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Topic:Genomics

Genomics in horses involves the study and analysis of the horse genome to understand genetic variations and their implications for equine health, performance, and breeding. This field encompasses the identification and mapping of genes associated with specific traits, diseases, and conditions in horses. Techniques such as whole-genome sequencing and genome-wide association studies (GWAS) are employed to explore genetic diversity and inheritance patterns among different horse breeds. Genomics provides insights into hereditary disorders, informs selective breeding practices, and aids in the development of personalized veterinary care. This page compiles peer-reviewed research studies and scholarly articles that explore the applications, methodologies, and findings of genomic research in equine science.
Modulation of stress and immune response by Amblyomin-X results in tumor cell death in a horse melanoma model.
Scientific reports    April 14, 2020   Volume 10, Issue 1 6388 doi: 10.1038/s41598-020-63275-2
Lichtenstein F, Iqbal A, de Lima Will SEA, Bosch RV, DeOcesano-Pereira C, Goldfeder MB, Chammas R, Trufen CEM, Morais KLP, de Souza JG, Natalino RJM....We have investigated Amblyomin-X-treated horse melanomas to better understand its mode of action through transcriptome analysis and the in vivo model. Amblyomin-X is a Kunitz-type homologous protein that selectively leads to the death of tumor cells via ER stress and apoptosis, currently under investigation as a new drug candidate for cancer treatment. Melanomas are immunogenic tumors, and a better understanding of the immune responses is warranted. Equine melanomas are spontaneous and not so aggressive as human melanomas are, as this study shows that the in vivo treatment of encapsulated hors...
Major inconsistencies of inferred population genetic structure estimated in a large set of domestic horse breeds using microsatellites.
Ecology and evolution    April 12, 2020   Volume 10, Issue 10 4261-4279 doi: 10.1002/ece3.6195
Funk SM, Guedaoura S, Juras R, Raziq A, Landolsi F, Luís C, Martínez AM, Musa Mayaki A, Mujica F, Oom MDM, Ouragh L, Stranger YM, Vega-Pla JL....STRUCTURE remains the most applied software aimed at recovering the true, but unknown, population structure from microsatellite or other genetic markers. About 30% of structure-based studies could not be reproduced (, 21, 2012, 4925). Here we use a large set of data from 2,323 horses from 93 domestic breeds plus the Przewalski horse, typed at 15 microsatellites, to evaluate how program settings impact the estimation of the optimal number of population clusters that best describe the observed data. Domestic horses are suited as a test case as there is extensive background knowledge on the his...
Whole genome detection of sequence and structural polymorphism in six diverse horses.
PloS one    April 9, 2020   Volume 15, Issue 4 e0230899 doi: 10.1371/journal.pone.0230899
Al Abri MA, Holl HM, Kalla SE, Sutter NB, Brooks SA.The domesticated horse has played a unique role in human history, serving not just as a source of animal protein, but also as a catalyst for long-distance migration and military conquest. As a result, the horse developed unique physiological adaptations to meet the demands of both their climatic environment and their relationship with man. Completed in 2009, the first domesticated horse reference genome assembly (EquCab 2.0) produced most of the publicly available genetic variations annotations in this species. Yet, there are around 400 geographically and physiologically diverse breeds of hors...
Gallop Racing Shifts Mature mRNA towards Introns: Does Exercise-Induced Stress Enhance Genome Plasticity?
Genes    April 9, 2020   Volume 11, Issue 4 410 doi: 10.3390/genes11040410
Cappelli K, Mecocci S, Gioiosa S, Giontella A, Silvestrelli M, Cherchi R, Valentini A, Chillemi G, Capomaccio S.Physical exercise is universally recognized as stressful. Among the "sport species", the horse is probably the most appropriate model for investigating the genomic response to stress due to the homogeneity of its genetic background. The aim of this work is to dissect the whole transcription modulation in Peripheral Blood Mononuclear Cells (PBMCs) after exercise with a time course framework focusing on unexplored regions related to introns and intergenic portions. PBMCs NGS from five 3 year old Sardinian Anglo-Arab racehorses collected at rest and after a 2000 m race was performed. Apart from d...
Animal domestication in the era of ancient genomics.
