Functional transcriptome analysis revealed upregulation of MAPK-SMAD signalling pathways in chronic heat stress in crossbred cattle.

Animal geneticists and breeders have the impending challenge of enhancing the resilience of Indian livestock to heat stress through better selection strategies. Climate change's impact on livestock is more intense in tropical countries like India where dairy cattle crossbreeds are more sensitive to heat stress. The main reason for this study was to find the missing relative changes in transcript levels in thermo-neutral and heat stress conditions in crossbred cattle through whole-transcriptome analysis of RNA-Seq data. Differentially expressed genes (DEGs) identified based on the minimum log twofold change value and false discovery rate 0.05 revealed 468 up-regulated genes and 2273 down-regulated significant genes. Functional annotation and pathway analysis of these significant DEGs were compared based on Gene Ontology (Biological process), Kyoto Encyclopedia of Genes and Genome (KEGG), and Reactome pathways using g: Profiler, ShinyGO v0.76, and iDEP.951 web tools. On finding network visualization, the most over-represented and correlated pathways were neuronal and sensory organ development, calcium signalling pathway, Mitogen-activated protein kinase (MAPK) and Smad signalling pathway, Ras-proximate-1, or Ras-related protein 1 (Rap 1) signalling pathway, apoptosis, and oxidative stress. Similarly, down-regulated genes were most expressed in mRNA processing, immune system, B-cell receptor signalling pathway, Nucleotide oligomerization domain (NOD)-like receptors (NLRs) signalling pathway and nonsense-mediated decay (NMD) pathway. The heat stress-responsive genes identified in this study will facilitate our understanding of the molecular basis for climate resilience and heat tolerance in Indian dairy crossbreeds.

Comparative assessment of the effective population size and linkage disequilibrium of Karan fries composite cattle revealed viable population dynamics.

Objective: Karan Fries (KF), a high-producing composite cattle was developed through crossing indicine Tharparkar cows with taurine bulls (Holstein Friesian, Brown Swiss, and Jersey), to increase the milk yield across India. This composite cattle population must maintain sufficient genetic diversity for long-term development and breed improvement in the coming years. The level of Linkage disequilibrium (LD) measures the influence of population genetic forces on the genomic structure and provides insights into the evolutionary history of populations, while the decay of LD is important in understanding the limits of genome-wide association studies for a population. Effective population size (Ne) which is genomically based on LD accumulated over the course of previous generations, is a valuable tool for evaluation of the genetic diversity and level of inbreeding. The present study was undertaken to understand KF population dynamics through the estimation of Ne and LD for the long-term sustainability of these breeds. Methods: The present study included 96 KF samples genotyped using Illumina HDBovine array to estimate the effective population and examine the LD pattern. The genotype data were also obtained for other crossbreds (Santa Gertrudis, Brangus, and Beefmaster) and Holstein Friesian cattle for comparison purposes. Results: The average LD between SNPs was r2 = 0.13 in the present study. LD decay (r2 = 0.2) was observed at 40 kb inter-marker distance, indicating a panel with 62765 SNPs was sufficient for genomic breeding value estimation in KF cattle. The pedigree-based effective population size (Ne) of KF was determined to be 78, while the Ne estimates obtained using LD-based methods were 52 (SNeP) and 219 (Genetic Optimization for Ne Estimation - GONE), respectively. Conclusion: KF cattle have an effective population size (Ne) exceeding the FAO's minimum recommended level of 50, which was desirable. The study also revealed significant population dynamics of KF cattle and increased our understanding of devising suitable breeding strategies for long-term sustainable development.

Rumen microbial diversity, enteric methane emission and nutrient utilization of crossbred Karan-Fries cattle (Bos taurus) and Murrah buffalo (Bubalus bubalis) consuming varied roughage concentrate ratio.

