Anuradha Bhardwaj

Microsatellite markers based genetic diversity and bottleneck studies in Zanskari pony

Genetic diversity in Zanskari pony breed was evaluated at 48 microsatellite loci using fifty adult, healthy and unrelated animals. Allele frequency data was used to detect genetic diversity and bottleneck. The estimated average number of alleles (±s.e.) was 8.5208±2.5010 with a total of 409 alleles. A high level of genetic diversity within this breed was observed in terms of number of alleles, observed heterozygosity (0.6763±0.1704), expected Levens heterozygosity (0.7724±0.795), expected Neis heterozygosity (0.7644±0.0787) and polymorphism information content (>0.5). In-breeding coefficient (F(is)) was 0.115±0.0209, suggesting moderately high in-breeding in Zanskari breed. Although analysis of bottleneck revealed no bottleneck in recent past but population of Zanskari ponies has decreased drastically and only a few thousand pure-bred animals are left. The information is useful for proposing effective population management strategies for future.

Phenotypic characterization of Indian equine breeds: a comparative study

Las poblaciones de las seis razas equinas registradas por laOficina Nacional India deRecursos Zoogeneticos han disminuido drasticamente a causa de la cria sin criterio y de su menor utilizacion. En este trabajo, se estudiaron 15 indices biometricos, junto con caracteristicas raciales tipicas, en 50 animales de cada raza, excepto en la raza Bhutia (35) para su caracterizacion fenotipica. En base a sus alzadas a la cruz, las razas Kathiawari y Marwari se clasificaron como �caballos�, mientras que las razas Zanskari, Manipuri, Bhutia y Spiti se clasificaron como �ponis�. La raza Marwari fue la de mayor alzada y la que era significativamente diferente a otras razas en la mayoria de los indices biometricos. La raza Spiti fue la que presento una menor alzada entre las seis poblaciones de caballos y ponis estudiadas. Se observo un moderado dimorfismo sexual en algunos de los indices biometricos en las diferentes razas. En las razas Marwari y Kathiawari, tanto los sementales como las yeguas, pueden girar las orejas en un angulo de 180°, quedando las puntas de las orejas en el centro, siendo una caracteristica tipica de estas dos razas. Este trabajo tiene como objetivo proporcionar informacion de referencia para la identificacion y comparacion de las distintas razas de equidos de la India, con el proposito de sensibilizar a genetistas y criadores acerca de la importancia de producir ejemplares tipicos, de su conservacion y para la mejora de la gestion de estos valiosos recursos geneticos.

Heterologous Expression and Characterization of Indian Sahiwal Cattle (Bos indicus) Alpha Inhibin

Inhibin is a non-steroidal glycoprotein hormone of gonadal origin with major action as negative feedback control of the production of FSH by the anterior pituitary gland. The physiological role of inhibin has led to the development of inhibin immunogens for fertility enhancement in farm animals. It is envisaged that a reduction of endogenous inhibin secretion would increase FSH concentrations and thus offers a potential for increasing the number of ovulatory follicles in the ovary. The present work was carried out to produce recombinant bovine (Indian Sahiwal Cattle; Bos indicus) alpha inhibin (bINH-α) in E. coli by optimizing its expression and purification in biologically active form and to study its immunological characterization. A bacterial protein expression vector system based on the phage T5 promoter was used. The bINH-α encoding gene was successfully cloned and expressed in E. coli and the purified recombinant bINH-α was characterized. Recombinant bINH-α (25 µg mL−1) immunized guinea pigs had a significant increase in litter size compared to the control group. These results indicate a role for recombinant bINH-α as a fecundity vaccine to enhance the ovulation rate and litter size in animals.

Mitochondrial DNA- a Tool for Phylogenetic and Biodiversity Search in Equines

It is imperative to assess the maternal lineage in order to achieve a broad picture of evolution, phylogenetic and genetic biodiversity within and among different breeds of livestock. In recent past, there has been a considerable advancement in sequencing of complete mammalian mtDNA molecules and their analysis. Most of the studies have focused on the mitochondrial D-loop region, the most variable part of mtDNA due to increased substitution rate than in the rest of the mtDNA genome which serves as a better genetic marker to assess the diversity. Mitochondrial DNA (mtDNA) possesses several favorable characteristics, including large quantity in the cell, small genome size, haploid, maternal inheritance with extremely low probability of paternal leakage, higher mutation rate than nuclear DNA, and amenable to change mainly through mutation rather than recombination. All these features make mtDNA a useful and one of the most frequently used markers in molecular systematic and has been widely employed to address questions of genetic diversity, population structure and population evolution of animals including equines. Many native breeds of horses as well as ponies were assessed for their genetic diversity and ancestry on the basis of studies on mitochondrial DNA to address the questions of evolution along with breed development and conservation.