Nature reviews. Genetics    April 7, 2020   Volume 21, Issue 8 449-460 doi: 10.1038/s41576-020-0225-0
Frantz LAF, Bradley DG, Larson G, Orlando L.The domestication of animals led to a major shift in human subsistence patterns, from a hunter-gatherer to a sedentary agricultural lifestyle, which ultimately resulted in the development of complex societies. Over the past 15,000 years, the phenotype and genotype of multiple animal species, such as dogs, pigs, sheep, goats, cattle and horses, have been substantially altered during their adaptation to the human niche. Recent methodological innovations, such as improved ancient DNA extraction methods and next-generation sequencing, have enabled the sequencing of whole ancient genomes. These gen...
Genome-wide association study for host genetic factors associated with equine herpesvirus type-1 induced myeloencephalopathy.
Equine veterinary journal    April 6, 2020   Volume 52, Issue 6 794-798 doi: 10.1111/evj.13261
Dunuwille WMB, YousefiMashouf N, Balasuriya UBR, Pusterla N, Bailey E.Equid herpesvirus (EHV-1) infections in horses can lead to equine herpesvirus myeloencephalopathy (EHM), characterised by neurological clinical signs. The sporadic occurrence of the disease in horse herds suggests a host genetic component. A recent study reported an association between the occurrence of EHM and genetic markers on horse chromosome 6 (ECA6). Objective: To investigate the association of EHM with genetic host factors, especially with reference to the association reported for ECA6. Methods: Genome-wide association study (GWAS) was conducted based on 94 horses that had EHV-1 infecti...
A De Novo MITF Deletion Explains a Novel Splashed White Phenotype in an American Paint Horse.
The Journal of heredity    April 4, 2020   Volume 111, Issue 3 287-293 doi: 10.1093/jhered/esaa009
Magdesian KG, Tanaka J, Bellone RR.Splashed white is a coat color pattern in horses characterized by extensive white patterning on the legs, belly, and face often accompanied by blue eyes and deafness. Three mutations in microphthalmia-associated transcription factor (MITF) and two mutations in Paired Box 3 (PAX3) have been identified that explain splashed white patterns (SW1-SW5). An American Paint Horse stallion with a splashed white phenotype and blue eyes, whose parents were not white patterned, was negative for the 5 known splashed white variants and other known white spotting alleles. This novel splashed white phenotype (...
Characterization and phylogenetic analysis of the first complete mitochondrial genome of Cylicocyclus radiatus.
Veterinary parasitology    April 3, 2020   Volume 281 109097 doi: 10.1016/j.vetpar.2020.109097
Hu L, Zhang M, Sun Y, Bu Y.Cylicocyclus spp. are common parasites in the intestine of equine animals. Hosts infected by these strongylid nematodes might face emaciation, anemia, and even death, all of which lead to significant economic losses. In the present study, the complete mitochondrial genome (mitogenome) of Cylicocyclus radiatus, the type species of the genus Cylicocyclus, was sequenced and characterized for the first time using next-generation sequencing technology. Results show that the mitogenome of C. radiatus is a double-stranded 13,836 bp circular molecule, containing 12 protein-coding genes, 22 tRNA genes,...
Metagenomic sequencing of clinical samples reveals a single widespread clone of Lawsonia intracellularis responsible for porcine proliferative enteropathy.
Microbial genomics    April 2, 2020   Volume 6, Issue 4 doi: 10.1099/mgen.0.000358
Bengtsson RJ, Wee BA, Yebra G, Bacigalupe R, Watson E, Guedes RMC, Jacobson M, Stadejek T, Archibald AL, Fitzgerald JR, Ait-Ali T.Lawsonia intracellularis is a Gram-negative obligate intracellular bacterium that is the aetiological agent of proliferative enteropathy (PE), a common intestinal disease of major economic importance in pigs and other animal species. To date, progress in understanding the biology of L. intracellularis for improved disease control has been hampered by the inability to culture the organism in vitro. In particular, our understanding of the genomic diversity and population structure of clinical L. intercellularis is very limited. Here, we utilized a metagenomic shotgun approach to directly sequenc...
Comparative analysis of CpG islands in equine infectious anemia virus strains.
