Genetic studies of heat stress regulation in goat during hot climatic condition.

Various direct and indirect environmental constraints have an impact on livestock performance. The physiological parameters, such as rectal temperature, heart rate, and respiratory rate, are the primary indicators of thermal stress. Under a stressed environment temperature humidity index (THI) had established as a vital measurement to identify the thermal stress in livestock. THI in association with climatic variations can define the environmental effect as stressful or comfortable for livestock. Goats are small ruminants that adapt to a wide range of ecological variations due to their anatomical and physiological characteristics. However, the productivity of animals declines at the individual level during thermal stress. Stress tolerance can be determined through genetic studies associated with at the cellular level using physiological as well as molecular approaches. Information on genetic association with thermal stress in goats is scanty, this severely affects their survival and hence productivity of livestock. The ever-increasing demand for food across the globe needs deciphering novel molecular markers as well as stress indicators that play a vital role in livestock improvement. This review represents an analysis of current knowledge of phenotypic differences during thermal stress and signifies the importance of physiological responses and their association at the cellular level in goats. The regulation of vital genes associated with thermal stress such as Aquaporins (AQP 0, 1, 2, 4, 5, 6, 8), aquaglyceroporins (AQP3, 7, 9, and 10) and super-aquaporins (AQP 11, 12); BAX inhibitors such as PERK (PKR like ER kinase), IRE 1(inositol-requiring-1); Redox regulating genes such as NOX; Transport of Na+ and K+ such as ATPase (ATP1A1) and several heat shock proteins have been implicated in heat-stress related adaptations have been elucidated. As these changes have a significant impact on production performance as well as on livestock productivity. Such efforts may help in the development of molecular markers and will assist the breeders to develop heat-tolerant goats with improved productivity.

Devising a selection strategy for the Jamunapari goat for improving lactation performance.

This study aimed to evaluate the genetic potential of the Jamunapari goat and formulate a selection strategy for improving lactation traits. The data set included 4049 phenotypic records for across parity milk yield at 90 days (MY90), 140 days (MY140), total milk yield (TMY), and lactation length (LL) obtained from the progeny of 83 sires and 1643 dams between the period 1990 and 2019. Animal model employing average information restricted maximum likelihood (AIREML) was used to estimate genetic parameters for milk yield traits and LL. The direct additive heritability estimates for across parity lactation traits that used repeatability model were 0.10 ± 0.03, 0.08 ± 0.03, and 0.12 ± 0.02 for MY90, MY140, and TMY, respectively, while it was low for LL (0.06 ± 0.02). The repeatability estimates were moderate ranging from 0.17 to 0.22 for milk yield traits and LL, indicating persistent performance over the parities. Animal permanent environment influence (c2) was significant in milk yield attributes, whereas direct maternal genetic effects were absent. As the early selection criteria based on first parity records are essential, we analyzed the data for the first parity separately and obtained moderate h2 estimates, viz., 0.26 ± 0.05, 0.16 ± 0.06, and 0.25 ± 0.06 for MY90, MY140, and TMY, respectively. These estimates augur further scope of selection in Jamunapari goats for higher milk yield. High and positive genetic correlation of MY90 with MY140 (0.97 ± 0.01) and TMY (0.91 ± 0.05) revealed the scope of using MY90 as the selection criterion. Based on these results, we recommend use of first parity MY90 as a single trait selection criterion for genetic improvement of all lactation traits in Jamunapari goat.

Molecular expression and identification of caprine estrogen receptor gene 1 for fertility status in bucks.

