Herwin Eding

Molecular tools and analytical approaches for the characterization of farm animal genetic diversity

Genetic studies of livestock populations focus on questions of domestication, within- and among-breed diversity, breed history and adaptive variation. In this review, we describe the use of different molecular markers and methods for data analysis used to address these questions. There is a clear trend towards the use of single nucleotide polymorphisms and whole-genome sequence information, the application of Bayesian or Approximate Bayesian analysis and the use of adaptive next to neutral diversity to support decisions on conservation.

Mitochondrial DNA D‐loop sequences suggest a Southeast Asian and Indian origin of Zimbabwean village chickens

This study sought to assess mitochondrial DNA (mtDNA) diversity and phylogeographic structure of chickens from five agro-ecological zones of Zimbabwe. Furthermore, chickens from Zimbabwe were compared with populations from other geographical regions (Malawi, Sudan and Germany) and other management systems (broiler and layer purebred lines). Finally, haplotypes of these animals were aligned to chicken sequences, taken from GenBank, that reflected populations of presumed centres of domestication. A 455-bp fragment of the mtDNA D-loop region was sequenced in 283 chickens of 14 populations. Thirty-two variable sites that defined 34 haplotypes were observed. In Zimbabwean chickens, diversity within ecotypes accounted for 96.8% of the variation, indicating little differentiation between ecotypes. The 34 haplotypes clustered into three clades that corresponded to (i) Zimbabwean and Malawian chickens, (ii) broiler and layer purebred lines and Northwest European chickens, and (iii) a mixture of chickens from Zimbabwe, Sudan, Northwest Europe and the purebred lines. Diversity among clades explained more than 80% of the total variation. Results indicated the existence of two distinct maternal lineages evenly distributed among the five Zimbabwean chicken ecotypes. For one of these lineages, chickens from Zimbabwe and Malawi shared major haplotypes with chicken populations that have a Southeast Asian background. The second maternal lineage, probably from the Indian subcontinent, was common to the five Zimbabwean chicken ecotypes, Sudanese and Northwest European chickens as well as purebred broiler and layer chicken lines. A third maternal lineage excluded Zimbabwean and other African chickens and clustered with haplotypes presumably originating from South China.

Objectives, criteria and methods for using molecular genetic data in priority setting for conservation of animal genetic resources

The genetic diversity of the worlds livestock populations is decreasing, both within and across breeds. A wide variety of factors has contributed to the loss, replacement or genetic dilution of many local breeds. Genetic variability within the more common commercial breeds has been greatly decreased by selectively intense breeding programmes. Conservation of livestock genetic variability is thus important, especially when considering possible future changes in production environments. The world has more than 7500 livestock breeds and conservation of all of them is not feasible. Therefore, prioritization is needed. The objective of this article is to review the state of the art in approaches for prioritization of breeds for conservation, particularly those approaches that consider molecular genetic information, and to identify any shortcomings that may restrict their application. The Weitzman method was among the first and most well-known approaches for utilization of molecular genetic information in conservation prioritization. This approach balances diversity and extinction probability to yield an objective measure of conservation potential. However, this approach was designed for decision making across species and measures diversity as distinctiveness. For livestock, prioritization will most commonly be performed among breeds within species, so alternatives that measure diversity as co-ancestry (i.e. also within-breed variability) have been proposed. Although these methods are technically sound, their application has generally been limited to research studies; most existing conservation programmes have effectively primarily based decisions on extinction risk. The development of user-friendly software incorporating these approaches may increase their rate of utilization.

Genetic diversity of Hungarian indigenous chicken breeds based on microsatellite markers

Six local chicken breeds are registered in Hungary and are regarded as Hungarian national treasures: Hungarian White, Yellow and Speckled, and Transylvanian Naked Neck White, Black and Speckled. Three Hungarian academic institutes have maintained these genetic resources for more than 30 years. The Hungarian Yellow, the Hungarian Speckled and the Transylvanian Naked Neck Speckled breeds were kept as duplicates in two separate subpopulations since time of formation of conservation flocks at different institutes. In this study, we investigated genetic diversity of these nine Hungarian chicken populations using 29 microsatellite markers. We assessed degree of polymorphism and relationships within and between Hungarian breeds on the basis of molecular markers, and compared the Hungarian chicken populations with commercial lines and European local breeds. In total, 168 alleles were observed in the nine Hungarian populations. The F(ST) estimate indicated that about 22% of the total variation originated from variation between the Hungarian breeds. Clustering using structure software showed clear separation between the Hungarian populations. The most frequent solutions were found at K = 5 and K = 6, respectively, classifying the Transylvanian Naked Neck breeds as a separate group of populations. To identify genetic resources unique to Hungary, marker estimated kinships were estimated and a safe set analysis was performed. We show that the contribution of all Hungarian breeds together to the total diversity of a given set of populations was lower when added to the commercial lines than when added to the European set of breeds.

