Mitigation of Heat Stress in Broiler Chickens during Hot Summer Seasons through Betaine Hydrochloride Osmoprotectant Intervention

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MONIKA, M. Gopi, M. Vispute, Nagesh Sonale, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3994204/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Modern poultry genotypes produce more body heat due to their greater metabolic activity and in tropical country; high environment temperature often worsens the situation. Heat stress is a major concern for the poultry industry as it adversely affects growth as well as immune functions. Nutritional interventions play an important role in ameliorating the negative effects of heat stress and will be more economical approach. In order to evaluate the role of osmolyte (betaine) as a heat ameliorating agent in broiler chicken, an experiment was conducted with gradient levels of 0, 0.5, 1.0 and 1.5 g/kg for a growing period of 42 days. Each group comprised 48 birds in six replicates of 8 each. All birds were maintained under standard managemental conditions. Thermo-Humidity Index value ranged 85–87 throughout the experimental period. Production performance, immunity, gut morphometry, hemato-biochemical and stress indicators were studied. Supplementation of betaine significantly improved production performance as well as breast meat yield (P < 0.05). Supplementation (1 and 1.5g/kg) significantly (P < 0.01) improved both the humoral and cellular immunity. Betaine supplemented groups also depict significant (P < 0.01) rise in hemoglobin and relative immune organ weight. Addition of betaine to basal diet significantly (P < 0.01) increased the villus length and also lowered H:L ratio, with better ALT and AST values than the control. The dose level of at 1 and 1.5g/kg improve all the parameters but villi structure was more improved in 1g/kg dose level and also it is economical than 1.5g/kg dose level. To conclude present study confirms that, the supplementation of betaine at 1g/kg levels could significantly improve overall growth performance, gut morphology and welfare in heat stressed broiler chickens. Betaine broilers growth performance heat stress immunity Figures Figure 1 Introduction Heat stress is a major issue in developing nations with tropical climates, particularly for broiler chickens because of their rapid growth and conventional open-shed housing. The most cost-effective and efficacious method for reducing heat stress for marginal farmers is nutritional management as opposed to housing management or automation. Because of its osmo-protectant, methyl donor, and amino acid sparing effects, the current investigation confirms that supplementing with 1g/kg of betaine can be the best strategy to control heat stress losses. Because of its effectiveness and low cost, our results can assist broiler farmers, industry, and even small-scale farmers. Ameliorating heat stress is the major challenge to the broiler industry especially in tropical regions. Among livestock, poultry is particularly sensitive to temperature associated environmental challenges especially heat stress. High ambient temperature has deleterious effect on productive performance in poultry which often results in economic losses. Heat stress is a result of disequilibrium between production and loss of body heat, where the former exceeds later. The major effects due to heat stress might be increased osmotic disturbances and elevated production of free radicals causing oxidative injuries at cellular level to ultimately imposing stress. In order to maintain homeostasis, physiological and behavioral responses are exhibited by birds, which compromises the performance, immunity and meat quality (Shakeri et al. 2018 ) Combating heat stress without compromising production performance is a difficult task. Efforts are under way to ameliorate the negative effects of heat stress through dietary interventions to achieve optimum performance. As a by-product of sugar beet processing, betaine is commercially available as a feed additive. Betaine is a trimethyl derivative of the amino acid glycine, a metabolite of plant and animal tissues which may provide a strategic solution for these challenges (Akhavan-Salamat and Ghasemi 2016 ). Protection of cells against osmotic inactivation is a principal biological function of betaine in living beings. Plants and bacteria also produce and accumulate betaine to cope with salt imbalances and thermal stress. Natural betaine has two roles in nutrition as protection of cells against osmotic inactivation and as a methyl donation via . Transmethylation (Alahgholi et al. 2014 ) As an osmo-protective agent, betaine accumulates inside the cell without and maintaining normal functions. As an osmolyte agent it protects the intestinal cell proteins and enzymes from high ambient temperature. Betaine keeps cells hydrated and maintains ion balance to improve thermotolerance (Abudabos et al. 2016 ). Betaine involves in transmethylation process, wherein methyl groups (–CH 3 ) containing a single carbon atom is donated, to synthesize critical metabolites and regulate gene expression through DNA methylation (Chand et al. 2017 ). Transmethylation helps in nucleic acid, protein and choline synthesis. From nutritional aspect, betaine can replace other methyl donors such as methionine up to 15–20%, total sulphur containing amino acids up to 10% of and almost all of choline. By decreasing the requirement of these essential and costly nutrients, betaine effectively minimizes feed cost. Besides, betaine has also been shown to improve embryonic and post-birth survival rate of piglets (Van Wettere et al., 2012 ). Betaine also offers several other beneficial effects such as to improve hatchability and chick weight; spare choline and methionine; improve gut health and function; alleviate heat stress; increases carcass and breast meat yield; reduces coccidiosis lesions (additive effect in combo with ionophores); reduce wet litter condition; improve immune response etc. High blood homocysteine level increases risk of chick mortality, betaine significantly counters homocysteine (Sakomura et al. 2013 ; He et al. 2015 ). Betaine as a methyl donor provides its labile methyl groups for the synthesis of several metabolically active substances such as creatine and carnitine. Enhanced methionine availability after dietary supplementation of betaine resulting in improved animal performance (Rao et al., 2011 ). To address the complex challenges posed by heat stress in broiler chickens, particularly in tropical regions, a comprehensive understanding of the physiological responses and nutritional interventions is imperative. High ambient temperatures disrupt the delicate balance between heat production and dissipation, leading to adverse effects on poultry performance and economic losses. The intricate interplay of osmotic disturbances and heightened free radical production at the cellular level exacerbates the stress imposed by heat. Efforts to counteract heat stress without compromising production performance are underway, with a focus on dietary interventions. Betaine, derived from sugar beet processing, emerges as a promising solution due to its multifaceted roles as an osmo-protectant, methyl donor, and amino acid sparing agent. Within cells, betaine accumulates, preserving normal functions and safeguarding proteins and enzymes in intestinal cells from the impact of high ambient temperatures. Its involvement in the transmethylation process contributes to the synthesis of critical metabolites and the regulation of gene expression through DNA methylation. From a nutritional standpoint, betaine proves to be a versatile substitute for other costly methyl donors, effectively reducing the requirements for essential nutrients such as methionine, sulphur-containing amino acids, and choline. The economic benefits extend beyond feed cost reduction, as betaine demonstrates positive effects on embryonic and post-birth survival rates in piglets. The study emphasizes the multifaceted benefits of betaine supplementation, including improvements in hatchability, chick weight, gut health, immune response, carcass and breast meat yield, and the reduction of coccidiosis lesions. Additionally, betaine counters the detrimental effects of high blood homocysteine levels, mitigating the risk of chick mortality. As a methyl donor, betaine contributes labile methyl groups for the synthesis of metabolically active substances, further enhancing animal performance. Against this backdrop, the present study evaluates the efficacy of post-hatch betaine supplementation on broiler performances reared under higher environmental temperatures, providing valuable insights for the sustainable management of heat stress in poultry farming. Material and methods Birds, experimental design and diet One hundred and ninety numbers of day-old broilers (CARIBRO-VISHAL) were divided into four treatments groups and fed with diets contained 0 (control), 0.5, 1.0, and 1.5g betaine/kg feed. Each group consisted of 48 birds in six replicates (8 birds). The experimental diets (starter and finisher) were formulated as per the broiler’s requirement (22 and 19.5% CP, respectively). All the birds were raised under uniform standard managemental conditions of brooding and vaccination. Where the basal diet of Broiler chicken was shown as control in Table 1 . Table 1 Ingredients and nutrient composition of basal diets for broiler chicken Ingredients (g/kg) Pre-starter Starter Finisher Maize 571 579 635 Soybean meal (46% CP) 342 330 288 Fish meal (48% CP) 30 32 20 Vegetable oil 24 29 28 Limestone powder 9.0 9.0 8.0 Di-calcium phosphate 14 12 12 DL-Methionine 2.0 1.0 1.0 L-lysine HCl 1.0 0.0 0.0 † Trace mineral Premix 1.0 1.0 1.0 ǂ Vitamin Premix 2.0 2.0 2.0 ¶ Vitamin B complex 1.5 1.5 1.5 Choline Chloride 1.0 1.0 1.0 Salt 3.0 3.0 3.0 Toxin binder 1.0 1.0 1.0 Nutrient composition Metabolizable energy (ME) (Kcal/kg) 3000 3050 3100 Crude protein, % 23.0 21.9 20.0 Lysine, % 1.30 1.19 0.99 Methionine, % 0.52 0.49 0.45 Calcium, % 1.00 1.00 1.00 Available phosphorus, % 0.45 0.41 0.41 Temperature-humidity index (THI) measurement The study was carried out during hot summer season in order to simulate the field conditions of heat stress to broilers. The temperature humidity index (THI) was calculated as per the formula suggested by Zulovich and DeShazer (1990). Ambient temperature and Relative humidity were measured using dry-bulb and wet-bulb temperatures. According to Zulovich and DeShazer, 1990, THI ranges were classified as comfort (THI 81) zones, used to assess the potential indices of heat stress for poultry. The Temperature Humidity Index (THI) was recorded throughout the experimental period (42 days) and were in the ranged from 85 to 87. Production performance Body weight (g) was measured in different growth phases (14–21, 21–42, 14–42 and 0–42 days) during the experimental period to ascertain body weight gain (BWG) during each phase. Weighed quantities of respective diets were offered adlib on daily basis to all the replicates of each dietary regimen. Replicate wise residues were weighed on weekly basis to ascertain weekly as well as overall feed consumed. From the body weight gain and feed consumption, the feed to gain conversion ratio was derived. Evaluation of immune response A. Humoral immune response The microtiter hemagglutination (HA) procedure was followed to determine antibody titre concentration in chickens against 1% Sheep RBCs (SRBC) (Siegel and Gross 1980). The 1% SRBC suspension was prepared in Alserver’s solution and subsequently injected intravenously (0.1 ml) into 12 randomly selected healthy birds per group (one male and one female broilers in each replicate) on 28th day post-hatch. Serum was harvested 5 days post-injection and HA antibody titre was determined against antigen (SRBC). HA titre was expressed in terms of Log 2 value of highest dilution wherein reaction was positive against SRBCs. B. In-vivo cell mediated immune response (CMIR) The CMIR was studied at four weeks of age by intra-dermal injection of 0.1 ml (1mg/ml) of phytohemagglutinin type-P (PHA-P) at 3rd and 4th inter-digital space of the right foot and 0.1 ml of sterile PBS in left foot (control) (Cheng and Lamont, 1988). The foot web index (mm) was calculated as the difference between the swelling in the right and left foot before (R1 and L1) and after (R2 and L2) 24 hours of injection. The formula for CMIR calculation is (R2-R1) - (L2-L1). where, L1 – fold thickness before injection in left inter-digital space L2 – fold thickness after PBS injection in left inter-digital space R1 – fold thickness before injection in right inter-digital space R2 – fold thickness after PHA-p injection in right inter-digital space. C. Relative weight of lymphoid organs Weights of lymphoid organs such as bursa of Fabricius, spleen and thymus (g) were recorded at 28 and 42 days of age and expressed as percentage of pre-slaughter live weight. Histomorphometry of intestinal mucosa Histomorphometry measure viz ., villus height (VH), villus width (VW), and ratio of villus height to villus width (VH: VW) were measured at 28 and 42 days of age. After identification with help of Meckel's diverticulum at jejuno-ileal junction, a piece (0.5–1.0 cm) of jejunum was collected from 12 healthy birds per treatment. Collected segments were preserved in 10% formalin solution after carefully removing intestinal contents without damaging the mucosa, until processing for histological slide preparation. Further, slides were stained with hematoxylin and eosin stain following by mounting with coverslip. Mounted slides were examined under microscope under low power (10×). The measurement of villus height (VH), villus width (VW) was carried out by using Primo Star 3.0 imaging software, Zeiss, Germany. VH: VW ratio was calculated by formula method. Hematological attributes and serum enzymes Blood samples were collected (~ 1ml) from the jugular vein using 26-gauge needle in K3- EDTA tubes on 2, 4 and 6 weeks of age to determine hematological attributes. Autoanalyzer (Abacus Junior Vet5, Diatron, USA) was used to determine hemoglobin (Hb) levels. A. Determination of H/L ratio Immediately after blood collection, thin blood smear was prepared on clean glass slides. After drying, the smears were fixed with methanol and stained with Giemsa stain. After drying, smears were examined under microscope at 100× objective with oil immersion. 