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Topic:Genetics

Genetics in horses encompasses the study of hereditary traits and the genetic makeup that influences various characteristics and health conditions in equine populations. This field involves the analysis of genes and their functions, inheritance patterns, and the impact of genetic variations on traits such as coat color, performance ability, and susceptibility to diseases. Research in equine genetics employs techniques such as genome mapping, sequencing, and genetic testing to identify specific genes and mutations associated with these traits. This page gathers peer-reviewed research studies and scholarly articles that explore the genetic basis of equine traits, the methodologies used in genetic research, and the implications for breeding, health management, and conservation of horse breeds.
Cone excitation ratios correlate with color discrimination performance in the horse (Equus caballus).
Journal of comparative psychology (Washington, D.C. : 1983)    November 23, 2006   Volume 120, Issue 4 438-448 doi: 10.1037/0735-7036.120.4.438
Hall CA, Cassaday HJ, Vincent CJ, Derrington AM.Six horses (Equus caballus) were trained to discriminate color from grays in a counterbalanced sequence in which lightness cues were irrelevant. Subsequently, the pretrained colors were presented in a different sequence. Two sets of novel colors paired with novel grays were also tested. Performance was just as good in these transfer tests. Once the horse had learned to select the chromatic from the achromatic stimulus, regardless of the specific color, they were immediately able to apply this rule to novel stimuli. In terms of the underlying visual mechanisms, the present study showed for the ...
Proteolytic maturation of replicase polyprotein pp1a by the nsp4 main proteinase is essential for equine arteritis virus replication and includes internal cleavage of nsp7.
The Journal of general virology    November 14, 2006   Volume 87, Issue Pt 12 3473-3482 doi: 10.1099/vir.0.82269-0
van Aken D, Zevenhoven-Dobbe J, Gorbalenya AE, Snijder EJ.The positive-stranded RNA genome of the arterivirus Equine arteritis virus (order Nidovirales) encodes the partially overlapping replicase polyproteins pp1a (1727 aa) and pp1ab (3175 aa). Previously, three viral proteinases were reported to cleave these large polyproteins into 12 non-structural proteins (nsps). The chymotrypsin-like viral main proteinase residing in nsp4 is responsible for eight of these cleavages. Processing of the C-terminal half of pp1a (the nsp3-8 region) was postulated to occur following either of two alternative proteolytic pathways (the 'major' and 'minor' pathways). He...
Analysis of peripheral blood mononuclear cells gene expression in endurance horses by cDNA-AFLP technique.
Research in veterinary science    November 13, 2006   Volume 82, Issue 3 335-343 doi: 10.1016/j.rvsc.2006.08.009
Cappelli K, Verini-Supplizi A, Capomaccio S, Silvestrelli M.The knowledge of molecular mechanisms of stress response in athlete horses can allow us to plan an appropriate and high-grade training to obtain better performance and to preserve horse welfare. It is well known that excessive muscular exercise can lead to a number of responses which may be associated with modification of the mRNA levels for a number of metabolic genes such as those involved in the immune response. In the present study cDNA-AFLP technique was applied to Arab endurance horses under stressing conditions to visualise variations of transcriptional profiles; 49 transcript derived f...
Pathogenicity and immunogenicity of equine herpesvirus type 1 mutants defective in either gI or gE gene in murine and hamster models.
The Journal of veterinary medical science    November 7, 2006   Volume 68, Issue 10 1029-1038 doi: 10.1292/jvms.68.1029
Tsujimura K, Yamanaka T, Kondo T, Fukushi H, Matsumura T.To develop a live vaccine for equine herpesvirus type 1 (EHV-1), two EHV-1 mutants containing no heterogeneous DNA, DeltagI and DeltagE, were constructed with deletions in the open reading frame of either glycoprotein I (gI) or E (gE), respectively. In equine cell culture, deletion mutants formed smaller plaques than the parental and revertant viruses, but the one-step growth patterns of the deletion mutants and the parental strain were approximately the same. These results suggest that both gI and gE contribute to the ability of EHV-1 to spread directly from cell-to-cell, but that these glyco...
A single amino acid difference within the alpha-2 domain of two naturally occurring equine MHC class I molecules alters the recognition of Gag and Rev epitopes by equine infectious anemia virus-specific CTL.
