DA Distance Research Articles

Microsatellite loci 분석을 통한 한우 지역 브랜드간 유전적 다양성의 비교

ABSTRACT Genotype data of eleven microsatellites typed in 713 Hanwoo breed (Korean cattle) animals were used to estimate the genetic diversities and relationship of nine brand populations distributed in six regional areas. Size of microsatellite markers decided using GeneMapper Software (v.4.0) after analyze in kinds of ABI machine of name of 3130 using ABI Stock Markers TM kit. Frequencies of microsatellites markers were used to estimate heterozygosities, gene diversities and genetic distances. Microsatellite markers showed high polymorphism and 163 alleles were detected from the genotyping of eleven microsatellite markers. Allele frequency results were resemblant to each other in case of most markers and this between each brand where several alleles shared the majority mean. Genetic distances between populations were obtained using Nei’s DA distance method. Expected heterozygosity between each brand population was estimated very analogously. Average expected heterozygosity (0.765) of whole Hanwoo populations had higher diversities than other species of Europe, North American continent and spanish breeds. Genetic distances (0.0413) between ChoneBuk (JS) and ChoneNam (SJ) was nearest than distances between other populations. Genetic distances between KyongGi (AS, YP) and KyongNam (NH) showed far distance than other populations by 0.0493 and 0.0489 each respectively. ChoneNam (SJ) showed near genetic distance relatively with other areas' populations. In the UPGMA tree that is made based on DA distance matrix, some brand populations ramified to area. Because this became evenly genetic because gene flow of Hanwoo was frequent comparatively between each area by stock bull selection of country unit and their sperms (KPN) supply system that is done till today. It can be considered that this effect is by transfer of calf between near geographical areas. Each individuals were not ramified to different group and were spread evenly in phylogenetic dendrogram about all Hanwoo of each brand.(

Empirical Tests of the Reliability of Phylogenetic Trees Constructed With Microsatellite DNA

Microsatellite DNA loci or short tandem repeats (STRs) are abundant in eukaryotic genomes and are often used for constructing phylogenetic trees of closely related populations or species. These phylogenetic trees are usually constructed by using some genetic distance measure based on allele frequency data, and there are many distance measures that have been proposed for this purpose. In the past the efficiencies of these distance measures in constructing phylogenetic trees have been studied mathematically or by computer simulations. Recently, however, allele frequencies of 783 STR loci have been compiled from various human populations. We have therefore used these empirical data to investigate the relative efficiencies of different distance measures in constructing phylogenetic trees. The results showed that (1) the probability of obtaining the correct branching pattern of a tree (PC) is generally highest for DA distance; (2) FST*, standard genetic distance (DS), and FST/(1-FST) give similar PC-values, FST* being slightly better than the other two; and (3) (deltamu)2 shows PC-values much lower than the other distance measures. To have reasonably high PC-values for trees similar to ours, at least 30 loci with a minimum of 15 individuals are required when DA distance is used.

The analysis of genetic diversity and differentiation of six Chinese cattle populations using microsatellite markers

A total of 321 individuals from six cattle populations of four species in a bovine subfamily in China were studied using 12 pairs of microsatellite markers. The genetic diversities within and between populations were calculated. The phylogenetic trees were constructed by (δμ) 2 and DA distances, and the divergence times between populations were estimated by (δμ) 2. Altogether, 144 microsatellite alleles were detected including 24 private alleles and nine shared alleles. Chinese Holstein had the largest number of private alleles (10), whereas, Bohai black and Buffalo had the smallest number of private alleles (2). Chinese Holstein showed the highest genetic variability. Its observed number of alleles ( Na), mean effective number of alleles (MNA), and mean heterozygosity ( He) were 7.7500, 4.9722, and 0.7719, respectively, whereas, the Buffalo and Yak showed low genetic variability. In the phylogenetic trees, Luxi and Holstein grouped first, followed by Bohai and Minnan. Yak branched next and buffalo emerged as the most divergent population from other cattle populations. Luxi and Bohai were estimated to have diverged 0.039–0.105 million years ago (MYA), however, buffalo and Holstein diverged 0.501–1.337 MYA. The divergence time of Yak versus Minnan, Holstein and buffalo was 0.136–0.363, 0.273–0.729, and 0.326–0.600 MYA, respectively.