Virus genes    April 1, 2020   Volume 56, Issue 3 339-346 doi: 10.1007/s11262-020-01749-1
Liu Q, Yu YY, Wang HY.Increasing evidence suggests that DNA methylation has key roles in the replication of retroviruses, including lentiviruses, and pathogenesis of diseases. However, the precise characteristics of CpG islands are not known for many retroviruses. In this study, we compared the distribution of CpG islands among strains of equine infectious anemia virus (EIAV), a lentivirus in the family Retroviridae and a model for HIV research. We identified CpG islands in 32 full-length EIAV genomic sequences obtained from the GenBank database using MethPrimer. Only one CpG island, from 100 to 120 bp, was identi...
Identification of genes required for the fitness of Streptococcus equi subsp. equi in whole equine blood and hydrogen peroxide.
Microbial genomics    March 31, 2020   Volume 6, Issue 4 e000362 doi: 10.1099/mgen.0.000362
Charbonneau ARL, Taylor E, Mitchell CJ, Robinson C, Cain AK, Leigh JA, Maskell DJ, Waller AS.The availability of next-generation sequencing techniques provides an unprecedented opportunity for the assignment of gene function. subspecies is the causative agent of strangles in horses, one of the most prevalent and important diseases of equids worldwide. However, the live attenuated vaccines that are utilized to control this disease cause adverse reactions in some animals. Here, we employ transposon-directed insertion-site sequencing (TraDIS) to identify genes that are required for the fitness of in whole equine blood or in the presence of HO to model selective pressures exerted by th...
Analysis of the miRNA transcriptome during testicular development and spermatogenesis of the Mongolian horse.
Reproduction, fertility, and development    March 27, 2020   Volume 32, Issue 6 582-593 doi: 10.1071/RD19133
Li B, He X, Zhao Y, Bai D, Li D, Zhou Z, Manglai D.Numerous studies have shown that microRNAs (miRNAs) are essential for testicular development and spermatogenesis. In order to further characterise these physiological processes, three immature and three mature testes of the Mongolian horse were collected and six libraries were established. Using small RNA sequencing technology, 531 mature miRNAs were identified, including 46 novel miRNAs without previously ascribed functions. Among the 531 miRNAs, 421 were expressed in both immature and mature libraries, 65 miRNAs were found solely in immature testis libraries and 45 miRNAs were found solely i...
Genetic Differentiation of the Two Types of Polish Cold-blooded Horses Included in the National Conservation Program.
Animals : an open access journal from MDPI    March 24, 2020   Volume 10, Issue 3 542 doi: 10.3390/ani10030542
Gurgul A, Jasielczuk I, Semik-Gurgul E, Pawlina-Tyszko K, Szmatoła T, Polak G, Bugno-Poniewierska M.The current role of the horse as a companion animal resulted in a decrease of interest in breeding and usage of draft horses. This meant that the population of cold-blooded horses in Poland has been dramatically reduced during the last decades. To avoid impoverishment of the gene pool of the local horse population, a conservation program was established which involves draft horses and other local horse breeds. The draft horses bred in Poland can be subdivided in a few horse types of which the most widespread and consolidated are Sztumski and Sokólski horses. These two subpopulations are pheno...
Complete genome analysis of Glutamicibacter creatinolyticus from mare abscess and comparative genomics provide insight of diversity and adaptation for Glutamicibacter.
Gene    March 17, 2020   Volume 741 144566 doi: 10.1016/j.gene.2020.144566
Santos RG, Hurtado R, Gomes LGR, Profeta R, Rifici C, Attili AR, Spier SJ, Mazzullo G, Morais-Rodrigues F, Gomide ACP, Brenig B, Gala-García A....Bacteria of the genusGlutamicibacterare considered ubiquitous because they can be found in soil, water and air. They have already been isolated from different habitats, including different types of soil, clinical samples, cheese and plants. Glutamicibacter creatinolyticus is a Gram-positive bacterium important to various biotechnological processes, however, as a pathogen it is associated to urinary tract infections and bacteremia. Recently,Glutamicibacter creatinolyticusLGCM 259 was isolated from a mare, which displayed several diffuse subcutaneous nodules with heavy vascularization. In this s...
Gut Microbiomes of Endangered Przewalski’s Horse Populations in Short- and Long-Term Captivity: Implication for Species Reintroduction Based on the Soft-Release Strategy.