Genetic Diversity and Structure Analysis of Donkey Population Clusters in Different Indian Agro-climatic Regions

This study conducted with a panel of 24 polymorphic microsatellites revealed high number of alleles and heterozygosity in all the Indian donkey clusters available in different agro-climatic regions. All the markers are highly polymorphic as reflected from high allele number and heterozygosity, thus suitable for genotyping studies. Genetic diversity within each donkey population as well as between populations of different regions was also quite high indicating no extinction threat to population. The genetic differentiation suggested that 89.59% genetic variation corresponds to difference among individuals and 10.41% is attributed to differences among population clusters. However conservation and preservations of donkeys is important as their overall population is decreasing rapidly in country. Even the 24 microsatellites utilized for individual assignment did not reveal 100% assignment of donkeys to their clustered population. Breed relationship analysis revealed closeness among Leh and Bihar donkeys which seems to be unique as geographically these populations are at distant places and mixing of these donkeys cluster is not feasible. Closeness of donkey population from Haryana, Rajasthan, Gujarat and Baramati regions may be due to sharing of common breeding tract and movement of donkeys in these agro-climatic regions with their owners during harsh and odd environmental conditions for their survival. Population structure analysis revealed that donkey population from seven regions can be divided in two main clusters: first cluster having donkey population from Bihar and leh while second cluster included donkeys from Gujarat, Rajasthan, Spiti. Population from Baramati and Haryana had more than 50% individual population as admixed while Spiti donkeys had no admixed individuals. The current study aimed to provide insight into the genetic relationships and diversity between various indigenous donkey populations will offer a valuable reference for rational strategies in breed assignment to local non-descript donkeys, donkey conservation and breeding programs.

Molecular characterization, modeling, in silico analysis of equine pituitary gonadotropin alpha subunit and docking interaction studies with ganirelix

Equine pituitary gonadotropins (eLH, eFSH, eCG) are heterodimeric glycoprotein hormones with alpha (α) and beta (β) subunits. It is responsible for maintenance of pregnancy in mares during early gestation and fairly valuable for inducing superovulation in animals other than equines. The alpha subunit is common, while beta subunit is species-specific in all glycoprotein hormones. In the present investigation, molecular cloning and in silico characterization including homology modeling and molecular docking analysis of the equine chorionic gonadotropin (eCG) alpha subunit was carried out for gaining structural and functional insights into the eCG alpha subunit and its possible interaction with ganirelix, a gonadotropin-releasing hormone (GnRH) antagonist. The equine chorionic gonadotropin (eCG) alpha subunit expressed in pituitary gland was selected, amplified from total RNA, cloned and sequenced. The in silico analyses were made for homology modelling, structural details, epitope identification and chromosomal localization. Molecular docking studies of eCG alpha were undertaken with a drug ganirelix which is used to control ovulation and has antagonistic activity against GnRH. The protein sequence corresponding to selected open reading frame (ORF) was 99–100% similar with domesticated horse, Przewalski’s horse, and 92–93% with Burchell’s zebra and donkey. Molecular docking studies revealed the possible interaction of eCG alpha with ganirelix. The possible drug-macromolecule interactions were visualized between eCG alpha and ganirelix. The study will provide structural insight into unique sites and an alternate route of gonadotropin suppression applicable to assisted reproductive technologies.

Biochemical profiles of Indian donkey population located in six different agro-climatic zones

To establish normal values of blood biochemical indices for different indigenous local donkey population available in various agro-climatic zones, blood samples were collected from 233 adult and apparently healthy donkeys. The samples were analysed for metabolites (albumin, total serum protein, direct bilirubin, total bilirubin, cholesterol, HDL cholesterol, urea, uric acid, triglyceride, creatinine) and minerals (calcium, phosphorus) to evaluate significant difference within and between populations. Confidence limit of each biochemical indices showed a close range as compared to their actual range observed under varied geographic areas. All the metabolites and minerals showed significant variations in their levels within and between donkey populations which could possibly be due to the differences in the nutritional status of donkeys, their managemental aspects and biochemical metabolism. In agro-climatic zone 1 (Spiti and Leh areas), having low vegetation cover with poor nutritious grasses for a limited period, levels of most of the biochemical profiles in donkey populations belonging to these areas were significantly lower than those in other zones (VI, IX, XII, XIV). This study indicated that normal biochemical values of different indices for a particular population should not be used as such for disease prognosis, diagnosis and therapeutic monitoring of other donkey population belonging to other agro-climatic zone having different nutritional and managemental practices.

Assessment of demographic bottleneck in Indian horse and endangered pony breeds

Bottleneck study of any continuously decreasing population is important and crucial issue in its conservation strategies including the analysis of simulated and real populations (Williamson-Natesan 2005; Busch et al. 2007). A bottleneck in a population can increase the rate of inbreeding, loss of genetic variation, fixation of deleterious alleles, thereby reducing evolutionary potential of animals to adapt to new selective pressures, such as climatic change or shift in available resources and increasing the probability of population extinction (Frankham 1995). The genetic changes caused by a bottleneck in a population’s effective size can lower the possibility of population’s persistence (Vrijenhoek 1994; Newman and Pilson 1997). Various endangered or threatened populations have been reported to have low levels of genetic variations (Vrijenhoek 1994; Gibbs et al. 1998). However, all the populations that have been reduced in size did not show quantifiable lower levels of genetic diversity (Waldman et al. 1998) which also necessitates the assessment of bottlenecks with molecular marker for their conservation and evolutionary genetics. India is bestowed with a rich biodiversity of equids in the form of two horses (Marwari and Kathiawari) and four endangered pony breeds (Bhutia, Spiti, Manipuri and Zanskari) besides indigenous donkeys and wild asses (Gupta et al. 2012a,b; 2014). Overall population of these breeds, specially endangered pony breeds has declined in most of the pockets in their home tracts (less than 1000) which is due to their decreased utility and increased modernization of transport system even in hilly and difficult terrains (Gupta et al. 2012a, b). It is expected that bottleneck might have taken