Estrogen and its receptors are essential for sexual development and reproduction. Oestrogen receptor alpha (ERα) is a nuclear receptor activated by the hormone oestrogen. In male, ERα is encoded by the gene ESR1 (oestrogen receptor1) responsible for better fertility. ESR1 is involved in the reabsorption of luminal fluid during the transit of spermatozoa from the testis to the head of the epididymis which is important for their survival and maturation during epididymal storage. The absence of ESR1 leads to reduced epididymal sperm content, reduced sperm motility and fertilizing ability. The present study was undertaken to investigate the expression and presence of ESR1 gene in fertile and low-fertile male goat breeds. We identified ESR1 gene through various molecular tools. Genotyping was carried out by high resonance melting analysis using Roche Light Cycler 480(LC-480) system and found three different genotypes. Genotypic frequency-AA (blue-0.67), BB(Red-0.2), AB(Green-0.08) with allele frequency A(0.71 and B (0.29). The predominance of this gene in head of epididymis in fertile bucks was confirmed by SDS-PAGE, Western blotting and immunohistochemistry. From the results, we corroborated that the present study provides a useful and effective way to predict male fertility in goat breeds, which in turn increases the percentage of fertility in flock leading to more number of offspring in a kidding season.

Symmetry and Correlation Effects on Band Structure Explain the Anomalous Transport Properties of (111) LaAlO_{3}/SrTiO_{3}.

The interface between the two insulating oxides SrTiO_{3} and LaAlO_{3} gives rise to a two-dimensional electron system with intriguing transport phenomena, including superconductivity, which are controllable by a gate. Previous measurements on the (001) interface have shown that the superconducting critical temperature, the Hall density, and the frequency of quantum oscillations, vary nonmonotonically and in a correlated fashion with the gate voltage. In this Letter we experimentally demonstrate that the (111) interface features a qualitatively distinct behavior, in which the frequency of Shubnikov-de Haas oscillations changes monotonically, while the variation of other properties is nonmonotonic albeit uncorrelated. We develop a theoretical model, incorporating the different symmetries of these interfaces as well as electronic-correlation-induced band competition. We show that the latter dominates at (001), leading to similar nonmonotonicity in all observables, while the former is more important at (111), giving rise to highly curved Fermi contours, and accounting for all its anomalous transport measurements.

Differential expression and characterization of ATP1A1 exon17 gene by high resolution melting analysis and RT-PCR in Indian goats.

The investigation was carried out to analyse the genetic polymorphism and gene expression of ATP1A1 gene in four different Indian goat breeds by using high resolution melting (HRM) and real time-PCR. ATPase is electro-genic ion pump which is maintains the balance of sodium and potassium ions in animal cells. The transport of Na+& K+ is variable at cellular level during extreme hot period. Therefore, susceptible and tolerant animals were selected based on the physiological responses during hot period. Blood samples were collected from individuals, DNA was isolated. The 300 bp fragment of ATP1A1 gene was amplified by PCR and HRM genotyping was performed. The melting curves were analysed, differential temperature-shift plot showed three different genotypes in all the analysed samples. Out of the 135 samples, the distribution percentages were 55.56% (AA/blue), 33.33% (AC/red) and 11.11% (CC/green). The sequence variation revealed a SNP at 143rd position (A>C). The nucleotide diversity was 0.695 ± 0.403, 0.732 ± 0.424, 0.662 ± 0.433 and 0.687 ± 0.398 in Barbari, Jamunapari, Jakharna and Sirohi, respectively. The respiration rate (RR) was significantly different (P < 0.05) between AA and AC (t = 1.875, df = 38) genotype and heart rate (HR) was significantly different (P < 0.05) between AA and CC genotype. The relative expression pattern of ATP1A1 in SNP variants and non-variants animal tissues showed 19.09 and 6.93 fold higher than control (non-variant), respectively. Jamunapari showed higher fold value of ATP1A1 gene in comparison to Barbari, Jakharna and Sirohi. However, the heat stress-susceptible phenotype had significantly higher gene expression than stress-tolerant in all the breeds. The variation may be used as a marker for selection on the basis of physiological parameters and expression of ATP1A1 gene in goats indicating the specificity of expression in each tissue.

Genetic analysis of growth parameters and survival potential of Jamunapari goats in semiarid tropics.