Frontiers in microbiology    March 12, 2020   Volume 11 363 doi: 10.3389/fmicb.2020.00363
Tang L, Li Y, Srivathsan A, Gao Y, Li K, Hu D, Zhang D.Captivity maybe the only choice for survival of many endangered vertebrates, and understanding its broad effects is important for animal management and conservation, including breeding endangered species for subsequent release. Extreme environmental changes during captivity may influence survival ability in the wild. Captivity decreases gut bacterial diversity in a wide range of animals. However, most studies directly compare animals living in captivity with those in the wild, and there is a lack of understanding of effects of gradient shift in lifestyle during species reintroduction based on ...
Complete Genome Sequence of Mycoplasma felis Strain Myco-2, Isolated from an Equine Tracheal Wash Sample in Japan.
Microbiology resource announcements    February 27, 2020   Volume 9, Issue 9 doi: 10.1128/MRA.00057-20
Kinoshita Y, Niwa H, Uchida-Fujii E, Nukada T.Mycoplasma felis causes conjunctivitis in cats and respiratory diseases in horses. We report here the complete genome assembly of equine Mycoplasma felis strain Myco-2, which was isolated from an ill horse in Japan.
The effect of different antimicrobial treatment regimens on the faecal shedding of ESBL-producing Escherichia coli in horses.
Veterinary microbiology    February 26, 2020   Volume 243 108617 doi: 10.1016/j.vetmic.2020.108617
Schoster A, van Spijk JN, Damborg P, Moodley A, Kirchgaessner C, Hartnack S, Schmitt S.Fourth-generation cephalosporins can select for extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae in horses, but it is unknown to what extent this occurs compared to penicillin/gentamicin combination treatment. The objective was to evaluate the effect of different antimicrobial treatments on faecal shedding and diversity of ESBL-producing Escherichia coli (ESBL-EC) in horses. Upon hospital admission, 86 horses in need of antimicrobial treatment or prophylaxis were randomly allocated to receive penicillin and gentamicin (PG) or cefquinome (CEF). Untreated horses were included a...
SNPs analysis of height traits in Ningqiang pony.
Animal biotechnology    February 24, 2020   Volume 32, Issue 5 566-572 doi: 10.1080/10495398.2020.1728288
Bai H, Lu H, Wang L, Wang S, Zeng W, Zhang T.Ningqiang pony is one of the five pony breeds in China and is listed as a rare species. It is, therefore, meaningful to conduct stature research on this breed for its efficient reproduction, utilization and protection. This study is based on four SNP variants of adjacent LCORL/NCAPG, HMGA2, ZFAT and LASP1 genes, which have been found to be associated with horse height, and then, the 4 SNP loci in 22 Ningqiang ponies were analyzed. The results showed that there were genetic variations at the four loci in Ningqiang pony, there was a SNP on LCORL/NCAPG, HMGA2 and ZFAT gene all including T allele ...
Comparative Transcriptomics Analysis of Testicular miRNA from Cryptorchid and Normal Horses.
Animals : an open access journal from MDPI    February 21, 2020   Volume 10, Issue 2 doi: 10.3390/ani10020338
Han H, Chen Q, Gao Y, Li J, Li W, Dang R, Lei C.In the biological process of testicular spermatogenesis, the expression and interaction of many genes are regulated by microRNAs (miRNAs). However, comparisons of miRNA expression between descended testes (DTs) and undescended testes (UDTs) are rarely done in horses. In this study, we selected two UDTs (CKY2b and GU4b) from Chakouyi (CKY) and Guanzhong (GU) horses and eight DTs (GU1-3, CKY1, CKY3, CKY2a, GU4a, and GU5). Three groups were compared to evaluate expression patterns of testicular miRNA in stallion testes. Group 1 compared normal CKY horses and GU horses (CKY1 and CKY3 vs. GU1-3). G...
Identification of animal hosts of Fort Sherman virus, a New World zoonotic orthobunyavirus.
