efficiency of broilers, resulting in profitable broiler production [\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e]. The leaf meal can be a cost-effective alternative to traditional protein sources, as it is often available at a lower price.\u003c/p\u003e\u003cp\u003eThe presence of health-promoting substances such as vitamins, carotenoids, phenols, flavonoids, and minerals in leaf meals is responsible for their good health qualities [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]. Along with protein, most leaf meals have high ash content, making them a rich source of minerals for poultry [\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e]. Cauliflower leaf meal (CLM) is rich also rich in essential amino acids and would offer enough of these nutrients for poultry and animal diets [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. Brassicas are known for their antioxidant properties, attributed to the presence of polyphenols and flavonoids [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Efforts have been stepped up to find an alternative to antibiotics for in-feed use. It has been found that using leaf meals improves the health and physiological conditions of broilers on farms. As a result, leaf meal could be used as an alternative to in-feed antibiotics in broiler chickens [\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e]. Among the active substances, phenols have been shown to aid in the development of immunological organs and boost the immune system of chicks [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. Phenols may also function as antimicrobials and antioxidants, protecting broilers' immune systems from the harmful effects of reactive oxygen species [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eOne of the most pleiotropic intesrleukins generated in areas of damage or infection is interleukin 6. High levels of IL-6 have been associated with an increased risk of disease and mortality in broiler chickens [\u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e72\u003c/span\u003e]. The HSP70 is the most conserved and well-studied member of the HSP family, and its expression is altered by a wide range of substances or stressors. The primary inducer of HSP-related genes is the high ambient temperature [\u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e77\u003c/span\u003e]. HSP70 gene expression has been extensively researched and utilised as a marker for heat stress in chickens. With this above brief background, the study was planned to investigate the effect of the inclusion of CLM on the thyroid hormone profile, immunity, heat stress, and antioxidant status of broiler chickens.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003e\u003cb\u003eEthics statement\u003c/b\u003e\u003c/p\u003e\u003cp\u003e The experimental procedures carried out in the present study were approved by the Institutional Animal Ethics Committee (IAEC). The guidelines of the \u0026lsquo;committee for the purpose of control and supervision of experiments on animals (CPCSEA) 2012\u0026rsquo; established under the \u0026lsquo;Prevention of Cruelty to Animals Act 1960\u0026rsquo; of the Indian Penal Code were followed.\u003c/p\u003e\u003cp\u003e\u003cb\u003ePreparation of Cauliflower leaf meal\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe cauliflower leaves waste was collected from the nearby vegetable market, in Delapeer, Bareilly, India. These wastes were chopped using a chaff cutter. The chopped waste is then sun-dried on a tarpaulin followed by milling using an attrition mill and stored in an airtight container.\u003c/p\u003e\u003cp\u003e\u003cb\u003eProximate analysis and determination of the chemical composition of cauliflower leaf meal\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe proximate analysis was carried out as per the method suggested by AOAC, (1990). The Crude Protein (CP) was estimated by the Kjeldahl method by taking a pooled sample, ether extract using soxhlet ether extraction apparatus, and crude fibre was estimated by acid (2.04N H2SO4) digestion followed by alkali (2.5 N NaOH) digestion, expressed as a difference in oven-dry weight and weight after ashing. The total ash (TA) was determined by ashing the samples in a muffle furnace at 550˚C after decarbonisation. Total phosphorus (TP) was estimated by the calorimetric method using phospho molybdo-vanadate reagent [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e] and Calcium as per the methods of [\u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e]. Glucosinolates and the total phenolic content were estimated by the method suggested by Abaza \u003cem\u003eet al.\u003c/em\u003e [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e] and Makkar \u003cem\u003eet al.\u003c/em\u003e [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e], respectively. Gross energy (kcal/kg) was estimated by using Gallenkamp adiabatic bomb calorimeter. The metabolisable energy was calculated by multiplying the gross energy value by a factor of 0.82 [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003cb\u003eExperimental birds\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe experiment was conducted for a total duration of 42 days. 240 straight run day old CARI BRO VISHAL chicks were allotted to six groups in a completely randomized design, fourty in each group. Each group had five replicates and each replicate had eight birds. These birds were fed increasing level of cauliflower leaf meal (CLM) at 5, 7.5, 10, 12.5 and 15% from the day old.\u003c/p\u003e\u003cp\u003e\u003cb\u003eFeeds and feeding\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe ingredients and nutrient composition of the basal experimental diet are given in the Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. The BIS (2007) feeding standards were followed to fulfill the nutrient requirements of broilers. As per BIS standard, the birds were fed three types of diet during the 42-day trial \u003cem\u003ei.e.\u003c/em\u003e, pre-starter (0\u0026ndash;7 days), starter (8\u0026ndash;21 days), and finisher (22\u0026ndash;42 days). The T1 group of birds was fed a control diet and T2, T3, T4, T5 \u0026amp; T6 groups were fed CLM included diet by partial replacement of maize and soya bean meal in the experimental diet at the level of 50, 75, 100, 125, and 150 g/kg, respectively. The diet of all the groups were made iso-caloric and iso-nitrogenous.\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\u003eIngredient and chemical composition of the control experimental diets\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\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\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eIngredients (g/kg)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBroiler pre-starter (0\u0026ndash;7 days)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eBroiler starter (8\u0026ndash;21 days)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eBroiler finisher (22\u0026ndash;42 days)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eMaize\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e534.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e579.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e656.4\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eSBM\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e410\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e364.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e279\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eOil\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e28\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e36\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eLimestone\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e11\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eDCP\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eMethionine\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.6\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eLysine HCL\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eSalt\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eMineral mix*\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eVitamin premix**\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eCoccidiostat\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eToxin binder\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"4\" nameend=\"c4\" namest=\"c1\"\u003e\u003cp\u003e\u003cb\u003eAnalysed (% DM)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eCP\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e22.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e22.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e20.3\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eCF\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2.61\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e3.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.7\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eEE\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.56\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e3.