Development of amplified fragment length polymorphism (AFLP) markers for the identification of Cholistani cattle

The identification issue of livestock can be resolved by using molecular identification tools that are acceptable to preserve and maintain pure breeds worldwide. The application of a molecular identification methodology is more important for developing nations, e.g., Pakistan, where uncontrolled crossbreeding has become a common practice and the import of exotic animals and germplasm is ever increasing. This presents a risk to local breeds as also stated by the FAO. Therefore, the current study was designed to develop standard molecular markers for Cholistani cattle to ascertain their purity for breeding purpose. In this study 50 and 48 unrelated males were sampled for Cholistani and each crossbred cattle, respectively. Candidate molecular markers present in Cholistani but absent in crossbred cattle and vice versa were detected using the amplified fragment length polymorphism (AFLP) method. Eleven markers were developed and were converted to single nucleotide polymorphism (SNP) markers for genotyping. The allele frequencies in both breeds were determined for discrimination ability using polymerase-chain-reaction–restriction-fragment-polymorphism (PCR-AFLP). The probability of identifying the Cholistani breed was 0.905 and the probability of misjudgment was 0.073 using a panel of markers. The identified markers can ascertain the breed purity and are likely to extend the facility for breed purity testing before entering into a genetic improvement program in the country.</p

Initial genomic characterization of Italian, Egyptian and Pakistani goat breeds

Selection and breeding practices in goats have differed greatly among countries and populations. These processes, together with natural selection and regional drift, have shaped the phenotypic variability of goat breeds (Kim et al., 2015). The availability of improved genomic analysis tools for this species may provide useful information on the history of selection, adaptation and differentiation of goats from different areas of the world, that can be evaluated by the study of gene frequencies and length of the Runs of Homozigosity (contiguous length of homozygous genotypes, ROH; Purfield et al., 2012). In current study, we examined using a goat medium density SNP chip animals from three different countries: Egypt (with lack of selection scheme), Italy (with several standardized breeds; Nicoloso et al., 2015) and Pakistan (with several breeds showing peculiar phenotypes) to produce a genomic landscape of goats breeds in these countries. A total of 1,123 animals of 39 different populations, and 48,895 SNP markers were analyzed. Genotypes were imputed on a country-based approach, and markers without known position in the genome were excluded from the analysis. MDS and ADMIXTURE plots confirmed the good differentiation among populations from the three countries. Runs of Homozygosity (ROH) were performed for each country and population allowed the detection of genomic regions with high homozygosity levels, common in at least two out of three sampling areas. These results provide new insights into goat genome structure within and among breeds and countries. The detection of conserved regions with different lengths may explain recent selection strategies or adaptation to different, extreme environmental conditions. The research was funded by INNOVAGEN project. Support by Iowa State University and the Ensminger funds for AE and AT as well as support by the Fulbright Foundation for AE are gratefully acknowledged. Sampling from Pakistan was funded by PAK-USAID project

The association of two single nucleotide polymorphisms (SNPs) in growth hormone (GH) gene with litter size and superovulation response in goat-breeds

Two active mutations (A 781 G and A 1575 G) in growth hormone (GH) gene, and their associations with litter size (LS), were investigated in both a high prolificacy (Matou, n = 182) and a low prolificacy breed (Boer, n = 352) by using the PCR-RFLP method. Superovulation experiments were designed in 57 dams, in order to evaluate the effect of different genotypes of the GH gene on superovulation response. Two genotypes (AA and AB, CC and CD) in each mutation were detected in these two goat breeds. Neither BB nor DD homozygous genotypes were observed. The genotypic frequencies of AB and CC were significantly higher than those of AA and CD. In the third parity, Matou dams with AB or CC genotypes had significantly larger litter sizes than those with AA and CD (p < 0.05). On combining the two loci, both Matou and Boer dams with ABCD genotype had the largest litter sizes when compared to the other genotypes (p < 0.05). When undergoing like superovulation treatments, a significantly higher number of corpora lutea and ova, with a lower incidence of ovarian cysts, were harvested in the AB and CC genotypes than in AA and CD. These results show that the two loci of GH gene are highly associated with abundant prolificacy and superovulation response in goat breeds