Journal of immunology (Baltimore, Md. : 1950)    November 4, 2006   Volume 177, Issue 10 7377-7390 doi: 10.4049/jimmunol.177.10.7377
Mealey RH, Lee JH, Leib SR, Littke MH, McGuire TC.Although CTL are critical for control of lentiviruses, including equine infectious anemia virus, relatively little is known regarding the MHC class I molecules that present important epitopes to equine infectious anemia virus-specific CTL. The equine class I molecule 7-6 is associated with the equine leukocyte Ag (ELA)-A1 haplotype and presents the Env-RW12 and Gag-GW12 CTL epitopes. Some ELA-A1 target cells present both epitopes, whereas others are not recognized by Gag-GW12-specific CTL, suggesting that the ELA-A1 haplotype comprises functionally distinct alleles. The Rev-QW11 CTL epitope is...
Comparative aspects of somatic cell nuclear transfer with conventional and zona-free method in cattle, horse, pig and sheep.
Theriogenology    November 1, 2006   Volume 67, Issue 1 90-98 doi: 10.1016/j.theriogenology.2006.09.011
Lagutina I, Lazzari G, Duchi R, Turini P, Tessaro I, Brunetti D, Colleoni S, Crotti G, Galli C.Nuclear transfer (NT) is a complex procedure that requires considerable technical skills. Over the years attempts have been made to simplify the micromanipulations involved and to make the procedure more user-friendly. A significant step forwards has been the development of the zona-free NT methods. We have used zona-free NT with mechanical aspiration of the metaphase plate as a mean of enucleation, in a comparative approach with the conventional nuclear transfer zona-enclosed method in cattle, horse, sheep and pig. The absence of the zona considerably facilitates the enucleation step and sign...
Effect of cumulus morphology and maturation stage on the cryopreservability of equine oocytes.
Reproduction (Cambridge, England)    October 31, 2006   Volume 132, Issue 5 759-769 doi: 10.1530/rep.1.01156
Tharasanit T, Colleoni S, Lazzari G, Colenbrander B, Galli C, Stout TA.Oocyte cryopreservation is a potentially valuable way of preserving the female germ line. However, the developmental competence of cryopreserved oocytes is presently poor. This study investigated whether the morphology of the cumulus complex surrounding an immature equine oocyte and/or the oocyte's stage of maturation affect its cryopreservability. Compact (Cp) and expanded (Ex) cumulus oocyte complexes (COCs) were vitrified either shortly after recovery (germinal vesicle stage, GV) or after maturation in vitro (IVM); cryoprotectant-treated and -untreated non-frozen oocytes served as controls....
Lymphoma, erythrocytosis, and tumor erythropoietin gene expression in a horse.
Journal of veterinary internal medicine    October 27, 2006   Volume 20, Issue 5 1251-1255 doi: 10.1892/0891-6640(2006)20[1251:leateg]2.0.co;2
Koch TG, Wen X, Bienzle D.No abstract available
Cyclooxygenase expression in the early stages of equine laminitis: a cytologic study.
Journal of veterinary internal medicine    October 27, 2006   Volume 20, Issue 5 1191-1196 doi: 10.1892/0891-6640(2006)20[1191:ceites]2.0.co;2
Blikslager AT, Yin C, Cochran AM, Wooten JG, Pettigrew A, Belknap JK.Recent reports indicate increased amounts of mRNA from inflammation-related genes in the prodromal stage of laminitis. Objective: Cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) undergo distinct patterns of expression in equine laminae in the developmental stage (DEV) and acute clinical stage (LAM) of laminitis. Methods: Horses selected from an outbred population were placed into 1 of 4 groups: DEV (n = 5), CON-3h (control group for DEV, n = 5), LAM (n = 5) and CON-10h (control group for LAM, n = 5). Methods: Laminar and skin samples were obtained from (1) animals either undergoing leuko...
Molecular characterization of the equine collagen, type IX, alpha 2 (COL9A2) gene on horse chromosome 2p16–>p15.