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.9\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eAIA\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.28\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.78\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.58\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"4\" nameend=\"c4\" namest=\"c1\"\u003e\u003cp\u003e\u003cb\u003eCalculated\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eME (kcal/kg)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3017.88\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e3106.4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3198.8\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eE:P ratio\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e131.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e140.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e160.2\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eAP (g/kg)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e4.5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eLys (g/kg)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e10.8\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eMet (g/kg)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e4.5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"4\"\u003eSBM soya bean meal; DCP digestible crude protein; CP crude protein; CF crude fibre; EE ether extract; AIA acid insoluble ash; AP available phosphorus; Lys lysine; Met methionine\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"4\"\u003e*Mineral mix supplied mg/kg diet: Mg 300, Mn 55, I 0.4, Fe 56, Zn 30, Cu 4.2.\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"4\"\u003e**Vitamin premix supplied per kg diet: Vit. A 8250 IU, Vit. D\u003csub\u003e3\u003c/sub\u003e 2400 IU, Vit. E 15 mg, Vit. K 1 mg, Vit. B\u003csub\u003e1\u003c/sub\u003e 6 mg, Vit. B\u003csub\u003e12\u003c/sub\u003e 8 mg, Niacin 12 mg, Pyridoxin 1.6 mg, Choline chloride 500 mg.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eHousing and management\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe chicks of different experimental groups were reared in cages with a wire mesh bottom. The electrically heated battery brooders were used to brood the chicks. 23 hrs lighting program was practiced. The experimental birds were fed in the morning every day. \u003cem\u003eAd-libitum\u003c/em\u003e feed and water were made available throughout the day. All the birds were reared under uniform and standard management practices.\u003c/p\u003e\u003cp\u003e\u003cb\u003eThyroid hormone profile\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThyroid hormone profile namely, T3, T4 and TSH was estimated by Quanti Microlisa\u0026reg; (J. Mitra and Co. Pvt. Ltd) based on the principle of competitive ELISA. Microwells are coated with respective anti-antibodies. Sample is added to microwell followed by addition of enzyme conjugate. Incubation is followed by a washing step to remove unbound compounds. The colour reaction is started by addition of substrate and stopped after a definite time. The colour intensity is proportional to the concentration of hormone in the sample.\u003c/p\u003e\u003cp\u003e\u003cb\u003eWeight of immune organs\u003c/b\u003e\u003c/p\u003e\u003cp\u003eAt the end of forty two days of experimental period, 10 birds from each dietary group (2 birds per replicate) were randomly selected and sacrificed after 17 hrs of fasting. Spleen, bursa, and thymus were collected and weighed. The weight of immune organs was expressed as % live weight (Pre-slaughter live weight) as shown below.\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\n$$\\:Spleen\\:\\left(\\%\\right)=\\frac{Weight\\:of\\:spleen\\:\\left(g\\right)}{\\text{P}\\text{r}\\text{e}\\:\\text{s}\\text{l}\\text{a}\\text{u}\\text{g}\\text{h}\\text{t}\\text{e}\\text{r}\\:\\text{l}\\text{i}\\text{v}\\text{e}\\:\\text{w}\\text{e}\\text{i}\\text{g}\\text{h}\\text{t}\\:\\left(\\text{g}\\right)}\\times\\:100$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equb\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equb\" name=\"EquationSource\"\u003e\n$$\\:Bursa\\:\\left(\\%\\right)=\\frac{Weight\\:of\\:bursa\\:\\left(g\\right)}{\\text{P}\\text{r}\\text{e}\\:\\text{s}\\text{l}\\text{a}\\text{u}\\text{g}\\text{h}\\text{t}\\text{e}\\text{r}\\:\\text{l}\\text{i}\\text{v}\\text{e}\\:\\text{w}\\text{e}\\text{i}\\text{g}\\text{h}\\text{t}\\:\\left(\\text{g}\\right)}\\times\\:100$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equc\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equc\" name=\"EquationSource\"\u003e\n$$\\:Thymus\\:\\left(\\%\\right)=\\frac{Weight\\:of\\:thymus\\:\\left(g\\right)}{\\text{P}\\text{r}\\text{e}\\:\\text{s}\\text{l}\\text{a}\\text{u}\\text{g}\\text{h}\\text{t}\\text{e}\\text{r}\\:\\text{l}\\text{i}\\text{v}\\text{e}\\:\\text{w}\\text{e}\\text{i}\\text{g}\\text{h}\\text{t}\\:\\left(\\text{g}\\right)}\\times\\:100$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eCell mediated immunity\u003c/b\u003e\u003c/p\u003e\u003cp\u003eUsing a method recommended by Cheng and Lamont (1988), the CMI response to the Phytohemagglutinin-P (PHA-P) mitogen was evaluated on day 27 of the experiment. PHA-P (1.0 mg/ml of PBS, 100microgram /bird) was administered intradermally to right foot web of ten birds in each treatment using a tuberculin syringe. Foot web of selected birds was cleaned using the cotton before injection. The left foot web of same bird was given 0.1 ml of freshly prepared autoclaved PBS serving as a control. The mean skin thickness of the right and left foot webs of the injected birds of each treatment was measured using a micrometre at 0 hours and 24 hours after PHA-P mitogen injection. The difference in thickness of the control foot web at 0 and 24 hours was subtracted from the difference in thickness of the mitogen-injected foot web to calculate the foot web index (FWI).\u003c/p\u003e\u003cp\u003eFWI (mm) = (R2 - R1) - (L2 - L1)\u003c/p\u003e\u003cp\u003eWhere,\u003c/p\u003e\u003cp\u003eR\u003csub\u003e1\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Thickness (millimetre) before injection of PHA-P injection\u003c/p\u003e\u003cp\u003eR\u003csub\u003e2\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Thickness (millimetre) after 24 hours of PHA-P injection\u003c/p\u003e\u003cp\u003eL\u003csub\u003e1\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Thickness (millimetre) before injection of PBS solution\u003c/p\u003e\u003cp\u003eL\u003csub\u003e2\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Thickness (millimetre) after 24 hours of PBS injection\u003c/p\u003e\u003cp\u003e\u003cb\u003eHumoral immune response\u003c/b\u003e\u003c/p\u003e\u003cp\u003eAs a measure of antibody titre against 1% SRBC suspension, humoral immunity was assessed. 10 birds in each treatment which were not involved in cell mediated immunity test were injected 1% SRBC via intravenous route on 29th day of the experiment. After five days post-inoculation, about 2.0 ml of blood was collected in serum vacutainers from immunized birds. The separated serum was used for microtiter haemagglutination (HA) test following a method reported by Siegel and Gross [\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003cb\u003eEstimation of serum IL6 and HSP70\u003c/b\u003e\u003c/p\u003e\u003cp\u003eTo evaluate the serum IL6 and HSP70, blood was collected on 21st day and 42nd day from 10 birds/treatment in serum vacutainers. The separated serum was transferred to 2ml Eppendorf tubes and stored at -20\u003csup\u003eo\u003c/sup\u003eC until analysed. Amount of IL6 in serum was quantified using Fine Test\u0026reg; Chicken Interleukin-6 ELISA kit (Catalogue no ECH0046). Likewise, serum HSP70 using Fine Test\u0026reg; Chicken HSP70 ELISA kit (Catalogue no ECH0072).\u003c/p\u003e\u003cp\u003e\u003cb\u003eGene expression\u003c/b\u003e\u003c/p\u003e\u003cp\u003eGene expression study of IL6 gene in spleen and HSP 70 gene in liver was done at 21st and 42nd day of experiment, respectively. Four birds per treatment were randomly chosen and sacrificed to collect 100 mg of spleen and liver samples (following standard protocol) from each bird. Collected samples were immediately transferred to nuclease-free, 2.0 ml Eppendorf tubes that contained 1.0 ml of RNAprotect\u0026reg; reagent. The tissues were homogenised using an automated homogenizer (Polytron) on a high setting for 30 to 60 seconds. The RNA was isolated from the homogenised samples using the TRIzol\u0026trade; reagent. Isolated RNA samples were checked for purity in a Nano drop UV spectrophotometer (Nano Drop 1000\u0026reg;, Thermo-Scientific, Singapore).The concentration of each RNA sample was calculated after obtaining the O.D. at 260/280. Revert Aid\u0026trade; first-strand cDNA synthesis kit (K 1622, Thermo Fisher Scientific, U.S.) was used to create the first-strand cDNA from the appropriate amount of RNA using random hexamer primers. Already reported primers (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) were used to quantify IL6 and HSP70 genes. mRNA expression folds of target genes were quantified relative to GAPDH and 28s RNA as housekeeping genes respectively. The 2-\u003csup\u003eΔΔ\u003c/sup\u003eCt method was used to analyse the real-time PCR data. Relative expression of genes IL-6 and HSP-70 in spleen and liver tissues respectively were quantified by a real-time PCR detection system (QIAquant 96\u0026reg; thermocycler) using 2X SYBR green master mix (RR420A, Takara TB Green\u0026trade; Premix Ex Taq\u0026trade;). All molecular reactions were performed by using a nuclease-free 8 tube-strips containing optically clear flat caps (Axygen Scientific\u0026reg;, Inc. USA).