100 cells, i.e, heterophils and/or lymphocytes counted in each smear to calculate H/L ratio (Gross and Siegel 1983 ). B. Estimation of serum ALT and AST levels The serum ALT (alanine amino transferase) and AST (aspartate amino transferase) values were determined for twelve healthy birds per treatment using standard kits (Reitman and Frankel, 1957 ) from M/s Coral Clinical Systems Ltd. India. Carcass traits After starvation for 12 h, twelve birds per treatment were weighed and sacrificed following standard practice of throat slitting. Following proper bleeding and defeathering, carcass traits viz ., blood loss, individual organ weights (liver, heart, gizzard, and intestine), carcass cut-up parts, cooking yield, eviscerated yield and dressing percentage were determined. Statistical analysis Data generated from different treatments were analyzed for their statistical significance by the method of using SPSS 20.0 software package. The data obtained as percentage were subjected to arcsine transformation before analysis. Variables having different observations were analyzed following Duncan’s multiple range test. Results Production performance The results obtained for growth for the period 14–21, 21–42, 14-42- and 0-42-day’s growth phase is presented in Table 2 . Body weight gain was significantly (P < 0.01) higher in betaine supplemented groups than control. For overall trial period (0–42 days) also the body weights of supplemented groups were higher than the control. In the present study, significant positive impact (P < 0.001) of betaine was observed on feed intake in all phases, except during 21–42 days of age. Treatment group supplemented with 0.5g betaine consumed (3010.8g) higher feed during 0–42 days period. Supplemental betaine (1.0 and 1.5g) significantly (P < 0.001) improved the FCR (1.88) compared with the rest of the treatments. Table 2 Effect of betaine supplementation on production performance of broiler chickens reared under heat stress. Parameter Phase (Days) Control 0.5g 1.0g 1.5g SEm Significance Body weight gain (g) 14–21 171.6 a 249.7 b 260.2 b 261.1 b 13.191 P < 0.001 21–42 1097.7 1126.4 1097.9 1099.0 6.927 NS 14–42 1269.3 a 1376.1 b 1358.1 b 1360 b 14.967 P < 0.001 0–42 1475.7 a 1582.8 b 1564.8 b 1570 b 14.747 P < 0.001 Feed intake (g) 14–21 287.6 a 415.8 b 432.6 b 434.5 b 21.682 P < 0.01 21–42 2211.1 2256.1 2172.3 2175 16.323 NS 14–42 2498.0. a 2671.9 b 2604.8 b 2610 b 24.364 P < 0.01 0–42 2837.1 a 3010.8 c 2942.7 b 2945 b 23.795 P < 0.001 FCR 14–21 1.67 b 1.66 a 1.66 a 1.66 a 0.004 P < 0.01 21–42 2.01 b 2.00 b 1.98 a 1.98 a 0.005 P < 0.01 14–42 1.97 c 1.94 b 1.92 a 1.91 a 0.007 P < 0.001 0–42 1.92 c 1.90 b 1.88 a 1.88 a 0.006 P < 0.001 Values bearing different superscripts within a row differed significantly (P < 0.05); FCR – Feed Conversion Ratio; SEm – Standard Error mean; NS – Non-Significant Immune response and immune organs weight The results describing cell mediated and humoral immunity of birds are presented in the Table 3 . Significant differences were observed for CMI (foot pad index) and HA titer against SRBC. Betaine supplemented groups exhibited significantly better (P < 0.001) immunity for CMI (0.39 mm) and HA titer (9.38) than control group. The weights of lymphoid organs measured at 28 and 42 days of age are furnished in Table 3 . Immune organs (bursa of Fabricius, spleen and thymus) are important components of the immune system where cells of immune system attain maturity and are stored, if needed. Betaine supplementation significantly (P < 0.001) influenced the lymphoid organs (thymus, spleen, bursa) weight at both 28 and 42 days of age. Table 3 Effect of betaine supplementation on immunity and immune organs weight (g) of broiler chickens reared under heat stress . Parameter Sex Control 0.5g 1.0g 1.5g Mean SEm Prob. CMI (foot web index in mm) Male 0.27 0.37 0.41 0.42 0.35 N 0.020 NS S Female 0.25 0.34 0.38 0.37 0.32 M 0.016 NS I Mean 0.26 a 0.35 b 0.39 c 0.39 c 0.33 0.013 P < 0.001 T HIR (Log 2 HA Titer) Male 7.25 9.00 9.50 9.45 8.58 0.398 NS S Female 7.00 8.50 9.25 9.30 8.25 0.411 NS I Mean 7.13 a 8.75 b 9.38 b 9.37 b 8.42 0.282 P < 0.01 T 28d Lymphoid wt. (% of live wt.) Thymus Male 0.37 0.43 0.44 0.46 0.41 0.019 NS S Female 0.32 0.41 0.44 0.45 0.39 0.023 NS I Mean 0.34 a 0.42 ab 0.44 b 0.44 b 0.40 0.015 P < 0.05 T Bursa of Fabricius Male 0.18 0.21 0.26 0.28 0.22 0.015 NS S Female 0.16 0.20 0.24 0.22 0.20 0.016 NS I Mean 0.17 a 0.21 b 0.25 b 0.25 b 0.21 0.011 P < 0.01 T Spleen Male 0.14 0.19 0.20 0.20 0.18 0.011 NS S Female 0.16 0.16 0.21 0.21 0.18 0.012 NS I Mean 0.15 a 0.18 ab 0.21 b 0.21 b 0.18 0.008 P < 0.05 T 42d Lymphoid wt. (% of live wt.) Thymus Male 0.28 0.30 0.26 0.25 0.28 0.008 NS S Female 0.27 0.30 0.29 0.30 0.29 0.010 NS I Mean 0.27 0.30 0.28 0.28 0.28 0.006 NS T Bursa of Fabricius Male 0.16 0.19 0.31 0.31 0.22 0.024 NS S Female 0.16 0.17 0.30 0.30 0.21 0.024 NS I Mean 0.16 a 0.18 b 0.31 c 0.31 c 0.22 0.016 P < 0.001 T Spleen Male 0.17 0.23 0.21 0.22 0.21 0.011 NS S Female 0.16 0.22 0.19 0.22 0.19 0.013 NS I Mean 0.17 a 0.22 b 0.20 ab 0.22 b 0.20 0.008 P < 0.05 T Values bearing different superscript viz. abc (Treatment), and MN (Sex), differed significantly (P < 0.05); NS = Non significant; mm - millimeter Gut morphology The morphometrics of the villus height and width and their ratios at 21 and 42 days of post-hatch is presented in Fig. 1 (A) and (B). Dietary betaine supplementation resulted in significantly (P < 0.05) higher villus height and better VH:VW ratio, therefore, positively altered intestinal epithelial morphometry, whilst, simultaneous reduction in the intestinal villus height and villus height to crypt depth ratio was observed in the control group. Hematological attributes and serum enzymes Results for blood parameters viz ., hemoglobin, H/L ratio, ALT and AST levels are presented in Table 4 . Dietary betaine supplementation significantly (P < 0.001) altered the blood hematology and serum enzyme profile at 28 and 42 days. Hemoglobin levels were significantly (P < 0.001) higher (14.44 g%) in betaine supplemented groups than control. On the contrary, H/L ratio was significantly (P < 0.001) reduced upon betaine supplemented groups (0.34) than the control group at both 28th and 42nd day of post-hatch. ALT and AST values were significantly (P < 0.001) higher in betaine supplemented birds compare to the control groups, at 28th and 42nd days of age. Table 4 Effect of betaine supplementation on hematology and serum biochemical parameters of broiler chickens reared under heat stress. Parameter Sex Control 0.5g 1g 1.5g Mean SEm Prob. 28 days Hemoglobin (g%) Male 12.73 14.11 14.48 15.50 13.77 0.301 NS S Female 12.47 14.05 14.41 13.39 13.64 0.336 NS I Mean 12.60 a 14.08 b 14.44 b 14.44 b 13.71 0.219 P < 0.001 T H:L ratio Male 0.43 0.35 0.32 0.36 0.37 M 0.016 P < 0.05 S Female 0.44 0.36 0.36 0.35 0.39 N 0.014 NS I Mean 0.43 b 0.36 a 0.34 a 0.35 a 0.38 0.011 P < 0.001 T AST (IU/L) Male 58.87 70.54 78.01 80.25 69.14 M 3.315 P < 0.01 S Female 68.88 75.71 98.50 86.20 81.03 N 4.082 NS I Mean 63.88 a 73.12 b 88.26 c 83.22 c 75.08 2.855 P < 0.001 T ALT (IU/L) Male 1.40 X 2.36 Y 2.74 Z 2.84 Z 2.17 M 0.171 P < 0.001 S Female 1.49 X 2.41 Y 3.01 Z 2.99 Z 2.30 N 0.190 P < 0.01 I Mean 1.44 a 2.38 b 2.88 C 2.91 c 2.23 0.126 P < 0.001 T 42 days Hemoglobin (g%) Male 13.33 14.81 15.25 15.35 14.47 0.396 NS S Female 13.13 14.15 14.63 14.70 13.97 0.289 NS I Mean 13.23 a 14.48 b 14.94 b 15.02 b 14.22 0.245 P < 0.01 T H:L ratio Male 0.42 0.34 0.34 0.34 0.37 0.014 NS S Female 0.42 0.35 0.34 0.34 0.37 0.013 NS I Mean 0.42 b 0.35 a 0.34 a 0.34 a 0.37 0.009 P < 0.001 T AST (IU/L) Male 146.39 158.86 170.53 160.10 158.59 N 4.948 P < 0.001 S Female 46.45 X 97.11 Y 112.86 Y 110.25 85.47 M 9.012 P < 0.01 I Mean 96.42 a 127.99 b 141.69 b 135.18 b 122.03 9.132 P < 0.001 T ALT (IU/L) Male 5.16 X 8.63 Y 9.53 Z 9.55 Z 7.77 N 0.572 P < 0.001 S Female 1.09 X 1.52 Y 1.74 Z 1.70 Z 1.45 M 0.084 P < 0.001 I Mean 3.12 a 5.07 b 5.63 c 5.62 c 4.61 0.717 P < 0.001 T Values bearing different superscript viz. abc (Treatment), MN (Sex), XYZ (Interaction) differed significantly (P < 0.05); NS = Non significant Carcass traits At 42nd day of age, dietary betaine improved the breast muscle yield of heat stressed broiler birds (Table 5 ). Betaine supplemented groups exhibited significantly higher (P < 0.05) breast muscle weight (17.36g) than the other groups. The other carcass parameters were not affected by the betaine supplementation during heat stress. Table 5 Effect of betaine (Bet) supplementation on carcass yield (g) of broiler chickens reared under heat stress. Parameter Control 0.5g 1.0g 1.5g Mean SEm Prob. Breast 14.95 a 15.84 ab 17.36 b 17.36 b 16.05 0.411 P < 0.05 Drumstick 10.52 10.74 10.31 10.41 10.52 0.162 NS Thigh 9.92 10.01 10.11 10.00 10.01 0.134 NS Back 16.23 16.98 16.62 16.60 16.61 0.346 NS Wings 9.06 9.10 8.90 8.92 9.02 0.226 NS Neck 3.95 4.42 5.04 5.12 4.47 0.224 NS Intestine 5.23 5.22 5.22 5.30 5.22 0.126 NS Liver 2.30 2.34 2.20 2.30 2.28 0.084 NS Gizzard 2.57 2.70 2.56 2.66 2.61 0.065 NS Heart 0.50 0.48 0.53 0.53 0.50 0.015 NS Ready to cook yield 71.95 73.78 74.94 75.04 73.56 0.827 NS Eviscerated Yield 66.58 68.26 69.64 69.44 68.16 0.798 NS Blood loss 3.31 3.30 3.32 3.32 3.31 0.147 NS Values bearing different superscript within a row differed significantly (P < 0.05); NS = Non significant; SEm – Standard Error mean Discussion Production performance The amelioration of negative effects of heat stress in broiler chickens could be beneficial in terms of performance, immune response and sustainability of farming activity. Dietary betaine supplementation has been observed to increase the live weight of the broiler chickens (Sakomura et al. 2013 ; Sayed and Downing 2015 ). The growth promoting effect of betaine could be attributed to its ability to equilibrate the water and ion concentration within the cells which is crucial for cell integrity and health of the birds (Saeed et al. 2017 ). Further betaine enhances the growth performance of the broilers by augmenting the stress through up scaling the levels of short-chain fatty acid (SCFA) and the blood electrolytes (Akhavan-Salamat and Ghasemi 2016 ). The thermal stress affects satiety center in brain which suppresses feed intake in stressed birds, which negatively affects intestinal development leading to poor nutrient absorption and growth performance (Farag and Alagawany 2018 ). However, in the present study, the feed consumption was higher in the betaine supplemented groups when compared to the un supplemented control. Studies have indicated significantly higher feed intake in birds upon betaine feeding (1.5 g/kg) as compared to control group (Sakomura et al. 2013 ; Awad et al. 2014 ). Improvement in weight gain and feed intake of broilers might be attributed to methyl donating activity of betaine. Besides this, betaine protects the intestinal epithelia from dehydration and osmotic disorders, to enhance the growth and feed consumption (Chand et al. 2017 ). Betaine supplementation (0.75 g/kg) in diet improves greatly the feed efficiency of the broiler birds (El-Hussein et al. 2007 ). The osmo-protective role of betaine could be a possible underlying cause for improving of FCR in heat stressed Cobb broilers (Sakomura et al. 2013 ). Feeding 1.5 and 2.0% of dietary betaine increases the feed consumption and BWG and reduced the FCR of fast-growing broilers under thermal stress (Chand et al. 2017 ). Osmo-protective property of betaine also aids in protecting the intestinal cell proteins and enzymes from environmental stressors thereby counteracting performance losses compare to the control groups. Immune response The release of nitric oxide from heterophils and macrophages and cytokine regulation plays a crucial role in enhancing immune response in betaine supplemented birds (Klasing et al. 2002 ; Zhang et al. 2016 ) Betaine supplementation improved overall immunocompetence and antibody titer against NDV, respectively upon betaine supplementation during heat stress (Farooqi et al. 2005; Zhan et al. 2006 and Chand et al. 2017 ). Consistent thermal stress reduces the weight of immune organs and subsequently compromises the immunocompetence. However, our findings revealed that the weight of lymphoid organs was significantly greater in betaine supplemented birds (1.0 and 1.5g) than control groups. This reveals that betaine can combat the adverse physiological responses originated from thermal stress. Gut morphology The deterioration effect of heat stress on the intestinal mucosa in broiler chickens has been well documented (Quinteiro-Filho et al. 2010 ; Liu et al. 2019). It was further hypothesized that the mechanism of homeostasis in animals fails when the ambient temperature exceeds its thermoneutral zone, resulting in vasodilation and increased peripheral blood flow as a response to heat stress. As a result, very little blood is supplied to the intestine, causing cellular hypoxia in intestine resulting epithelial shedding, which further leads to a deeper crypts and shorter villi (Liu et al. 2019). The methyl donating ability of betaine could exhibit positive response in heat stressed broilers through