Cytogenetic and genome research    October 27, 2006   Volume 115, Issue 2 107-114 doi: 10.1159/000095229
Boneker C, Kuiper H, Drögemüller C, Chowdhary BP, Distl O.The mammalian collagen, type IX, alpha 2 gene (COL9A2) encodes the alpha-2 chain of type IX collagen and is located on horse chromosome 2p16-->p14 harbouring a quantitative trait locus for osteochondrosis. We isolated a bacterial artificial chromosome (BAC) clone containing the equine COL9A2 gene and determined the complete genomic sequence of this gene. Cloning and characterization of equine COL9A2 revealed that the equine gene consists of 32 exons spanning approximately 15 kb. The COL9A2 transcript encodes a single protein of 688 amino acids. Thirty two single nucleotide polymorphisms (SNPs)...
Allele frequency and likely impact of the glycogen branching enzyme deficiency gene in Quarter Horse and Paint Horse populations.
Journal of veterinary internal medicine    October 27, 2006   Volume 20, Issue 5 1207-1211 doi: 10.1892/0891-6640(2006)20[1207:afalio]2.0.co;2
Wagner ML, Valberg SJ, Ames EG, Bauer MM, Wiseman JA, Penedo MC, Kinde H, Abbitt B, Mickelson JR.Glycogen Branching Enzyme Deficiency (GBED), a fatal condition recently identified in fetuses and neonatal foals of the Quarter Horse and Paint Horse lineages, is caused by a nonsense mutation in codon 34 of the GBE1 gene, which prevents the synthesis of a functional GBE protein and severely disrupts glycogen metabolism. The aims of this project were to determine the mutant GBE1 allele frequency in random samples from the major relevant horse breeds, as well as the frequency with which GBED is associated with abortion and early neonatal death using the tissue archives from veterinary diagnosti...
Molecular typing and anti-microbial susceptibility of clinical isolates of Streptococcus equi ssp. zooepidemicus from equine bacterial endometritis.
Journal of veterinary medicine. B, Infectious diseases and veterinary public health    October 26, 2006   Volume 53, Issue 9 451-454 doi: 10.1111/j.1439-0450.2006.01001.x
Luque I, Fernández-Garayzábal JF, Blume V, Maldonado A, Astorga R, Tarradas C.The anti-microbial susceptibility and genetic diversity of 65 strains of Streptococcus equi ssp. zooepidemicus (Sez) isolated from mares presenting clinical signs of endometritis was determined by disk agar diffusion and pulsed field gel electrophoresis (PFGE) methods, respectively. Overall, Sez isolates were susceptible to beta-lactams, enrofloxacin, trimethoprim-sulfamethoxazole and gentamicin. These anti-microbials could be recommended as empiric anti-microbial therapy in cases of endometritis caused by Sez. Pulsed field gel electrophoresis typing revealed a great genetic diversity (56 diff...
Equine infectious anaemia in Ireland: characterisation of the virus.
The Veterinary record    October 24, 2006   Volume 159, Issue 17 570 doi: 10.1136/vr.159.17.570-a
Mooney J, Flynn O, Sammin D.No abstract available
Mass spectrometry studies of demetallation of haemin by recombinant horse L chain apoferritin and its mutant (E 53,56,57,60 Q).
FEBS letters    October 24, 2006   Volume 580, Issue 26 6275-6280 doi: 10.1016/j.febslet.2006.10.034
de Val N, Herschbach H, Potier N, Dorsselaer AV, Crichton RR.An essential difference between eukaryotic ferritins and bacterioferritins is that the latter contain naturally, in vivo haem as Fe-protoporphyrin IX. This haem is located in a hydrophobic pocket along the 2-fold symmetry axes and is liganded by two Met 52. However, in in vivo studies, a cofactor has been isolated in horse spleen apoferritin similar to protoporphyrin IX; in in vitro experiments, it has been shown that horse spleen apoferritin is able to interact with haem. Studies of haemin (Fe(III)-PPIX) incorporation into horse spleen apoferritin have been carried out, which show that the me...
Genetic relatedness and pathogenicity of equine herpesvirus 1 isolated from onager, zebra and gazelle.
