Bhukya Prakash

Development and Validation of a Simple, Sensitive, Second Antibody Format Enzyme Immunoassay (EIA) for LH Determination in Mithun (Bos Frontalis) Plasma

Abstract The objective of this study was to develop and validate a simple and highly sensitive enzyme immunoassay (EIA) for LH determination in mithun plasma on microtitreplates using the biotin‐streptavidin amplification system and the second antibody coating technique. Biotin was coupled to LH and used to bridge between streptavidin‐peroxidase and immobilized antiserum in competitive assay. The EIA was carried out directly in 20 µL mithun plasma. The LH standards ranging from 6.25 pg/well/20 µL to 400 pg/well/20 µL were prepared in hormone free plasma collected from a mithun on day 3 post calving. The sensitivity of EIA procedure was 6.25 pg/well LH, which corresponds to 0.31 ng/mL plasma; the 50 percent relative binding sensitivity was seen at 100 pg/well/20 µL. Plasma volumes for the EIA viz. 10 and 20 µL did not influence the shape of standard curve even though a slight drop in the OD450 was seen with higher plasma volumes. A parallelism test was carried out to compare the endogenous mithun plasma LH with bovine LH standards. It showed good parallelism with the bovine standard curve. For the biological validation of the assay, 3 mithuns were used. These were administered 10 µg i.v., with a synthetic analogue of GnRH (Buserelin‐Acetate, Intervet, India) and blood samples were collected at 15 min intervals using indwelling jugular catheter beginning 1 h prior to GnRH injection till 8 h post injection. In all animals, sharp increases in LH concentrations were recorded post GnRH administration, which confirms the biological validation of the EIA. In conclusion, the EIA developed for LH determination in mithun blood plasma is sufficiently reliable, economical, and sensitive enough to estimate LH in all physiological variations in mithun.

Effect of Supplementing Organic Selenium on Performance, Carcass Traits, Oxidative Parameters and Immune Responses in Commercial Broiler Chickens

An experiment was conducted to determine the effect of supplementing various concentrations (0, 100, 200, 300, or 400 μg/kg diet) of organic Se on growth performance, carcass traits, oxidative stress, and immune responses in commercial broiler chickens reared in open-sided poultry house under tropical climatic conditions. Each diet was fed ad libitum to eight replicates consisting of six birds in each pen from 1 to 42 d of age. Body weight gain and feed efficiency, and relative weight of liver, abdominal fat and ready to cook yields were not affected (p>0.05) by organic Se supplementation to broiler diets. Lipid peroxidation in plasma decreased, while activities of glutathione peroxidase and glutathione reductase in plasma increased (p<0.01) linearly with Se concentration in diet. The ratios between heterophyls and lymphocytes and relative weight of lymphoid organs (bursa, spleen, and thymus), and antibody production to Newcastle disease vaccination were not affected (p>0.05) by Se supplementation to broiler diets. However, the cell-mediated immunity (lymphocyte proliferation ratio) increased (p<0.01) linearly with dietary Se concentration. The results of the present study indicate that the supplementation of Se did not influence body weight and feed efficiency. However, supplementation of Se increased antioxidant status and lymphocyte proliferation in broiler chickens.

Development and Validation of a Sensitive Radioimmunoassay for Progesterone Estimation in Unextracted Mithun (Bos frontalis) Plasma

Abstract The objective of the study was to develop and validate a simple, reliable, and highly sensitive radioimmunoassay (RIA) for progesterone determination in mithun (Bos frontalis) plasma. The RIA was carried out in 20 µL unextracted mithun plasma. The progesterone standards ranging from 2 to 500 pg/20 µL/tube were prepared in charcoal‐treated hormone‐free plasma. The sensitivity of the RIA procedure was 2 pg progesterone/20 µL/tube, which corresponds to 0.1 ng/mL; the 50 percent relative binding sensitivity was seen at 32 pg/20 µL/tube. Plasma volumes for the RIA, viz. 10 and 20 µL, did not influence the shape of standard curve, even though a slight drop in the counts was seen with higher plasma volumes. For the biological validation of the assay, three cyclic, three in early pregnancy, and two in late pregnancy mithuns were used. Blood samples collected at weekly intervals for 42 days, from all animals, were assayed for plasma progesterone. The peak level of progesterone was registered at day 14 (day 21 of sampling) of the estrous cycle and the lowest at estrus; the progesterone concentrations increased and decreased gradually as sampling time advanced, in early and late pregnancy, respectively, which confirm the biological validation of the RIA. The RIA avoids the troublesome and laborious plasma extraction procedures. In conclusion, the RIA developed for progesterone determination in mithun blood plasma is sufficiently reliable, simple, and sensitive enough to estimate progesterone in all physiological variations in mithun.

Effect of dietary inclusion of toasted guar (Cyamopsis tetragonoloba) meal as a source of protein on performance of White Leghorn layers.