Dietary mix and host species have both been shown to have a significant impact on rumen microbial diversity, enteric methane emission and animal performance. The goal of this study was to see how the roughage concentrate ratio 70:30 (Low concentrate; LC) vs 40:60 (High concentrate; HC) and the host species crossbred cattle vs buffalo affected rumen microbial diversity, enteric methane emissions and nutrient utilization. Dry matter intake (kg/d) and dry matter percent digestibility were considerably (p < 0.05) higher in the HC ration and buffalo compared to LC ration and crossbred cattle, respectively. Both dietary mix and host species had a substantial (p < 0.05) impact on intake of various nutrients, including organic matter (OM), crude protein (CP), ether extract (EE), neutral detergent fiber (NDF), and acid detergent fiber (ADF). Increased concentrate proportion in the ration improved nitrogen balance, resulting in increased average daily gain and considerably reduced methane (g/d) output (p < 0.05). Furthermore, 16S rRNA genes were sequenced using Oxford Nanopore Technology (ONT) and subsequently annotated using the Centrifuge workflow to uncover ruminal bacterial diversity. Firmicutes was considerably (p < 0.01) greater in the LC diet, whereas, Bacteroidetes was higher in the HC ration. Genus Prevotella dominated all rumen samples, and buffalo fed LC ration had significantly (p < 0.01) higher Oscillospira abundance. At the species level, simple sugar-utilizing bacteria such as Prevotella spp. and Selenomonas ruminantium predominated in the crossbred cattle, but fibrolytic bacteria such as Oscillospira guilliermondii were statistically (p < 0.01) more abundant in the buffalo. Overall, dietary mix and host species have both been shown to have a significant impact on rumen microbial diversity, enteric methane emission and animal performance, however, host species remained a major driving force to change ruminal community composition as compared to roughage concentrate ratio under similar environmental conditions.

Effect of non-genetic factors on linear type traits in Karan fries (Bos taurus × Bos indicus) and Sahiwal (Bos indicus) cows in sub-tropical climatic conditions of India.

The focus of present study was to find out the effect of non-genetic factors on linear type traits in Karan Fries and Sahiwal cows reared at an organized farm of northern India. The present study was conducted on Lactating Karan Fries (N = 123) and Sahiwal (N = 133) cows maintained at Livestock Research Center of ICAR- National Dairy Research Institute, Karnal, India during the period of 2017-2019. Total eight udder morphometric traits and seven teat morphometric traits were measured. The linear model including fixed effects of season, parity and stage of lactation was used for the analysis. In Karan Fries cows, linear type traits were significantly affected by parity and stage of lactation, while in Sahiwal cows linear type traits were significantly affected by season, parity and stage of lactation. Udder depth (UD) and udder circumference (UC) were significantly (p < 0.05) affected by season, parity and stage of lactation in Sahiwal cattle, while in Karan Fries cattle udder length (UL) and shortest distance from rear teat ends to floor (SDR) were significantly (p < 0.01) affected by parity and stage of lactation. The results pertaining to present study indicated that season, parity and stages of lactation were important sources of variation for most of linear type traits. Adjustment of data for these effects is necessary to reduce known differences between animals and to obtain reliable estimates of the traits.

Association of udder type traits with single nucleotide polymorphisms in Sahiwal (Bos indicus) and Karan Fries (Bos taurus × Bos indicus) cattle.

Udder structure of milking cows is not merely vital to display the visual characteristics, but also important for high milk output and low mastitis risk incidence as well. The present study measured different udder type traits in Sahiwal (Bos indicus) and Karan Fries (Bos taurus × Bos indicus) and investigated their association with single nucleotide polymorphisms in Vitamin D Receptor and Protein Tyrosine Phosphatase, Receptor Type, R genes. GG genotype of SNP rs454303072 was found to have wider rear udder, larger udder circumference, longer distance between fore-rear teats and left-right teats in Karan Fries cattle. Whereas, in Sahiwal cattle, AA genotype of this SNP was found to be associated with the higher and wider rear udder, larger udder circumference and wider udder. AA genotype of SNP rs382671389 was found to be associated with longer fore teat in Karan Fries cattle. The TT and CC genotype of SNP rs435289107 was associated with udder type traits in Karan Fries and Sahiwal cattle respectively. These results suggest that BTA 5 harbors genomic regions associated with udder traits in Bos indicus and Bos indicus x Bos taurus cattle.

