Genetic Analysis of Three Lithuanian Native Horse Breeds

This study was the first to analyze the polymorphic characteristics of a wide range of genetic markers in three Lithuanian horse breeds. Genetic variation at seven red blood cell alloantigen (blood group) loci, 10 biochemical genetic loci and 16 microsatellites was examinated in 31 Zemaitukai, 30 large-type Zemaitukai and 24 Lithuanian Heavy Draught horse and compared to that of other domestic horse breeds. Genetic variability of the Zemaitukai breeds based upon blood groups and biochemical loci was higher than average for domestic horse breeds. For microsatellite loci, the variability of the large-type Zemaitukai and the Heavy Draught breeds also was high compared to the mean variation for domestic breeds; however, for the Zemaitukai breed, variation levels were very near the average for horses.     

Genetic analysis of three South African horse breeds

Genetic variability at 7 blood-group and 10 biochemical genetic loci was examined in 3 South African horse breeds, the Nooitgedacht, Boerperd and Basuto Pony. Observed heterozygosity for these breeds was intermediate for domestic horses, with the highest heterozygosity in the Boerperd and the lowest in the Basuto Pony. The 3 breeds show greater genetic similarity to each other than to other domestic horse breeds. Compared to other breeds, the South African breeds show greater genetic similarity to breeds such as the Thoroughbred, Holstein, Trakehner and Hanovarian and also to North American breeds such as the Saddlebred, Standardbred and Morgan Horse.     

Molecular characterization and differentiation of five horse breeds raised in Algeria using polymorphic microsatellite markers

In this study, genetic analyses of diversity and differentiation were performed on five horse breeds raised in Algeria (Barb, Arab‐Barb, Arabian, Thoroughbred and French Trotter). All microsatellite markers were highly polymorphic in all the breeds. A total of 123 alleles from 14 microsatellite loci were detected in 201 horses. The average number of alleles per locus was the highest in the Arab‐Barb horses (7.86) and lowest in the thoroughbred breed (5.71), whereas the observed and expected heterozygosities per breed ranged from 0.71 (Thoroughbred) to 0.752 (Barb) and 0.71 (Thoroughbred) to 0.77 (Arab‐Barb), respectively. The genetic differentiation between the breeds was significant (p < 0.01) based on the infinitesimal model (F_ST). Three different approaches for evaluating the genetic relationships were applied. Genetic distances, the factorial correspondence analysis and structure analysis showed that a significant amount of genetic variation is maintained in the native horse populations and the other breeds. The Barb and Arab‐Barb breeds seem to be the most genetically related and support the decision to consider the breeds as same population.     

Molecular genetic studies of the "Einsiedler" horse population

In this study it was investigated whether the "Einsiedler" warmblood horse, a historically old horse population from central Switzerland (Abbey of Einsiedeln), is distinguishable from micellaneous horse breeds, using molecular genetic techniques. The breeding history of Einsiedler horses is characterised by systematic line breeding through the dams. Therefore, two Einsiedler dam lines (N = 28), going back to the middle of the 19th century according to pedigree entries, were the focus of the survey. Random samples of diverse warmblood horse populations, but also samples from more distinct types of horse breeds, served as comparison populations (N = 52).Variation in the mitochondrial genome appeared to be only partially informative to demarcate the studied horses, as horses of distinct breeds may share identical mtDNA sequence fragments. Both dam lines revealed haplotypes commonly found in Iberian horse breeds. This is to take as an indication on the genetic origin of Einsiedler horses. Furthermore, the Klima dam line held a homologous mtDNA sequence fragment with E. ferus przewalskii. Therefore, this seems to be a phylogenetically old haplotype. The analysis of microsatellite loci revealed that horses from the two Einsiedler dam lines were in fact distinguishable from more distinct types of horses, but not from closely related European warmblood horse breeds and English thoroughbred.     

Genetic characterisation of the Uruguayan Creole horse and analysis of relationships among horse breeds

The genetic variability within the Uruguayan Creole horse and its relationship to a group of geographically or historically related breeds (Spanish Pure-bred, Barb, Quarter horse, Paso Fino, Peruvian Paso, Arabian and Thoroughbred horse), was evaluated using 25 loci (seven of blood groups, nine of protein polymorphisms and nine microsatellites) analyzed on a total of 145 Uruguayan Creole horses. In this study, blood group and protein polymorphism variants that are considered to be breed markers of Spanish Pure-bred and Barb horses were detected in the Creole breed. Conversely, some microsatellites and protein polymorphisms alleles were found uniquely in the Creole horse. American horse breeds together with Barb and Arabian horses clearly formed a separate cluster from the Spanish pure-bred and Thoroughbred breeds, as shown by an UPGMA dendrogram based on Nei's standard genetic distance. Data in this study provided evidence for considerable genetic variation within Uruguayan Creole horses and of a distinctive breed profile. Both traits were most likely inherited from the XVIth century Spanish horses, more closely related to Barb than to Spanish Pure-bred.     