proliferation of gut epithelia (Burkholder et al. 2008 ; Wang et al. 2020 ). Besides, its osmo-protective and antioxidant effect, betaine improves the gut environment; and alleviates heat stress and intestinal oxidative damage respectively. Role of betaine in maintaining gut villi integrity and promoting nutrient digestibility and absorption in broilers has been demonstrated (Eklund et al. 2005 ). Overall, betaine favorably impacted the morphometry and functioning of intestine and boosted the overall performance of birds. Hematological attributes and serum enzymes Lesser H/L ratio is indicative of properly functioning immune system and overall welfare of birds. Elevated H/L ratio in control group (0.42 and 0.43) at 28th and 42nd day may be a consequence of hyper-activation of the hypothalamic–pituitary–adrenal (HPA) axis. Under chronic thermal stress, H/L ratio may get elevated owing to hyper secretion of inflammatory cytokines that stimulates the hypothalamic production of corticotrophin releasing hormones, which consequentially declines growth rate and immune-competence (Ghasemi et al. 2014 ). Declined H/L ratio upon dietary inclusion of betaine in broiler chickens has been reported (Nofal et al. 2015; Ghasemi and Nari 2020 ). Carcass traits Dietary betaine linearly influenced semi-eviscerated carcass weight, eviscerated carcass weight, and breast muscle yield in broilers (Shakeri et al. 2018 ; Liu et al. 2019). The findings indicated that supplementary betaine has a great potential in improving the breast muscle yield by stimulating skeletal muscle deposition and water holding. Other carcass traits such as eviscerated carcass yield, ready to cook yield and cut-up parts were not influenced by the supplementation of betaine in heat stress broiler chickens. On the basis of our results, it can be concluded that betaine supplementation at the level of 1 g/kg feed can be incorporated in the basal diet of the broiler birds to ameliorate the negative effects heat stress without compromising the production performance of the broiler birds Declarations Ethics approval The present study was carried out at the ICAR- Central Avian Research Institute, Izatnagar, U.P. India, with due approval from the Institute’s Animal Ethical Committee (Approval No. 452/01/ab/CPCSEA). Betaine, as betaine HCl (feed grade, 96% purity) was obtained from Indian Trading Bureau Private Limited, Kolkata, India. Declaration of interest None. Financial support statement The study is not funded by any other outside agency; it was carried out with help of ICAR-CARI in house facilities. ** Data of this experiment is available. Data will be provided on request Authors contributions: Dr Rokade J.J. and Dr M. Monika executed the experiment in farm and also collected parameters. Vispute M taken care of feed formulation. Dr Nagesh Sonale taken care of data compilation and statistical analysis of it. Dr M. Monika, Dr Rokade J.J and Dr. Wadajkar Prasad contributed in writing of the manuscript. Acknowledgments The authors are greatly thankful to Indian Council of Agriculture Research and Director, ICAR-Central Avian Research Institute, Izatnagar for the provision of facilities to conduct this experiment. References Abudabos, A.M. et al., 2016. The effect of phytogenic feed additives to substitute in-feed antibiotics on growth traits and blood biochemical parameters in broiler chicks challenged with Salmonella typhimurium, Environmental Science and Pollution Research, 23, 24151–24157 Akhavan-Salamat, H., and Ghasemi, H.A., 2016. Alleviation of chronic heat stress in broilers by dietary supplementation of betaine and turmeric rhizome powder: dynamics of performance, leukocyte profile, humoral immunity, and antioxidant status, Tropical Animal Health and Production, 48, 181–188 Alahgholi, M. et al., 2014. Effect of betaine as an osmolyte on broiler chickens exposed to different levels of water salinity, Archives Animal Breeding, 57, 1–12 Awad, A.L., Fahim, H.N., and Ibrahim, A.F., 2014. Effect of dietary betaine supplementation on productive and reproductive performance of domyati ducks under summer conditions, Egyptian Poultry Science Journal, 34, 453–474 Burkholder, K.M. et al., 2008. Influence of Stressors on Normal Intestinal Microbiota, Intestinal Morphology, and Susceptibility to Salmonella Enteritidis Colonization in Broilers, Poultry Science, 87, 1734–1741 Chand, N. et al., 2017. Effect of Betaine Supplementation on the Performance and Immune Response of Heat Stressed Broilers, Pakistan Journal of Zoology, 49. Eklund, M. et al., 2005. Potential nutritional and physiological functions of betaine in livestock, Nutrition Research Reviews, 18, 31–48. El-Hussein. et al., 2007. Response of Broilers Performance to Dietary Betaine and Folic Acid at Different Methionine Levels, International Journal of Poultry Science, 6, 515–523. Farag, M.R., and Alagawany, M., 2018. Physiological alterations of poultry to the high environmental temperature, Journal of Thermal Biology, 76, 101–106. Ghasemi, H.A., and Nari, N., 2020. Effect of supplementary betaine on growth performance, blood biochemical profile, and immune response in heat-stressed broilers fed different dietary protein levels, Journal of Applied Poultry Research, 29, 301–313. Ghasemi, H.A., Kasani, N., and Taherpour, K., 2014. Effects of black cumin seed (Nigella sativa L.), a probiotic, a prebiotic and a synbiotic on growth performance, immune response and blood characteristics of male broilers, Livestock Science, 164, 128–134. Gross, W.B., and Siegel, H.S., 1983. Evaluation of the heterophil/lymphocyte ratio as a measure of stress in chickens, Avian diseases, 27, 972–9. He, S. et al., 2015. Effects of dietary betaine on growth performance, fat deposition and serum lipids in broilers subjected to chronic heat stress, Animal science journal, 86, 897–903. Klasing, K.C. et al., 2002. Dietary Betaine Increases Intraepithelial Lymphocytes in the Duodenum of Coccidia-Infected Chicks and Increases Functional Properties of Phagocytes, The Journal of Nutrition, 132, 2274–2282. Liu, Yuan, Sun, Balasubramanian, Zhao, and An, 2019. Effects of Dietary Betaine on Growth Performance, Digestive Function, Carcass Traits, and Meat Quality in Indigenous, Yellow-Feathered Broilers under Long-Term Heat Stress, Animals, 9, 506. Quinteiro-Filho, W.M. et al., 2010. Heat stress impairs performance parameters, induces intestinal injury, and decreases macrophage activity in broiler chickens, Poultry Science, 89, 1905–1914. Rao, S.V.R. et al., 2011. Effect of Supplementing Betaine on Performance, Carcass Traits and Immune Responses in Broiler Chicken Fed Diets Containing Different Concentrations of Methionine, Asian-Australasian Journal of Animal Sciences, 24, 662–669. Reitman, S., and Frankel, S., 1957. A Colorimetric Method for the Determination of Serum Glutamic Oxalacetic and Glutamic Pyruvic Transaminases, American Journal of Clinical Pathology, 28, 56–63. Saeed, M. et al., 2017. Reconsidering betaine as a natural anti-heat stress agent in poultry industry: a review, Tropical Animal Health and Production, 49, 1329–1338. Sakomura, N. et al., 2013. Effect of dietary betaine supplementation on the performance, carcass yield, and intestinal morphometrics of broilers submitted to heat stress. Brazilian Journal of Poultry Science, 15, 105–112. Sayed, M.A.M., and Downing, J., 2015. Effects of dietary electrolyte balance and addition of electrolyte–betaine supplements in feed or water on performance, acid–base balance and water retention in heat-stressed broilers, British Poultry Science, 56, 195–209. Shakeri, M. et al., 2018. Betaine and Antioxidants Improve Growth Performance, Breast Muscle Development and Ameliorate Thermoregulatory Responses to Cyclic Heat Exposure in Broiler Chickens, Animals, 8, 162 Shakeri, M. et al., 2018. Betaine and Antioxidants Improve Growth Performance, Breast Muscle Development and Ameliorate Thermoregulatory Responses to Cyclic Heat Exposure in Broiler Chickens, Animals, 8(10),162. Van Wettere, W.H.E.J., Herde, P., and Hughes, P.E., 2012. Supplementing sow gestation diets with betaine during summer increases litter size of sows with greater numbers of parities, Animal Reproduction Science, 132, 44–49 Wang, Y. et al., 2020. Metabolite profile based on 1H NMR of broiler chicken breasts affected by wooden breast myodegeneration, Food Chemistry, 310, 125852 Zhan, X.A. et al., 2006. Effects of methionine and betaine supplementation on growth performance, carcase composition and metabolism of lipids in male broilers, British Poultry Science, 47, 576–580 Zhang, M. et al., 2016. Antioxidant Mechanism of Betaine without Free Radical Scavenging Ability, Journal of Agricultural and Food Chemistry, 64, 7921–7930 Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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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-3994204","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":276076262,"identity":"4b56a5b9-c6a9-471e-9f1b-06c23b2fdebf","order_by":0,"name":"Jaydip Rokade","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA60lEQVRIiWNgGAWjYBACxgYGBgkIk/kAkJCQIUELG1sCSAsPUTZBtfAYgCjCWpjbzxje+FBxJ7Ffvufzqxs1FjwM7IePbsDrsJ4cY8sZZ54lzmzj3WadcwzoMJ60tBt4tTTkmEnzth02NjjGu804hw2oRYLHDL+W/jdm0n//HTa2P8bzzDjnHzFaZgBtYWw4LGfAxsP8OLeNKC3Pii17jj2TkziWZsac2yfBw0bIL4b9yRtv/Ki5w8PffPjx55xvdXL87IeP4dfSwAGKjgMgNhs4gtjwKQcBeQb2BzAtzB8IqR4Fo2AUjIKRCQDTjEiSgF5XvgAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0001-8202-5751","institution":"Central Avian Research Institute","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Jaydip","middleName":"","lastName":"Rokade","suffix":""},{"id":276076263,"identity":"45f9bc40-dc8a-4895-9733-626550a563dd","order_by":1,"name":"M. MONIKA","email":"","orcid":"","institution":"Indian Agricultural Research Institute","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"M.","middleName":"","lastName":"MONIKA","suffix":""},{"id":276076264,"identity":"bf85cdb8-55a4-4272-97cb-06ed27a42478","order_by":2,"name":"M. Gopi","email":"","orcid":"","institution":"National Institute of Animal Nutrition and Physiology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"M.","middleName":"","lastName":"Gopi","suffix":""},{"id":276076265,"identity":"91d600df-a022-41f4-a91a-ae2cee08f922","order_by":3,"name":"M. Vispute","email":"","orcid":"","institution":"Central Avian Research Institute","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"M.","middleName":"","lastName":"Vispute","suffix":""},{"id":276076266,"identity":"fa00b5ec-7487-4983-8b42-f9f538b6e6e3","order_by":4,"name":"Nagesh Sonale","email":"","orcid":"","institution":"Central Avian Research Institute","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Nagesh","middleName":"","lastName":"Sonale","suffix":""},{"id":276076267,"identity":"40c8b2ce-a0bc-4ad5-9e4f-203c577b8954","order_by":5,"name":"WADAJKAR PRASAD","email":"","orcid":"","institution":"Central Avian Research Institute","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"WADAJKAR","middleName":"","lastName":"PRASAD","suffix":""}],"badges":[],"createdAt":"2024-02-27 15:00:05","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3994204/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3994204/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":52074162,"identity":"509aebff-00c5-4f71-9c38-1d3bd33e09dc","added_by":"auto","created_at":"2024-03-06 09:03:38","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":185587,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(A) and (B)\u003c/strong\u003e. Effect of betaine supplementation on gut morphometry of broiler chickens reared under heat stress. \u003cstrong\u003eVH – Villus Height; VW- Villus Weight\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-3994204/v1/c176d01ffdfea04ce732421e.png"},{"id":52269409,"identity":"6887773f-396c-49fa-97aa-9cfc1846e7b4","added_by":"auto","created_at":"2024-03-08 13:16:24","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":657548,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3994204/v1/5b5c0077-1213-4426-b093-e6e2ccc4ddb4.pdf"}],"financialInterests":"","formattedTitle":"\u003cp\u003eMitigation of Heat Stress in Broiler Chickens during Hot Summer Seasons through Betaine Hydrochloride Osmoprotectant Intervention\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eHeat stress is a major issue in developing nations with tropical climates, particularly for broiler chickens because of their rapid growth and conventional open-shed housing. The most cost-effective and efficacious method for reducing heat stress for marginal farmers is nutritional management as opposed to housing management or automation. Because of its osmo-protectant, methyl donor, and amino acid sparing effects, the current investigation confirms that supplementing with 1g/kg of betaine can be the best strategy to control heat stress losses. Because of its effectiveness and low cost, our results can assist broiler farmers, industry, and even small-scale farmers.