Udder and teat morphometry in relation to clinical mastitis in dairy cows.

The present study was undertaken to explore the association of occurrence of clinical mastitis with udder and teat morphometry in cattle. The relationship of nine udder and eight teat morphometric traits with the occurrence of clinical mastitis was analysed using a binomial logistic regression model. A significant association of the udder and teat morphometric traits with the occurrence of mastitis was observed both in Karan Fries and Sahiwal cattle. The rear udder height, rear udder width, udder width and fore udder attachment were found to be the important udder morphometric traits associated with the occurrence of mastitis. In both groups, mastitic animals had broader rear udder width, lower rear udder height and loose fore udder attachment with lengthy and thicker teats. The inclusion of udder and teat morphometric traits in the selection and breeding program of Karan Fries and Sahiwal cattle may help in selection of animals for mastitis resistance.

Genetic polymorphism in HSPB6 gene and their association with heat tolerance traits in Indian Karan Fries (Bos taurus x Bos indicus) cattle.

Heat shock proteins (HSPs) act as a chaperone activity ensuring the folding, unfolding, and refolding of denatured proteins, which help in a protective role during thermal stress in dairy cattle. This study aimed to detect genetic variations of the HSPB6 gene and to determine their association with heat tolerance traits in Karan Fries cattle. Five single nucleotide polymorphisms (SNPs) (SNP 1-5) were reported in the Karan Fries cattle, which included three transitions viz. SNP1-g.161G > A, SNP2-g.436G > A, and SNP4-g.2152A > G and two transversions viz. SNP3-g.1743C > G, SNP5-g.2417A > T. The association analysis revealed that the three SNPs loci i.e., SNP1-g.161G > A, SNP2-g.436G > A, and SNP3-g.1743C > G were significantly (p < 0.01) associated with the respiration rate (RR) and rectal temperature (RT) (°C) traits. Furthermore, in the case of heat tolerance coefficient (HTC) trait was found significantly associated (p < 0.01) with SNPs loci i.e., SNP1-g.161G > A, SNP2-g.436G > A, and SNP3-g.1743C > G. The Hap 4 (GACAT) was found to more adaptable than cattle of other haplotypes as reflected by lower values of RR, RT and HTC. This study provides the first association analyses between the SNPs and haplotypes of HSPB6 gene and heat tolerance traits in Karan Fries cattle, which could be used as effective SNP markers in genetic selection for heat tolerance in cattle breeding program.

Molecular characterization of coat color gene in Sahiwal versus Karan Fries bovine.

BACKGROUND: Melanocortin-1-receptor gene (MC1R) plays a significant role in signaling cascade of melanin production. In cattle, the coat colors, such as red and black, are an outcome of eumelanin and pheomelanin pigments, respectively. The coat colors have become critical factors in the animal selection process. This study is therefore aimed at the molecular characterization of reddish-brown coat-colored Sahiwal cattle in comparison to the black and white-colored Karan Fries. RESULTS: The Sequence length of the MC1R gene was 954 base pairs in Sahiwal cattle. The sequences were examined and submitted to GenBank Acc.No. MG373575 to MG373605. Alignment of both (Sahiwal and Karan Fries) protein sequences by applying ClustalO multiple sequence alignment programs revealed 99.8-96.8% sequence similarity within the bovine. MC1R gene phylogenetic studies were analyzed by MEGA X. The gene MC1R tree, protein confines, and hereditary difference of cattle were derived from Ensemble Asia Cow Genome Browser 97. One unique single-nucleotide polymorphism (c.844C>A) (SNP) was distinguished. Single amino acid changes were detected in the seventh transmembrane structural helix region, with SNP at p.281 T>N of MC1R gene in Karan Fries cattle. CONCLUSIONS: In this current research, we first distinguished the genomic sequence of the MC1R gene regions that showed evidence of coat variation between Indian indigenous Sahiwal cattle breed correlated with crossbreed Karan Fries. These variations were found in the Melanocortin 1 receptor coding regions of the diverse SNPs. The conclusions of this research provide new insights into understanding the coat color variation in crossbreed compared to the Indian Sahiwal cattle.