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003ePrimers (forward and reverse) used for real-time assay\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=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" 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\u003eS. No\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGene\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSequence (5\u0026rsquo;-3\u0026rsquo;)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eAnnealing\u003c/p\u003e\u003cp\u003eTemperature\u003c/p\u003e\u003cp\u003e(˚C)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eLength (bp)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eAccession no\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eIL6\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eF-AAATCCCTCCTCGCCAATCT\u003c/p\u003e\u003cp\u003eR-CCCTCACGGTCTTCTCCATAAA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e61\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e106\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eNM204628.1\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eHSP70\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eF-ACGACCAGGGGAACCGCACCAC\u003c/p\u003e\u003cp\u003eR-GGCTTGCCACCCTCGTTCACCAC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e56\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e74\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eHM587997\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eGAPDH\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eF-CCGTCCTCTCTGGCAAAGTCC\u003c/p\u003e\u003cp\u003eR-AGCCCCAGCCTTCTCCATG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e58\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e266\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eNM204305\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e4\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e28S RNA\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eF-CAGGTGCAGATCTTGGTGGTAGTA\u003c/p\u003e\u003cp\u003eR- GCTCCCGCTGGCTTCTCC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e58\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e273\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eJN639848\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"6\"\u003eIL6 Interleukin 6; HSP70 Heat shock protein 70; GAPDH Glyceraldehyde 3-phosphate dehydrogenase; RNA Ribonucleic acid; F Forward; R Reverse\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eTemperature humidity index (THI)\u003c/b\u003e\u003c/p\u003e\u003cp\u003eTHI was calculated using the formula suggested by Habeeb \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]Z. THI\u0026thinsp;=\u0026thinsp;db\u0026deg; F\u0026ndash;[(0.55\u0026ndash;0.55 x RH) (db\u0026deg; F\u0026ndash;58)].Where, db\u0026deg; F\u0026thinsp;=\u0026thinsp;dry bulb temperature (in Fahrenheit) and RH\u0026thinsp;=\u0026thinsp;relative humidity percentage (RH%)/100.THI is classified as: \u0026lt;82\u0026thinsp;=\u0026thinsp;absence of heat stress, 82 to \u0026lt;\u0026thinsp;84\u0026thinsp;=\u0026thinsp;moderate heat stress, 84 to \u0026lt;\u0026thinsp;86\u0026thinsp;=\u0026thinsp;severe heat stress, and 86 and more\u0026thinsp;=\u0026thinsp;very severe heat stress.\u003c/p\u003e\u003cp\u003e\u003cb\u003eAntioxidant status\u003c/b\u003e\u003c/p\u003e\u003cp\u003eSerum superoxide dismutase was quantified using Amplite\u0026trade; colorimetric superoxide dismutase assay kit by AAT Bioquest\u0026reg; (Catalogue no 11305).Serum catalase enzyme using SIGMA-ALDRICH\u0026reg; catalase assay kit (Catalogue no CAT100).Serum glutathione peroxidase enzyme using Cayman\u0026reg; glutathione peroxidase assay kit (Item no. 703102).Total antioxidant capacity of serum is evaluated by ferric reducing ability of plasma (FRAP) as suggested by Benzie \u003cem\u003eet al.\u003c/em\u003e, 1999 and 2,2-diphenyl-1-picrylhydrazyl (DPPH) by Brand \u003cem\u003eet al.\u003c/em\u003e, [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e].\u003c/p\u003e\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eStatistical Analysis\u003c/h2\u003e\u003cp\u003eThe experimental data were analysed using the SPSS computer package (IBMSPSS software version 26.0, SPSS Inc., Chicago, USA) adopting standard statistical procedures of Snedecor and Cochran, [\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e]. The parameters were analysed by one-way ANOVA using the general linear model and Tukey post-hoc analysis. Significance was declared at P\u0026thinsp;\u0026lt;\u0026thinsp;0.05 unless otherwise stated.\u003c/p\u003e\u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cb\u003eChemical composition of cauliflower leaf meal\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe chemical composition of cauliflower leaf meal (CLM) was given in the Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. The dry matter (DM), crude protein (CP), ether extract (EE), crude fibre (CF), nitrogen-free extract (NFE), total ash (TA), and acid insoluble ash (AIA)content of the fresh cauliflower waste were 13.6, 22.3, 6.93, 9.47, 49.76, 11.54 and 1.37 percent, respectively. Similarly, the calcium (Ca), phosphorus (P), total phenolics (TP), and glucosinolates were 3.60, 0.6, 7.68, and 0.042 percent respectively. The gross energy (GE) and metabolisable energy (ME) content of CLM was 2560 kcal/kg and 2099 kcal/kg respectively.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eChemical composition (%) of the cauliflower leaf meal\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"2\"\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\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eParticulars\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\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\u003cb\u003eCP\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e22.3\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eEE\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.93\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eCF\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e9.47\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eNFE\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e49.76\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTA\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e11.54\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eAIA\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.37\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eCa\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.61\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eP\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.28\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTPC\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e7.68\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eGlucosinolates\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.042\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eGross Energy (kcal/kg)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2560\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eMetabolisable energy (kcal/kg)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2099\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"2\"\u003eCP crude protein; EE ether extract; CF crude fibre; NFE nitrogen free extract; TA total ash; AIA acid insoluble ash; Ca calcium; P phosphorus; TPC total phenolic content\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eThyroid hormone profile\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe thyroid hormone profiles (T3, T4, and TSH) of experimental group of birds fed CLM in the present investigation were comparable with the control group of birds on the 21st and 42nd day of the experiment (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\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 feeding cauliflower leaf meal on the thyroid hormone profile\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=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" 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\u003eTreatment\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"3\" nameend=\"c4\" namest=\"c2\"\u003e\u003cp\u003e21st day\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"3\" nameend=\"c7\" namest=\"c5\"\u003e\u003cp\u003e42nd day\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eT3 (ng/ml)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eT4 (\u0026micro;g/ml)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eTSH (\u0026micro;IU/ml)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eT3 (ng/ml)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eT4 (\u0026micro;g/ml)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eTSH (\u0026micro;IU/ml)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e1.12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e19.61\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0.86\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1.99\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e21.79\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.25\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e1.21\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e17.70\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0.91\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2.08\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e19.55\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.52\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e1.