Abstract Two experiments were conducted to study the effect of including toasted (120°C/35 min) guar meal (GM, Cyamopsis tetragonoloba) in the diet on performance and egg shell quality of White Leghorn (WL) layers. Totals of 2376 and 2816 layer chickens (Babcock, BV 300) were randomly distributed into 27 and 32 replicates with 88 birds each in Experiments 1 and 2, respectively. Three diets in Experiment 1 (0, 50 and 100 g GM) and 4 diets in Experiment 2 (0, 50, 100 and 150 g GM/kg) were prepared having similar concentrations of energy and protein. Each diet was fed ad libitum to 9 and 8 replicates, respectively, in Experiments 1 (from 53 to 68 weeks) and 2 (35 to 46 weeks of age). Compared to soya bean meal (SBM) GM contained similar concentrations of protein, but was deficient in all essential amino acids except arginine, which was 70% higher than in SBM. Total non-starch polysaccharide (NSP) content in GM (166 g/kg) was lower than that of SBM (179 g/kg). Amongst different NSP fractions, GM contained higher levels of arabans, xylans, mannans and glucans compared to SBM. The galactomannan gum content in GM was 46 g/kg. Egg production (EP), body weight (BW), food intake (FI), food efficiency (FE) and egg quality (shell weight, shell per cent, shell thickness, Haugh unit score, egg density and egg breaking strength) parameters were not affected by incorporating GM up to 100 g/kg diet in Experiment 1. However, egg weight (EW) and egg mass (EM) were reduced significantly in groups fed on 100 g/kg diet. In Experiment 2, EP and FE were not affected by incorporating GM up to 100 g/kg, but were reduced at 150 g/kg diet. FI, EW, BW and egg quality parameters were not affected by incorporating toasted GM up to 150 g/kg diet. Based on the results of both experiments, it is concluded that toasted GM can be included in WL layer diets up to 100 g/kg without affecting EP, FE, EW, EM, Haugh unit score, BW and egg shell quality parameters.

Effect of feeding Lagerstroemia speciosa and conventional fodder based rations on nutrient utilization, ruminal metabolites and body weight gain in mithun (Bos frontalis)

The aim of the present study was to evaluate the effect of feeding green fodder, rice straw and concentrate-based total mixed rations (TMR) on dry matter (DM) intake (DMI), nutrient utilization, rumen fermentation patterns and body weight (BW) gain (BWG) in mithun (Bos frontalis) calves. In a randomized block design, male mithun calves (n = 18, 8-10 months of age, 121 +/- 2 kg BW) were randomly divided into three experimental equal groups (six animals in each group) and fed isonitrogenous TMRs ad libitum for 120 days. The TMR(1) contained 30% Napier grass and 30% rice straw, TMR(2) contained 60% rice straw and TMR(3) contained 30% tree leaves (Lagerstroemia speciosa) and 30% rice straw (DM basis). All the TMRs contained 40% concentrate mixture (DM basis). The results indicated that the BWG, DMI and feed conversion efficiency were significantly (p < 0.01) increased with the inclusion of green fodder in TMRs. The apparent digestibility of DM, crude protein, ether extract, crude fibre and nitrogen free extract were also improved significantly (p < 0.01) with the inclusion of green fodder in TMRs. The higher concentration of total nitrogen and total volatile fatty acid in rumen liquor, but low ruminal pH were evident in animals fed green fodder supplemented TMRs. An increased (p < 0.01) molar proportion of acetic acid was evident in animals fed rice straw-based TMR. In contrast, the molar proportion of propionic and butyric acids were significantly (p < 0.01) higher in animals fed green fodder supplemented TMRs. On the basis of higher DMI and higher daily BWG, it is concluded that Napier grass and L. speciosa tree leaves may be incorporated upto 30% (DM basis) in TMR of growing mithuns for feeding in complete confinement system.

Rumen microbial variation and nutrient utilisation in mithun (Bos frontalis) under different feeding regimes

The aim of the study was to investigate the effect of feeding different diets on fermentation, enzyme activities and microbial population in the rumen fluid of mithun (Bos frontalis). In a randomized block design, 20 male mithun (6-8 months of age, 152 ± 12.6 kg body weight) were randomly divided into four experimental groups (n = 5/group) and fed experimental diets ad libitum for 180 days. The diet R1 contained tree foliages (TF), R2 comprised of 50% concentrate mixture (CM) and 50% TF, R3 contained 50% CM and 50% rice straw, and R4 contained 50% CM, 25% TF and 25% rice straw. Rumen liquor was collected at 0 and 180 days of the experiment for estimation of different ruminal parameters and a digestion trial was conducted at the end of the experiment. Rumen fluid was analysed for pH, ammonia nitrogen (NH3 -N), total-N, ruminal enzymes, short chain fatty acid (SCFA) and microbial profile. The relative quantification of ruminal microbes was carried out with real-time PCR using bacteria as the house keeping gene. The dry matter intake, nutrients digestibility, body weight gain, NH3 -N, total-N, carboxymethyl cellulase, avicelase, xylanase, amylase, protease and molar proportion of butyrate were (p < 0.05) higher in mithun fed R2 , R3 and R4 compared to those fed R1 diet. In contrast, increased (p < 0.05) ruminal pH, molar proportion of acetate and, acetate to propionate ratio was recorded in mithun fed only TF than those fed concentrate supplemented diets. Similarly, an increase (p < 0.05) in the population of Fibrobacter succinogenes, Ruminococcus flavefaciens and total bacteria were evident in mithun fed R2 , R3 and R4 compared to those fed R1 . Therefore, it is concluded that TF 25% and/or rice straw 25% along with CM 50% may be fed to the growing mithun for improved rumen ecology, nutrient utilization and thus better performance under stall fed system.