Goats are adaptable to varied farming environments and contribute significantly towards sustainable livestock production and food security. Selective breeding of goats for improved performance is an important component of sustainable production. The objectives of the present study were to determine the most appropriate models of analysis for growth traits, to estimate genetic parameters, survival potential of kids from birth to 12 months of age and to estimate genetic trends for growth traits of Jamunapari kids at different ages. Genetic parameter estimates were obtained from 6590 records generated between 1982 and 2012 from 5922 animals with a pedigree covering over 13 generations. The most parsimonious model for early growth traits included permanent environmental effects due to the dam (PE) and litter effects. Similarly, the most appropriate model for early average daily gain (ADG) between birth and 3 or 6 months also included PE and litter effects. The estimates of heritability for survival from birth to 12 months of age ranged from 0.10 to 0.43. The estimates of heritability for ADG varied from 0.04 to 0.41. In general, higher estimates of heritability were observed when a sire model was fitted in the random effect. There was no genetic variation observed for survival between birth and 3 months of age. However, heritability estimates of between 0.18 and 0.39 were observed for survivability during post-weaning period to 12 months of age. The genetic trend at 9 months of age and 12 months of age was 0.144 kg 0.189 kg per year respectively. The genetic trend at all the ages was positive during the study period.

Link between the Superconducting Dome and Spin-Orbit Interaction in the (111) LaAlO_{3}/SrTiO_{3} Interface.

We measure the gate voltage (V_{g}) dependence of the superconducting properties and the spin-orbit interaction in the (111)-oriented LaAlO_{3}/SrTiO_{3} interface. Superconductivity is observed in a dome-shaped region in the carrier density-temperature phase diagram with the maxima of superconducting transition temperature T_{c} and the upper critical fields lying at the same V_{g}. The spin-orbit interaction determined from the superconducting parameters and confirmed by weak-antilocalization measurements follows the same gate voltage dependence as T_{c}. The correlation between the superconductivity and spin-orbit interaction as well as the enhancement of the parallel upper critical field, well beyond the Chandrasekhar-Clogston limit, suggest that superconductivity and the spin-orbit interaction are linked in a nontrivial fashion. We propose possible scenarios to explain this unconventional behavior.

Estimation of genetic parameters and genetic trends for milk yield traits in Jamunapari goats in semiarid tropics.

The Jamunapari goats are reared as dairy animals in semi-arid conditions, therefore it is necessary to evaluate the genetic potential for the genetic improvement of milk production traits. The data comprised of 2217 phenotypic records for milk yield at 90 days (MY90) and 140 days (MY140), total milk yield (TMY) and lactation length (LL) obtained from the progeny of 173 sires and 446 dams during the period 1990-2013. The data were analysed using mixed linear models exploring random effects due to direct additive, maternal and permanent environment variance due to animal. The most appropriate genetic models for milk yield traits were those that included permanent environment effects due to the animal. The direct additive heritability estimates were 0.15 ± 0.05, 0.26 ± 0.07, 0.25 ± 0.08 for MY90, MY140 and TMY, respectively. The additive heritability estimate for LL was low and non-significant at 0.02 ± 0.03. The repeatability estimates were moderate to high ranging from 0.68 to 0.73 for milk yield traits. The repeatability for lactation length was 0.20 ± 0.03. Maternal variances were low ranging from 0.03 for MY90 to 0.13 for TMY. There was an increase in mean milk yield of 0.25, 0.70 and 0.72 kg/year respectively at 90 and 140 days, and for TMY. Genetic trends and phenotypic trends for MY90, MY140 and TMY were positive and indicated significant improvement in milk traits due to selective breeding.

Molecular expression of caprine estrogen receptor gene 1 in reproductive and non-reproductive tissues.