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e17.75\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0.87\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2.16\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e18.80\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.28\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT4\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e1.82\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e17.77\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0.94\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1.78\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e19.07\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.19\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT5\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e1.25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e18.08\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0.98\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2.05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e18.88\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.20\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT6\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e1.45\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e17.85\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0.91\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2.22\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e19.04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.18\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eSEM\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0.452\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0.283\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0.025\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.259\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.452\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.058\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eP-value\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0.851\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0.242\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0.779\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.745\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.289\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.618\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"7\"\u003eT1 Control diet; T2 (50 mg/kg CLM); T3 (75 mg/kg CLM); T4 (100 mg/kg CLM); T5 (125 mg/kg CLM); T6 (150 mg/kg CLM); CLM Cauliflower; T3 Triiodothyronine; T4 Thyroxine; TSH Thyroid stimulating hormone; SEM standard error mean.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eWeight of immune organs\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe weight of different immune organs in the current investigation was presented in the Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e.The weight of the spleen was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher in T6 group as compared to the control group. The weight of the thymus was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher in the T6 group of birds as compared to the control group. The weight of bursa of experimental group of birds fed CLM was comparable with the control group of birds.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eEffect of feeding cauliflower leaf meal on the weight of immune organs\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\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=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colspan=\"3\" nameend=\"c4\" namest=\"c2\"\u003e\u003cp\u003eLive weight (%)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTreatment\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSpleen\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eBursa\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eThymus\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.245\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0.146\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.282\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.250\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0.140\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.300\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.248\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0.148\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.313\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT4\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.258\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0.144\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.343\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\u003eT5\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.271\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0.158\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.354\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\u003eT6\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.320\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0.151\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.440\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eSEM\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.041\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0.010\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.017\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eP-value\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.038\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0.517\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.029\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"4\"\u003eT1 Control diet; T2 (50 mg/kg CLM); T3 (75 mg/kg CLM); T4 (100 mg/kg CLM); T5 (125 mg/kg CLM); T6 (150 mg/kg CLM); CLM Cauliflower leaf meal; SEM standard error mean\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"4\"\u003e\u003csup\u003eabc\u003c/sup\u003eMean with superscript in a column differ significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05)\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eCell-mediated and Humoral immune response\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe cell-mediated immune (CMI) response of the experimental group of birds fed CLM in the current investigation was comparable with the control group (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). The humoral immune response of experimental group of birds against sheep RBCs (SRBCs) was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher in T6 group when compared with the control group of birds (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eEffect of feeding cauliflower leaf meal on cell-mediated immunity, humoral immunity, serum IL6 levels, and relative expression of IL6 gene\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=\"char\" char=\".\" 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\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eTreatment\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eFWI (mm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eHA titre (log2)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e\u003cp\u003eIL6 ELISA (pg/ml)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eIL6 Fold change\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e21st day\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003e42nd