Evolutionary Relationships Among Prehistoric Human Populations: An Evaluation of Relatedeness Patterns Based on Facial Morphometric Data Using Molecular Data

The study of evolutionary relationships among human populations is fundamental to inferring processes that determine their structure and history. Among the different data types used to infer such relationships, molecular data, particularly nuclear and mitochondrial DNA, are preferred because of their high heritability and the low probability of changes during development. However, although the reliability of relatedness patterns based on other traits is discussed, except in unusual circumstances most prehistoric populations remain within the domain of morphological study. Therefore the primary goal of this study is to test the reliability of relatedness patterns constructed on the basis of craniometric data on a regional scale. In particular, we analyze samples from populations belonging to the Chaco, Pampa, and Patagonia regions of South America for which craniometric and molecular data are available. We compare a strongly supported relatedness pattern based on molecular data with the results obtained through landmark-based and semilandmark-based facial data. The matrices based on Euclidean distance for morphometric data and DA distances for molecular data were used to perform principal coordinates analyses and to obtain reticulograms. Finally, a principal components analysis of all individuals was performed with morphometric data. The results indicate that ordination analyses yield slightly different results depending on the morphometric data used. However, the reticulograms obtained with both landmark-based and semilandmark-based data allow the separation of the Chubut samples from the Chaco samples, with the Pampa sample in between the others; this pattern is congruent with molecular-based analyses. As a consequence, our results indicate that facial morphometric data allow the inference of the structure and history of the prehistoric populations for the studied region.

Microsatellite DNA Analysis of Wild Hops, Humulus lupulus L.

To study the relationships and genetic diversity among wild hops, Humulus lupulus, we analyzed 133 samples of wild hops collected from Europe, Asia and North America using polymorphism on 11 microsatellite loci. Although only three primers showed bands in Japanese hops, all other samples showed polymorphic bands at most loci. There were no duplicate genotypes among samples of European, Chinese and North American hops, and each individual hop could be distinguished completely. The phylogenetic tree constructed from DA distance with the UPGMA method showed a large cluster comprised of European hops, although Russian hops from the Caucasus and Altai regions were separate from the European cluster. Chinese and North American samples gave distinct clusters suggesting genetic differentiation. This study has indicated that hop microsatellite DNA is differentiated, and is dependent upon the origin in regions of Europe, Asia and North America.

STR Markers for Kinship Analysis

The analysis of short tandem repeats is a widely used method to estimate relatedness between closely related populations or individuals. The AmpFlSTR PCR Amplification Kit has 15 highly variable autosomal markers of tetranucleotide repeats and is principally made to identify individuals and first- or second-degree relatives. However, in many studies one is searching for individuals who are related through more than one generation. We wanted to test whether the amplification kit can also be used to identify more distantly related individuals. Therefore we compared 16 different methods that calculate genetic distance with regard to each method's ability to cluster more distantly related individuals from two test families. Among all the tested methods Nei et al.'s (1983) DA distance performed well in clustering family members within a group of unrelated individuals for a broad range of scenarios. However, second-degree relatives were difficult to cluster with any of the examined methods when other family members were absent. With a simulation we further estimated how many markers would actually be needed to detect a certain degree of relatedness. According to this simulation, one would need at least 123 independent microsatellite markers to detect third-degree relatives with 90% probability. In conclusion, the 15 STR markers in the amplification kit are suitable for detecting only very closely related individuals or entire families.

Genetic variation analysis of the Bali street dog using microsatellites

BackgroundApproximately 800,000 primarily feral dogs live on the small island of Bali. To analyze the genetic diversity in this population, forty samples were collected at random from dogs in the Denpasar, Bali region and tested using 31 polymorphic microsatellites. Australian dingoes and 28 American Kennel Club breeds were compared to the Bali Street Dog (BSD) for allelic diversity, heterozygosities, F-statistics, GST estimates, Nei's DA distance and phylogenetic relationships.ResultsThe BSD proved to be the most heterogeneous, exhibiting 239 of the 366 total alleles observed across all groups and breeds and had an observed heterozygosity of 0.692. Thirteen private alleles were observed in the BSD with an additional three alleles observed only in the BSD and the Australian dingo. The BSD was related most closely to the Chow Chow with a FST of 0.088 and also with high bootstrap support to the Australian dingo and Akita in the phylogenetic analysis.ConclusionsThis preliminary study into the diversity and relationship of the BSD to other domestic and feral dog populations shows the BSD to be highly heterogeneous and related to populations of East Asian origin. These results indicate that a viable and diverse population of dogs existed on the island of Bali prior to its geographic isolation approximately 12,000 years ago and has been little influenced by domesticated European dogs since that time.