Polymorphism in DNA repair gene BRCA1 associated with clinical mastitis and production traits in indigenous dairy cattle.

The present study was conducted on Karan Fries cows to identify SNPs in breast cancer 1 (BRCA1) gene and their association with inflammation of mammary gland and production traits. Chi square analysis showed significant association of both SNPs i.e. G43737229T (P < 0.01) and G43761121A (P < 0.05) with incidence of clinical mastitis. Logistic modeling showed that GG genotype cows of G43737229T SNP was less susceptible to mastitis in comparison to TT genotype cows as the odds ratio of TT against GG for the non-incidence of mastitis was less i.e. 0.23. Whereas, cows with genotype AA of G43761121A SNP were less prone to clinical mastitis in comparison to GG genotype cows. G-T haplotype and GGTT combined genotype exhibited maximum mastitis incidences. Least squares analysis revealed significant association of G43737229T SNP with total milk yield (P < 0.01), 305 days milk yield (P < 0.01) and lactation length (P < 0.05). The TT and GT genotyped Karan Fries cows were associated with higher milk yields during first and second lactation, as well as with first lactation length. These observations on SNPs, genotypes and combined genotypes of BRCA1 gene and their differential association with the incidence of mastitis and production traits can be utilized as an aid to selection for simultaneous improvement of both antagonistic traits.

Effect of supplementation with rumen-protected choline and green tea extract on production performance of transition Karan Fries cows.

AIM: The main objective of this study was to estimate the effect of supplementation of rumen-protected choline (RPC) and green tea extract (GTE) on production parameters in transition Karan Fries (KF) cows. MATERIALS AND METHODS: The present experiment was carried out on 32 pregnant KF cows. In the control group, cows were fed basal diet. In T1, each cow was fed RPC (55 g/day), in T2 - GTE (3 g/d), and in T3- RPC + GTE (55+3) g/day along with basal diet. The duration of the experiment was 30 days before calving to 60 days after parturition. RESULTS: Feeding of both RPC and GTE significantly increased milk yield (p≤0.01), 4% fat corrected milk yield (p≤0.01), milk fat (p≤0.01), and total solid content (p≤0.05) than control. There was no significant difference (p≥0.05) in milk protein, lactose, and solids not fat (SNF) content among the groups. CONCLUSION: Supplementing RPC and GTE in combination improved milk yield and fat content of the milk without altering protein, lactose content of the milk in transition KF cows.

Effect of betaine supplementation on growth performance, nutrient intake and expression of IGF-1 in Karan Fries heifers during thermal stress.

Heat stress hampers nutrient utilisation and production of animals, and dietary betaine supplementation can mitigate the adverse effects of heat stress on animals and improve their productivity. The present study was conducted to explore the effects of betaine supplementation on the growth performance of eighteen growing Karan Fries (KF) heifers having similar age and body conditions. The experiment was carried out on three groups (n = 6) of KF heifers viz. control, treatment I (betaine supplemented at 25  g/d/animal), and treatment II (betaine supplemented at 50  g/d/animal). The experiment lasted for eight months covering the three major seasons of Indian tropical conditions viz. hot-dry (temperature humidity index, THI = 83), hot-humid (THI = 85) and thermoneutral season (THI = 73). Blood samples were collected at fortnightly intervals and analysed for plasma growth hormone (GH; competitive ELISA) and total insulin-like growth factor 1 (IGF-1; Sandwich ELISA), as well as expression of IGF-I in peripheral blood mononuclear cells (PBMC) using real-time polymerase chain reaction. Betaine supplementation resulted in significant (p < 0.05) increase in dry matter intake, feed conversion efficiency, body weight gain, plasma GH and IGF-1 levels during all seasons. The concentrations of plasma IGF-1 and the mRNA expression of IGF-1 were higher (p < 0.01) in treatment I as compared to other groups during all seasons. Betaine supplementation at 25  g/d/animal was more cost-effective in improving growth performance of heat-stressed heifers as compared to 50  g/d/animal. The study suggests that the betaine protects intestinal integrity, enhances nutrient utilisation during heat stress and improves growth performance of growing heifers.