day\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0.283\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e18.98\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e171.71\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e287.81\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0.410\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e19.29\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e144.57\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e284.00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.96\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0.355\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e19.06\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e120.57\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e278.29\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.10\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT4\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0.321\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e25.33\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e76.00\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e276.86\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.18\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\u003eT5\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0.372\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e26.57\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e68.76\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e259.72\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.27\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\u003eT6\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0.390\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e31.22\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e70.42\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e193.52\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.77\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eSEM\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0.015\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.055\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e9.340\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e10.08\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.076\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eP-value\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0.182\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.003\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.049\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.012\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"6\"\u003eT1 Control diet; T2 (50 mg/kg CLM); T3 (75 mg/kg CLM); T4 (100 mg/kg CLM); T5 (125 mg/kg CLM); T6 (150 mg/kg CLM); CLM Cauliflower leaf meal; FWI Foot web index; HA Haemagglutination; IL6 Interleukin 6; SEM standard error mean\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"6\"\u003e\u003csup\u003eabc\u003c/sup\u003eMean with superscript in a column differ significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eSerum IL-6 levels\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe serum interleukin-6 (IL6) levels on 21st \u0026amp; 42nd day of the experiment were presented in the Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e. In the current investigation, the serum IL-6 level (pg/ml) on the 21st day was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) lower in T4, T5 and T6 group of birds. On the 42nd day, the serum IL 6 was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) lower in T6 group and the serum IL6 levels in the other experimental group of birds fed CLM were comparable with the control group.\u003c/p\u003e\u003cp\u003e\u003cb\u003eIL-6 gene expression study\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe results of relative expression of IL6 gene were presented in the Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e. The relative expression of IL6 gene was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher in T6 group of birds as compared to the control group.\u003c/p\u003e\u003cp\u003e\u003cb\u003eTemperature humidity index\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe weekly temperature humidity index (THI) during the experimental were 80.69, 82.01, 83.02, 85.20, 85.61, and 89.32 during the 1st, 2nd, 3rd, 4th, 5\u003csup\u003eth,\u003c/sup\u003e and 6th week of the experiment, respectively. The weekly temperature humidity index (THI) was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher during 4th, 5th, and 6th week of the experiment.\u003c/p\u003e\u003cp\u003e\u003cb\u003eHSP 70 gene expression study\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe results of relative expression of HSP70 gene was presented in the Table\u0026nbsp;\u003cspan refid=\"Tab7\" class=\"InternalRef\"\u003e7\u003c/span\u003e. The relative expression of HSP70 gene was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) lower in T6 group of birds as compared to the control group.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab7\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 7\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eEffect of feeding cauliflower leaf meal on relative expression HSP70 gene and serum HSP70 levels\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\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=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e\u003cp\u003eSerum HSP 70 (ng/ml)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTreatment\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHSP70 Fold change\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003e21st day\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e42nd day\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1.187\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.227\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.93\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1.1617\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.193\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\u003eT3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.79\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1.1718\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.172\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\u003eT4\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.81\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1.163\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.155\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\u003eT5\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.68\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1.1383\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.093\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT6\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.52\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1.137\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.077\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eSEM\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.048\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0.009\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.014\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eP value\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.025\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0.557\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.005\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"4\"\u003eT1 Control diet; T2 (50 mg/kg CLM); T3 (75 mg/kg CLM); T4 (100 mg/kg CLM); T5 (125 mg/kg CLM); T6 (150 mg/kg CLM); CLM Cauliflower leaf meal; HSP70 Heat shock protein 70; SEM Standard error mean\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"4\"\u003e\u003csup\u003eabc\u003c/sup\u003eMean with superscript in a column differ significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eSerum HSP 70 levels\u003c/b\u003e\u003c/p\u003e\u003cp\u003eOn the 21st day, the serum HSP70 levels of the experimental group of birds fed CLM were comparable with the control group however, on the 42nd day of the experiment, the mean serum levels of HSP 70 were significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) lower in T5 and T6 groups than T2, T3, T4, and the control group (Table\u0026nbsp;\u003cspan refid=\"Tab7\" class=\"InternalRef\"\u003e7\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cb\u003eAntioxidant status\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe results pertaining to the concentration of serum antioxidant enzymes and total antioxidant capacity of serum was presented in the Table\u0026nbsp;\u003cspan refid=\"Tab8\" class=\"InternalRef\"\u003e8\u003c/span\u003e. The serum SOD levels were significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher in the T6 group of birds. Catalase level was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher in the T4, T5 and T6 group of birds. The serum GPx levels in the experimental group of birds were comparable with the control group. The serum FRAP values were significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher in T5 and T6 group as compared to the control group of birds (Table\u0026nbsp;\u003cspan refid=\"Tab8\" class=\"InternalRef\"\u003e8\u003c/span\u003e). The serum DPPH values were significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05)higher in the T6 group of birds and the DDPH values of the other experimental group of birds fed CLM were comparable with the control group (Table\u0026nbsp;\u003cspan refid=\"Tab8\" class=\"InternalRef\"\u003e8\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab8\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 8\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eEffect of feeding cauliflower leaf meal on the antioxidant status\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=\"char\" char=\".