Kisspeptin stimulates oestradiol biosynthesis by upregulating steroidogenic transcripts and proliferation markers in the bubaline granulosa cells in vitro.

Kisspeptin (Kp), an upstream regulator of GnRH release, is essential for the development and function of reproductive axis. Previously, we demonstrated the localization of Kp and its receptor (Kiss1r) in the active follicle in the bubaline ovary. Present study aimed to determine the effect of Kp on granulosa cell (GCs) functions, especially oestradiol (E2 ) and progesterone (P4 ) production, and differential expression of genes regulating the proliferation, apoptosis and steroidogenesis in the buffalo. The ovaries with 6-10 mm size follicles obtained from the cyclic buffaloes after slaughtering were used for isolation of GCs for in vitro study. The primary GCs culture was treated with Kp (0, 10, 50 and 100 nM) and incubated for 48 h. Production of E2 and P4 was estimated in the culture supernatant by ELISA. The expression of gonadotropin receptors (FSHR and LHR), steroidogenic genes (STAR, 3ß-HSD, CYP19A1), proliferation marker (PCNA), apoptotic factors (CASP3 and BCL2) and Kp signalling molecule (extracellular signal-regulated kinase 1/2, ERK1/2 and p-ERK1/2) was studied in the GCs by qPCR. Significant E2 production was found in the Kp 50 and 100 nM groups (p < .05), whereas P4 production was reduced in Kp 100 nM group (p < .05). There was concomitant upregulation of FSHR, ERK1/2, STAR and CYP19A1 in the Kp 100 nM treated GCs. In addition, Kp at 100 nM stimulated the proliferation of GCs by upregulating the expression of BCL2 (5.0 fold) and PCNA (94.9 fold). Further, high immunoreactivity of p-ERK1/2 was observed in the Kp-treated GCs. It was concluded that Kp at 100 nM concentration stimulated E2 production by upregulating the steroidogenic pathway through ERK1/2, STAR and CYP19A1 and modulating PCNA and BCL2 expressions in the GCs. Further experiments are warranted using Kp antagonist in different combinations to establish the signalling pathway in Kp-mediated steroidogenesis in the GCs for developing strategies to control ovarian functions.

Genetic analysis of first lactation and lifetime performance traits in composite Vrindavani cattle: important considerations for higher milk production.

The present study was aimed to evaluate the influence of non-genetic factors on several first lactation and lifetime performance traits and elucidate their genetic parameters in an organized Vrindavani cattle population. Data on eight first-lactation and thirteen lifetime traits were collected on 2400 cows with pedigree records that were reared during 33-year period (1989-2021). The first-lactation traits included age at first calving (AFC), total milk yield (FTMY), standard milk yield (FSMY305), peak yield (FPY), lactation length (FLL), dry period (FDP), service period (FSP) and calving interval (FCI). Whereas, the lifetime traits mainly included total lifetime milk yield (TLMY), total standard milk yield (TSMY), number of lactations completed (NL), total lactation length (TLL), herd life (HL), productive life (PL), average milk yield per day of herd life (TLMY/HL), average milk yield per day of productive life (TLMY/PL), average milk yield per day of productive life (TLMY/TLL). Other lifetime production traits included average service period (ASP), average dry period (ADP), average calving interval (ACI) and unproductive days (UD). The heritability estimates of first-lactation traits ranged between 0.026 and 0.228 and were found to be low for AFC (0.180 ± 0.042), FCI (0.191 ± 0.125), FSMY305 (0.145 ± 0.061), FTMY (0.165 ± 0.048), FDP (0.052 ± 0.049) and FSP (0.026 ± 0.033); however, FLL (0.229 ± 0.044) and FPY (0.202 ± 0.046) showed moderate heritability. Positive phenotypic correlation (p < 0.001) was revealed among FTMY, TLMY, TLL, HL and PL. The AFC produced a significant effect (p < 0.05) on several traits i,e, TLL, TLMY/HL, FSMY305, FPY, TLMY, HL and TLMY/PL. Lower AFC was associated with higher TLMY, TLL and TLMY/HL; while FSMY305, FPY, HL and TLMY/PL were higher in heifers that calved late in their life. The results revealed that AFC may be optimized with first lactation and lifetime traits for this population.

