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.     

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.     

Analysis of genetic diversity and the determination of relationships among western Mediterranean horse breeds using microsatellite markers

The distribution of genetic diversity and the genetic relationships among western Mediterranean horse breeds were investigated using microsatellite markers. The examined sample included seven Spanish and three Italian local horse breeds and populations, plus a Spanish Thoroughbred outgroup. The total number of animals examined was 682 (on average 62 animals per breed; range 20–122). The microsatellite marker set analysed provided 128 alleles (10.7 alleles per locus). Within‐breed genetic diversity was always high (>0.70), with breeds contributing about 8% of the total genetic variability. The mean molecular coancestry of the entire population examined was 0.205, Losino being the breed that contributed most. In addition to Nei's standard and Reynolds’ genetic distances, pair‐wise kinship distance and molecular coancestry were estimated. Remarkably similar breed rankings were obtained with all methods. Clustering analysis provided an accurate representation of the current genetic relationships among the breeds. Determining coancestry is useful for analysing genetic diversity distribution between and within breeds and provides a good framework for jointly analysing molecular markers and pedigree information. An integrated analysis was undertaken to obtain information on the population dynamics in western Mediterranean native horse populations, and to better determine conservation priorities.     

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.     

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.     

Genealogical analyses in open populations: the case of three Arab-derived Spanish horse breeds

This research assesses the genetic composition of three Arab-derived Spanish horse breeds as an example to highlight the major shortcomings related to genealogical analyses in open populations and to propose approaches useful to deal with this task. The studbooks of three Spanish Arab (SA)-derived horse breeds, Spanish Anglo-Arab (dAA), Hispano-Arab (dHA) and Spanish Sport Horse (dSSH) and those of their parental breeds SA, Spanish Purebred (SPB) and Thoroughbred (TB), totalling 211 754 individuals, were available. The genealogies of the dAA, dHA and dSSH were analysed not only using the corresponding studbook (breed exclusive dataset) but also including the genealogies of the founders from parental breeds (completed dataset). Coancestry analyses revealed that the present SA-derived populations share more genes with the Arab than with the other parental breeds. Effective population size was computed by accounting for migration rates to obtain an equivalent closed-population effective size (_eq N _e) of 39.2 for the dAA, 56.3 for dHA and 114.1 for dSSH. The essayed methodologies were useful for characterising populations involving migration. The consequences of the management of the analysed breeds are discussed. The results emphasize the need to include the complete genealogies of the individuals to attain reliable genealogical parameters.     

Comparison of plasma biochemical parameters in Thoroughbred and Purebred Arabian horses during the same-intensity exercise

The objective of the present study was to compare changes of blood parameters induced by the same work and performed in the same conditions in two racehorse breeds, Thoroughbred and Purebred Arabian. The effect of moderate-intensity exercise was studied in 20 stallions--ten Thoroughbreds, aged 2-3 years and the same number of Purebred Arabians, 3-4 years old. All the horses were administrated the same effort test consisting in 1200 m gallop at a speed typical of the daily training sessions. Three jugular venous blood samples were collected for each horse: at rest, just after the end of the gallop and after 30-minute rest. In the gathered blood, a hemoglobin (Hb) concentration was determined as well as plasma level of glucose (Glc), triacylglycerols (TG), glycerol, free fatty acids (FFA), total plasma proteins (TP) and the activity of creatine kinase (CK), lactate dehydrogenase (LDH) and aspartate aminotransferase (AST). In the Arabian horses, an increase in levels of TP, glycerol, FFA and CK activity measured just after exercise was higher than that in Thoroughbreds. Similarly, after a 30-minute rest, a post-exercise rise of TP, AST, glycerol and FFA proved to be higher in the Arabian horses compared to that in the Thoroughbreds. Only TG plasma concentration measured 30 minutes following the effort was significantly lower in the Arabian horses than in Thoroughbreds. It can be concluded that the Thoroughbred horses adapted better to the effort test applied in this study as compared to the Purebred Arabian horses. The parameters related to lipid metabolism proved to be the most sensitive indicators of breed differences in relation to moderate-intensity exercise.     

Genetic structure of Norwegian horse breeds

The evaluation of the genetic structure of four native Norwegian horse breeds was investigated using 35 genetic markers including nine biochemical loci and 26 microsatellites. The proportion of genetic variation measured as heterozygosity and the number of alleles indicated high variation in Fjord Horse and Coldblooded Trotter, but somewhat reduced variation in Døle Horse. No clear signs of bottleneck effects were found in either of the breeds, but significant inbreeding estimates in Nordland/Lyngen Horse may indicate subdivision of this breed. Significant population differentiation was detected between all breeds, also between the recently diverged Døle Horse and Coldblooded Trotter. The principal component analysis supports the close genetic relationship between these two breeds.     

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.     