Major histocompatibility complex locus DRA polymorphism in the endangered Sorraia horse and related breeds

The major histocompatibility complex (MHC) genes play well‐defined roles in eliciting immune responses and combating infectious diseases. This genetic system is among the most polymorphic. The extent of genetic variation within a population has been directly correlated with fitness for many traits. The MHC class II locus DRA polymorphism was analysed in the endangered Sorraia horse, two other Portuguese and four New World horse breeds considered to be historically close to the Sorraia. Comparison of the Sorraia with other breeds demonstrated less MHC variation among Sorraia horses. If DRA polymorphism provides greater disease resistance, selective breeding to increase MHC polymorphism may increase fitness of this population.     

Segment-based coancestry,additive relationship and genetic variance within and between the Norwegian and the Swedish Fjord horse populations

ABSTRACT

The Fjord horse originates from Norway but forms a global population due to several small populations in foreign countries. There exists no information about the additive relationship and the genetic variance between these subpopulations. By collecting blood samples from Norwegian and Swedish Fjord horses, a sample of 311 Norwegian and 102 Swedish horses gave 485,918 SNPs available for analysis. Their inbreeding coefficients were calculated and compared to the pairwise coancestry and the shared genomic segments. The effective population size was almost similar with the two methods in the Norwegian Fjord horse population (63 and 71), but very different in the Swedish population (269 and 1136) and unprecise due to a much smaller number of observations. The study showed that coancestry from shared genomic segments can be used to estimate additive genetic relationship and genetic variation within and between the global populations of the Fjord horse.     

利用微卫星分析康西草原马匹种群的遗传多样性和群体结构

旨在揭示康西草原马匹的品种来源以及群体遗传结构。本研究采集了当地60匹健康状况良好、体重适中、3～6岁之间不区分公母马的血液样品,试验分为6个种群并以纯血马等4个国外马品种、1个我国地方马品种作为对照， 通过设计引物并提取基因组DNA结合微卫星扩增法对染色体上12个微卫星位点的等位基因进行检测，并对各种群遗传多样性参数、群体分化指数、瓶颈效应以及群体结构进行分析。试验按照群体个数为依据分为6组，每组1个重复，每个重复以该组样本量为标准。分析6个种群马的遗传参数结果表明，其中共检测到221个等位基因,平均Shannon指数为1.691；有效等位基因数(Ne)为3.649～5.397；期望杂合度(He)为0.681～0.798；观测杂合度(Ho)为0.632～0.780；平均多态信息含量(PIC)为0.643～0.772。这些结果表明，6个群体都具有很高的遗传多样性。通过微卫星DNA位点的连锁不平衡及哈迪-温伯格平衡分析发现， 大多数位点的P值都大于校正值(P=0.006 5)，表示较多位点都处于哈迪-温伯格及连锁平衡状态。各群体间群体分化系数(Fst)表明，群体间分化程度较低(Fst<0.15)。基于Nei’s遗传距离与地理距离之间Mantel相关性检验显示二者之间呈现正相关性(R=0.502)，但未达到显著水平(P=0.310)；各马匹种群瓶颈效应分析表明，P值极显著(P_(IAM)<0.01)说明该地区的马群体受到过瓶颈效应的影响且该马群体历史上数量有大规模的减少。通过 FCA因子分析、基于Nei’s标准遗传距离UPGMA树状图以及Structure贝叶斯聚类分析发现，该地区马种群与纯血马和温血马聚为一类，说明该种群最有可能主要来源于温血马和纯血马。这些结果揭示，康西草原地区马匹具有较高的遗传多样性，该地马群体的血统与纯血马和温血马相近，这为当地马遗传资源评估以及开发利用打下了坚实的基础。     

马生长激素基因多态性与体尺指标之间的关联性分析

为研究广西德保矮马、百色马和新疆伊犁马的生长激素(GH)基因多态性与体尺指标之间的相关性,本实验利用PCR-SSCP技术分析3个品种277匹马GH基因5′侧翼区、第3外显子、第4外显子、第5外显子的遗传多态性。结果表明:只有GH基因第5外显子出现多态性,表现为AA、BB和AB 3种基因型,其他片段未发现多态性。群体遗传学分析表明,等位基因A和B的基因频率相等;Hardy-Weinberg平衡检验显示,3个品种马均处于非平衡状态;它们的多态信息含量均为中度多态。统计分析表明,GH基因第5外显子的基因型与3个品种马体尺指标之间无显著相关性。     