During the last decades, physiological effects of oestrogens have been increasingly explored by scientists and biotechnologists. Estrogens exert a wide range of effects on a large variety of cell types. Oestrogen and its receptors are essential for sexual development and reproduction. Estrogen receptor alpha is a nuclear receptor activated by the hormone oestrogen. In male, ERα is encoded by the gene estrogen receptor gene 1 (ESR1), responsible for better fertility. The ESR1 is involved in the reabsorption of luminal fluid during the transit of spermatozoa from the testis to the head of the epididymis which is important for their survival and maturation during epididymal storage. The absence of ESR1 leads to reduced epididymal sperm content, reduced sperm motility and fertilizing ability. Therefore, this is a good startby to study the expression pattern of estrogen receptor 1 gene in high-fertile (G1) and low-fertile (G2) bucks of Jamunapari and Barbari breeds identified on the basis of seminal quality traits and fertility trials. RNA was extracted from the tissues by TRIzol method. The identification and expression pattern of caprine ESR1 gene was analysed by real-time PCR (Roche LC-480). Our work shows that the relative quantification by RT-PCR indicates more fold in head of epididymis as compared to spleen of caprine ESR1 gene. Furthermore, the RT-PCR indicated that fertile bucks of Jamunapari breed have more fold value as compared to Barbari breed in respect of reproductive organ.

Safety evaluation of Simarouba glauca seed fat.

Simarouba glauca DC is a tree of the family Simaroubaceae, which grows well up to 1,000 m above sea level in all types of well-drained soils (pH 5.5 to 8.0) and in places with 250 to 2,500 mm annual rainfall. The seed oil has been extracted both by mechanical expelling and solvent extraction. The fatty acid composition and iodine value of the oil indicate that it possesses saturated (40.8-42.6%), monounsaturated (52.9-55.0%), and polyunsaturated (2.5-3.4%) fatty acid in ratios close to that of palm oil. These characteristics are suitable for its use as edible oil. Acute oral toxicity and safety evaluation in a 13-week feeding trial on albino rats showed that the oil is comparable to groundnut oil in all the parameters.

Genetic variation and population structure in Jamunapari goats using microsatellites, mitochondrial DNA, and milk protein genes.

Jamunapari, a dairy goat breed of India, has been gradually declining in numbers in its home tract over the years. We have analysed genetic variation and population history in Jamunapari goats based on 17 microsatellite loci, 2 milk protein loci, mitochondrial hypervariable region I (HVRI) sequencing, and three Y-chromosomal gene sequencing. We used the mitochondrial DNA (mtDNA) mismatch distribution, microsatellite data, and bottleneck tests to infer the population history and demography. The mean number of alleles per locus was 9.0 indicating that the allelic variation was high in all the loci and the mean heterozygosity was 0.769 at nuclear loci. Although the population size is smaller than 8,000 individuals, the amount of variability both in terms of allelic richness and gene diversity was high in all the microsatellite loci except ILST 005. The gene diversity and effective number of alleles at milk protein loci were higher than the 10 other Indian goat breeds that they were compared to. Mismatch analysis was carried out and the analysis revealed that the population curve was unimodal indicating the expansion of population. The genetic diversity of Y-chromosome genes was low in the present study. The observed mean M ratio in the population was above the critical significance value (Mc) and close to one indicating that it has maintained a slowly changing population size. The mode-shift test did not detect any distortion of allele frequency and the heterozygosity excess method showed that there was no significant departure from mutation-drift equilibrium detected in the population. However, the effects of genetic bottlenecks were observed in some loci due to decreased heterozygosity and lower level of M ratio. There were two observed genetic subdivisions in the population supporting the observations of farmers in different areas. This base line information on genetic diversity, bottleneck analysis, and mismatch analysis was obtained to assist the conservation decision and management of the breed.

Differential expression and regulation of HSP70 gene during growth phase in ruminants in response to heat stress.