\u003c/p\u003e \u003cp\u003eAmeliorating heat stress is the major challenge to the broiler industry especially in tropical regions. Among livestock, poultry is particularly sensitive to temperature associated environmental challenges especially heat stress. High ambient temperature has deleterious effect on productive performance in poultry which often results in economic losses. Heat stress is a result of disequilibrium between production and loss of body heat, where the former exceeds later. The major effects due to heat stress might be increased osmotic disturbances and elevated production of free radicals causing oxidative injuries at cellular level to ultimately imposing stress. In order to maintain homeostasis, physiological and behavioral responses are exhibited by birds, which compromises the performance, immunity and meat quality (Shakeri et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) Combating heat stress without compromising production performance is a difficult task. Efforts are under way to ameliorate the negative effects of heat stress through dietary interventions to achieve optimum performance. As a by-product of sugar beet processing, betaine is commercially available as a feed additive. Betaine is a trimethyl derivative of the amino acid glycine, a metabolite of plant and animal tissues which may provide a strategic solution for these challenges (Akhavan-Salamat and Ghasemi \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Protection of cells against osmotic inactivation is a principal biological function of betaine in living beings. Plants and bacteria also produce and accumulate betaine to cope with salt imbalances and thermal stress. Natural betaine has two roles in nutrition as protection of cells against osmotic inactivation and as a methyl donation \u003cem\u003evia\u003c/em\u003e. Transmethylation (Alahgholi et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2014\u003c/span\u003e)\u003c/p\u003e \u003cp\u003eAs an osmo-protective agent, betaine accumulates inside the cell without and maintaining normal functions. As an osmolyte agent it protects the intestinal cell proteins and enzymes from high ambient temperature. Betaine keeps cells hydrated and maintains ion balance to improve thermotolerance (Abudabos et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Betaine involves in transmethylation process, wherein methyl groups (\u0026ndash;CH\u003csub\u003e3\u003c/sub\u003e) containing a single carbon atom is donated, to synthesize critical metabolites and regulate gene expression through DNA methylation (Chand et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Transmethylation helps in nucleic acid, protein and choline synthesis. From nutritional aspect, betaine can replace other methyl donors such as methionine up to 15\u0026ndash;20%, total sulphur containing amino acids up to 10% of and almost all of choline. By decreasing the requirement of these essential and costly nutrients, betaine effectively minimizes feed cost. Besides, betaine has also been shown to improve embryonic and post-birth survival rate of piglets (Van Wettere et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2012\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eBetaine also offers several other beneficial effects such as to improve hatchability and chick weight; spare choline and methionine; improve gut health and function; alleviate heat stress; increases carcass and breast meat yield; reduces coccidiosis lesions (additive effect in combo with ionophores); reduce wet litter condition; improve immune response etc. High blood homocysteine level increases risk of chick mortality, betaine significantly counters homocysteine (Sakomura et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; He et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Betaine as a methyl donor provides its labile methyl groups for the synthesis of several metabolically active substances such as creatine and carnitine. Enhanced methionine availability after dietary supplementation of betaine resulting in improved animal performance (Rao et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). To address the complex challenges posed by heat stress in broiler chickens, particularly in tropical regions, a comprehensive understanding of the physiological responses and nutritional interventions is imperative. High ambient temperatures disrupt the delicate balance between heat production and dissipation, leading to adverse effects on poultry performance and economic losses. The intricate interplay of osmotic disturbances and heightened free radical production at the cellular level exacerbates the stress imposed by heat. Efforts to counteract heat stress without compromising production performance are underway, with a focus on dietary interventions. Betaine, derived from sugar beet processing, emerges as a promising solution due to its multifaceted roles as an osmo-protectant, methyl donor, and amino acid sparing agent. Within cells, betaine accumulates, preserving normal functions and safeguarding proteins and enzymes in intestinal cells from the impact of high ambient temperatures. Its involvement in the transmethylation process contributes to the synthesis of critical metabolites and the regulation of gene expression through DNA methylation.\u003c/p\u003e \u003cp\u003eFrom a nutritional standpoint, betaine proves to be a versatile substitute for other costly methyl donors, effectively reducing the requirements for essential nutrients such as methionine, sulphur-containing amino acids, and choline. The economic benefits extend beyond feed cost reduction, as betaine demonstrates positive effects on embryonic and post-birth survival rates in piglets. The study emphasizes the multifaceted benefits of betaine supplementation, including improvements in hatchability, chick weight, gut health, immune response, carcass and breast meat yield, and the reduction of coccidiosis lesions. Additionally, betaine counters the detrimental effects of high blood homocysteine levels, mitigating the risk of chick mortality. As a methyl donor, betaine contributes labile methyl groups for the synthesis of metabolically active substances, further enhancing animal performance.\u003c/p\u003e \u003cp\u003eAgainst this backdrop, the present study evaluates the efficacy of post-hatch betaine supplementation on broiler performances reared under higher environmental temperatures, providing valuable insights for the sustainable management of heat stress in poultry farming.\u003c/p\u003e"},{"header":"Material and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n\u003ch2\u003eBirds, experimental design and diet\u003c/h2\u003e\n\u003cp\u003eOne hundred and ninety numbers of day-old broilers (CARIBRO-VISHAL) were divided into four treatments groups and fed with diets contained 0 (control), 0.5, 1.0, and 1.5g betaine/kg feed. Each group consisted of 48 birds in six replicates (8 birds). The experimental diets (starter and finisher) were formulated as per the broiler\u0026rsquo;s requirement (22 and 19.5% CP, respectively). All the birds were raised under uniform standard managemental conditions of brooding and vaccination. Where the basal diet of Broiler chicken was shown as control in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab1\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eIngredients and nutrient composition of basal diets for broiler chicken\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eIngredients (g/kg)\u003c/div\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003ePre-starter\u003c/div\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eStarter\u003c/div\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eFinisher\u003c/div\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eMaize\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e571\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e579\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e635\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eSoybean meal (46% CP)\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e342\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e330\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e288\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eFish meal (48% CP)\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e30\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e32\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e20\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eVegetable oil\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e24\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e29\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e28\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eLimestone powder\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e9.0\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e9.0\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e8.0\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eDi-calcium phosphate\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e14\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e12\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e12\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eDL-Methionine\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e2.0\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.0\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.0\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eL-lysine HCl\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.0\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e0.0\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e0.0\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e\u003csup\u003e\u0026dagger;\u003c/sup\u003eTrace mineral Premix\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.0\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.0\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.0\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e\u003csup\u003eǂ\u003c/sup\u003eVitamin Premix\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e2.0\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e2.0\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e2.0\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e\u003csup\u003e\u0026para;\u003c/sup\u003eVitamin B complex\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.5\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.5\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.5\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eCholine Chloride\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.0\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.0\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.0\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eSalt\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e3.0\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e3.0\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e3.0\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eToxin binder\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.0\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.0\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.0\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"4\" align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e\u003cspan class=\"Italic\"\u003eNutrient composition\u003c/span\u003e\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eMetabolizable energy (ME) (Kcal/kg)\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e3000\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e3050\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e3100\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eCrude protein, %\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e23.0\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e21.9\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e20.0\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eLysine, %\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.30\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.19\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e0.99\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eMethionine, %\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e0.52\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e0.49\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e0.45\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eCalcium, %\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.00\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.00\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e1.00\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003eAvailable phosphorus, %\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e0.45\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e0.41\u003c/div\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cdiv class=\"SimplePara\"\u003e0.41\u003c/div\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\n\u003ch2\u003eTemperature-humidity index (THI) measurement\u003c/h2\u003e\n\u003cp\u003eThe study was carried out during hot summer season in order to simulate the field conditions of heat stress to broilers. The temperature humidity index (THI) was calculated as per the formula suggested by Zulovich and DeShazer (1990). Ambient temperature and Relative humidity were measured using dry-bulb and wet-bulb temperatures. According to Zulovich and DeShazer, 1990, THI ranges were classified as comfort (THI\u0026thinsp;\u0026lt;\u0026thinsp;70), alert (THI 70\u0026ndash;75), danger (THI 76\u0026ndash;81), and emergency (THI\u0026thinsp;\u0026gt;\u0026thinsp;81) zones, used to assess the potential indices of heat stress for poultry. The Temperature Humidity Index (THI) was recorded throughout the experimental period (42 days) and were in the ranged from 85 to 87.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\n\u003ch2\u003eProduction performance\u003c/h2\u003e\n\u003cp\u003eBody weight (g) was measured in different growth phases (14\u0026ndash;21, 21\u0026ndash;42, 14\u0026ndash;42 and 0\u0026ndash;42 days) during the experimental period to ascertain body weight gain (BWG) during each phase. Weighed quantities of respective diets were offered \u003cem\u003eadlib\u003c/em\u003e on daily basis to all the replicates of each dietary regimen. Replicate wise residues were weighed on weekly basis to ascertain weekly as well as overall feed consumed. From the body weight gain and feed consumption, the feed to gain conversion ratio was derived.