Breed Differences in Dopamine Receptor D4 Gene (DRD4) in Horses

Genetic polymorphisms in genes related to neurotransmitters or hormones affect personality or behavioral traits in many animal species including humans. In domestic animals, the allele frequency of such genes has been reported to be different among breeds and it may account for breed differences in behavior. In this study, we investigated breed differences in horses in the dopamine receptor D4 gene (DRD4), which has been reported to affect horse personality. We collected samples from seven horse breeds including those native to Japan and Korea, and compared the sequence of the DRD4 exon3 region among these breeds. We found that there were two types of polymorphisms (VNTR and SNPs) in the exon3 region, and some of them seemed to be breed-specific. In addition, we found that the allele frequency of G292A, reported to be associated with horse personality, differed greatly between native Japanese horses and Thoroughbred horses. The frequency of the A allele which is associated with low curiosity and high vigilance, was much lower in native Japanese horses (Hokkaido, 0.03; Taishu, 0.08) than in Thoroughbreds (0.62). This difference may account for breed differences in personality or behavioral traits. Further studies of the function of these polymorphisms and their effect on behavior are indicated.     

The use of foal and studbook traits in the breeding programmes of Finnhorse and Standardbred trotters

Genetic correlations for body measurements and subjectively scored traits between foals and studbook horses were estimated using bivariate linear mixed models. Observations for nine foal and eleven studbook traits in Finnhorses on 6529 foals and 6596 studbook horses and in Standardbred trotters on 3069 foals and 2112 studbook horses were available from the Finnish horse breeding shows. The number of sires with progeny in both foal and studbook data was 203 in Finnhorse and 145 in Standardbred trotters. Estimates of heritability for body measurements in foals and studbook horses using univariate models were high in both breeds (0.41-0.84). Heritability estimates for subjectively scored traits using univariate models were generally higher for foals (0.08-0.46) than for studbook horses (0.06-0.21) in both breeds. Genetic correlations between foals and studbook horses for body measurements were highly positive ranging from 0.74 to 0.96 in Finnhorses and from 0.79 to 0.99 in Standardbred trotters. Low to highly positive genetic correlations between foals and studbook horses for subjectively scored traits were obtained in Finnhorse trotters, whereas in Standardbred trotters genetic correlations for subjectively scored traits varied from moderately negative to highly positive. Higher estimates of heritability for foal traits and generally high genetic correlations between the foal and studbook traits indicate that an early selection for conformation traits would be efficient in the breeding programmes.     

Mitochondrial Profiles of the East Bulgarian and the Pleven Horse Breeds

It is well known that horse breeding in Bulgaria is a cultural heritage in Bulgaria, dating from prehistoric and historic times. Until now, molecular data on Bulgarian horses from the plain regions of the country were not available. Therefore, for the first time, we have collected genetic information about some modern horse breeds from the plain regions in Bulgaria. A total of 50 horses originating from different families from two different breeds were investigated: the first one was the Pleven horse (n = 11, breeding in the Danubian Plain), and the second one was the East Bulgarian horse breed (n = 39, Shumen district). These breeds were genotyped according to the mitochondrial D-loop region. The results showed that the Pleven horse particularly carries the European haplogroup L (45.45%), followed by the Middle East haplogroup C (27.27%). In contrast to the Pleven horse, the East Bulgarian horse breed revealed almost equal frequencies of the European specific haplogroup L (33.33%) and the Central Asiatic haplogroup Q (35.90%). Analyses of these two horse breeds revealed a specific genetic profile, but it is obvious that the East Bulgarian horse showed an unusual, mixed profile—a massive admixture with the Asiatic-type haplogroup Q and a high haplogroup diversity. In conclusion, the differences in genetic structure of the two plain horse breeds may be explained with the various horse breeds involved in their formation.     