Transboundary and emerging diseases    February 16, 2020   Volume 67, Issue 4 1433-1441 doi: 10.1111/tbed.13499
de Oliveira Filho EF, Carneiro IO, Ribas JRL, Fischer C, Marklewitz M, Junglen S, Netto EM, Franke CR, Drexler JF.An orthobunyavirus termed Fort Sherman virus (FSV) was isolated in 1985 from a febrile US soldier in Panama, yet potential animal reservoirs remained unknown. We investigated sera from 192 clinically healthy peri-domestic animals sampled in northeastern Brazil during 2014-2018 by broadly reactive RT-PCR for orthobunyavirus RNA, including 50 cattle, 57 sheep, 35 goats and 50 horses. One horse sampled in 2018 was positive (0.5%; 95% CI, 0.01-3.2) at 6.2 × 103 viral RNA copies/mL. Genomic comparisons following virus isolation in Vero cells and deep sequencing revealed high identity of translat...
Selection in Australian Thoroughbred horses acts on a locus associated with early two-year old speed.
PloS one    February 12, 2020   Volume 15, Issue 2 e0227212 doi: 10.1371/journal.pone.0227212
Han H, McGivney BA, Farries G, Katz LM, MacHugh DE, Randhawa IAS, Hill EW.Thoroughbred horse racing is a global sport with major hubs in Europe, North America, Australasia and Japan. Regional preferences for certain traits have resulted in phenotypic variation that may result from adaptation to the local racing ecosystem. Here, we test the hypothesis that genes selected for regional phenotypic variation may be identified by analysis of selection signatures in pan-genomic SNP genotype data. Comparing Australian to non-Australian Thoroughbred horses (n = 99), the most highly differentiated loci in a composite selection signals (CSS) analysis were on ECA6 (34.75-34.85 ...
X chromosome aneuploidy and micronuclei in fertile mares.
Theriogenology    February 10, 2020   Volume 147 34-38 doi: 10.1016/j.theriogenology.2020.02.020
Pieńkowska-Schelling A, Kaul A, Schelling C.Abnormalities of chromosomes are an important and well documented cause of disorders of sexual development, fertility problems and congenital anomalies in mammals. Detection of low-level 63,X/64,XX mosaicism during routine cytogenetic evaluation is a challenge because its clinical significance is not yet fully clear. This study describes the prevalence and levels of 63,X mosaicism for a cohort of fertile mares and compares the results with eight problem mares for which no clinical cause of sub-fertility was found. The study design allowed for the analysis of micronuclei which are biomarkers of...
A Genome-Wide Association Analysis in Noriker Horses Identifies a SNP Associated With Roan Coat Color.
Journal of equine veterinary science    February 5, 2020   Volume 88 102950 doi: 10.1016/j.jevs.2020.102950
Grilz-Seger G, Reiter S, Neuditschko M, Wallner B, Rieder S, Leeb T, Jagannathan V, Mesarič M, Cotman M, Pausch H, Lindgren G, Velie B, Horna M....The roan coat color in horses is characterized by dispersed white hair and dark points. This phenotype segregates in a broad range of horse breeds, while the underlying genetic background is still unknown. Previous studies mapped the roan locus to the KIT gene on equine chromosome 3 (ECA3). However, this association could not be validated across different horse breeds. Performing a genome-wide association analysis (GWAS) in Noriker horses, we identified a single nucleotide polymorphism (SNP) (ECA3:g.79,543.439 A > G) in the intron 17 of the KIT gene. The G -allele of the top associated SNP...
Methyl-coenzyme M Reductase (MCR) Receptor as Potential Drug Target for Inhibiting Methanogenesis in Horses Using Moringa oleifera L.: An in Silico Docking Study.
Journal of equine veterinary science    February 5, 2020   Volume 88 102949 doi: 10.1016/j.jevs.2020.102949
Khusro A, Aarti C, Salem AZM, Pliego AB, Rivas-Caceres RR.Methane (CH) emission from nonruminant livestock, particularly equines, is a colossal burden for veterinarians worldwide. In view of this, the present context was investigated to predict the antimethanogenic attributes of Moringa oleifera L. associated phytocomponents by targeting methyl-coenzyme M reductase (MCR) receptor in horses using in silico tools. Initially, the pharmacokinetics and ADME (absorption, distribution, metabolism, and excretion) properties of 26 phytocomponents were analyzed using Lipinski's rule of five and Swiss ADME tool, respectively. Among all the tested phytocomponent...