Heat shock proteins regulate the physiological mechanism of heat stress adaptation at cellular level. The present investigation was carried out to analyse the HSP70 gene regulation in various growth stage in ruminants in peripheral blood mononuclear cells (PBMCs). The relationship between HSP gene expression and thermotolerance in age-specific manner in ruminants has not been analysed. Therefore m-RNA HSP70 expression level was examined in different age groups of Jamunpari goat during hot climatic conditions. The experiment was carried out in 32 animals of Jamunapari goat belonging to the age groups of 3-months, 9-months, 12-months, and adults (2-3 year). Total RNA was isolated from peripheral blood mononuclear cells. The physiological response such as rectal temperature (RT), respiration rate (RR) and heart rate (HR) was used as indicator to heat stress. Temperature Humidity Index (THI) was used as an indicator of severity of environmental stress. The THI range varied from 82.00-92.08 during experimental period. The m-RNA HSP70 expression level at 9-month age of animals was up-regulated and significantly higher than other age groups. It was observed that the level of HSP70 transcripts in PBMCs was highest at 9-month age group, and age-related decline in HSP70 expression was observed in adult age. Based on the physiological response, the contrasting heat-stress phenotypes were recognised as heat stress susceptible (HSS) and heat stress tolerant (HST) individuals and the expression of m-RNA HSP70 was analysed at different ages in response to chronic heat stress. The differential mRNA expression of HSS individuals at 3 and 9-month of age showed the highest fold expression than HST. Age and phenotype had significant effect (p < 0.01) on the crossing point (CP) value. The m-RNA HSP70 gene expression in different age groups was correlated with heat stress tolerance and this could be used as biomarker for breeders to analyse the HSP response in -vivo in ruminants.

Genetic parameters for direct and maternal effects on post-weaning body measurements of Muzaffarnagari sheep in India.

Variance components and genetic parameters were estimated for post-weaning (i.e., at 6, 9, and 12 months of age) body measurements in Muzaffarnagari sheep maintained at the Central Institute for Research on Goats, Makhdoom, Mathura, India over a period of 29 years (1976 through 2004). Records of 2,965 lambs descended from 162 rams and 1,213 ewes were used in the study. Analyses were carried out by REML fitting an animal model and ignoring or including maternal genetic or permanent environmental effects. Six different animal models were fitted for all traits. The best model was chosen after testing the improvement of the log-likelihood values. Direct heritability estimates were inflated substantially for all traits when maternal effects were ignored. Moderate estimates of direct heritability for body length (0.11-0.15), height at withers (0.14-0.19), and heart girth (0.14-0.24) of lambs were observed at post-weaning stages of growth. Results suggest that only direct additive genetic effects were important for body measurements at post-weaning stages of growth, and hence, modest rates of genetic progress were possible for post-weaning body measurements.

Post translational modifications of milk proteins in geographically diverse goat breeds.

Goat milk is a source of nutrition in difficult areas and has lesser allerginicity than cow milk. It is leading in the area for nutraceutical formulation and drug development using goat mammary gland as a bioreactor. Post translational modifications of a protein regulate protein function, biological activity, stabilization and interactions. The protein variants of goat milk from 10 breeds were studied for the post translational modifications by combining highly sensitive 2DE and Q-Exactive LC-MS/MS. Here we observed high levels of post translational modifications in 201 peptides of 120 goat milk proteins. The phosphosites observed for CSN2, CSN1S1, CSN1S2, CSN3 were 11P, 13P, 17P and 6P, respectively in 105 casein phosphopeptides. Whey proteins BLG and LALBA showed 19 and 4 phosphosites respectively. Post translational modification was observed in 45 low abundant non-casein milk proteins mainly associated with signal transduction, immune system, developmental biology and metabolism pathways. Pasp is reported for the first time in 47 sites. The rare conserved peptide sequence of (SSSEE) was observed in αS1 and αS2 casein. The functional roles of identified phosphopeptides included anti-microbial, DPP-IV inhibitory, anti-inflammatory and ACE inhibitory. This is first report from tropics, investigating post translational modifications in casein and non-casein goat milk proteins and studies their interactions.

Characterization of casein gene complex and genetic diversity analysis in Indian goats.