α-Synuclein structures from fluorescence energy-transfer kinetics: Implications for the role of the protein in Parkinson's disease

Parkinson's disease is associated with the deposition and accumulation of alpha-synuclein fibrils in the brain. A30P and A53T mutations have been linked to the early-onset familial disease state. Time-resolved tryptophan fluorescence energy-transfer measurements have been used to probe the structures of pseudo-wild-type and mutant (A30P) alpha-synucleins at physiological pH (7.4), in acidic pH (4.4) solutions, and in the presence of SDS micelles, a membrane mimic. Fluorescent donor-energy acceptor (DA) distance distributions for six different tryptophan/3-nitro-tyrosine pairs reveal the presence of compact, intermediate, and extended conformations of the protein. CD spectra indicate that the protein develops substantial helical structure in the presence of SDS micelles. DA distributions show that micelles induce compaction in the N-terminal region and expansion of the acidic C terminus. In acidic solutions, there is an increased population of collapsed structures in the C-terminal region. Energy-transfer measurements demonstrate that the average DA distances for the W4-Y19 and Y19-W39 pairs are longer in one of the two disease-related mutants (A30P).

Genetic variation and relationships of eighteen Chinese indigenous pig breeds.

Chinese indigenous pig breeds are recognized as an invaluable component of the world's pig genetic resources and are divided traditionally into six types. Twenty-six microsatellite markers recommended by the FAO (Food and Agriculture Organization) and ISAG (International Society of Animal Genetics) were employed to analyze the genetic diversity of 18 Chinese indigenous pig breeds with 1001 individuals representing five types, and three commercial breeds with 184 individuals. The observed heterozygosity, unbiased expected heterozygosity and the observed and effective number of alleles were used to estimate the genetic variation of each indigenous breed. The unbiased expected heterozygosity ranged between 0.700 (Mashen) and 0.876 (Guanling), which implies that there is an abundant genetic variation stored in Chinese indigenous pig breeds. Breed differentiation was shown by fixation indices (FIT, FIS, and FST). The FST per locus varied from 0.019 (S0090) to 0.170 (SW951), and the average FST of all loci was 0.077, which means that most of the genetic variation was kept within breeds and only a little of the genetic variation exists between populations. The Neighbor-Joining tree was constructed based on the Nei DA (1978) distances and one large cluster with all local breeds but the Mashen breed, was obtained. Four smaller sub-clusters were also found, which included two to four breeds each. These results, however, did not completely agree with the traditional type of classification. A Neighbor-Joining dendrogram of individuals was established from the distance of – ln(proportions of shared alleles); 92.14% of the individuals were clustered with their own breeds, which implies that this method is useful for breed demarcation. This extensive research on pig genetic diversity in China indicates that these 18 Chinese indigenous breeds may have one common ancestor, helps us to better understand the relative distinctiveness of pig genetic resources, and will assist in developing a national plan for the conservation and utilization of Chinese indigenous pig breeds.

Distribution of HLA-A alleles in eight ethnic groups from Pakistan.

The extreme polymorphism found at some loci of the HLA system has made it an invaluable tool for population genetic analyses. In this study eight diverse ethnic groups from Pakistan were analyzed at the HLA-A locus using sequence specific primers for polymerase chain reaction (PCR-SSP) and then further typed to the allele level using a two-stage sequence specific oligonucleotide probe (SSOP) strategy. Four of these ethnic groups (Burusho, Hazara, Kalash, Pathan) were from the north and four (Baloch, Brahui, Sindhi and Parsi) were from the south of Pakistan. Nine alleles were identified as unique to a particular ethnic group within Pakistan. Maximum variation was seen in the HLA-A*02 allele family for which 11 alleles were detected in the eight Pakistani ethnic groups. The alleles that showed significant variation between the Pakistani ethnic groups include A*0101, A*0206, A*0209, A*0207, A*0217, A*1101, A*2402/09 N/11 N, A*2902, A*3301 and A*3001. A phylogenetic tree based on DA distances for HLA-A allele frequencies separated the Pakistani populations from other world populations and also separated the only Dravidian speaking population of Pakistan, the Brahui, from the remaining Indo-European speaking ethnic groups of Pakistan.