Read-depth based approach on whole genome resequencing data reveals important insights into the copy number variation (CNV) map of major global buffalo breeds.

BACKGROUND: Elucidating genome-wide structural variants including copy number variations (CNVs) have gained increased significance in recent times owing to their contribution to genetic diversity and association with important pathophysiological states. The present study aimed to elucidate the high-resolution CNV map of six different global buffalo breeds using whole genome resequencing data at two coverages (10X and 30X). Post-quality control, the sequence reads were aligned to the latest draft release of the Bubaline genome. The genome-wide CNVs were elucidated using a read-depth approach in CNVnator with different bin sizes. Adjacent CNVs were concatenated into copy number variation regions (CNVRs) in different breeds and their genomic coverage was elucidated. RESULTS: Overall, the average size of CNVR was lower at 30X coverage, providing finer details. Most of the CNVRs were either deletion or duplication type while the occurrence of mixed events was lesser in number on a comparative basis in all breeds. The average CNVR size was lower at 30X coverage (0.201 Mb) as compared to 10X (0.013 Mb) with the finest variants in Banni buffaloes. The maximum number of CNVs was observed in Murrah (2627) and Pandharpuri (25,688) at 10X and 30X coverages, respectively. Whereas the minimum number of CNVs were scored in Surti at both coverages (2092 and 17,373). On the other hand, the highest and lowest number of CNVRs were scored in Jaffarabadi (833 and 10,179 events) and Surti (783 and 7553 events) at both coverages. Deletion events overnumbered duplications in all breeds at both coverages. Gene profiling of common overlapped genes and longest CNVRs provided important insights into the evolutionary history of these breeds and indicate the genomic regions under selection in respective breeds. CONCLUSION: The present study is the first of its kind to elucidate the high-resolution CNV map in major buffalo populations using a read-depth approach on whole genome resequencing data. The results revealed important insights into the divergence of major global buffalo breeds along the evolutionary timescale.

Allelic to genome wide perspectives of swine genetic variation to litter size and its component traits.

Litter size is a complex and sex limited trait that depends on various biological, managemental and environmental factors. Owing to its low heritability it is inefficaciously selected by traditional methods. However, due to higher heritability of ovulation rate and embryo survival, selection based on component traits of litter size is advocated. QTL analysis and candidate gene approach are among the various supplementary/alternate strategies for selection of litter size. QTL analysis is aimed at identifying genomic regions affecting trait of interest significantly. Candidate gene approach necessitates identification of genes potentially affecting the trait. There are various genes that significantly affect litter size and its component traits viz. ESR, LEP, BF, IGFBP, RBP4, PRLR, CTNNAL1, WNT10B, TCF12, DAZ, and RNF4. These genes affect litter size in a complex interacting manner. Lately, genome wide association study (GWAS) have been utilized to unveil the genetic and biological background of litter traits, and elucidate the genes governing litter size. Favorable SNPs in these genes have been identified and offers a scope for inclusion in selection programs thereby increasing breeding efficiency and profit in pigs. The review provides a comprehensive coverage of investigations carried out globally to unravel the genetic variation in litter size and its component traits in pigs, both at allelic and genome wide level. It offers a current perspective on different strategies including the profiling of candidate genes, QTLs, and genome wide association studies as an aid to efficient selection for litter size and its component traits.

Genome wide copy number variations using Porcine 60K SNP Beadchip in Landlly pigs.