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEvaluation of immune response\u003c/strong\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\n\u003ch2\u003eA. Humoral immune response\u003c/h2\u003e\n\u003cp\u003eThe microtiter hemagglutination (HA) procedure was followed to determine antibody titre concentration in chickens against 1% Sheep RBCs (SRBC) (Siegel and Gross 1980). The 1% SRBC suspension was prepared in Alserver\u0026rsquo;s solution and subsequently injected intravenously (0.1 ml) into 12 randomly selected healthy birds per group (one male and one female broilers in each replicate) on 28th day post-hatch. Serum was harvested 5 days post-injection and HA antibody titre was determined against antigen (SRBC). HA titre was expressed in terms of Log 2 value of highest dilution wherein reaction was positive against SRBCs.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\n\u003ch2\u003eB. In-vivo cell mediated immune response (CMIR)\u003c/h2\u003e\n\u003cp\u003eThe CMIR was studied at four weeks of age by intra-dermal injection of 0.1 ml (1mg/ml) of phytohemagglutinin type-P (PHA-P) at 3rd and 4th inter-digital space of the right foot and 0.1 ml of sterile PBS in left foot (control) (Cheng and Lamont, 1988). The foot web index (mm) was calculated as the difference between the swelling in the right and left foot before (R1 and L1) and after (R2 and L2) 24 hours of injection.\u003c/p\u003e\n\u003cp\u003eThe formula for CMIR calculation is (R2-R1) - (L2-L1).\u003c/p\u003e\n\u003cp\u003ewhere,\u003c/p\u003e\n\u003cp\u003eL1 \u0026ndash; fold thickness before injection in left inter-digital space\u003c/p\u003e\n\u003cp\u003eL2 \u0026ndash; fold thickness after PBS injection in left inter-digital space\u003c/p\u003e\n\u003cp\u003eR1 \u0026ndash; fold thickness before injection in right inter-digital space\u003c/p\u003e\n\u003cp\u003eR2 \u0026ndash; fold thickness after PHA-p injection in right inter-digital space.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\n\u003ch2\u003eC. Relative weight of lymphoid organs\u003c/h2\u003e\n\u003cp\u003eWeights of lymphoid organs such as bursa of Fabricius, spleen and thymus (g) were recorded at 28 and 42 days of age and expressed as percentage of pre-slaughter live weight.\u003c/p\u003e\n\u003cdiv id=\"Sec9\" class=\"Section3\"\u003e\n\u003ch2\u003eHistomorphometry of intestinal mucosa\u003c/h2\u003e\n\u003cp\u003eHistomorphometry measure \u003cem\u003eviz\u003c/em\u003e., villus height (VH), villus width (VW), and ratio of villus height to villus width (VH: VW) were measured at 28 and 42 days of age. After identification with help of Meckel's diverticulum at jejuno-ileal junction, a piece (0.5\u0026ndash;1.0 cm) of jejunum was collected from 12 healthy birds per treatment. Collected segments were preserved in 10% formalin solution after carefully removing intestinal contents without damaging the mucosa, until processing for histological slide preparation. Further, slides were stained with hematoxylin and eosin stain following by mounting with coverslip. Mounted slides were examined under microscope under low power (10\u0026times;). The measurement of villus height (VH), villus width (VW) was carried out by using Primo Star 3.0 imaging software, Zeiss, Germany. VH: VW ratio was calculated by formula method.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec10\" class=\"Section3\"\u003e\n\u003ch2\u003eHematological attributes and serum enzymes\u003c/h2\u003e\n\u003cp\u003eBlood samples were collected (~\u0026thinsp;1ml) from the jugular vein using 26-gauge needle in K3- EDTA tubes on 2, 4 and 6 weeks of age to determine hematological attributes. Autoanalyzer (Abacus Junior Vet5, Diatron, USA) was used to determine hemoglobin (Hb) levels.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n\u003ch2\u003eA. Determination of H/L ratio\u003c/h2\u003e\n\u003cp\u003eImmediately after blood collection, thin blood smear was prepared on clean glass slides. After drying, the smears were fixed with methanol and stained with Giemsa stain. After drying, smears were examined under microscope at 100\u0026times; objective with oil immersion. 100 cells, i.e, heterophils and/or lymphocytes counted in each smear to calculate H/L ratio (Gross and Siegel \u003cspan class=\"CitationRef\"\u003e1983\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n\u003ch2\u003eB. Estimation of serum ALT and AST levels\u003c/h2\u003e\n\u003cp\u003eThe serum ALT (alanine amino transferase) and AST (aspartate amino transferase) values were determined for twelve healthy birds per treatment using standard kits (Reitman and Frankel, \u003cspan class=\"CitationRef\"\u003e1957\u003c/span\u003e) from M/s Coral Clinical Systems Ltd. India.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\n\u003ch2\u003eCarcass traits\u003c/h2\u003e\n\u003cp\u003eAfter starvation for 12 h, twelve birds per treatment were weighed and sacrificed following standard practice of throat slitting. Following proper bleeding and defeathering, carcass traits \u003cem\u003eviz\u003c/em\u003e., blood loss, individual organ weights (liver, heart, gizzard, and intestine), carcass cut-up parts, cooking yield, eviscerated yield and dressing percentage were determined.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n\u003ch2\u003eStatistical analysis\u003c/h2\u003e\n\u003cp\u003eData generated from different treatments were analyzed for their statistical significance by the method of using SPSS 20.0 software package. The data obtained as percentage were subjected to arcsine transformation before analysis. Variables having different observations were analyzed following Duncan\u0026rsquo;s multiple range test.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eProduction performance\u003c/h2\u003e \u003cp\u003eThe results obtained for growth for the period 14\u0026ndash;21, 21\u0026ndash;42, 14-42- and 0-42-day\u0026rsquo;s growth phase is presented in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. Body weight gain was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) higher in betaine supplemented groups than control. For overall trial period (0\u0026ndash;42 days) also the body weights of supplemented groups were higher than the control. In the present study, significant positive impact (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) of betaine was observed on feed intake in all phases, except during 21\u0026ndash;42 days of age. Treatment group supplemented with 0.5g betaine consumed (3010.8g) higher feed during 0\u0026ndash;42 days period. Supplemental betaine (1.0 and 1.5g) significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) improved the FCR (1.88) compared with the rest of the treatments.\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\u003e\u003cb\u003eEffect of betaine supplementation on production performance of broiler chickens reared under heat stress.\u003c/b\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eParameter\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePhase (Days)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.5g\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.0g\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.5g\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eSEm\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eSignificance\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003eBody weight gain (g)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14\u0026ndash;21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e171.6\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e249.7\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e260.2\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e261.1\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e13.191\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e21\u0026ndash;42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1097.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1126.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1097.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1099.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e6.927\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14\u0026ndash;42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1269.3\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1376.1\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1358.1\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1360\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e14.967\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0\u0026ndash;42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1475.7\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1582.8\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1564.8\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1570\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e14.747\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003eFeed intake\u003c/p\u003e \u003cp\u003e(g)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14\u0026ndash;21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e287.6\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e415.8\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e432.6\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e434.5\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e21.682\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e21\u0026ndash;42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2211.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2256.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2172.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2175\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e16.323\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14\u0026ndash;42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2498.0.\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2671.9\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2604.8\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2610\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e24.364\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0\u0026ndash;42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2837.1\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3010.8\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2942.7\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2945\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e23.795\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003eFCR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14\u0026ndash;21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.67\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.66\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.66\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.66\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.004\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e21\u0026ndash;42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.01\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.98\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.98\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14\u0026ndash;42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.97\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.94\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.92\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.91\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.007\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0\u0026ndash;42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.92\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.90\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.88\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.88\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.006\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"8\"\u003eValues bearing different superscripts within a row differed significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05); FCR \u0026ndash; Feed Conversion Ratio; SEm \u0026ndash; Standard Error mean; NS \u0026ndash; Non-Significant\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003eImmune response and immune organs weight\u003c/h2\u003e \u003cp\u003eThe results describing cell mediated and humoral immunity of birds are presented in the Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. Significant differences were observed for CMI (foot pad index) and HA titer against SRBC. Betaine supplemented groups exhibited significantly better (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) immunity for CMI (0.39 mm) and HA titer (9.38) than control group. The weights of lymphoid organs measured at 28 and 42 days of age are furnished in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. Immune organs (bursa of Fabricius, spleen and thymus) are important components of the immune system where cells of immune system attain maturity and are stored, if needed. Betaine supplementation significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) influenced the lymphoid organs (thymus, spleen, bursa) weight at both 28 and 42 days of age.\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\u003e\u003cb\u003eEffect of betaine supplementation on immunity and immune organs weight (g) of broiler chickens reared under heat stress\u003c/b\u003e.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eParameter\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSex\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.5g\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.0g\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.5g\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eSEm\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eProb.