\" 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\u003eTreatment\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSOD (U/ml)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eCatalase\u003c/p\u003e\u003cp\u003e(\u0026micro;mol/min/ml)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eGPx (nmol/min/ml)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eFRAP (mM/ml)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eDPPH (% Antioxidant activity)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e66.16\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e11.24\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6.64\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e2.822\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e14.82\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e67.43\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e11.51\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e5.09\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e3.157\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e15.03\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e69.05\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e11.67\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e5.10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e3.177\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e16.07\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT4\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e68.64\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e11.72\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e4.97\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e3.147\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e15.38\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eT5\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e70.34\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12.29\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e4.25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e3.388\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e17.19\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\u003eT6\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e73.88\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12.54\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e5.82\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e3.444\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e18.66\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eSEM\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.625\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.113\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0.662\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.045\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.637\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eP-value\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.003\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0.169\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.042\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"6\"\u003eT1 Control diet; T2 (50 mg/kg CLM); T3 (75 mg/kg CLM); T4 (100 mg/kg CLM); T5 (125 mg/kg CLM); T6 (150 mg/kg CLM); CLM Cauliflower leaf meal; SOD Super oxide dismutase; GPx Glutathione peroxidase; FRAP Ferric reducing ability of plasma; DPPH2,2-diphenylpicrylhydrazyl; SEM Standard error mean\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"6\"\u003e\u003csup\u003eabc\u003c/sup\u003eMean with superscript in a column differ significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003e\u003cb\u003eChemical composition of cauliflower leaf meal\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe dry matter (DM) content of cauliflower leaves in the present experiment was similar to the DM content reported by Wadhwa and Bakshi [\u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e73\u003c/span\u003e] and (2016) however, it was found to be higher than values reported by Sharma and Prasad [\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e] and Raza \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e].The crude protein (CP) content of CLM in the current investigation, was very close to the value reported by Mahgoub \u003cem\u003eet al.\u003c/em\u003e [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e] and Gupta and Wagle [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. However, the CP content was lower than the value reported by Wani \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e75\u003c/span\u003e] and higher than the value reported by Mogra \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e] and Bakshi \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Similarly, the ether extract (EE) content of CLM was higher than the EE content reported earlier [28, 48 and 11). The crude fibre (CF) content of CLM (9.47%) was comparable to the values reported by Gupta and Wagle [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e] and lower than the value reported by Mogra \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. The nitrogen-free extract (NFE) of CLM was comparable to the value reported by Wani \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e75\u003c/span\u003e] and Ahmed \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. The total ash (TA) content of CLM was consistent with the findings of Ahmed \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e] \u0026amp; Gupta and Wagle [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e] and lower than the value reported by Mogra \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. The acid insoluble ash (AIA) of the CLM was higher than the reports of Gupta and Wagle [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]and it may be due to different ways of collection and drying. The calcium (Ca) content of CLM was comparable to the value reported by Revathi \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e] and higher than the value reported by Ahmed \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. The phosphorus (P) content of CLM was also comparable to the value reported by Revathi \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e] and higher than the value reported by Ahmed \u003cem\u003eet al\u003c/em\u003e. (2013).The total phenolic content (TPC) of CLM in the present experiment was higher than the values reported by Abul-Fadl [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e] and Wu \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e76\u003c/span\u003e]. However, the TPC values in the current study were lower than those reported by Collado-Alvarado \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. The glucosinolates content of CLM in the present investigation was comparable to the value reported by Gupta and Wagle [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e] \u0026amp; Drabińska \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e] and slightly lower than the levels reported by Abul-Fadl [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e] \u0026amp; Chandra [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003cb\u003eThyroid hormone profile\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe thyroid hormone profile of birds in the current investigation were consistent with the findings of Rabie \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e] who reported that the feeding canola meal (\u003cem\u003eBrassica napus\u003c/em\u003e) upto 15% in broilers did not affect the plasma thyroid hormone profile. Further, Abdel-Moneim \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e] reported that the serum T3 concentration was unaffected in the Japanese quail fed a diet containing full-fat canola seeds at 150g/kg. Kermanshahi and Pour [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e] reported no significant difference in thyroid hormone profile of broilers fed rapeseed meal upto 20% level. Similarly, Pena [\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e] reported that there was no significant difference in serum T3, and T4 hormone levels of laying hens fed Broccoli Stems and Leaves Meal upto 15% level.\u003c/p\u003e\u003cp\u003e\u003cb\u003eWeight of immune organs\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe higher weight of immune organs in the 15% CLM fed group of birds in the current experiment corroborated with that of Ao and Kim [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e] who reported higher relative weight of the spleen in broilers fed grape seed extract rich in polyphenols. The similar findings were also reported by Omar \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e] who reported a higher weight of thymus in broilers fed onion extract rich in polyphenols. The improved humoral immune response against sheep red blood cells (SRBC) seen in this experiment may be co-related with the increased weight of immune organs. Similar co-relation between greater spleen, thymus, and other immune organ weight and higher antibody titre against SRBCs was also reported by Kabir \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. Increased weight of immune organs may be attributed due to the proliferation of lymphocytes. When challenged with an antigen, lymphocytes respond by creating lymphokines, proliferating their antigen-specific lymphocyte clones, and enhancing immune responses [\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e]. Yang \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e78\u003c/span\u003e] reported an increased splenic lymphocyte proliferation response and spleen weight in broilers supplemented with quercetin (phenolic compound).