In the present study, Porcine 60K SNP genotype data from 69 Landlly pigs were used to explore Copy Number Variations (CNVs) across the autosomes. A total of 386 CNVs were identified using Hidden Markov Model (HMM) in PennCNV software, which were subsequently aggregated to 115 CNV regions (CNVRs). Among the total detected CNVRs, 58 gain, 49 were loss type while remaining 8 events were both gain and loss types. Identified CNVRs covered 12.5 Mb (0.55%) of Sus scrofa reference 11.1 genome. Comparison of our results with previous investigations on pigs revealed that approximately 75% CNVRs were novel, which may be due to differences in genetic background, environment and implementation of artificial selection in Landlly pigs. Functional annotation and pathway analysis showed the significant enrichment of 267 well-annotated Sus scrofa genes in CNVRs. These genes were involved in different biological functions like sensory perception, meat quality traits, back fat thickness and immunity. Additionally, KIT and FUT1 were two major genes detected on CNVR in our population. This investigation provided a comprehensive overview of CNV distribution in the Indian porcine genome for the first time, which may be useful for further investigating the association of important quantitative traits in Landlly pigs.Highlights115 CNVRs were identified in 69 Landlly pig population.Approximately 75% detected CNVRs were novel for Landlly population.Significant enrichment of 267 well-annotated Sus scrofa genes observed in these CNVRs.These genes were involved in different biological functions like sensory perception, meat quality traits, back fat thickness and immunity.Comprehensive CNV map in the Indian porcine genome developed for the first time.

Estimation of microsatellite-based autozygosity and its correlation with pedigree inbreeding coefficient in crossbred cattle.

In countries where farming is largely subsistence, no pedigree records of farm animals are maintained at farmers' herd and scientific mating plans are not observed which leads to the accumulation of inbreeding and loss of production potential. Microsatellites have been widely used as reliable molecular markers to measure inbreeding. We attempted to correlate autozygosity estimated from microsatellite data with the inbreeding coefficient (F) calculated from pedigree data in Vrindavani crossbred cattle developed in India. The inbreeding coefficient was calculated from the pedigree of ninety-six Vrindavani cattle. Animals were further grouped into three groups viz. acceptable/low (F: 0-5%), moderate (F: 5-10%) and high (F: ≥10%), based on their inbreeding coefficients. The overall mean of the inbreeding coefficient was found to be 0.070 ± 0.007. A panel of twenty-five bovine-specific loci were chosen for the study according to ISAG/FAO. The mean FIS, FST, and FIT values were 0.0548 ± 0.025, 0.012 ± 0.001 and 0.0417 ± 0.025, respectively. There was no significant correlation between the FIS values obtained and the pedigree F values. The locus-wise individual autozygosity was estimated using the method-of-moments estimator (MME) formula for locus-specific autozygosity. The autozygosities ascribing to CSSM66 and TGLA53 were found to be significantly (p < .01 and p < .05, respectively) correlated with pedigree F values.

Genetic group and swim-up processing affect SYBR Green real-time PCR-based porcine semen sex ratio in Landrace and its crossbred boars.

Determination of factors affecting sex ratio is important while considering application of sex ratio enrichment approach. Present study aimed to design a SYBR Green qPCR-based method for measurement of primary sex ratio and to evaluate different factors (genetic group, sire, spermiogenic cycle and processing layer) affecting boar sperm sex ratio. The qPCR was based on relative copy number analysis of sex chromosome-specific single copy gene fragments with an autosomal gene as reference and was evaluated using DNA dilution series from pigs with numerically normal karyotype. The sex ratio was estimated from genomic DNA samples isolated from boar semen collected from different genetic groups at different time points and different processing layers. The X chromosome frequencies of semen samples revealed significant effect of genetic group. However, significant variation was observed neither within same genetic group nor between ejaculates of different spermatogenic cycles. Among the processing techniques studied, swim-up technique produced a significant X sperm enrichment in comparison to control whereas, Percoll density gradient failed to show any significant difference among layers. The lower layer in swim-up technique was found to contain higher proportion of X sperms. The designed qPCR is found to be an easy, less time-consuming method and does not require high end laboratory facilities or the specialized expertise. The lower layer of swim-up processing has a scope for X sperm enrichment in boar semen with proper validation.