Archives of virology    October 19, 2006   Volume 152, Issue 2 245-255 doi: 10.1007/s00705-006-0855-3
Ibrahim ES, Kinoh M, Matsumura T, Kennedy M, Allen GP, Yamaguchi T, Fukushi H.Equine herpesvirus 1 was isolated from an onager in 1985, a zebra in 1986 and a Thomson's gazelle in 1996 in USA. The genetic relatedness and pathogenicity of these three viruses were investigated based on the nucleotide sequences of the glycoprotein G (gG) gene, experimental infection in hamsters, and comparison with horse isolates. The gG gene sequences of EHV-1 from onager and zebra were identical. The gG gene sequences of the gazelle isolate showed 99.5% identity to those of onager and zebra isolates. The gG gene sequences of EHV-1 isolated from horses were 99.9-100% identical and 98, 98 a...
Alobar holoprosencephaly, mobile proboscis and trisomy 13 in a fetus with maternal gestational diabetes mellitus: a 2D ultrasound diagnosis and review of the literature.
Archives of gynecology and obstetrics    October 18, 2006   Volume 275, Issue 5 385-387 doi: 10.1007/s00404-006-0264-6
Capobianco G, Cherchi PL, Ambrosini G, Cosmi E, Andrisani A, Dessole S.Alobar holoprosencephaly is a rare and severe brain malformation due to early arrest in brain cleavage and rotation. Methods: We report a congenital anomalous fetus with alobar holoprosencephaly, prenatally diagnosed by two-dimensional (2D) sonography at the 40 weeks of gestation. The mother was affected by gestational diabetes mellitus and was obese (BMI > 30 kg/m(2)). 2D Ultrasound depicted the cerebral malformation, cyclopy, proboscis, cardiac defects (atrial septal defect, hypoplastic left heart, anomalous communication between right ventricle and aorta) and extremities defects. The new...
Multiple endocrine neoplasia-like syndrome in a horse.
The Veterinary record    October 17, 2006   Volume 159, Issue 16 530-532 doi: 10.1136/vr.159.16.530
Germann SE, Rütten M, Derungs SB, Feige K.No abstract available
Developmental competence of equine oocytes and embryos obtained by in vitro procedures ranging from in vitro maturation and ICSI to embryo culture, cryopreservation and somatic cell nuclear transfer.
Animal reproduction science    October 17, 2006   Volume 98, Issue 1-2 39-55 doi: 10.1016/j.anireprosci.2006.10.011
Galli C, Colleoni S, Duchi R, Lagutina I, Lazzari G.Development of assisted reproductive technologies in horses has been relatively slow compared to other domestic species, namely ruminants and pigs. The scarce availability of abattoir ovaries and the lack of interest from horse breeders and breed associations have been the main reasons for this delay. Progressively though, the technology of oocyte maturation in vitro has been established followed by the application of ICSI to achieve fertilization in vitro. Embryo culture was initially performed in vivo, in the mare oviduct or in the surrogate sheep oviduct, to achieve the highest embryo devel...
Expression and bioactivity of a single chain recombinant equine luteinizing hormone (reLH).
Theriogenology    October 17, 2006   Volume 67, Issue 2 311-320 doi: 10.1016/j.theriogenology.2006.06.013
Jablonka-Shariff A, Roser JF, Bousfield GR, Wolfe MW, Sibley LE, Colgin M, Boime I.To study structure-activity relationships and the role of equine gonadotropins in the normal and pathophysiology of equine reproduction, the availability of purified hormones is essential. Previous expression studies in transfected CHO cells showed inefficient assembly of the human and bovine alpha and beta subunits, resulting in low levels of recombinant LH. The ability to express a single chain bearing genetically linked alpha and beta subunits bypasses this rate-limiting assembly step. A chimera was constructed by overlap PCR in which the carboxy terminal end of the eLHbeta subunit was gene...
Isolation of equine herpesvirus-5 from blood mononuclear cells of a gelding. Bell SA, Balasuriya UB, Nordhausen RW, MacLachlan NJ.Horses are commonly infected by herpesviruses, but isolation of equine herpesvirus-5 (EHV-5) has only infrequently been reported. We describe the isolation and characterization of a strain of EHV-5 from the blood mononuclear cells of a healthy adult horse in California. The virus was initially identified by EHV-5 specific polymerase chain reaction (PCR), and it caused lytic infection of cultured rabbit kidney cells only after repeated serial passage. Virions with characteristic herpesvirus morphology were readily demonstrated in cell culture lysate by transmission electron microscopy. A portio...
Hepatitis E virus infection in work horses in Egypt.
Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases    October 12, 2006   Volume 7, Issue 3 368-373 doi: 10.1016/j.meegid.2006.07.007
Saad MD, Hussein HA, Bashandy MM, Kamel HH, Earhart KC, Fryauff DJ, Younan M, Mohamed AH.Hepatitis E virus (HEV) is an important cause of hepatitis among young Egyptian adults with high seroprevalence rates seen in both rural areas of the Nile Delta and in suburban Cairo. Because natural antibodies to HEV have been detected in animals and zoonotic transmission is postulated, we surveyed work horses in Cairo for evidence of HEV exposure and viremia. Sera from 200 Cairo work horses were tested by ELISA for the presence of IgG anti-HEV antibody revealed a seropositivity of 13%. Among 100 samples processed for detection of viral genome by means of nested polymerase chain reaction (N-P...
Generation of superoxide anion by equine spermatozoa as detected by dihydroethidium.
Theriogenology    October 12, 2006   Volume 67, Issue 3 580-589 doi: 10.1016/j.theriogenology.2006.07.021
Burnaugh L, Sabeur K, Ball BA.Low-level production of the superoxide anion (O2*-) is an important signal transduction event in sperm function including capacitation; however, excessive production of O2*- can be detrimental to sperm function. The objective of this study was to assess dihydroethidium (DHE) as a probe for O2*- in equine spermatozoa. Ejaculated spermatozoa were separated by centrifugation over a Percoll gradient (40:80), and loaded with DHE (2.0 microM) as well as with calcein-acetoxymethylester (CAM, 7.8 nM) to determine cell viability. In Experiment 1, cells were incubated with the xanthine-xanthine oxidase ...
Individual-based assessment of population structure and admixture in Austrian, Croatian and German draught horses.
Heredity    October 11, 2006   Volume 98, Issue 2 114-122 doi: 10.1038/sj.hdy.6800910
Druml T, Curik I, Baumung R, Aberle K, Distl O, Sölkner J.All over Europe, the number of draught horses has decreased drastically during the last 50 years. As a prerequisite for efficient management decisions, we analysed the conservation status in Austrian (Noriker Carinthia - NC, Noriker Salzburg - NS), Croatian (Croatian Coldblood - C, Posavina horse - P) and German (Altmaerkisch Coldblood - A, Black Forest horse - BF, Mecklenburg Coldblood - M, Rhenish German Draught horse - R, Saxon Thuringa Coldblood - ST, Schleswig Draught horse - Sch, South German Coldblood - SG) draught horses (434) using multilocus genotypic information from 30 (effectively...
Cell growth characteristics and differentiation frequency of adherent equine bone marrow-derived mesenchymal stromal cells: adipogenic and osteogenic capacity.
Veterinary surgery : VS    October 10, 2006   Volume 35, Issue 7 601-610 doi: 10.1111/j.1532-950X.2006.00197.x
Vidal MA, Kilroy GE, Johnson JR, Lopez MJ, Moore RM, Gimble JM.To characterize equine bone marrow (BM)-derived mesenchymal stem cell (MSC) growth characteristics and frequency as well as their adipogenic and osteogenic differentiation potential. Methods: In vitro experimental study. Methods: Foals (n=3, age range, 17-51 days) and young horses (n=5, age range, 9 months to 5 years). Methods: Equine MSCs were harvested and isolated from sternal BM aspirates and grown up to passage 10 to determine cell-doubling (CD) characteristics. Limit dilution assays were performed on primary and passaged MSCs to determine the frequency of colony-forming units with a fibr...
A missense mutation in PMEL17 is associated with the Silver coat color in the horse.
BMC genetics    October 9, 2006   Volume 7 46 doi: 10.1186/1471-2156-7-46
Brunberg E, Andersson L, Cothran G, Sandberg K, Mikko S, Lindgren G.The Silver coat color, also called Silver dapple, in the horse is characterized by dilution of the black pigment in the hair. This phenotype shows an autosomal dominant inheritance. The effect of the mutation is most visible in the long hairs of the mane and tail, which are diluted to a mixture of white and gray hairs. Herein we describe the identification of the responsible gene and a missense mutation associated with the Silver phenotype. Results: Segregation data on the Silver locus (Z) were obtained within one half-sib family that consisted of a heterozygous Silver colored stallion with 34...