Genomic polymorphism in Uruguayan Creole cattle using RAPD and microsatellite markers

Uruguayan Creole cattle inhabit areas that cannot sustain conventional farming. They have adapted to fragile environments and are influenced only by natural selection. In this study, random amplified polymorphic DNA (RAPD) and microsatellite (MS) markers were used to analyse Creole cattle genome polymorphism. A comparative analysis using the RAPD technique was performed in pooled DNA of three cattle breeds (Holstein Friesian, Creole and Hereford) in order to evaluate their amplification patterns. A primary screening of RAPD primers allowed us to select and use those with higher percentage of GC base composition. A total of 215 loci ranging between 300 and 2500 bp were amplified. Bandsharing frequency (BSF) among breeds showed that less related fingerprints were observed between Creole and Hereford cattle (0.77), while the highest similarity frequency corresponded to Holstein Friesian compared to Hereford (0.81). Specific RAPD bands were identified in the three DNA pools and they were tested in every individual of each breed. It may be possible to isolate and sequence these bands to create breed-specific molecular markers. The identification of multiple alleles of the MSCYP 21 in Creole cattle with an heterozygosity of He = 0.846 supported the variability of this genetic resource. The use of molecular markers such as RAPD s and microsatellites is proposed to establish genetic distance among American Creole cattle and possibly related ancestral Iberian breeds.     

Genetic Diversity and Population Structure of Arabian Horse Populations Using Microsatellite Markers

Understanding the genetic diversity and the relationships among the show Arabian horse populations is a current issue for breeders and professionals. This study aimed to define the relationship among the Desert breed, the Straight Egyptian, and the Polish Arabian populations by considering the historical background of their origin and to verify their genetic diversity. All selected samples were related to Arabian show activities. One hundred forty four hair samples were collected from horses at stud farms having notoriety in the breeding of Arabians from different geographic regions. A set of 17 microsatellites markers for parentage control were used for genotyping. Genetic diversity among and between these populations were evaluated using several statistical methods. All the microsatellites were informative and the marker set analyzed provided 145 alleles. The average number of alleles per locus was 6.52, 6.35, and 7 for the Desert breed, Straight Egyptian, and Polish Arabian, respectively. The high genetic diversity observed within the three populations (0.63-0.71) was associated with a high number of effective alleles. Desert breed and Polish Arabian populations appeared the closest, whereas the Egyptian population was more distant. The significant positive inbreeding coefficient F_IS found in Desert breed, Straight Egyptian, and Polish Arabian horses (0.09, 0.14, and 0.11, respectively) confirmed the deficit of heterozygosity observed in these populations. These results suggested that the three populations have high levels of gene flow or share the same origin and have a recent divergence. This study may highlight the risk of the loss of gene diversity in these populations and help to implement appropriate breeding programs to preserve genetic diversity.     

Phylogenetic relationships of German heavy draught horse breeds inferred from mitochondrial DNA D-loop variation

Summary We analysed a 610-bp mitochondrial (mt)DNA D-loop fragment in a sample of German draught horse breeds and compared the polymorphic sites with sequences from Arabian, Hanoverian, Exmoor, Icelandic, Sorraia and Przewalski's Horses as well as with Suffolk, Shire and Belgian horses. In a total of 65 horses, 70 polymorphic sites representing 47 haplotypes were observed. The average percentage of polymorphic sites was 11.5% for the mtDNA fragment analysed. In the nine different draught horse breeds including South German, Mecklenburg, Saxon Thuringa coldblood, Rhenisch German, Schleswig Draught Horse, Black Forest Horse, Shire, Suffolk and Belgian, 61 polymorphic sites and 24 haplotypes were found. The phylogenetic analysis failed to show monophyletic groups for the draught horses. The analysis indicated that the draught horse populations investigated consist of diverse genetic groups with respect to their maternal lineage.     

Polysaccharide storage myopathy in Morgan, Arabian, and Standardbred related horses and Welsh-cross ponies

Polysaccharide storage myopathy is an equine neuromuscular disorder characterized by accumulation of glycogen-related polysaccharide inclusions within skeletal muscle fibers. The pathologic criteria for diagnosis of this disorder are somewhat controversial; however, periodic acid-Schiff-positive, amylase-resistant inclusions are considered pathognomonic. Although these inclusions are most often found in affected horses related to the Quarter Horse, draft horse, and Warmblood breeds, this report describes these characteristic inclusions in muscle of five horses from nonrelated breeds (two Morgans, one Arabian, one Arabian x Thoroughbred, and one Standardbred) and two Welsh cross ponies. Affected horses had histories of recurrent exertional rhabdomyolysis, and one developed progressive weakness leading to increased recumbency. The affected ponies were part of an unrelated research project and had no apparent clinical signs.     