Evaluation of sexed semen-based artificial insemination in Tharparkar cattle under organized farm condition.

Sexed semen facilitates additional female calf production for the expansion of a herd at a faster rate and also curtails the surplus production of unwanted male calves. A study was conducted to evaluate the performance of sexed semen in indigenous Tharparkar cows based on 114 artificial inseminations (AI) performed at natural oestrus using two protocols i.e., single AI (n = 48) and double AI (n = 66). Overall, the first service conception rate (CR) was significantly higher in double (53.0%) than single (33.3%) AI protocol. The odds ratio of conception rate in double AI was 2.26 (χ2 = 4.4, df = 1, p = .04) with respect to single AI. The time that elapsed since the detection of oestrus to insemination was also analysed. In a single AI protocol, the CR was higher (p < .05) at 16 h (54.6%) than insemination at 8 h (27.0%) following the onset of oestrus. Yet, the CR using double AI protocol did not differ (p = .73) significantly when AIs were performed either at 8 h and 24 h (51.9%) or 16 h and 24 h (57.1%) post onset of oestrus. Besides, like the single AI protocol, the parity of the animals also influenced the CR, being higher in heifers (n = 22) than those of parous (n = 92) cows (72.73 vs. 40.43%, χ2 = 7.48, df = 1, p = .006) in the present study. The odds ratio of conception in heifers was 3.93 with respect to parous cows. Overall, the birth of female calf was 91.7%. In conclusion, the present study indicates a future promise of the sexed semen for the production of more female offspring from Tharparkar cattle.

An intelligent model for predicting the dressed weight of pigs using morphometric measurements.

Determining the slaughter weight of pigs is crucial to the profitability of swine production farms. Unfortunately, in developing countries, the basic infrastructure for weight measurement may not always be available, affecting farmers' income. This study presents a machine learning-based approach to determine the dressed weight of pigs using four morphometric dimensions: paunch girth (PG), heart girth (HG), body length and wither height, which can be measured in situ. Different neural network model structures were constructed taking LM, GDX and BR training algorithms, tansigmoid/logsigmoid hidden layer transfer functions and 5-30 hidden layer neurons (HLNs). Results showed that LM training algorithm with logsigmoidal transfer function and 20 HLNs resulted in 99.8% accuracy in determining the pig dressed weight. Further, the number of morphometric parameters as inputs was gradually reduced and it was found that 99% accuracy can still be achieved using just PG and HG, thereby reducing the measurement time.

Bayesian analysis reveals the influence of maternal effect on pre-weaning body weights in Landlly piglets.

The present study was undertaken to estimate the (co)variance components and genetic parameters of body weights recorded in Landlly piglets from birth to weaning at weekly intervals (w0 to w6). The data pertained to body weights of 2462 piglets, born to 91 sires and 159 dams across different generations during a 7-year period from 2014 to 2020. Five animal models (I-V), differentiated by inclusion or exclusion of maternal effects with or without covariance between maternal and direct genetic effects, were fitted on the data using the Bayesian algorithm. The analyses were implemented by Gibbs sampling in the BLUPF90 program and Markov chain Monte Carlo (MCMC) methodology was used to draw samples of posterior distribution pertaining to (co)variance components. Based on deviance information criteria (DIC), model V with inclusion of direct additive genetic, direct maternal genetic and permanent environmental effect of dam as random factors along with covariance between direct additive and maternal effects best fitted the data on pre-weaning traits (w0 to w5). Whereas, model I incorporating only the direct additive genetic effect best fitted the weaning weight (w6) data in Landlly piglets. The posterior mean estimates of direct heritability under the best models for W0 to W6 were 0.13, 0.19, 0.29, 0.13, 0.26, 0.32 and 0.46, respectively. Inclusion of the maternal component helped in better partitioning of variance for different body weights in Landlly piglets. The maternal heritability ranged from 0.06 to 0.14, while the litter heritability ranged from 0.11 to 0.15 for pre-weaning weights (W0 to W5) under the best-fit models. The influence of maternal environment was greater than maternal genetic effect from birth to 4th week of age. The results implied that variations in body weight of Landlly pigs were genetically controlled to moderate levels (especially w2 and w4) with contributions from direct additive and maternal genotype that can be exploited by designing efficient breeding programmes.