Genomic selection of Nili-Ravi buffalo: A choice for buffalo breeders

Among three well documented breeds of buffalo dairy breeds in Pakistan, Nili-Ravi is the best milk producer owing to its characteristic of disease and parasitic resistance, and better convertor of roughages into useful products than cattle. A selection program to enhance the genetic potential for milk production of Nili-Ravi using progeny testing program is going on. Traditional progeny testing program has made a remarkable improvement in the genetic potential of dairy cattle in the developed world. However, this program faces severe implementation issues in buffalo improvement due to limitation of resources and basic infrastructure. Simulated studies have shown the potential of genomic selection in shortening generation interval and increasing the accuracy of selection (especially young bulls) that can bring a relatively rapid genetic improvement. The current study intends to explore the application of genomic selection in a typical buffalo breeding perspective using Nili-Ravi in Pakistan as an example. The assumed size of the training population for genomic selection was 15860 present with BRI, Pattoki. Our calculations indicated that genomic selection can reduce the generation intervals in the male to male selection pathway from 9.5 years down to 3.3 years. It can result in almost 2 times increase in response to selection compared to that in a progeny testing program. Furthermore, it reduced the costs of proving bulls by 88%. The present study suggests the initiation of the program of genomic selection for Nili-Ravi in Pakistan and may serve as an example for other developing countries. The findings of the current study may encourage the researchers and policy makers to use the genomic selection for improvement in the productivity of dairy buffalo of developing countries

Evolution of hypothalamus-pituitary growth axis among fish, amphibian, birds and mammals

Hypothalamus-pituitary growth axis (HP growth axis) regulates animal growth and development in pre-natal and post natal life governed by many factors. However, until recently, the evolutionary history of this axis among lineages is not understood. Aim of the present study was to understand the major events in evolution and evolutionary history and trend of HP growth axis. The diversity among Homo sapience, Mus musculus, Rattus norvegicus, Gallus gallus, Danio rerio and Xenopus laevis was determined for genes involved in HP growth axis in current study. Sequences of HP growth axis genes were retrieved from NCBI (http://www.ncbi.nlm.nih.gov/). Nucleotide diversity using Kimura?s two-parameter method; codon-based test of positive selection using the Nei-Gojobori; equality of evolutionary rate with Tajima's relative rate test and phylogenetic history using the RelTime method were estimated in MEGA6. Estimates of the coefficients of evolutionary differentiation based on nucleotides and amino acids substitution patterns of HP growth axis genes showed contrasting evolutionary patterns among the lineages. The results demonstrated that although these genes might have crucial functional roles in each of the species, however, their sequence divergence did not necessarily reflect similar molecular evolution among the species. Codon-based test of positive selection revealed that Human vs Mouse, Chicken vs Rat, Human vs Rat and Mouse vs Rat had similar and higher non synonymous substitutions (P &gt; 0.05). Higher rate of non-synonymous substitutions at similar orthologs level among species indicated a similar positive selection pressure in these species. Results for relative rate test assessed with the chi-squared test showed difference on unique mutations among lineages at synonymous and non synonymous sites except Chicken vs Mouse, Human vs Mouse, Chicken vs Rat, Human vs Rat and Mouse vs Rat. This indicated that the mutagenic process that generates substitutional mutation is taking place at approximately the same rate at synonymous and non-synonymous sites these lineages. Moreover, despite of common ancestry, our results indicate a different divergent time among genes of these species. This is the first demonstration that variable rates of molecular evolution may be present within HP growth axis genes among different species. This difference could be of interest for comparative genomics analysis and physiological genes functions identification among tho comparative genomics, evolution rate, HP growth axis, positive selection se species whose HP growth axis is not explored.</jats:p