\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eCMI\u003c/p\u003e \u003cp\u003e(foot web index in mm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.35\u003csup\u003eN\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.020\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eS\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.32\u003csup\u003eM\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.016\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eI\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.26\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.35\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.39\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.39\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.013\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eHIR\u003c/p\u003e \u003cp\u003e(Log\u003csub\u003e2\u003c/sub\u003e HA Titer)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e9.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e8.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.398\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eS\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e9.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e8.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.411\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eI\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7.13\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8.75\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9.38\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e9.37\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e8.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.282\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.01\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"9\" nameend=\"c9\" namest=\"c1\"\u003e \u003cp\u003e28d Lymphoid wt. (% of live wt.)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eThymus\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.019\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eS\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.023\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eI\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.34\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.42\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.44\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.44\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.015\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.05\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eBursa of Fabricius\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.015\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eS\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.016\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eI\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.17\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.21\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.25\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.25\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.011\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.01\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eSpleen\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.011\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eS\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.012\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eI\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.15\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.18\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.21\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.21\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.008\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.05\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"9\" nameend=\"c9\" namest=\"c1\"\u003e \u003cp\u003e42d Lymphoid wt. (% of live wt.)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eThymus\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.008\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eS\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.010\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eI\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.006\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eBursa of Fabricius\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.024\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eS\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.024\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eI\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.16\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.18\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.31\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.31\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.016\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eSpleen\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.011\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eS\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.013\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eI\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.17\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.22\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.20\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.22\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.008\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.05\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"9\"\u003eValues bearing different superscript viz. \u003csup\u003eabc\u003c/sup\u003e(Treatment), and \u003csup\u003eMN\u003c/sup\u003e(Sex), differed significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05); NS\u0026thinsp;=\u0026thinsp;Non significant; mm - millimeter\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eGut morphology\u003c/h2\u003e \u003cp\u003eThe morphometrics of the villus height and width and their ratios at 21 and 42 days of post-hatch is presented in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e (A) and (B). Dietary betaine supplementation resulted in significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher villus height and better VH:VW ratio, therefore, positively altered intestinal epithelial morphometry, whilst, simultaneous reduction in the intestinal villus height and villus height to crypt depth ratio was observed in the control group.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003eHematological attributes and serum enzymes\u003c/h2\u003e \u003cp\u003eResults for blood parameters \u003cem\u003eviz\u003c/em\u003e., hemoglobin, H/L ratio, ALT and AST levels are presented in Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e. Dietary betaine supplementation significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) altered the blood hematology and serum enzyme profile at 28 and 42 days. Hemoglobin levels were significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) higher (14.44 g%) in betaine supplemented groups than control. On the contrary, H/L ratio was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) reduced upon betaine supplemented groups (0.34) than the control group at both 28th and 42nd day of post-hatch. ALT and AST values were significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) higher in betaine supplemented birds compare to the control groups, at 28th and 42nd days of age.\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 betaine supplementation on hematology and serum biochemical parameters of broiler chickens reared under heat stress.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eParameter\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSex\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.5g\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1g\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.5g\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eSEm\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eProb.\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"9\" nameend=\"c9\" namest=\"c1\"\u003e \u003cp\u003e28 days\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eHemoglobin (g%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e12.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e14.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e15.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e13.77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.301\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eS\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e12.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e14.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e13.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e13.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.336\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eI\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e12.60\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.08\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e14.44\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e14.44\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e13.71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.219\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eH:L ratio\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.37\u003csup\u003eM\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.016\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.05\u003csup\u003eS\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.39\u003csup\u003eN\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.014\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eI\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.43\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.36\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.34\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.35\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.011\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eAST (IU/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e58.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e70.54\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e78.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e80.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e69.14\u003csup\u003eM\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.315\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.01\u003csup\u003eS\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e68.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e75.71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e98.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e86.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e81.03\u003csup\u003eN\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e4.082\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eI\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e63.88\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e73.12\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e88.26\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e83.22\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e75.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2.855\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eALT (IU/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.40\u003csup\u003eX\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.36\u003csup\u003eY\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.74\u003csup\u003eZ\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.84\u003csup\u003eZ\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.17\u003csup\u003eM\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.171\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003csup\u003eS\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.49\u003csup\u003eX\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.41\u003csup\u003eY\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.01\u003csup\u003eZ\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.99\u003csup\u003eZ\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.30\u003csup\u003eN\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.190\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.01\u003csup\u003eI\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.44\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.38\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.88\u003csup\u003eC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.91\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.126\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"9\" nameend=\"c9\" namest=\"c1\"\u003e \u003cp\u003e42 days\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eHemoglobin (g%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e13.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.81\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e15.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e15.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e14.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.396\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eS\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e13.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e14.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e14.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e13.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.289\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eI\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e13.23\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.48\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e14.94\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e15.02\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e14.