Accurate assessment of body weights using morphometric measurements in Landlly pigs.

The aim of the study was to evaluate 16 novel morphometric traits of Landlly piglets at weaning (6 weeks) and post weaning (8 weeks) stage and to predict corresponding body weight from the measurements. A total of 279 Landlly piglets (n = 279, 75% Landrace + 25% Ghurrah crosses) were enrolled in this study. Body length, heart girth, paunch girth, height at wither, height at back, rump width, thigh circumference, neck circumference, and body depth had high correlation coefficients (0.8-0.97) with body weight at both the stages. Stepwise regression showed that body length and heart girth contributed most in prediction of both body weights while height at wither for body weight at 6 weeks and neck circumference for body weight at 8 weeks was the next highest contributing trait. Akaike's information criterion, Bayesian information criteria, adjusted R2, concordance correlation coefficient, bias correction factor, modeling efficiency, and coefficient of model determination were used to determine the most appropriate model for the prediction of body weight. Model containing body length and heart girth was fitted best to data for prediction of body weight at both weaning and post weaning stage with adjusted R2 values of 0.94 and 0.96, respectively. Hence, 2 different models were proposed for accurately predicting body weight in Landlly pigs at 6 and 8 weeks.

Population structure and admixture analysis in Frieswal crossbred cattle of India - a pilot study.

The present study was aimed to assess parameters related to genetic diversity, population structure and admixture in the Frieswal crossbred cattle of India. A total of three datasets were analyzed during this study. Dataset A (n = 80) consisted of data on two purebred populations, i.e., Shorthorn (n = 35) and Brahman (n = 25) and one crossbred strain Santa Gertrudis (n = 20). The dataset B (n = 71) consisted of data on three populations that included Holstein-Friesian (n = 30), Sahiwal (n = 27) and Frieswal (n = 14) cattle. The dataset C included data on all the six breeds under study. Dataset C was used to assess the indices of population structure and genetic diversity of different breeds prior to and after LD pruning. For Frieswal cattle strain, the proportion of polymorphic SNPs and MAF levels was 84.54% and 0.24, respectively. Frieswal strain maintained appreciable genetic diversity with observed heterozygosity measure of 0.414. PCA plots for three datasets depicted effective stratification of different breeds in the respective datasets. The genomic clustering levels of Sahiwal and Holstein-Friesian were found to be 98.45 and 99.89%, respectively, while the admixture of Frieswal was estimated to be about 61.5 and 38.5% from Holstein-Friesian and Sahiwal breeds, respectively.

Distribution analysis of single nucleotide polymorphisms related to feet and legs and their association with lameness in Vrindavani cattle.

Lameness is one of the most serious economic problems in dairy animals. The present study was undertaken to identify the SNPs associated with lameness in Vrindavani, a synthetic crossbred cattle developed in India. A total of 187 cows were screened for lameness at Cattle and Buffalo Farm, Indian Veterinary Research Institute, Izatnagar, India and 16.04% prevalence was noticed. The effect of age, body weight, parity, and peak yield was not significant on lameness score. Heritability estimate of lameness score was low (0.07 ± 0.01). A total of 90 Vrindavani cows were genotyped by PCR-RFLP for 10 SNPs that were reported to have association with feet and legs in cattle. Five SNPs displayed significant departure from Hardy Weinberg equilibrium. Varying levels of heterozygosity, polymorphic information content, and allelic diversity were noticed at different SNPs. Odds ratio and least squares analysis revealed a significant association (P < 0.05) of three SNPs with lameness in the herd. Cows with AG, TT, and GG genotypes respectively at rs41632254, rs41603160, and rs41636945 loci were less prone to lameness. These genetic markers would certainly aid in development of an early detection system for lameness.