Genetic diversity of north-east Asian cattle based on microsatellite data.

In order to assess the genetic variability and population structure of north-east Asian cattle, 13 microsatellite loci were analysed for a total of 200 individuals including Korean, Chinese, Japanese Black and European Holstein cattle. Observed and expected heterozygosity, two estimators (F(ST) and G(ST)) of gene differentiation, and Nei's DA distance were evaluated. Based on expected mean heterozygosity, the lowest genetic diversity was exhibited in Japanese Black cattle (H(E)=0.471), and the highest in Chinese cattle (H(E)=0.744). Korean cattle revealed a relatively high degree of genetic diversity (H(E)=0.728). Average proportion of genetic variation because of interpopulation subdivision among north-east Asian cattle varied between 10.9 and 9.9%, depending on the estimator used. N-J tree based on Nei's DA genetic distance showed that Korean and Chinese cattle are closely related, whereas Japanese Black cattle are clearly distinct from the other two populations, forming a north-east Asian outgroup.

Patterns of genetic diversity at the nine forensically approved STR loci in the Indian populations.

Genetic diversity at the nine short tandem repeat (STR) loci, which are universally approved and widely used for forensic investigations, has been studied among nine Indian populations with diverse ethnic, linguistic, and geographic backgrounds. The nine STR loci were profiled on 902 individuals using fluorescent detection methods on an ABI377 System, with the aid of an Amp-F1 Profiler Plus Kit. The studied populations include two upper castes, Brahmin and Kayastha; a tribe, Garo, from West Bengal; a Hindu caste, Meitei, with historical links to Bengal Brahmins; a migrant group of Muslims; three tribal groups, Naga, Kuki and Hmar, from Manipur in northeast India; and a middle-ranking caste, Golla, who are seminomadic herders from Andhra Pradesh. Gene diversity analysis suggests that the average heterozygosity is uniformly high (>0.8) in the studied populations, with the coefficient of gene differentiation at 0.050 +/- 0.0054. Both neighbor-joining (NJ) and unweighted pair group method with arithmetic mean (UPGMA) trees based on DA distances bring out distinct clusters that are consistent with ethnic, linguistic, and/or geographic backgrounds of the populations. The fit of the Harpending and Ward model of regression of average heterozygosity on the gene frequency centroid is found to be good, and the observed outliers are consistent with the population structure and history of the studied populations. Our study suggests that the nine STR loci, used so far mostly for forensic investigations, can be used fruitfully for microevolutionary studies as well, and for reconstructing the phylogenetic history of human populations, at least at the local level.

Genetic variability in East Asian dogs using microsatellite loci analysis.

An analysis of eight microsatellite loci in 213 animals was performed to define the genetic structure and variability of 11 East Asian native dog populations. Allele diversity, observed heterozygosities, expected heterozygosities, F-statistics, G(ST) estimates, number of migrants per generation (Nm), and Nei's DA distance were calculated. Expected mean heterozygosities of Asian native dogs varied within a range of 0.310-0.718 with a mean value of 0.580. In a sample of 11 Asian dogs, the highest genetic diversity was exhibited in the Korean native dogs and the lowest in the Shiba, the Japanese native dog. All populations except the Kishu and Akita showed statistically significant deviation from Hardy-Weinberg equilibrium at more than one locus. After corrections for multiple significance tests, deviations over all loci were statistically significant in 7 of 11 dog populations, meaning that Asian dogs are genetically subdivided (global F(ST) = 0.154). Despite the locus-specific deviations, statistically significant departures from the Hardy-Weinberg equilibrium reflect deviations in the direction of heterozygote deficit, the global F(IS) being 0.072. In the neighbor-joining and unweighted pair group method with arithmetic mean (UPGMA) dendrograms based on Nei's DA distance, the Korean native breeds (the Sapsaree and the Jindo) were grouped together, then with the Eskimo dog. The two Japanese native dogs (the Hokkaido and the Akita) also clustered together, with moderate bootstrap support. In spite of some deviation, the three-dimensional scattergram based on principal components supported the conclusions suggested by the dendrograms based on Nei's DA distance. From these two analyses, the Korean native dogs formed the closest groups and then showed a close relationship to the Eskimo dogs, reflecting the fact that the Korean native dogs might be originated from dogs in the northern part of Far East Asia.