\u003c/p\u003e\u003cp\u003e\u003cb\u003eCell-mediated and Humoral immune response\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe cell mediated immune response of experimental group of birds recorded in the current investigation was consistent with the findings of Kumar \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e] who reported that there was no significant difference in CMI response of broilers fed pomegranate peel extract rich in polyphenols during the summer. Similar findings were reported by Rao \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e] where he reported that there was no significant difference in cell mediated immune response to PHA-P in broilers fed \u003cem\u003eMoringa olefera\u003c/em\u003e leaf meal and pomegranate peal meal at 1000mg/kg \u0026amp; 500mg/kg of basal diet respectively. In contrast, to the present findings, Gopi \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e] reported that pigeons fed polyphenols isolated from pomegranate peels had better cell mediated immune response against intradermal inoculation of PHA-P. Dumaup and Ampode [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e] also reported significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher CMI in broiler chickens fed diets containing water hyacinth meal.\u003c/p\u003e\u003cp\u003eThe humoral immune response of experimental group of birds recorded in the current investigation were consistent with the findings of Pourhossein \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e], who reported the enhanced immune response against SRBCs in birds fed sweet orange peel extract rich in polyphenols. Emadi and Kermanshahi [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e] reported that the broiler chicks fed turmeric rhizome powder rich in polyphenols upto 0.75% level had a higher antibody titre against SRBCs. In further support of these findings, Gholamrezaie \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e] reported that broiler chickens supplemented with \u003cem\u003eArtemsia annua\u003c/em\u003e extract and leaf powder (contains flavonoids and quercetin) at 1.5% level increased total antibody titre against SRBC in the experimental groups when compared with the control group. Increased antibody titre against SRBC in the current study is suggestive of increase in humoral immunity of experimental birds when compared with the control group.\u003c/p\u003e\u003cp\u003e\u003cb\u003eSerum IL-6 levels\u003c/b\u003e\u003c/p\u003e\u003cp\u003eIL6 is a pleiotropic cytokine, and its concentration is used as an indicator to study the inflammatory status of animals [\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e]. In the present investigation, the results were in line with the reports of Sun \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e], who reported that the supplementation of a polyphenol-rich \u003cem\u003eYucca schidigera\u003c/em\u003e extract at 300mg/kg of basal diet in broiler chickens, resulted in lower serum IL-6 concentrations when compared with the control group. Adeyemi \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e] also reported lower serum IL6 levels in broiler chickens fed diets supplemented with \u003cem\u003eAnacardium occidentale\u003c/em\u003e leaf powder upto 4g/kg. In further support of these findings, Liu \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e] reported that the grape seed extract rich in polyphenols reduced inflammation in mice fed a high-fat diet by significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) lowering serum pro-inflammatory cytokine (IL6) concentrations. Similarly, Kilany \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e] reported that the there was a significant decrease in serum Il6 levels of broilers fed a diet containing 5% \u003cem\u003eMoringa olefera\u003c/em\u003e leaf meal and 0.5% sweet basil.\u003c/p\u003e\u003cp\u003eThe findings of the present investigation were consistent with the results of Nootash \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e], who reported that the Interleukin-6 gene expression in the spleen of rainbow trout was upregulated on feeding green tea extract (\u003cem\u003eCamellia sinensis\u003c/em\u003e) rich in polyphenol at a rate of 500 mg/kg. Further, Shekarabi \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e] reported that the transcription levels of interleukin\u003cem\u003e-6\u003c/em\u003e genes were up-regulated in the fish fed dandelion flower extract (DFE) at 3 and 4 g/kg of basal diet. In further support of this study, Awad \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e] reported that the IL-6 gene was up-regulated in the spleen of broiler chickens fed rutin a bioactive chemical found in apple and tea leaves at a rate of 1g/kg diet. Similarly, Khalid \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e] reported significantly increased IL6 gene expression in rabbits fed \u003cem\u003eMoringa olefera\u003c/em\u003e leaf powder at 200 mg/kg body weight.\u003c/p\u003e\u003cp\u003e\u003cb\u003eTemperature humidity index\u003c/b\u003e\u003c/p\u003e\u003cp\u003eDuring the last 3 weeks of the experimental trial the THI was in severe to very severe heat stress. The effects of heat stress on laying hens can be compared between various study conditions by utilising THI, which takes into account both ambient temperature and humidity [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. El-Tarabany, [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e] reported that, Japanese quail exposed to heat stress showed decreased immunological and production characteristics as well as negative welfare consequences.\u003c/p\u003e\u003cp\u003e\u003cb\u003eHSP 70 gene expression study\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe heat shock transcription factor activity or concentration can be increased at high temperatures to cause the synthesis of HSP70 mRNA [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. The HSP70 is the most conserved and well-studied members of the HSP family, and their expression is altered by a wide range of substances or stressors. It is reported that the primary inducer of HSP-related gene expression is high ambient temperature [\u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e77\u003c/span\u003e]. The results of the present investigation were consistent with the findings of Hajati \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e], who reported that the broilers supplemented with grape seed extract rich in polyphenols and vitamin C during chronic heat stress conditions resulted in a down-regulation of HSP-70 gene expression in the liver and heart when compared with the control group. Further, Sugito \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e] reported the downregulation of HSP-70 gene in the kidney of broilers fed with salix extract under heat stress conditions. Wan \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e74\u003c/span\u003e] reported that the mRNA expression levels of HSP70 was decreased in breast muscle of broilers supplemented with \u003cem\u003eArtemisia annua\u003c/em\u003e L. when compared with the control group. Similarly, Song \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e] reported significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) decreased HSP70 mRNA expression in the intestine of broilers supplemented with \u003cem\u003eArtemisia annua\u003c/em\u003eat 1g/kg of diet.\u003c/p\u003e\u003cp\u003e\u003cb\u003eSerum HSP 70 levels\u003c/b\u003e\u003c/p\u003e\u003cp\u003eWhen analysing the broiler's response to heat stress, it is crucial to take oxidative stress into account [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. Bird's maintaining heat homeostasis under stressful conditions results in higher reactive oxygen species (ROS) concentrations. It is reported that the body begins producing heat shock proteins to defend itself from ROS' damaging effects on the cells [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. The results of the present findings were consistent with the findings of Hassan and Asim [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e], who reported that the broilers supplemented with vitamin C and acetylsalicylic acid had lower serum HSP70 levels than controls when raised in heat stress conditions (40\u0026deg;C for 4 hours per day). Hu \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e] reported that the serum HSP70 levels were significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) reduced in broilers fed the diet supplemented with phloretin (a type of natural phenol that is found in apple tree leaves) at 200 mg/kg. In further support of these findings, Zhang \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR79\" class=\"CitationRef\"\u003e79\u003c/span\u003e] and Liu \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e] reported the significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) decrease in HSP70 levels in broilers fed a diet supplemented with the plant antioxidants like curcumin and resveratrol. The reduced serum HSP70 levels in treatment groups may also be attributed to the fact that the polyphenols present in the CLM will scavenge the ROS which is also one of the factors responsible for HSP production as suggested by Mahmoud \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003cb\u003eAntioxidant status\u003c/b\u003e\u003c/p\u003e\u003cp\u003eBroilers require higher antioxidant capacity since low antioxidant capacity has been linked to cellular damage, immune system degeneration, and metabolic abnormalities [\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e]. The results of present investigation were similar with the Drazbo \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e] who reported that there was a significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) increase in serum SOD levels of turkey fed rape seed meal at a 15% level. Wan \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e74\u003c/span\u003e] reported that the broilers fed \u003cem\u003eArtemisia annua L.