Detection of Single-Nucleotide Polymorphism in AGPAT6 Gene, Associated with Milk Fat Content, using Tetra-Primer ARMS PCR-Based Assay, in Karan Fries Breeding Bulls.

BACKGROUND: The bovine AGPAT6 gene is one of the potential candidate genes governing milk fat synthesis. OBJECTIVES: Identification of single nucleotide polymorphisms (SNP) in the targeted region of AGPAT6 gene and their effect on expected breeding values (EBV) of first lactation milk production traits viz. fat %, fat yield and 305 days milk yield in Karan Fries (KF) breeding bulls were sought. MATERIALS AND METHODS: A tetra-primer ARMS PCR technique was adapted to genotype an SNP, g.36,175,805C>T located on 5' flanking region of AGPAT6 gene. The relationship between EBV of milk production traits and polymorphic locus of AGPAT6 gene was assessed. RESULTS: Three kinds of genotype (CC, CT, and TT) with respect to g.36,175,805C>T SNP locus were observed. The identified SNP had significant (P<0.05) influence on EBV of fat % (EBV-FP). The KF bulls with CC and CT genotype had comparatively higher EBV-FP than the bulls with the TT genotype. The substitution of "C" allele by "T" allele led to a decrease of 0.0045 % in the EBV-FP. CONCLUSION: The identified SNP was significantly associated with EBV-FP, thus it may be utilized as a molecular marker for developing marker-assisted selection strategy to enhance the milk fat content in KF cattle population.

Molecular characterization and SNP identification in HSPB6 gene in Karan Fries (Bos taurus x Bos indicus) cattle.

Heat shock proteins (HSPs) act as molecular chaperones which are preferentially transcribed in response to severe perturbations of the cellular homeostasis such as heat stress. The present study was undertaken for molecular characterization and detection of genetic polymorphisms of HSPB6 gene in 100 Karan Fries Cattle. HSPB6 gene was mapped on Bos taurus autosome 18 (BTA 18), comprising three exons and two introns. Four sets of primers for HSPB6 gene were designed using Primer3 software (version 0.4.0). For detecting single nucleotide polymorphisms (SNPs), sequence data was analyzed using BioEdit software (version 7.2). Comparative sequence analysis of HSPB6 gene showed five nucleotide polymorphisms, which included three transitions viz. g.161A > G, g.436G > A and g.2152A > G and two transversions viz. g.1743C > G and g.2417A > T compared to B. taurus (NCBI GenBank: AC_000175.1). HSPB6 gene of Karan Fries cattle exhibited a high percentage of nucleotide identity (47.0-100.0%) with the corresponding mammalian homologue. The present study indicated a high degree of genetic variability in the HSPB6 gene in the Karan Fries cattle populations.

Effect of feed supplement and additives on stress mitigation in Karan Fries heifers.