Genetic parameters of pre-weaning weights in crossbred piglets using multi-trait animal model.

Variances and genetic parameters (heritability and correlation) were estimated for pre-weaning weights from birth to 6 weeks of age at weekly intervals in crossbred (Landrace X Desi) pigs. Data were analysed using multi-trait animal model incorporating dam's age (covariable), parity of sow, sex of piglet, season of birth and year of birth as fixed effects and direct additive genetic, maternal genetic and permanent litter as random effects. The estimates of direct additive genetic and maternal genetic heritability were 0.18 ± 0.04 and 0.14 ± 0.04; 0.73 ± 0.06 and 0.62 ± 0.06; 0.64 ± 0.06 and 0.37 ± 0.05; 0.42 ± 0.05 and 0.30 ± 0.05; 0.37 ± 0.04 and 0.33 ± 0.05; 0.43 ± 0.04 and 0.34 ± 0.06 and 0.47 ± 0.04 and 0.36 ± 0.05 for birth weight and subsequent weights at weekly intervals up to 6 weeks, respectively. The permanent litter effect ranged from 0.02 ± 0.01 (birth weight) to 0.11 ± 0.04 (weight at first week). The estimates of correlation between direct additive genetic component and maternal genetic component (i.e. ram) were moderate to high but all negative. ram ranged from - 0.79 ± 0.15 (weight at first week) to - 0.49 ± 0.26 (birth weight). The direct additive genetic and maternal genetic correlations between birth weight and weaning weight were 0.59 ± 0.04 and 0.62 ± 0.13, respectively, which revealed sufficient scope for selection based on birth weight itself. Early weights were found to be highly heritable with due influence of maternal and permanent litter effects, and thus, appropriate selection interventions may be aimed at this stage for favourable changes at subsequent ages.

Livestock and poultry farm wastewater treatment and its valorization for generating value-added products: Recent updates and way forward.

Livestock and poultry wastewater poses a potent risk for environmental pollution accelerating disease load and premature deaths. It is characterized by high chemical oxygen demand, biological oxygen demand, suspended solids, heavy metals, pathogens, and antibiotics, among other contaminants. These contaminants have a negative impact on the quality of soil, groundwater, and air, and is a potential hazard to human health. Depending on the specific characteristics of wastewater, such as the type and concentration of pollutants present; several physical, chemical and biological strategies have been developed for its treatment. This review aims at providing comprehensive overview of the profiling of livestock wastewater from the dairy, swine and poultry sub-sectors along with the biological, physico-chemical, AI-based and integrated treatment methodologies, and valorisation for the generation of value-added products such as bioplastics, biofertilizers, biohydrogen and microalgal-microbial fuel cells. Additionally, future perspectives for efficient and sustainable wastewater treatment are contemplated.

Early-Life Intervention of Lactoferrin and Probiotic in Suckling Piglets: Effects on Immunoglobulins, Intestinal Integrity, and Neonatal Mortality.