Morphometry and abnormalities of the feet of Kaimanawa feral horses in New Zealand

Objective The present study investigated the foot health of the Kaimanawa feral horse population and tested the hypotheses that horses would have a large range of foot morphology and that the incidence of foot abnormality would be significantly high. Procedures Abnormality was defined as a variation from what the two veterinarian assessors considered as optimal morphology and which was considered to impact negatively on the structure and/or function of the foot. Fifteen morphometric variables were measured on four calibrated photographic views of all four feet of 20 adult Kaimanawa feral horses. Four morphometric variables were measured from the lateromedial radiographs of the left forefoot of each horse. In addition, the study identified the incidence of gross abnormality observed on the photographs and radiographs of all 80 feet. Results There was a large variation between horses in the morphometric dimensions, indicating an inconsistent foot type. Mean hoof variables were outside the normal range recommended by veterinarians and hoof care providers; 35% of all feet had a long toe conformation and 15% had a mediolateral imbalance. Abnormalities included lateral (85% of horses) and dorsal (90% of horses) wall flares, presence of laminar rings (80% of horses) and bull-nose tip of the distal phalanx (75% of horses). Both hypotheses were therefore accepted. Conclusions The Kaimanawa feral horse population demonstrated a broad range of foot abnormalities and we propose that one reason for the questionable foot health and conformation is lack of abrasive wearing by the environment. In comparison with other feral horse populations in Australia and America there may be less pressure on the natural selection of the foot of the Kaimanawa horses by the forgiving environment of the Kaimanawa Ranges. Contrary to popular belief, the feral horse foot type should not be used as an ideal model for the domestic horse foot.     

Analysis of pedigree and conformation data to explain genetic variability of the horse breed Franches-Montagnes

Franches‐Montagnes is the only native horse breed in Switzerland, therefore special efforts should be made for ensuring its survival. The objectives of this study were to characterize the structure of this population as well as genetic variability with pedigree data, conformation traits and molecular markers. Studies were focused to clarify if this population is composed of a heavy‐ and a light‐type subpopulation. Extended pedigree records of 3‐year‐old stallions (n = 68) and mares (n = 108) were available. Evaluations of body conformation traits as well as pedigree data and molecular markers did not support the two‐subpopulation hypothesis. The generation interval ranged from 7.8 to 9.3 years. The complete generation equivalent was high (>12). The number of effective ancestors varied between 18.9 and 20.1, whereof 50% of the genetic variability was attributed to seven of them. Genetic contribution of Warmblood horses ranged from 36% to 42% and that of Coldblood horses from 4% to 6%. The average inbreeding coefficient reached 6%. Inbreeding effective population size was 114.5 when the average increase of the inbreeding coefficient per year since 1910 was taken. Our results suggest that bottleneck situations occurred because of selection of a small number of sire lines. Promotion of planned matings between parents that are less related is recommended in order to avoid a reduction of the genetic diversity.     

Genetic characterization of the endangered Kiso horse using 31 microsatellite DNAs

In order to contribute to conservation of the endangered Kiso horse, we clarified their genetic information using 31 microsatellite DNAs, and genotyped 125 horses, 83% of the existing breed. First, we clarified the current status of the horses. The horses were confirmed to have experienced rapid loss of population causing a bottleneck, and their effective population size was much smaller than their census size. Moreover, the number of alleles (6.3), observed heterozygosity (0.674), and expected heterozygosity (0.662) were in the same range as other endangered horses all over the world. Therefore, although their inbreeding level was not so severe (F(is): -0.017), the Kiso horse is surely one of the endangered. Second, we obtained genetic information of individuals. This information allowed us to understand the genetic distance of individuals, and might help in development of a reproductive strategy concerning the genetic distance between the mating pairs. Moreover, there appeared to be 4 subpopulations of Kiso horse, and this result was in good agreement with their historical background. Third, we confirmed that the parentage test for identification using the 31 microsatellite DNAs was highly reliable (probability of exclusion: 0.999999993). This identification increases the reliability of stud certification, and is also helpful for effective management. Understanding the genetic diversity within the population and the relationships among individuals is important to ensuring effective management for maintenance of genetic variation, and this study may help in conservation of the endangered Kiso horse.     

The legacy of Columbus in American horse populations assessed by microsatellite markers

Criollo horse populations descend from horses brought from the Iberian Peninsula over the period of colonization (15th to 17th century). They are spread throughout the Americas and have potentially undergone genetic hybridization with other breeds in the recent past. In this study, 25 autosomal microsatellites were genotyped in 50 horse breeds representing Criollo populations from 12 American countries (27 breeds), breeds from the Iberian Peninsula (19), one breed each from France and Morocco and two cosmopolitan horse breeds (Thoroughbred and Arabian). The genetic relationships among breeds identified five clusters: Celtic; Iberian; North American with Thoroughbred influence; most Colombian breeds; and nearly all other Criollo breeds. The group of “all other Criollo breeds” had the closest genetic relationship with breeds originating from the Iberian Peninsula, specifically with the Celtic group. For the whole set of Criollo breeds analysed, the estimated genetic contribution from other breeds was approximately 50%, 30% and 20% for the Celtic, Iberian and Arab-Thoroughbred groups, respectively. The spatial distribution of genetic diversity indicates that hotspots of genetic diversity are observed in populations from Colombia, Ecuador, Brazil, Paraguay and western United States, possibly indicating points of arrival and dispersion of Criollo horses in the American continent. These results indicate that Criollo breeds share a common ancestry, but that each breed has its own identity.     