AIM: The objective of this study was to evaluate the effects of protected fat plus yeast, niacin, zinc, and chromium dietary supplementation on the reduction of heat stress in Karan Fries (KF) heifers during hot humid months. MATERIALS AND METHODS: The basal ration for both the control and treatment groups was the same, containing maize as green fodder and concentrate mixture. However, the treatment group was supplemented with protected fat (2.5% of dry matter intake [DMI]), yeast (10 g/animal/day), niacin (6 g/animal/day), zinc (40 mg/kg DMI), and chromium (1.5 mg/kg DMI). RESULT: The overall mean value of afternoon rectal temperature for control and treatment group was 103.17±0.09 and 102.72±0.10°F, respectively, and was significantly (p<0.01) lower in the treatment group. The overall mean value of afternoon respiration rate for control and treatment group was 76.35±0.56 and 73.13±0.58 breaths/min, respectively, and was also significantly (p<0.01) lower in the treatment group. The overall mean value of afternoon pulse rate for control and treatment group was 97.09±0.63 and 94.67±0.67 beats/minute, respectively, and was also significantly (p<0.01) lower in the treatment group. Finally, the mean cortisol concentration for control and treatment group was 3.94±0.05 ng/ml and 3.70±0.06 ng/ml, respectively, and was significantly (p<0.01) lower in the treatment group. CONCLUSION: The present study shows that supplementation with the above feed additives could serve as a heat stress abatement strategy in growing KF heifers during extreme conditions in summer months.

Expression of HSP70 genes in skin of zebu (Tharparkar) and crossbred (Karan Fries) cattle during different seasons under tropical climatic conditions.

Skin is most important environmental interface providing a protective envelope to animals. It's always under the influence of both internal and external stressors. Heat shock proteins (HSP) are highly conserved stress proteins which play crucial roles in environmental stress tolerance and thermal adaptation. Present study was planned to observe the relative mRNA expression of inducible (HSP70.1 and HSP70.2) and constitutive (HSP70.8) HSP in skin of zebu (Tharparkar) and crossbred (Karan Fries) cattle during different seasons. Skin biopsies were collected from rump region of each animal, aseptically during winter, spring and summer season. Quantitative real time polymerase chain reaction was performed to examine the gene expression of constitutive (HSP70.8) and inducible (HSP70.1 and HSP70.2) HSP in skin of both the breeds during different seasons. Present study observed higher expression of both constitutive and inducible HSP genes in both the breeds during summer and winter than spring season, but magnitude of increase was higher during summer than winter. During summer season, expression pattern of HSPs in skin showed breed differences, where constitutive HSP expression was higher in Tharparkar than Karan Fries and that of inducible HSP was higher in Karan Fries than Tharparkar. Hence, present study suggested that HSP may be conveniently used as biomarkers for assessing protective response of skin against heat stress in zebu and crossbred cattle. Variation in expression between breeds is associated with their heat tolerance and thermal adaptability. In summary, skin of zebu cattle (Tharparkar) is more resistant to summer stress than crossbred (Karan Fries), providing greater protection against heat stress during summer season. Superior skin protective mechanism of zebu (Tharparkar) than crossbred (Karan-Fries) cattle against heat stress may contribute to superior adaptability of zebu cattle to tropical climatic conditions than crossbreed.

Expression of heat-shock protein 72 mRNA in relation to heart rate variability of Sahiwal and Karan-Fries in different temperature-humidity indices.

AIM: To investigate the effect of temperature-humidity index (THI) on the expression pattern of heat-shock protein 72 (HSP72) mRNA of Sahiwal and Karan-Fries (KF) cattle in different THIs. MATERIALS AND METHODS: Five different periods of a year were selected based on combinations of Tmax/Tmin, viz., P1: <20°C/<10°C; P2: >20°C/<10°C, P3: <30°C/<15°C; P4: >35°C/<20°C, and P5: >35°C/>20°C. The THI was calculated from the records of temperature and relative humidity in different periods. Heart rate variability (HRV) was calculated from electrocardiogram records in different periods. HSP72 mRNA expression was estimated by reverse transcription polymerase chain reaction. RESULTS: The THI recorded during P1, P2, P3, P4, and P5 were 55.5, 60.3, 70.1, 74.5, and 79.0, respectively. THI in P4 and P5 were stressful to animals. HSP72 mRNA expression increased during cold stress in P1 in Sahiwal and heat stress in P4 and P5 in both Sahiwal and KF. Sahiwal maintained increased HSP72 mRNA expression longer than KF without causing a significant change in HRV. CONCLUSION: Both low THI in winter and high THI in summer increased HSP72 mRNA of Sahiwal and KF without significant change in HRV. Thermotolerance of Sahiwal could be due to the maintenance of higher HSP72 expression longer than KF in prolonged heat stress in summer.