Genetic link between Asians and native Americans: evidence from HLA genes and haplotypes

We have been studying polymorphisms of HLA class I and II genes in East Asians including Buryat in Siberia, Mongolian, Han Chinese, Man Chinese, Korean Chinese, South Korean, and Taiwan indigenous populations in collaboration with many Asian scientists. Regional populations in Japan, Hondo-Japanese, Ryukyuan, and Ainu, were also studied. HLA-A, -B, and -DRB1 gene frequencies were subjected to the correspondence analysis and calculation of DA distances. The correspondence analysis demonstrated several major clusters of human populations in the world. “Mongoloid” populations were highly diversified, in which several clusters such as Northeast Asians, Southeast Asians, Oceanians, and Native Americans were observed. Interestingly, an indigenous population in North Japan, Ainu, was placed relatively close to Native Americans in the correspondence analysis. Distribution of particular HLA-A, -B, -DRB1 alleles and haplotypes was also analyzed in relation to migration and dispersal routes of ancestral populations. A number of alleles and haplotypes showed characteristic patterns of regional distribution. For example, B39-HR5-DQ7 (B∗3901-DRB1∗1406-DQB1∗0301) was shared by Ainu and Native Americans. A24-Cw8-B48 was commonly observed in Taiwan indigenous populations, Maori in New Zealand, Orochon in Northeast China, Inuit, and Tlingit. These findings further support the genetic link between East Asians and Native Americans. We have proposed that various ancestral populations in East Asia, marked by different HLA haplotypes, had migrated and dispersed through multiple routes. Moreover, relatively small genetic distances and the sharing of several HLA haplotypes between Ainu and Native Americans suggest that these populations are descendants of some Upper Paleolithic populations of East Asia.

Genetic diversity in Spanish donkey breeds using microsatellite DNA markers

Genetic diversity at 13 equine microsatellite loci was compared in five endangered Spanish donkey breeds: Andaluza, Catalana, Mallorquina, Encartaciones and Zamorano-Leonesa. All of the equine microsatellites used in this study were amplified and were polymorphic in the domestic donkey breeds with the exception of HMS1, which was monomorphic, and ASB2, which failed to amplify. Allele number, frequency distributions and mean heterozygosities were very similar among the Spanish donkey breeds. The unbiased expected heterozygosity (HE) over all the populations varied between 0.637 and 0.684 in this study. The low GST value showed that only 3.6% of the diversity was between breeds (P < 0.01). Significant deviations from Hardy-Weinberg equilibrium were shown for a number of locus-population combinations, except HMS5 that showed agreement in all analysed populations. The cumulative exclusion probability (PE) was 0.999 in each breed, suggesting that the loci would be suitable for donkey parentage testing. The constructed dendrogram from the DA distance matrix showed little differentiation between Spanish breeds, but great differentiation between them and the Moroccan ass and also with the horse, used as an outgroup. These results confirm the potential use of equine microsatellite loci as a tool for genetic studies in domestic donkey populations, which could also be useful for conservation plans.

Genetic distance and classification of domestic animals using genetic markers

Genetic distance is used to classify and elucidate the evolutionary relationship between populations, such as species, which have been diverging for a long period. The properties of genetic distance measures in livestock classification have hitherto not been investigated. We used simulation studies to compare the accuracy of eight genetic distance measures to classify recently diverged populations. The effect of varied population sizes from generation to generation were also taken into account. Comparatively better results are obtained from Nei’s standard distance with two alleles at each locus and from Nei’s DA distance with four alleles at each locus under various conditions. However, the difference in accuracy among distance measures was small and very high correlations were shown. The number of alleles at each locus had a greater effect on accuracy than the number of loci or type of distance measures. It indicates that highly polymorphic markers, such as DNA microsatellite, are powerful tool to improve the accuracy in classification of domestic animals. The six distance measures had a linear relationship with generation number at constant population size. None of the distance measures, however, showed a linear relationship when population size varied. The effect of fluctuating population size on the average distance was obvious over the first few generations but declined when the number of generations was increased.