Heterogeneity of antimicrobial susceptibility testing results for sulfamethoxazole/trimethoprim obtained from clinical equine Staphylococcus aureus isolates using different methods.
Veterinary microbiology    February 1, 2020   Volume 242 108600 doi: 10.1016/j.vetmic.2020.108600
Scholtzek AD, Hanke D, Eichhorn I, Walther B, Lübke-Becker A, van Duijkeren E, Köck R, Schwarz S, Feßler AT.Based on antimicrobial susceptibility testing (AST), correct classifications as susceptible, intermediate or resistant are challenging for some antimicrobial agent-bacterial species combinations. In this study, we investigated 19 equine Staphylococcus aureus isolates for their susceptibility to the combination sulfamethoxazole/trimethoprim (SXT) by using broth microdilution (BMD), agar disk diffusion (DD) and automated test systems. To elucidate the presence of the corresponding genetic resistance properties among the isolates, whole genome sequence analysis was performed and the genomes were ...
ASlive: a database for alternative splicing atlas in livestock animals.
BMC genomics    January 30, 2020   Volume 21, Issue 1 97 doi: 10.1186/s12864-020-6472-9
Liu J, Tan S, Huang S, Huang W.Alternative splicing is an important biological process whose precision must be tightly regulated during growth and development. Although there are species, disease (e.g. cancers), or study specific databases available in many organisms, no database exists in livestock animals specifically tailored for alternative splicing. Methods: We present in this study the development and implementation of a database for alternative splicing atlas in livestock animals (ASlive.org). Using publicly available RNASeq data sets across many tissues, cell types, and biological conditions totaling 28.6 T bases,...
Mapping the bacterial ecology on the phyllosphere of dry and post soaked grass hay for horses.
PloS one    January 27, 2020   Volume 15, Issue 1 e0227151 doi: 10.1371/journal.pone.0227151
Moore-Colyer M, Longland A, Harris P, Zeef L, Crosthwaite S.Soaking hay fodder to reduce dust and soluble carbohydrate (WSC) contents prior to feeding is common practice among horse owners. Soaking can increase bacteria load in hay but no information exists on how this process alters the bacteria profile, which could pose a health risk or digestive challenge, to horses by introducing foreign bacteria into the gastrointestinal tract and so altering the normal profile. The current objectives were to map the bacterial profile of 3 different hays and determine how soaking alters this with the aim of improving best practice when feeding stabled horses. A Pe...
Genetic diversity within and between British and Irish breeds: The maternal and paternal history of native ponies.
Ecology and evolution    January 27, 2020   Volume 10, Issue 3 1352-1367 doi: 10.1002/ece3.5989
Winton CL, McMahon R, Hegarty MJ, McEwan NR, Davies-Morel MCG, Morgan C, Nash DM.The UK and Ireland have many native pony breeds with historical and cultural importance as well as being a source of uncharacterized genetic diversity. However, there is a lack of comprehensive research investigating their genetic diversity and phylogenetic interrelationships. Many studies contain a limited number of pony breeds or small sample sizes for these breeds. This may result in erroneous grouping of pony breeds that otherwise have intricate interrelationships with each other and are not evaluated correctly when placed as a token subset of a larger dataset. This is the first study that...
The case of an arctic wild ass highlights the utility of ancient DNA for validating problematic identifications in museum collections.
Molecular ecology resources    January 22, 2020   Volume 20, Issue 5 1182-1190 doi: 10.1111/1755-0998.13130
Vershinina AO, Kapp JD, Baryshnikov GF, Shapiro B.Museum collections are essential for reconstructing and understanding past biodiversity. Many museum specimens are, however, challenging to identify. Museum samples may be incomplete, have an unusual morphology, or represent juvenile individuals, all of which complicate accurate identification. In some cases, inaccurate identification can lead to false biogeographic reconstructions with cascading impacts on paleontological and paleoecological research. Here, we analyzed an unusual Equid mandible found in the Far North of the Taymyr peninsula that was identified morphologically as Equus hemionu...
Mitochondrial Profiles of the East Bulgarian and the Pleven Horse Breeds.
Journal of equine veterinary science    January 22, 2020   Volume 88 102933 doi: 10.1016/j.jevs.2020.102933
Hristov P, Yordanov G, Vladov V, Neov B, Palova N, Radoslavov G.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 bre...
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