Genetic Characterization of Mangalarga Marchador Breed Horses Based on Microsatellite Molecular Markers

The Mangalarga Marchador (MM) horse breed has expressive importance in the Brazilian economy. Thus, the aim of this study was to investigate diversity in the MM breed. A database with a total of 3,193 genotyped horses was used (MM, n = 2,829; Andalusian – AND, n = 67; Pure Blood Lusitano – LUS, n = 43; English Thoroughbred – THO, n = 54; Arabian – ARA, n = 99; Campolina – CAM, n = 61; and Mangalarga – MAN, n = 40) for 13 microsatellite. Diversity parameters were estimates, such as mean number of alleles (Nm_a) and the number of rare alleles (AR), expected heterozygosity (He), F statistics, genetic distances, Hardy-Weinberg equilibrium test (HWE), population structure, and others. The Nm_a was 10.85, the AR was prevalent in the MM, and the He was 0.7402. In MM, the values of Fis (−0.0195), Fit (0.0566), Fst (0.0748), and deviations of HWE were observed. The genetic distances of the ARA and THO breeds with the other breeds were greater than the distances between the Brazilian breeds and between these and the breeds in the Iberian Peninsula. The population structure indicated that MM was substructured, yet there were some more genetically defined breeding farms. The genetic diversity is satisfactory for MM conservation, but the population is substructure, and parameters indicate moderate gene flow and the existence, though few, of crosses with other horse breeds. Immediate implementation of a genetic breeding program is required, especially seeking to conserve the structure of the MM breed as a well-defined genetic entity.     

Epidemiological features of 5009 cases of equine cryptorchism

Data from 16 North American veterinary university teaching facilities, 5009 cryptorchid horses, were analysed using relative risk methodology. In five breeds (Thoroughbred, Standardbred, Morgan, Tennessee Walking horse and Arabian), cryptorchism was diagnosed significantly (P less than 0.05) less frequently than expected by their representation in the hospital population. Three breeds, (Percheron, American Saddle horse and American Quarterhorse), plus ponies (as a group) and crossbred horses were significantly over-represented within the series. The over-representation of Quarter-horses was evident at each veterinary teaching facility. Other developmental defects and testicular tumours diagnosed in cryptorchid horses were presented. Analysis of the frequency of diagnosis of second anomalies among cryptorchids showed testicular hypoplasia to occur significantly more often, and inguinal and umbilical hernias less often, than expected (P less than 0.05).     

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。     

Palmar/Plantar Process Fractures of the Proximal Phalanx In 15 Horses

Fifteen fractures of the palmar or plantar process of the proximal phalanx in 15 horses are described. Ten were articular and five were nonarticular. Two modes of therapy were used depending on the location of the fracture. Nonarticular fractures were treated with stall rest or reduction in exercise, and articular fractures were treated with either surgical removal or internal fixation of the fragment. All horses that were operated on were sound within 6 months after surgery. Follow-up information was obtained on all horses through client questionnaire and race records in the racing breeds (Standardbred and Thoroughbred) and client questionnaire alone is the nonracing breeds (Quarterhorse and Arabian). Fourteen horses returned to an equal or better level of performance than before injury; one horse returned to a lower level of performance.     

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.     

Humoral immune responses in the horse after intrathecal challenge with ovalbumin

Background : Diagnosis of neuro-inflammatory conditions in the horse can be challenging. Current methods include evaluation of cerebrospinal fluid (CSF) for inflammation and determination of specific antibody status. The antibody index (AI) and Goldman-Witmer coefficient (C-value) can be used to aid in the interpretation.
Hypothesis : The null hypothesis to be tested was that the AI and C-values do not change in horses with neuro-inflammation.
Animals : Twelve horses of various ages (3–17 years) and breeds (Thoroughbred, Thoroughbred cross, draft, and Arabian) were included in the study.
Methods : The study was designed as a prospective randomized study. All horses were immunized with ovalbumin in adjuvant, twice. Horses of Group 1 then were challenged by intrathecal (IT) injection of ovalbumin, whereas horses of Group 2 were challenged IM. The AI and C-values for ovalbumin and equine herpesvirus were calculated.
Results : The AI for ovalbumin increased up to 5.92 in horses after intrathecal challenge, and remained normal (<1) in horses challenged IM. The C-value for ovalbumin reached a peak of 7.48, whereas for equine herpesvirus it achieved a value of 2.69. The changes in ovalbumin C-value and AI were significantly different between days 20 and 30 in horses after intrathecal challenge at day 20 ( P = .002 and .0005, respectively).
Conclusions and Clinical Importance : The results confirm the value of the AI and C-value in the evaluation of neuro-inflammation in the horse.     

Detailed insight into genetic diversity of the Old Kladruber horse substructure in comparison to the Lipizzan breed

The aim of the present study was to analyse the genetic subdivision of the Old Kladruber horse population compared to the historically close Lipizzan breed and to estimate genetic relatedness between them. A set of 13 microsatellites was used for genotyping a total of 270 Old Kladruber horses representing grey and black colour varieties and 418 Lipizzan horses from Slovak and Slovenian studs. The proportion of obtained heterozygosity indicates no major loss of genetic diversity within them. At the individual level across analysed populations, the formation of clusters in respect to breed’s origin and particular studs was observed. The Wright’s F_ST and genetic distances indicated genetic segregation of both colour varieties at the intraspecific level of the Kladruber breed. Moreover, the membership probability outputs showed that the frequencies of alleles varied across the three main regions represented by both Old Kladruber varieties and Lipizzan, depending on breeding history and strategy of studs.