Milk protein polymorphism plays an important role in genetic diversity analysis, phylogenetic studies, establishing geographical diversity, conservation decision, and improving breeding goals. Milk protein polymorphism in Indian goat breeds has not been well studied; therefore, an investigation was carried out to analyze the genetic structure of the casein gene and milk protein diversity at six milk protein loci in nine Indian goat breeds/genetic groups from varied agro-climatic zones. Milk protein genotyping was carried out in 1098 individual milk samples by SDS-PAGE at alphaS1-CN (CSN1S1), beta-CN (CSN2), alphaS2-CN (CSN1S2), kappa-CN (CSN3), beta-LG, and alpha-LA loci. Indian goats exhibited alphaS1-casein A allele in higher frequency in the majority of breeds except Ganjam and local goats. The alphaS1-casein A allele frequencies varied from 0.45 to 0.77. A total of 16 casein haplotypes were observed in seven breeds and breed specific haplotypes were observed with respect to geographic region. The average number of alleles was lowest in Ganjam (1.66 +/- 0.81) and highest in Sirohi goats (2.50 +/- 1.05). Expected heterozygosity at six different loci demonstrated genetic diversity and breed fragmentation. Neighbor-Joining tree was built basing on Nei's distance. There was about 16.95% variability due to differences between breeds, indicating a strong subdivision. Principal component analysis was carried out to highlight the relationship among breeds. The variability among goat breeds was contributed by alphaS2-CN, beta-LG and alphaS1-CN. The Indian goats exhibited alphaS1-CN (CSN1S1) A allele in higher frequency in all the breeds indicating the higher casein yield in their milk.

Chiral superconductivity in the alternate stacking compound 4Hb-TaS2.

Van der Waals materials offer unprecedented control of electronic properties via stacking of different types of two-dimensional materials. A fascinating frontier, largely unexplored, is the stacking of strongly correlated phases of matter. We study 4Hb-TaS2, which naturally realizes an alternating stacking of 1T-TaS2 and 1H-TaS2 structures. The former is a well-known Mott insulator, which has recently been proposed to host a gapless spin-liquid ground state. The latter is a superconductor known to also host a competing charge density wave state. This raises the question of how these two components affect each other when stacked together. We find a superconductor with a T c of 2.7 Kelvin and anomalous properties, of which the most notable one is a signature of time-reversal symmetry breaking, abruptly appearing at the superconducting transition. This observation is consistent with a chiral superconducting state.

Characterization of exon 2 and intron 2 of leptin gene in Indian goats.

Leptin, the hormonal product of the leptin (obese) gene (LEP), has multiple physiological effects and plays a pivotal role in the control of body growth, immune function, and reproduction. LEP gene polymorphism has been analyzed in cattle, buffalo, and pigs, and the polymorphic pattern has been associated with energy balance, milk production, and live weight and fertility trait in different livestock species. The present study has been designed to analyze polymorphism in exon 2 and intron 2 region of leptin gene in Indian goats. Genotyping was carried out in 111 kids including 70 samples from Barbari and 41 samples from Jamunapari breeds. The amplified product of exon 2 and intron 2 region of leptin gene was 152 bp and 400 bp, respectively, in both breeds. Sequencing of the exon 2 and intron 2 region of leptin gene and restriction analysis were carried out to analyze the polymorphism in goats. Five major haplotypes were observed in exon 2 region and six major haplotypes observed in intron 2 region in both breeds. Restriction fragment analysis and sequence analysis confirmed the mutation at 60 bp position of exon 2 and 100 bp position in intron 2 in all the analyzed samples. Sequences of exon 2 region of goats were unique as compared to other livestock species in BLAST analysis.

Silencing of quinolinic acid phosphoribosyl transferase (QPT) gene for enhanced production of scopolamine in hairy root culture of Duboisia leichhardtii.

Scopolamine is a pharmaceutically important tropane alkaloid which is used therapeutically in the form of an anesthetic and antispasmodic drug. The present study demonstrates enhanced scopolamine production from transgenic hairy root clones of Duboisia leichhardtii wherein the expression of quinolinate phosphoribosyl transferase (QPT) gene was silenced using the QPT-RNAi construct under the control of CaMV 35 S promoter. The RNAi hairy roots clones viz. P4, P7, P8, and P12 showed the enhanced synthesis of scopolamine with significant inhibition of nicotine biosynthesis. Optimization of culture duration in combination with methyl jasmonate elicitor in different concentrations (50 µM-200 µM) was carried out. Maximum synthesis of scopolamine had obtained from HR clones P7 (8.84 ± 0.117 mg/gm) on the 30th day of cultivation. Conspicuously, elicitation with wound-associated hormone methyl jasmonate enhanced the yield of scopolamine 2.2 fold (19.344 ± 0.275 mg/gm) compared to the culture lacking the elicitor. The transgenic hairy roots cultures established with RNAi mediated silencing of quinolinate phosphoribosyl transferase gene provides an alternative approach to increase the yield of scopolamine in fulfilling the demand of this secondary metabolite.