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.245\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.01\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eH:L ratio\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.014\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eS\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.013\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eNS\u003csup\u003eI\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.42\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.35\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.34\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.34\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.009\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eAST (IU/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e146.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e158.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e170.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e160.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e158.59\u003csup\u003eN\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e4.948\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003csup\u003eS\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e46.45\u003csup\u003eX\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e97.11\u003csup\u003eY\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e112.86\u003csup\u003eY\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e110.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e85.47\u003csup\u003eM\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e9.012\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.01\u003csup\u003eI\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e96.42\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e127.99\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e141.69\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e135.18\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e122.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e9.132\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eALT (IU/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.16\u003csup\u003eX\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8.63\u003csup\u003eY\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9.53\u003csup\u003eZ\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e9.55\u003csup\u003eZ\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e7.77\u003csup\u003eN\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.572\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003csup\u003eS\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.09\u003csup\u003eX\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.52\u003csup\u003eY\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.74\u003csup\u003eZ\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.70\u003csup\u003eZ\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.45\u003csup\u003eM\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.084\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003csup\u003eI\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.12\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.07\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.63\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5.62\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e4.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.717\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"9\"\u003eValues bearing different superscript viz.\u003csup\u003eabc\u003c/sup\u003e(Treatment),\u003csup\u003eMN\u003c/sup\u003e(Sex),\u003csup\u003eXYZ\u003c/sup\u003e(Interaction) differed significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05); NS\u0026thinsp;=\u0026thinsp;Non significant\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003eCarcass traits\u003c/h2\u003e \u003cp\u003eAt 42nd day of age, dietary betaine improved the breast muscle yield of heat stressed broiler birds (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). Betaine supplemented groups exhibited significantly higher (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) breast muscle weight (17.36g) than the other groups. The other carcass parameters were not affected by the betaine supplementation during heat stress.\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 betaine (Bet) supplementation on carcass yield (g) of broiler chickens reared under heat stress.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"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 \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eParameter\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.5g\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.0g\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.5g\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eSEm\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eProb.\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBreast\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14.95\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15.84\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e17.36\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e17.36\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e16.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.411\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDrumstick\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10.74\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10.31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e10.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e10.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.162\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eThigh\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9.92\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e10.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e10.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.134\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBack\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e16.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e16.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e16.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e16.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.346\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWings\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e9.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e8.92\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e9.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.226\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNeck\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e4.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.224\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIntestine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e5.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.126\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLiver\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.084\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGizzard\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.065\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHeart\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.015\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eReady to cook yield\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e71.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e73.78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e74.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e75.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e73.56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.827\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEviscerated Yield\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e66.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e68.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e69.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e69.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e68.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.798\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBlood loss\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e3.31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.147\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"8\"\u003eValues bearing different superscript within a row differed significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05); NS\u0026thinsp;=\u0026thinsp;Non significant; SEm \u0026ndash; Standard Error mean\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cdiv id=\"Sec22\" class=\"Section2\"\u003e \u003ch2\u003eProduction performance\u003c/h2\u003e \u003cp\u003eThe amelioration of negative effects of heat stress in broiler chickens could be beneficial in terms of performance, immune response and sustainability of farming activity. Dietary betaine supplementation has been observed to increase the live weight of the broiler chickens (Sakomura et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Sayed and Downing \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). The growth promoting effect of betaine could be attributed to its ability to equilibrate the water and ion concentration within the cells which is crucial for cell integrity and health of the birds (Saeed et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Further betaine enhances the growth performance of the broilers by augmenting the stress through up scaling the levels of short-chain fatty acid (SCFA) and the blood electrolytes (Akhavan-Salamat and Ghasemi \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). The thermal stress affects satiety center in brain which suppresses feed intake in stressed birds, which negatively affects intestinal development leading to poor nutrient absorption and growth performance (Farag and Alagawany \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). However, in the present study, the feed consumption was higher in the betaine supplemented groups when compared to the un supplemented control. Studies have indicated significantly higher feed intake in birds upon betaine feeding (1.5 g/kg) as compared to control group (Sakomura et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Awad et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Improvement in weight gain and feed intake of broilers might be attributed to methyl donating activity of betaine. Besides this, betaine protects the intestinal epithelia from dehydration and osmotic disorders, to enhance the growth and feed consumption (Chand et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eBetaine supplementation (0.75 g/kg) in diet improves greatly the feed efficiency of the broiler birds (El-Hussein et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). The osmo-protective role of betaine could be a possible underlying cause for improving of FCR in heat stressed Cobb broilers (Sakomura et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Feeding 1.5 and 2.0% of dietary betaine increases the feed consumption and BWG and reduced the FCR of fast-growing broilers under thermal stress (Chand et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Osmo-protective property of betaine also aids in protecting the intestinal cell proteins and enzymes from environmental stressors thereby counteracting performance losses compare to the control groups.\u003c/p\u003e \u003cdiv id=\"Sec23\" class=\"Section3\"\u003e \u003ch2\u003eImmune response\u003c/h2\u003e \u003cp\u003eThe release of nitric oxide from heterophils and macrophages and cytokine regulation plays a crucial role in enhancing immune response in betaine supplemented birds (Klasing et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Zhang et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) Betaine supplementation improved overall immunocompetence and antibody titer against NDV, respectively upon betaine supplementation during heat stress (Farooqi et al. 2005; Zhan et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2006\u003c/span\u003e and Chand et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Consistent thermal stress reduces the weight of immune organs and subsequently compromises the immunocompetence. However, our findings revealed that the weight of lymphoid organs was significantly greater in betaine supplemented birds (1.0 and 1.5g) than control groups. This reveals that betaine can combat the adverse physiological responses originated from thermal stress.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec24\" class=\"Section2\"\u003e \u003ch2\u003eGut morphology\u003c/h2\u003e \u003cp\u003eThe deterioration effect of heat stress on the intestinal mucosa in broiler chickens has been well documented (Quinteiro-Filho et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Liu et al. 2019). It was further hypothesized that the mechanism of homeostasis in animals fails when the ambient temperature exceeds its thermoneutral zone, resulting in vasodilation and increased peripheral blood flow as a response to heat stress. As a result, very little blood is supplied to the intestine, causing cellular hypoxia in intestine resulting epithelial shedding, which further leads to a deeper crypts and shorter villi (Liu et al. 2019).\u003c/p\u003e \u003cp\u003eThe methyl donating ability of betaine could exhibit positive response in heat stressed broilers through proliferation of gut epithelia (Burkholder et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Wang et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Besides, its osmo-protective and antioxidant effect, betaine improves the gut environment; and alleviates heat stress and intestinal oxidative damage respectively. Role of betaine in maintaining gut villi integrity and promoting nutrient digestibility and absorption in broilers has been demonstrated (Eklund et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). Overall, betaine favorably impacted the morphometry and functioning of intestine and boosted the overall performance of birds.