The aim of this study was to determine the effects of early-life bovine lactoferrin and host specific probiotic interventions on growth performance, mortality, and concentrations of immunoglobulin A and immunoglobulin G and transforming growth factor beta 1 (a marker of intestinal integrity) in serum of neonatal piglets. A total of eight piglet litters from parity matched sows were randomly divided into four groups and assigned to one of the four interventions: control (sterile normal saline), bovine lactoferrin (100 mg bovine lactoferrin), probiotic (1 × 109 colony forming unit (cfu) of swine origin Pediococcus acidilactici FT28 probiotic), and bovine lactoferrin + probiotic (100 mg bovine lactoferrin and 1 × 109 CFU of P. acidilactici FT28 probiotic). All the interventions were given once daily through oral route for first 7 days of life. The average daily gain (p = 0.0004) and weaning weight (p < 0.0001) were significantly improved in the probiotic group. The piglet survivability was significantly higher in bovine lactoferrin and probiotic groups than control group in Log-rank (Mantel-Cox) test. The concentrations of immunoglobulin A on day 21 in bovine lactoferrin, probiotic, and bovine lactoferrin + probiotic groups increased significantly (p < 0.05). Immunoglobulin G concentrations on day 7 and 15 in bovine lactoferrin and bovine lactoferrin + probiotic groups and on day 15 in probiotic group were significantly (p < 0.05) elevated, whereas, the concentration of transforming growth factor-ß1 was significantly (p < 0.05) increased from day 7 to 21 in all the supplemented groups. In conclusion, the early-life bovine lactoferrin and P. acidilactici FT28 probiotic interventions reduced the mortality in the suckling piglets by promoting the systemic immunity and enhancing the intestinal integrity.

Effect of physical form of the therapeutic diet on the behaviour of crossbred calves experimentally infected with foot-and-mouth disease virus.

The oral lesions associated with foot-and-mouth disease (FMD) negatively affect animal behavior, which can adversely impact animal production and welfare. Physical form of a therapeutic diet (TD) can improve the feed intake and wellbeing during the acute phase of FMD. Accordingly, we tested the effect of two physical forms of a previously developed TD on the behavior of calves experimentally infected with FMD virus (FMDV). Crossbred Holstein Friesian male calves of 10-12 months (n = 12) were experimentally infected with a virulent strain of FMDV and were offered a TD enriched with 19% CP and 2.9 Mcal ME/kg for 11 days post-FMDV infection. One group received the TD in mash form (TDM) while the other (n = 6/group) received it in cooked form (TDC). A group of four calves served as uninfected control and were fed TDM. The time spent by the calves on certain behaviours was recorded in a pre-set form from 06:00-18:00 h for 10 days from day 2-11 post-FMDV infection. The data was divided into two sessions. Session 1 (06:00-13:00 h) represented after the offering of TD, while session 2 (13:01-18:00 h) represented the data after offering green fodder. Based on exploratory data analysis, data recorded from day 2-7 post-FMDV infection was included in the final analysis. Linear mixed model was used by fitting treatment, day and their interaction as fixed effects while calf as random effect. Orthogonal contrast was applied by comparing the infected TDM with other two groups. The results revealed that the cooked form of TD improved the ingestion time, resting time, sleeping time and licking time from day 2-7 post-FMDV infection as compared with the infected TDM group. Ingestive behaviour was better in the infected TDC than that of TDM group (p < 0.01). The sleeping time was significantly high in the infected groups as compared to the uninfected TDM group (p < 0.01) till day 6 post-FMDV infection. Daily activities such as licking, standing and resting differed significantly between the infected TDM and TDC groups in session 1, but not in session 2. Urination and defecation did not differ significantly between the infected TDM and TDC groups. It was concluded that cooked form of TD remediated the effects of infection with FMDV as evidenced by improvement in the behaviour of the calves.

Hematological and serum biochemical parameters and profiling of cytokine genes in lumpy skin disease in Vrindavani cattle.