Induction of immune response in mice with a DNA vaccine encoding outer membrane protein (omp31) of Brucella melitensis 16M.

Brucellosis causes serious economic losses to goat farmers by way of reproductive losses in the form of abortions and stillbirths. Nucleic acid vaccines provide an exciting approach for antigen presentation to the immune system. In this study, we evaluated the ability of DNA vaccine encoding the omp31 protein of Brucella melitensis 16M to induce cellular and humoral immune responses in mice. We constructed eukaryotic expression vectors called pTargeTomp31, encoding outer membrane protein (omp31) of B. melitensis 16M. pTargeTomp31 was injected intramuscularly three times, at 3-week intervals in groups of mice 6 weeks of age. pTargeTomp31 induced good antibody response in ELISA . pTargeTomp31 elicited a T-cell-proliferative response and also induced a strong gamma interferon production upon restimulation with either the omp31 antigen or B. melitensis 16M extract. We also demonstrate that animals immunized with this plasmid elicited a strong and long-lived memory immune response. Furthermore, pTargeTomp31 elicited a typical T-helper 1-dominated immune response in mice, as determined by immunoglobulin G isotype analysis. This vaccine also provided the moderate degree of protection to the mice. This study for the first time focuses on DNA immunization of a gene from B. melitensis. These results may lead to the development of a DNA-based vaccine for the control of brucellosis in goats.

Differential expression pattern of heat shock protein 70 gene in tissues and heat stress phenotypes in goats during peak heat stress period.

It has been established that the synthesis of heat shock protein 70 (Hsp70) is temperature-dependent. The Hsp70 response is considered as a cellular thermometer in response to heat stress and other stimuli. The variation in Hsp70 gene expression has been positively correlated with thermotolerance in Drosophila melanogaster, Caenorhabditis elegans, rodents and human. Goats have a wide range of ecological adaptability due to their anatomical and physiological characteristics; however, the productivity of the individual declines during thermal stress. The present study was carried out to analyze the expression of heat shock proteins in different tissues and to contrast heat stress phenotypes in response to chronic heat stress. The investigation has been carried out in Jamunapari, Barbari, Jakhrana and Sirohi goats. These breeds differ in size, coat colour and production performance. The heat stress assessment in goats was carried out at a temperature humidity index (THI) ranging from 85.36-89.80 over the period. Phenotyping for heat stress susceptibility was carried out by combining respiration rate (RR) and heart rate (HR). Based on the distribution of RR and HR over the breeds in the population, individual animals were recognized as heat stress-susceptible (HSS) and heat stress-tolerant (HST). Based on their physiological responses, the selected animals were slaughtered for tissue collection during peak heat stress periods. The tissue samples from different organs such as liver, spleen, heart, testis, brain and lungs were collected and stored at -70 °C for future use. Hsp70 concentrations were analyzed from tissue extract with ELISA. mRNA expression levels were evaluated using the SYBR green method. Kidney, liver and heart had 1.5-2.0-fold higher Hsp70 concentrations as compared to other organs in the tissue extracts. Similarly, the gene expression pattern of Hsp70 in different organs indicated that the liver, spleen, brain and kidney exhibited 5.94, 4.96, 5.29 and 2.63-fold higher expression than control. Liver and brain tissues showed the highest gene expression at mRNA levels as compared to kidney, spleen and heart. HST individuals had higher levels of mRNA level expression than HSS individuals in all breeds. The Sirohi breed showed the highest (6.3-fold) mRNA expression levels as compared to the other three breeds, indicating the better heat stress regulation activity in the breed.