Optimization of dry period in Karan Fries cow.

AIM: The objective of this study was to optimize dry period (DP) length that can maximize the production across adjacent lactations and overall lifetime yield. MATERIALS AND METHODS: Performance records with respect to DP spread over a period of 15-year in Karan Fries (KF) cattle maintained at Livestock Research Centre (National Dairy Research Institute), were collected for the study. Data of 681 KF cows were analyzed by least square technique to examine the effect of non-genetic factors on DP. Season of calving was classified into four seasons: Winter season (December-March), summer season (April-June), rainy season (July-September), and autumn season (October-November); period of calving into five periods: 1998-2000 (1-period), 2001-2003 (II-period), 2004-2006 (III-period), 2007-2009 (IV-period), and 2010-2012 (V-period), and parity into six parities, i.e., 1(st), 2(nd), 3(rd), 4(th), 5(th), and >6(th) parities to see the effect of non-genetic factors on DP. RESULTS: Period of calving, season of calving, and parity did not affect the DP significantly (p<0.05). The overall least square mean of DP was 67.93±2.12 days. For the optimization of DP with regard to milk productivity, analysis was carried out by class interval method. DP was classified into eight classes (<22, 23-45, 46-67, 68-89, 90-111, 112-133, 134-155, and >156 days), and optimum level was obtained at 46-67 days (3(rd) class) with the following respective milk yield (MY) of 305 daily MY (4016.44±43.68 kg), total MY (4704.21±61.51 kg), MY per day of lactation length (13.03±0.13 kg), and MY per day of calving interval (11.68±0.41 kg). CONCLUSION: From the study, it was concluded that this optimal DP length (46-67 days) is suitable for maximizing the production. Hence, one should aim to dry off pregnant cows to achieve a DP of appropriate length to enhance productivity in the next lactation, as very short and very long DP reduces the economic profitability in dairy animals.

Estimation of metabolic heat production and methane emission in Sahiwal and Karan Fries heifers under different feeding regimes.

AIM: The objective of this study was designed to estimate the metabolic heat production and methane emission in Sahiwal and Karan Fries (Holstein-Friesian X Tharparkar) heifers under two different feeding regimes, i.e., feeding regime-1 as per the National Research Council (NRC) (2001) and feeding regime-2 having 15% higher energy (supplementation of molasses) than NRC (2001). MATERIALS AND METHODS: Six (n = 6) healthy heifers of Sahiwal and Karan Fries with 18-24 months of age were selected from Indian Council of Agricultural Research-National Dairy Research Institute, Karnal. An initial 15 days was maintained under feeding regime-1 and feeding regime-2 as adaptation period; actual experiment was conducted from 16(th) day onward for next 15 days. At the end of feeding regimes (on day 15(th) and 16(th)), expired air and volume were collected in Douglas bag for two consecutive days (morning [6:00 am] and evening [4:00 pm]). The fraction of methane and expired air volume were measured by methane analyzer and wet test meter, respectively. The oxygen consumption and carbon dioxide production were measured by iWorx LabScribe2. RESULTS: The heat production (kcal/day) was significantly (p<0.05) higher in feeding regime-2 as compared to feeding regimen-1 in both breeds. The heat production per unit metabolic body weight was numerically higher in feeding regime-1 than feeding regime-2; however, the values were found statistically non-significant (p>0.05). The energy loss as methane (%) from total heat production was significantly (p<0.05) higher in feeding regime-1. The body weight (kg), metabolic body weight (W(0.75)), and basal metabolic rate (kcal/kg(0.75)) were significantly (p<0.05) higher in feeding regime-2 in both breeds. CONCLUSIONS: This study indicates that higher energy diet by supplementing molasses may reduce energy loss as methane and enhance the growth of Sahiwal and Karan Fries heifers.