Lumpy skin disease (LSD) is a notifiable re-emerging transboundary viral disease of bovines that inflicts heavy losses in affected livestock farms. Genetic variations contribute substantially to the inter-individual differences in the immune response against disease agents. The present study aimed to evaluate the genetic basis of differential immune response in Vrindavani cattle by comparing the hematological, biochemical and cytokine genes' expression profiles of LSD-affected and unaffected animals. After 21 days of the outbreak at the farm, the animals were grouped as affected (those who developed symptoms) and unaffected/healthy (those who did not). Standard hematological and biochemical parameters were evaluated in both the groups. Expression profiling of important Th1 (IL2, INFG and GMCSF) and Th2 (IL4, IL6 and IL10) cytokines was also performed via a relative quantification approach using real-time PCR. Erythrogram and leucogram analyses revealed significant differences in total leucocyte count (TLC: 14.18 ± 0.74 versus 11.38 ± 0.68 x103/µL), hemoglobin (Hb: 8.66 ± 0.42 versus 10.84 ± 0.17 g%) and percentage of neutrophils (46.40 ± 1.98 versus 35.40 ± 2.11%), lymphocytes (49.40 ± 1.99 versus 62.40 ± 1.86) and monocytes (4.20 ± 0.37 versus 2.40 ± 0.40) between the affected and healthy animals, respectively. The production of liver enzymes (SGOT and SGPT) was significantly higher in affected animals (74.18 ± 4.76 and 59.51 ± 2.75) when compared to the healthy counterparts (65.95 ± 9.18 and 39.21 ± 3.31). The expression profiling of Th1 and Th2 cytokines revealed significant differences between the two groups, except IL10. The expression of IL2, GMCSF and IL6 were upregulated in healthy animals while that of INFG, IL4 and IL10 were upregulated in LSD-affected animals. The highest abundance was observed for IL2 transcripts in healthy animals among all assessed cytokines with log2fold change of 1.61 as compared to affected counterparts. Overall, the immune response in healthy animals (after exposure to LSD virus) was predominated by the expression of Th1 cell proliferation and there was an increased production of pro-inflammatory cytokines as compared to the affected counterparts. The results revealed that the effective immune response to LSD in cattle consists of changes in hematological and biochemical parameters and altered expression profile of cytokines with enhanced phagocytosis and lymphocyte recruitment. Furthermore, optimal expression of Th1 cytokines is required for maintaining optimal health against infectious insult with LSD virus in cattle.

Efficacy of Microencapsulated Probiotic as Adjunct Therapy on Resolution of Diarrhea, Copper-Zinc Homeostasis, Immunoglobulins, and Inflammatory Markers in Serum of Spontaneous Rotavirus-Infected Diarrhoetic Calves.

The objective of this study was to assess the efficacy of a microencapsulated probiotic as an adjunct therapy in rotavirus-positive diarrhea of neonatal calves that received supportive treatment or supportive along with microencapsulated probiotic treatment, for 5 days. We examined whether microencapsulated Lactobacillus acidophilus NCDC15 probiotic treatment in rotavirus-infected diarrhoetic calves led to faster resolution of diarrhea, amelioration of zinc-copper imbalance, improved the immunoglobulin A and immunoglobulin G, and decreased the inflammatory markers in serum. Calves with rotavirus-positive diarrhea < 4-week age and fecal scores ≥ 2 were randomly assigned into two groups. The supportive along with microencapsulated probiotic treatment significantly (p < 0.05) increased zinc and immunoglobulin A concentrations and decreased copper, tumor necrosis factor-α, and nitric oxide level in serum on days 3 and 5 from pretreatment values; the immunoglobulin G concentration was elevated (p < 0.05) on day 5. The mean resolution time of abnormal fecal score was 5.3 and 3.3 days in supportive treatment and supportive along with microencapsulated probiotic groups, respectively, in log-rank Mantel-Cox test. The calves in the supportive along with microencapsulated probiotic treatment group had faster resolution of diarrhea than supportive treatment group in Dunn's multiple comparisons test. This study demonstrates that supportive treatment along with microencapsulated probiotic administered to naturally rotavirus-infected diarrhoetic calves at onset of diarrhea led to faster resolution of diarrhea, improved zinc and immunoglobulin levels, and decreased the inflammatory parameters in serum of rotavirus-infected diarrhoetic calves.