\u003c/p\u003e \u003cdiv id=\"Sec25\" class=\"Section3\"\u003e \u003ch2\u003eHematological attributes and serum enzymes\u003c/h2\u003e \u003cp\u003eLesser H/L ratio is indicative of properly functioning immune system and overall welfare of birds. Elevated H/L ratio in control group (0.42 and 0.43) at 28th and 42nd day may be a consequence of hyper-activation of the hypothalamic\u0026ndash;pituitary\u0026ndash;adrenal (HPA) axis. Under chronic thermal stress, H/L ratio may get elevated owing to hyper secretion of inflammatory cytokines that stimulates the hypothalamic production of corticotrophin releasing hormones, which consequentially declines growth rate and immune-competence (Ghasemi et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Declined H/L ratio upon dietary inclusion of betaine in broiler chickens has been reported (Nofal et al. 2015; Ghasemi and Nari \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec26\" class=\"Section3\"\u003e \u003ch2\u003eCarcass traits\u003c/h2\u003e \u003cp\u003eDietary betaine linearly influenced semi-eviscerated carcass weight, eviscerated carcass weight, and breast muscle yield in broilers (Shakeri et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Liu et al. 2019). The findings indicated that supplementary betaine has a great potential in improving the breast muscle yield by stimulating skeletal muscle deposition and water holding. Other carcass traits such as eviscerated carcass yield, ready to cook yield and cut-up parts were not influenced by the supplementation of betaine in heat stress broiler chickens.\u003c/p\u003e \u003cp\u003eOn the basis of our results, it can be concluded that betaine supplementation at the level of 1 g/kg feed can be incorporated in the basal diet of the broiler birds to ameliorate the negative effects heat stress without compromising the production performance of the broiler birds\u003c/p\u003e \u003c/div\u003e "},{"header":"Declarations","content":"\u003cdiv id=\"Sec27\" class=\"Section3\"\u003e \u003ch2\u003eEthics approval\u003c/h2\u003e \u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eThe present study was carried out at the ICAR- Central Avian Research Institute, Izatnagar, U.P. India, with due approval from the Institute\u0026rsquo;s Animal Ethical Committee (Approval No. 452/01/ab/CPCSEA). Betaine, as betaine HCl (feed grade, 96% purity) was obtained from Indian Trading Bureau Private Limited, Kolkata, India.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e\u003cp\u003e \u003ch2\u003eDeclaration of interest\u003c/h2\u003e \u003cp\u003eNone.\u003c/p\u003e \u003c/p\u003e\u003cp\u003e \u003ch2\u003eFinancial support statement\u003c/h2\u003e \u003cp\u003eThe study is not funded by any other outside agency; it was carried out with help of ICAR-CARI in house facilities.\u003c/p\u003e \u003cp\u003e** Data of this experiment is available. Data will be provided on request\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eAuthors contributions:\u003c/h2\u003e \u003cp\u003eDr Rokade J.J. and Dr M. Monika executed the experiment in farm and also collected parameters. Vispute M taken care of feed formulation. Dr Nagesh Sonale taken care of data compilation and statistical analysis of it. Dr M. Monika, Dr Rokade J.J and Dr. Wadajkar Prasad contributed in writing of the manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgments\u003c/h2\u003e \u003cp\u003eThe authors are greatly thankful to Indian Council of Agriculture Research and Director, ICAR-Central Avian Research Institute, Izatnagar for the provision of facilities to conduct this experiment.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbudabos, A.M. et al., 2016. The effect of phytogenic feed additives to substitute in-feed antibiotics on growth traits and blood biochemical parameters in broiler chicks challenged with Salmonella typhimurium, Environmental Science and Pollution Research, 23, 24151\u0026ndash;24157\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAkhavan-Salamat, H., and Ghasemi, H.A., 2016. Alleviation of chronic heat stress in broilers by dietary supplementation of betaine and turmeric rhizome powder: dynamics of performance, leukocyte profile, humoral immunity, and antioxidant status, Tropical Animal Health and Production, 48, 181\u0026ndash;188\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAlahgholi, M. et al., 2014. Effect of betaine as an osmolyte on broiler chickens exposed to different levels of water salinity, Archives Animal Breeding, 57, 1\u0026ndash;12\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAwad, A.L., Fahim, H.N., and Ibrahim, A.F., 2014. Effect of dietary betaine supplementation on productive and reproductive performance of domyati ducks under summer conditions, Egyptian Poultry Science Journal, 34, 453\u0026ndash;474\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBurkholder, K.M. et al., 2008. Influence of Stressors on Normal Intestinal Microbiota, Intestinal Morphology, and Susceptibility to Salmonella Enteritidis Colonization in Broilers, Poultry Science, 87, 1734\u0026ndash;1741\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChand, N. et al., 2017. Effect of Betaine Supplementation on the Performance and Immune Response of Heat Stressed Broilers, Pakistan Journal of Zoology, 49.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEklund, M. et al., 2005. Potential nutritional and physiological functions of betaine in livestock, Nutrition Research Reviews, 18, 31\u0026ndash;48.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEl-Hussein. et al., 2007. Response of Broilers Performance to Dietary Betaine and Folic Acid at Different Methionine Levels, International Journal of Poultry Science, 6, 515\u0026ndash;523.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFarag, M.R., and Alagawany, M., 2018. Physiological alterations of poultry to the high environmental temperature, Journal of Thermal Biology, 76, 101\u0026ndash;106.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGhasemi, H.A., and Nari, N., 2020. Effect of supplementary betaine on growth performance, blood biochemical profile, and immune response in heat-stressed broilers fed different dietary protein levels, Journal of Applied Poultry Research, 29, 301\u0026ndash;313.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGhasemi, H.A., Kasani, N., and Taherpour, K., 2014. Effects of black cumin seed (Nigella sativa L.), a probiotic, a prebiotic and a synbiotic on growth performance, immune response and blood characteristics of male broilers, Livestock Science, 164, 128\u0026ndash;134.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGross, W.B., and Siegel, H.S., 1983. Evaluation of the heterophil/lymphocyte ratio as a measure of stress in chickens, Avian diseases, 27, 972\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHe, S. et al., 2015. Effects of dietary betaine on growth performance, fat deposition and serum lipids in broilers subjected to chronic heat stress, Animal science journal, 86, 897\u0026ndash;903.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKlasing, K.C. et al., 2002. Dietary Betaine Increases Intraepithelial Lymphocytes in the Duodenum of Coccidia-Infected Chicks and Increases Functional Properties of Phagocytes, The Journal of Nutrition, 132, 2274\u0026ndash;2282.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiu, Yuan, Sun, Balasubramanian, Zhao, and An, 2019. Effects of Dietary Betaine on Growth Performance, Digestive Function, Carcass Traits, and Meat Quality in Indigenous, Yellow-Feathered Broilers under Long-Term Heat Stress, Animals, 9, 506.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eQuinteiro-Filho, W.M. et al., 2010. Heat stress impairs performance parameters, induces intestinal injury, and decreases macrophage activity in broiler chickens, Poultry Science, 89, 1905\u0026ndash;1914.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRao, S.V.R. et al., 2011. Effect of Supplementing Betaine on Performance, Carcass Traits and Immune Responses in Broiler Chicken Fed Diets Containing Different Concentrations of Methionine, Asian-Australasian Journal of Animal Sciences, 24, 662\u0026ndash;669.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eReitman, S., and Frankel, S., 1957. A Colorimetric Method for the Determination of Serum Glutamic Oxalacetic and Glutamic Pyruvic Transaminases, American Journal of Clinical Pathology, 28, 56\u0026ndash;63.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSaeed, M. et al., 2017. Reconsidering betaine as a natural anti-heat stress agent in poultry industry: a review, Tropical Animal Health and Production, 49, 1329\u0026ndash;1338.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSakomura, N. et al., 2013. Effect of dietary betaine supplementation on the performance, carcass yield, and intestinal morphometrics of broilers submitted to heat stress. Brazilian Journal of Poultry Science, 15, 105\u0026ndash;112.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSayed, M.A.M., and Downing, J., 2015. Effects of dietary electrolyte balance and addition of electrolyte\u0026ndash;betaine supplements in feed or water on performance, acid\u0026ndash;base balance and water retention in heat-stressed broilers, British Poultry Science, 56, 195\u0026ndash;209.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShakeri, M. et al., 2018. Betaine and Antioxidants Improve Growth Performance, Breast Muscle Development and Ameliorate Thermoregulatory Responses to Cyclic Heat Exposure in Broiler Chickens, Animals, 8, 162\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShakeri, M. et al., 2018. Betaine and Antioxidants Improve Growth Performance, Breast Muscle Development and Ameliorate Thermoregulatory Responses to Cyclic Heat Exposure in Broiler Chickens, Animals, 8(10),162.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVan Wettere, W.H.E.J., Herde, P., and Hughes, P.E., 2012. Supplementing sow gestation diets with betaine during summer increases litter size of sows with greater numbers of parities, Animal Reproduction Science, 132, 44\u0026ndash;49\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang, Y. et al., 2020. Metabolite profile based on 1H NMR of broiler chicken breasts affected by wooden breast myodegeneration, Food Chemistry, 310, 125852\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhan, X.A. et al., 2006. Effects of methionine and betaine supplementation on growth performance, carcase composition and metabolism of lipids in male broilers, British Poultry Science, 47, 576\u0026ndash;580\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhang, M. et al., 2016. Antioxidant Mechanism of Betaine without Free Radical Scavenging Ability, Journal of Agricultural and Food Chemistry, 64, 7921\u0026ndash;7930\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Betaine, broilers, growth performance, heat stress, immunity","lastPublishedDoi":"10.21203/rs.3.rs-3994204/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3994204/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eModern poultry genotypes produce more body heat due to their greater metabolic activity and in tropical country; high environment temperature often worsens the situation. Heat stress is a major concern for the poultry industry as it adversely affects growth as well as immune functions. Nutritional interventions play an important role in ameliorating the negative effects of heat stress and will be more economical approach. In order to evaluate the role of osmolyte (betaine) as a heat ameliorating agent in broiler chicken, an experiment was conducted with gradient levels of 0, 0.5, 1.0 and 1.5 g/kg for a growing period of 42 days. Each group comprised 48 birds in six replicates of 8 each. All birds were maintained under standard managemental conditions. Thermo-Humidity Index value ranged 85\u0026ndash;87 throughout the experimental period. Production performance, immunity, gut morphometry, hemato-biochemical and stress indicators were studied. Supplementation of betaine significantly improved production performance as well as breast meat yield (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Supplementation (1 and 1.5g/kg) significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) improved both the humoral and cellular immunity. Betaine supplemented groups also depict significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) rise in hemoglobin and relative immune organ weight. Addition of betaine to basal diet significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) increased the villus length and also lowered H:L ratio, with better ALT and AST values than the control. The dose level of at 1 and 1.5g/kg improve all the parameters but villi structure was more improved in 1g/kg dose level and also it is economical than 1.5g/kg dose level. To conclude present study confirms that, the supplementation of betaine at 1g/kg levels could significantly improve overall growth performance, gut morphology and welfare in heat stressed broiler chickens.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e","manuscriptTitle":"Mitigation of Heat Stress in Broiler Chickens during Hot Summer Seasons through Betaine Hydrochloride Osmoprotectant Intervention","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-03-06 09:03:33","doi":"10.21203/rs.3.rs-3994204/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"c0ae88f3-2752-4bea-a1cf-8d36b1a13f79","owner":[],"postedDate":"March 6th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-03-08T13:08:16+00:00","versionOfRecord":[],"versionCreatedAt":"2024-03-06 09:03:33","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-3994204","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3994204","identity":"rs-3994204","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}