Derivation and induction of the differentiation of animal ES cells as well as human pluripotent stem cells derived from fetal membrane.
Human cell    October 7, 2006   Volume 18, Issue 3 135-141 doi: 10.1111/j.1749-0774.2005.tb00003.x
Saito S, Yokoyama K, Tamagawa T, Ishiwata I.We succeeded in the derivation and maintenance of pluripotent embryonic stem (ES) cells from equine and bovine blastocysts. These cells expressed markers that are characteristics of mouse ES cells, namely, alkaline phosphatase, stage-specific embryonic antigen 1, STAT 3 and Oct 4. We confirmed the pluripotential ability of these cells, which were able to undergo somatic differentiation in vitro to neural progenitors and to endothelial or hematopoietic lineages. We were able to use bovine ES cells as a source of nuclei for nuclear transfer and we generated cloned cattle with a higher frequency ...
cDNA cloning and substrate specificity of equine tryptase, a possible mediator in equine heaves.
Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology    October 4, 2006   Volume 36, Issue 10 1303-1309 doi: 10.1111/j.1365-2222.2006.02571.x
Dacre KJ, McAleese SM, Knight P, McGorum BC, Pemberton AD.Mast cell mediators are believed to play a central role in inflammatory lung disorders such as human allergic and occupational asthma. Equine heaves is characterized by reversible neutrophilic airway inflammation and airway obstruction, primarily due to bronchospasm and mucus hypersecretion, following exposure of susceptible horses to organic stable dusts. As such, heaves shares many similarities with human occupational dust-induced asthma and therefore it is proposed that mast cells may also be implicated in the pathogenesis of heaves. Tryptase, a mast cell-specific proteinase, can be used as...
Mutation of the maturase lipoprotein attenuates the virulence of Streptococcus equi to a greater extent than does loss of general lipoprotein lipidation.
Infection and immunity    October 2, 2006   Volume 74, Issue 12 6907-6919 doi: 10.1128/IAI.01116-06
Hamilton A, Robinson C, Sutcliffe IC, Slater J, Maskell DJ, Davis-Poynter N, Smith K, Waller A, Harrington DJ.Streptococcus equi is the causative agent of strangles, a prevalent and highly contagious disease of horses. Despite the animal suffering and economic burden associated with strangles, little is known about the molecular basis of S. equi virulence. Here we have investigated the contributions of a specific lipoprotein and the general lipoprotein processing pathway to the abilities of S. equi to colonize equine epithelial tissues in vitro and to cause disease in both a mouse model and the natural host in vivo. Colonization of air interface organ cultures after they were inoculated with a mutant ...
Presence of new mecA and mph(C) variants conferring antibiotic resistance in Staphylococcus spp. isolated from the skin of horses before and after clinic admission.
Journal of clinical microbiology    September 27, 2006   Volume 44, Issue 12 4444-4454 doi: 10.1128/JCM.00868-06
Schnellmann C, Gerber V, Rossano A, Jaquier V, Panchaud Y, Doherr MG, Thomann A, Straub R, Perreten V.Because of the frequency of multiple antibiotic resistance, Staphylococcus species often represent a challenge in incisional infections of horses undergoing colic surgery. To investigate the evolution of antibiotic resistance patterns before and after preventative peri- and postoperative penicillin treatment, staphylococci were isolated from skin and wound samples at different times during hospitalization. Most staphylococci were normal skin commensals and belonged to the common coagulase-negative group. In some cases they turned out to be opportunistic pathogens present in wound infections. M...
Fracture rate in Thoroughbred racehorses is affected by dam age and parity.
Veterinary journal (London, England : 1997)    September 25, 2006   Volume 174, Issue 2 295-301 doi: 10.1016/j.tvjl.2006.07.023
Verheyen KL, Price JS, Wood JL.This study's aim was to determine the effects of dam age and parity on the rate of fracture in offspring in Thoroughbred racehorses in training for flat racing. It was hypothesised that first foals and those from older mares would have a higher fracture rate than subsequent foals and those from younger mares. A two-year observational cohort study collected data from eight trainers on 335 horses that were monitored since the start of their training as yearlings. Multivariable Poisson regression analyses showed that first foals had a significantly lower fracture rate than subsequent ones (RR=0.3...