\u003c/em\u003e leaves rich in polyphenols showed greater blood levels of SOD and catalase. Further, Hu \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e] reported that the serum catalase and glutathione peroxidase concentration were significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) increased in broilers fed the diet supplemented with phloretin (natural phenol in apple tree leaves) at 200 mg/kg diet. These results corroborated with the findings of Drazbo \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e], who reported significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher FRAP in the turkey fed a rape seed meal at a 15% level. Similar results were reported by Ognik \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e] in turkey hens fed a diet supplemented with the quercetin (phenolic compound). Llorach \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e], reported that the testing of a cauliflower by-product extract using FRAP method revealed a significant antioxidant potential. He also reported that there is a linear correlation between FRAP readings and the overall phenolic content of cauliflower leaves. These results are consistent with the findings of Wan \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e74\u003c/span\u003e] who reported the broilers fed \u003cem\u003eArtemisia annua L.\u003c/em\u003e leaves rich in polyphenols showed significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher serum DPPH levels when compared with the control. Llorach \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e] reported that an extract of cauliflower by-products effectively scavenges DPPH free radicals. Similarly, Seong \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e] reported the significantly higher DPPH free radical scavenging capacity of Chinese cabbage extract.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eTherefore, it is concluded that cauliflower leaf meal (CLM) inclusion in the diet upto 15% improves immunity, which is evident by increased immune organ weight, higher haemagglutination titre against sheep RBCs, and upregulation of IL6 gene in the spleen. CLM feeding also increases the antioxidant status and alleviates heat stress in broiler chickens.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003ch2\u003eCompeting Interests\u003c/h2\u003e\u003cp\u003eThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e\u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e\u003cp\u003eThe work has been done under the institute project entitled \u0026ldquo;Studies on the utilization of fruit and vegetable waste as an alternative feed\u0026rsquo;\u0026rsquo; and funded by Indian Veterinary Research Institute, Bareilly, India.\u003c/p\u003e\u003ch2\u003eAuthor Contributions\u003c/h2\u003e\u003cp\u003eTM is involved in conducting experiment, analysis, and writing the manuscript. SS and AB planned and executed of both experiments, edited manuscript, coordinated studies. GK and AB are involved in conducting experimental trial and analysis part. RM edited manuscript.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eCorresponding author should be contacted to get data available on request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eA.O.A.C. Methods of analysis. Association of Official Analytical Chemists. (15th Edn.) Washington, USA : 20\u0026ndash;44. (1990)\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eA.O.A.C. Official Methods of analysis. AOAC international. 18th edn., Virginia, USA (2000)\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAbaza, R.H., Blake, J.T.: Fisher. Oxalate determination: Analytical problems encountered with certain plant species. J. Association Official Anal. 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Poult. sci. \u003cb\u003e88\u003c/b\u003e(10), 2159\u0026ndash;2166 (2009). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3382/ps.2009-00165\u003c/span\u003e\u003cspan address=\"10.3382/ps.2009-00165\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"waste-and-biomass-valorization","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"wave","sideBox":"Learn more about [Waste and Biomass Valorization](http://link.springer.com/journal/12649)","snPcode":"12649","submissionUrl":"https://submission.nature.com/new-submission/12649/3","title":"Waste and Biomass Valorization","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Antioxidant, Cauliflower leaf meal, Immunity, Heat stress, Polyphenol, Interleukin 6, Heat shock protein 70","lastPublishedDoi":"10.21203/rs.3.rs-7145387/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7145387/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe present study was investigated effect of CLM on thyroid hormone profile, immunity, expression of IL6 \u0026amp; HSP70 gene and antioxidant status of broiler. 240 day-old chicks were distributed into 5 groups having 8 birds in each replicate. The diet was formulated as per BIS (2007) standards for T1, while T2, T3, T4, T5 and T6 were given diets substituted with CLM @ 50, 75, 100, 125, and 150 g/kg, respectively for 42 days. The weight of the spleen, thymus and humoral immune response was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher in T6 group. On 21st day, serum IL6 levels was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) lower in T4, T5, and T6. On 42nd day, it was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) lower in T6. Relative expression of the HSP70 gene in liver tissue was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) down regulated in T6. The serum SOD and catalase levels were significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher in T4, T5, and T6. Serum FRAP and DPPH values were significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher in T5 and T6. The thyroid hormone profile, weight of bursa, cell-mediated immune response and serum GPx levels were remained comparable. Hence, it is concluded that CLM inclusion in a diet up to 15% improves immunity and antioxidant and alleviates heat stress in chickens.\u003c/p\u003e","manuscriptTitle":"Exploring Nutritional Evaluation of Cauliflower (Brassica Oleracea Var Botrytis) Waste on the Thyroid Hormone, Antioxidant Status, Immunity and Expression of Interleukin 6 and HSP 70 Genes in Broilers","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-08-22 09:43:28","doi":"10.21203/rs.3.rs-7145387/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"","date":"2025-12-01T06:43:52+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-08-14T06:20:05+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"Waste and Biomass Valorization","date":"2025-08-09T06:17:59+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-07-17T18:45:14+00:00","index":"","fulltext":""},{"type":"submitted","content":"Waste and Biomass Valorization","date":"2025-07-17T02:19:09+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"waste-and-biomass-valorization","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"wave","sideBox":"Learn more about [Waste and Biomass Valorization](http://link.springer.com/journal/12649)","snPcode":"12649","submissionUrl":"https://submission.nature.com/new-submission/12649/3","title":"Waste and Biomass Valorization","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"23e5df8d-9044-4902-8706-303a8dc6f6eb","owner":[],"postedDate":"August 22nd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-02-16T15:59:41+00:00","versionOfRecord":{"articleIdentity":"rs-7145387","link":"https://doi.org/10.1007/s12649-026-03499-x","journal":{"identity":"waste-and-biomass-valorization","isVorOnly":false,"title":"Waste and Biomass Valorization"},"publishedOn":"2026-02-10 15:56:56","publishedOnDateReadable":"February 10th, 2026"},"versionCreatedAt":"2025-08-22 09:43:28","video":"","vorDoi":"10.1007/s12649-026-03499-x","vorDoiUrl":"https://doi.org/10.1007/s12649-026-03499-x","workflowStages":[]},"version":"v1","identity":"rs-7145387","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7145387","identity":"rs-7145387","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}