Estimation of Genetic Parameters for First Lactation Monthly Test-day Milk Yields using Random Regression Test Day Model in Karan Fries Cattle.

A single trait linear mixed random regression test-day model was applied for the first time for analyzing the first lactation monthly test-day milk yield records in Karan Fries cattle. The test-day milk yield data was modeled using a random regression model (RRM) considering different order of Legendre polynomial for the additive genetic effect (4th order) and the permanent environmental effect (5th order). Data pertaining to 1,583 lactation records spread over a period of 30 years were recorded and analyzed in the study. The variance component, heritability and genetic correlations among test-day milk yields were estimated using RRM. RRM heritability estimates of test-day milk yield varied from 0.11 to 0.22 in different test-day records. The estimates of genetic correlations between different test-day milk yields ranged 0.01 (test-day 1 [TD-1] and TD-11) to 0.99 (TD-4 and TD-5). The magnitudes of genetic correlations between test-day milk yields decreased as the interval between test-days increased and adjacent test-day had higher correlations. Additive genetic and permanent environment variances were higher for test-day milk yields at both ends of lactation. The residual variance was observed to be lower than the permanent environment variance for all the test-day milk yields.

Effect of in vitro zinc supplementation on HSPs expression and Interleukin 10 production in heat treated peripheral blood mononuclear cells of transition Sahiwal and Karan Fries cows.

The changing climatic scenario with apprehended rise in global temperature is likely to affect the livestock adversely vis-à-vis production and reproduction. This has prompted more focus in addressing the unfavorable effects of thermal stress in livestock system. Presuming that the trace element zinc is indispensible for cellular antioxidant system and immune function, the present study was designed to investigate the effect of zinc treatment on heat stress alleviation and immune modulation in peripheral blood mononuclear cells (PBMC) of indigenous and crossbred transition cows. Twelve cows, six each of Sahiwal and Karan Fries (KF) in their second parity with confirmed pregnancy were selected for the experiment. The blood samples were collected at -21, 0 and +21 days in relation to expected date of calving. The experiment was carried out in vitro after isolating PBMC from whole blood. The 48h cultured PBMC were subjected to assorted levels of exposures viz. 37°C, 42°C to impose heat stress and 42°C+zinc to alleviate heat stress and modulate immunity. The PBMC viability was 86%, 69% and 78%, respectively. The mRNA expression of heat shock proteins (HSP 40, 70 and 90α) and Interleukin-10 (IL-10) production varied between the two breeds vis-à-vis days and levels of exposure. The mRNA expression of HSP40 and HSP70 was significantly (P<0.05) higher in Karan Fries than the Sahiwal cows. Both the breeds showed maximum expression of HSP on the day of parturition, more so in KF than Sahiwal. There was a significant (P<0.05) difference in the HSP mRNA expression at different levels of exposure. Zinc treatment to heat stressed PBMC caused a significant (P<0.05) down regulation of HSP. For immune status, anti-inflammatory cytokine, IL-10 in the culture supernatant was accessed. The IL-10 was significantly (P<0.05) higher in Karan Fries (168.18±14.09pg/ml) than the Sahiwal cows (147.24±11.82pg/ml). The IL-10 concentration was highest on the day of calving. Zinc treatment reduced the IL-10 concentration. From the study, it could be concluded that the zinc supplementation in heat stressed PBMC can ameliorate thermal stress and modulate immune response which can act as a model for reducing heat stress during the periparturient period in tropical livestock.