TP53 polymorphisms and haplotypes in South Amerindians and neo-Brazilians

To evaluate the genetic diversity of Brazilian populations and contribute to the knowledge of their evolutionary history this study investigated three TP53 polymorphisms (BstUI and MspI RFLPs in exon 4 and intron 6, respectively, and a 16bp duplication in intron 3). The populations studied were: 114 Amerindians from five Brazilian Indian tribes (Gavião, Surui, Zoró, Wai-Wai and Xavante), 95 Euro-Brazilians and 70 Afro-Brazilians. The polymorphisms were all analysed using PCR amplifications. Gene frequencies and haplotype prevalences were calculated using the ARLEQUIN software. The genetic affinities of these groups with other world populations were estimated by the DA distance and neighbour joining method, using the NJBAFD computer program. Neo-Brazilians (immigrants from Europe and Africa) generally presented more variability than Amerindians, Afro-Brazilians being the most variable population. Among Amerindians, Gavião is the only group polymorphic for the three markers. Wai-Wai showed variability in BstUI and MspI RFLPs, while the other tribes were monomorphic for the 16bp A1 and MspI A2 alleles. A rare haplotype (1-2-1) was verified among the Wai-Wai. This haplotype was previously described in a Chinese sample only, but with low frequency. Therefore, either this combination was lost in the other tribes by genetic drift, recombination, or other factor, or it occurs in the Wai-Wai and Chinese by independent events. The Gavião also presented a haplotype (2-1-1) not observed in the other Amerindians; but since it is present in Euro- and Afro-Brazilians, its occurrence there is probably due to interethnic admixture. The relationships of several world populations obtained using TP53 indicates that this marker is very efficient in clustering populations of the same ethnic group.

Genetic distances and reconstruction of phylogenetic trees from microsatellite DNA.

Recently many investigators have used microsatellite DNA loci for studying the evolutionary relationships of closely related populations or species, and some authors proposed new genetic distance measures for this purpose. However, the efficiencies of these distance measures in obtaining the correct tree topology remains unclear. We therefore investigated the probability of obtaining the correct topology (PC) for these new distances as well as traditional distance measures by using computer simulation. We used both the infinite-allele model (IAM) and the stepwise mutation model (SMM), which seem to be appropriate for classical markers and microsatellite loci, respectively. The results show that in both the IAM and SMM CAVALLI-SFORZA and EDWARDS' chord distance (DC) and NEI et al.'s DA distance generally show higher PC values than other distance measures, whether the bottleneck effect exists or not. For estimating evolutionary times, however, NEI's standard distance and GOLDSTEIN et al.'s (delta mu)2 are more appropriate than other distances. Microsatellite DNA seems to be very useful for clarifying the evolutionary relationships of closely related populations.

Prediction of multiple genotypes in bovine embryos by PCR-RFLP analysis

Bovine leukocyte antigens (BoLA) are extensively used as markers for bovine disease and immunological traits. However, none of the BoLA genes in Southeast Asian breeds have been characterized by polymerase chain reaction (PCR)-sequence-based typing (SBT). Therefore, we sequenced exon 2 of the BoLA class II DRB3 gene from 1120 individual cows belonging to the Holstein, Sahiwal, Simbrah, Jersey, Brahman, and Philippine native breeds using PCR-SBT. Several cross-breeds were also examined. BoLA-DRB3 PCR-SBT identified 78 previously reported alleles and five novel alleles. The number of BoLA-DRB3 alleles identified in each breed from the Philippines was higher (71 in Philippine native cattle, 58 in Brahman, 46 in Holstein × Sahiwal, and 57 in Philippine native × Brahman) than that identified in breeds from other countries (e.g., 23 alleles in Japanese Black and 35 in Bolivian Yacumeño cattle). A phylogenetic tree based on the DA distance calculated from the BoLA-DRB3 allele frequency showed that Philippine native cattle from different Philippine islands are closely related, and all of them are closely similar to Philippine Brahman cattle but not to native Japanese and Latin American breeds. Furthermore, the BoLA-DRB3 allele frequency in Philippine native cattle from Luzon Island, located in the Northern Philippines was different from that in cattle from Iloilo, Bohol, and Leyte Islands, which are located in the Southern Philippines. Therefore, we conclude that Philippine native cattle can be divided into two populations, North and South areas. Moreover, a neutrality test revealed that Philippine native cattle from Leyte showed significantly greater genetic diversity, which may be maintained by balancing selection. This study shows that Asian breeds have high levels of BoLA-DRB3 polymorphism. This finding, especially the identification of five novel BoLA-DRB3 alleles, will be helpful for future SBT studies of BoLA-DRB3 alleles in East Asian cattle.