Effect of feeding higher concentrations of limiting amino acids on performance, slaughter variables and nitrogen retention in broiler chicken fed graded levels of toasted guar (Cyamopsis tetragonoloba) meal

ABSTRACT 1. An experiment was conducted to study the effect of supplementing higher concentrations (100% vs. 110%) of critical amino acids (CAA) on performance (body weight gain – BWG, feed efficiency – FE), slaughter variables and nitrogen retention in broiler chicken (1–6 weeks of age) fed graded levels of toasted guar meal (TGM) as a protein source in diets. 2. The TGM was included at five graded concentrations (0, 50, 100, 150 and 200 g/kg) in iso-caloric and iso-protein diets with either the recommended concentration (100%) of CAA (lysine, total sulphur amino acids, threonine, tryptophan and valine) or at 10% higher (110%) concentration. A metabolism trial of 3-day duration was conducted during 6th week of age to study nitrogen retention. 3. The TGM levels and CAA concentration at 21 or 42 d of age did not influence BWG, FI and FE. BWG was not affected with inclusion of TGM up to 100 g/kg in starter and overall production (1–42 d of age) phases. The FE improved with TGM supplementation during starter phase, while at the end of experiment (42 d), FE was depressed by inclusion of TGM in dose dependant manner. All performance variables improved with increase in concentration of CAA from 100% to 110%. 4. Breast meat weight improved and abdominal fat weight reduced with higher levels of CAA in diet. Retention of nitrogen reduced with increase in level of TGM in broiler diet. Increasing concentrations of CAA in diet improved nitrogen retention. 5. It was concluded that TGM could be incorporated up to 100 g/kg with 100% CAA and up to 150 g/kg with 110% CAA without affecting performance. Increasing CAA concentration (110%) in diets significantly improved BWG and FE (21 and 42 d), breast meat weight and nitrogen retention in broiler chicken.

Effect of supplementing graded concentrations of non-phytate phosphorus on performance, egg quality and bone mineral variables in White Leghorn layers

ABSTRACT 1. An experiment was conducted to determine optimal non-phytate phosphorus (NPP) concentrations for White Leghorn (WL) layers (22–72 weeks) fed diet containing 38 g Ca/kg. 2. Eight diets with graded concentrations (1.5–3.25 g/kg in increments of 0.25 g) of NPP were prepared. Each diet was fed to eight pen replicates containing 88 birds in each. Performance data was evaluated in three different phases (phase I-22–37 weeks, phase II-38–53 weeks and phase III-54–72 weeks). Optimum levels of NPP were determined by fitting a quadratic polynomial (QP) regression model. 3. Egg production (EP) was not affected (P = 0.059) by the concentration of NPP and interaction between NPP and diet phase was non-significant, indicating that the lowest concentration (1.5 g/kg diet) of NPP used in the study was adequate across the three phases. However, EP was influenced by phase (P < 0.001). 4. Optimum concentration of NPP for feed intake (FI) was estimated to be 1.5, 1.71 and 2.40 g/kg diet during phases I, II and III, respectively. FI per egg mass (EM) or feed efficiency (FE) responded quadratically with NPP and also differed significantly between phases. Optimum concentration of NPP for FE during phases I, II and III was 1.5, 2.56 and 2.32 g/kg diet, respectively. 5. Egg weight (EW), EM, shell weight and thickness were not affected by NPP concentration although all of these variables (except shell weight) were influenced by phases. 6. Breaking strength of tibia and Ca contents in tibia ash were not affected by the concentration of NPP, but bone ash and P contents in tibia ash were influenced (P < 0.001) by NPP. Predicted optimal concentrations of NPP for responses for tibia ash at 44 or 72 weeks, tibia ash P at 44 weeks and tibia ash P at 72 weeks were 1.55, 2.63 and 1.5 g/kg diet, respectively. 7. Based on the results, it was concluded that WL layers required 1.5 g, 2.63 g and 2.4 g, respectively/kg diet during phase I, II and III with the calculated daily intake of 137.3, 278.3 and 262 mg NPP/b/d.