Assessing genetic diversity of Vietnamese local chicken breeds using microsatellites

This study aimed to assess genetic diversity within and between nine Vietnamese local chicken breeds and two Chinese breeds included for comparison. Genotyping 29 microsatellites revealed high diversity of both Vietnamese and Chinese breeds. Cluster analysis using the STRUCTURE software suggested six clusters as the most likely grouping of the 11 breeds studied. These groups encompassed four homogeneous clusters, one formed by the two Chinese breeds and the other three representing a single breed each: the Mekong Delta breed Ac, the South Central Coast breed Choi, and the Red River Delta breed Dong Tao. The six remaining breeds formed two additional admixed clusters.

Global diversity and genetic contributions of chicken populations from African, Asian and European regions

Genetic diversity and population structure of 113 chicken populations from Africa, Asia and Europe were studied using 29 microsatellite markers. Among these, three populations of wild chickens and nine commercial purebreds were used as reference populations for comparison. Compared to commercial lines and chickens sampled from the European region, high mean numbers of alleles and a high degree of heterozygosity were found in Asian and African chickens as well as in Red Junglefowl. Population differentiation (FST ) was higher among European breeds and commercial lines than among African, Asian and Red Junglefowl populations. Neighbour-Net genetic clustering and structure analysis revealed two main groups of Asian and north-west European breeds, whereas African populations overlap with other breeds from Eastern Europe and the Mediterranean region. Broilers and brown egg layers were situated between the Asian and north-west European clusters. structure analysis confirmed a lower degree of population stratification in African and Asian chickens than in European breeds. High genetic differentiation and low genetic contributions to global diversity have been observed for single European breeds. Populations with low genetic variability have also shown a low genetic contribution to a core set of diversity in attaining maximum genetic variation present from the total populations. This may indicate that conservation measures in Europe should pay special attention to preserving as many single chicken breeds as possible to maintain maximum genetic diversity given that higher genetic variations come from differentiation between breeds.

Choice of breeding stock, preference of production traits and culling criteria of village chickens among Zimbabwe agro-ecological zones

Free ranging chickens reared by smallholder farmers represent genetic diversity suited for particular environments and shaped by the socio-economic and cultural values of the farming systems. This study sought to investigate the existence of chicken strains and evaluate the breeding goals and strategies used by village chicken farmers in Zimbabwe. A semi-structured questionnaire was administered to 97, 56, 70, 104 and 37 households randomly selected from five agro-ecological-zones I-V, respectively. Fifteen chicken strains mostly defined by morphological traits were reported in the five eco-zones. Production criteria such as body size, and fertility were highly ranked (ranging from 1.3 – 2.6) by farmers across all the eco-zones, while cultural traits were the least preferred production traits. As a common breeding practice, farmers chose the type of hens and cocks to retain for breeding purposes and these randomly mixed and mated with others from community flocks. Chicken body size was ranked the major determinant in choosing breeding animals followed by mothering ability, and fertility. More households culled chickens associated with poor reproductive performance, poor growth rates and those intolerant to disease pathogens. The focus on many negatively correlated production traits and the absence of farmer records compromises breeding strategies in these production systems.

Population structure and genetic diversity of Sudanese native chickens

The objectives of this study were to analyze genetic diversity and population structure of Sudanese native chicken breeds involved in a conservation program. Five Sudanese native chicken breeds were compared with populations studied previously, which included six purebred lines, six African populations and one Sudanese chicken population. Twenty-nine (29) microsatellite markers were genotyped individually in these five populations. Expected and observed heterozygosity, mean number of alleles per locus and inbreeding coefficient were calculated. A model based cluster analysis was carried out and a Neighbor net was constructed based on marker estimated kinships. Two hundred and one alleles were detected in all populations, with a mean number of 6.93 ± 3.52 alleles per locus. The mean observed and expected heterozygosity across 29 loci was 0.524 and 0.552, respectively. Total inbreeding coefficient (F IT ) was 0.069±0.112, while differentiation of subpopulations (F ST 0.026±0.049) was low indicating the absence of clear sub-structuring of the Sudanese native chicken populations. The inbreeding coefficient (F IS ) was 0.036±0.076. STRUCTURE software was used to cluster individuals to 2 ≤ k ≤ 7 assumed clusters. Solutions with the highest similarity coefficient were found at K=5 and K=6, in which Malawian, Zimbabwean, and purebred lines split from Sudanese gene pool. The six Sudanese native chicken populations formed one heterogeneous cluster. We concluded that Sudanese native chickens are highly diverse, and are genetically separated from Malawian, Zimbabwean chickens and six purebred lines. Our study reveals the absence of population sub-structuring of the Sudanese indigenous chicken populations. Key words: Genetic diversity, microsatellites, population structure, Sudanese native chickens.