Phylogenetic analysis of hindgut microbiota in Hokkaido native horses compared to light horses

The analysis of 16S rDNA sequence of bacteria in feces of Hokkaido native horses and light horses were performed to compare the hindgut microbiota between the two breeds. One hundred and four bacterial 16S rDNA clones (57 clones from four native horses and 47 clones from two light horses) were obtained. Only four sequences (3.8% of total sequences) showed 97% or more similarity to known species. The sequences were mainly affiliated with Cytophaga–Flavobacter–Bacteroides and low GC Gram‐positive bacteria (LGCGP). Proportion of LGCGP was higher in light horses. Other phyla including Verrucomicrobia, Spirochaetes and Archaea were detected only for native horses, suggesting high diversity of microbiota in native horses. In LGCGP, clusters related to known cellulolytic species were found only for native horses, while a cluster related to soluble sugar‐utilizing species was detected only for light horses. The library composition‐comparing software LIBSHUFF showed significant (P < 0.05) difference of fecal microbiota between the horse breeds. The number of Fibrobacter succinogenes‐related sequence and the frequency of detection of novel groups were found to be higher in native horses by selective amplification analysis. The results suggest that genetic diversity and population size of the F. succinogenes group are higher in the hindgut of native horses.     

蒙古家马和蒙古野马线粒体细胞色素b基因遗传多样性及系统进化分析

通过对马的线粒体细胞色素b（Cytb）基因序列的遗传变异分析,来探讨蒙古家马、蒙古野马和驴之间的亲缘关系和遗传多样性。对各类马和驴样本 mtDNA Cytb 基因进行PCR扩增和测序,运用现代分子生物学软件进行数据处理。结果共检测到15个单倍型,90个突变位点,单倍型多样性（Hd）为0.9356,核苷酸多样性（Pi）为0.01022,平均核苷酸差异数（K）为8.966,各群体间的遗传多样性相差不大。系统发育树和NETwork图中每一分支都至少包含2个群体的信息。4个类型蒙古家马总体上具有较高的遗传多态性,蒙古家马与蒙古野马之间的亲缘关系较近, 且出现数个分支,蒙古家马分布在各独立的进化支中,说明蒙古家马母系来源非常广泛或经过多次驯化。     

利用微卫星标记对宁强矮马和蒙古马遗传多样性的研究

通过8个微卫星位点的多态性检测对宁强矮马和蒙古马共83个个体进行了遗传多样性分析。计算各微卫星位点的等位基因频率、有效等位基因数(Ne)、多态信息含量(PIC)、杂合度(H)和F统计量。结果表明:8个微卫星座位中HMS2变异最大,HTG6变异最小;蒙古马的Ne、PIC、H的平均值均高于宁强马;总群体的平均遗传分化系数Fst为0.017(P<0.01),群体内近交系数Fis为0.171(P<0.01)。说明蒙古马和宁强矮马虽外形和性能上有很大差异,但在品种形成和进化上有着较近的遗传关系。     

Breeding or assisted reproduction? Relevance of the horse model applied to the conservation of endangered equids

Many wild equids are at present endangered in the wild. Concurrently, increased mechanization has pushed back the numbers of some old native horse breeds to levels that are no longer compatible with survival of the breed. Strong concerns arose in the last decade to preserve animal biodiversity, including that of rare horse breeds. Genome Resource Banking refers to the cryostorage of genetic material and is an approach for ex situ conservation, which should be applied in combination with in situ conservation programmes. In this review, we propose that, owing to the great reproductive similarity among the different members of the genus Equus, the domestic horse can be used to optimize cryopreservation and embryo production protocols for future application in wild equids. We will give this hypothesis a scientific underpinning by listing successful applications of epididymal sperm freezing, embryo freezing, intracytoplasmic sperm injection, oocyte vitrification and somatic cell nuclear transfer in domestic horses. Some ART fertilization methods may be performed with semen of very low quality or with oocytes obtained after the death of the mare.