Evaluation of the field application of PCR in the eradication of contagious equine metritis from Japan

The effectiveness of the polymerase chain reaction (PCR) as a field application test for the eradication of contagious equine metritis (CEM) was evaluated. Seven-thousands five-hundred and thirty-four genital swabs were collected from 4,026 Thoroughbred broodmares and stallions in Japan to test "high risk" horses as well as for general surveillance testing from 1998 to 2001. Bacterial isolation as well as PCR testing of original specimens and cultured specimens was performed for detection of Taylorella equigenitalis from genital swabs. As a result, T. equigenitalis was detected in 12 mares and 1 stallion by PCR, although the bacteria were isolated from only 2 of the PCR-positive mares. CEM-infected and carrier horses were treated by a combination of chemotherapy and surgery. Subsequent follow-up testing over a 3-year period did not detect T. equigenitalis. It was demonstrated that PCR testing was more sensitive than isolation as a method for the detection of T. equigenitalis from genital swabs of horses in the field. It was therefore suggested that a combination of PCR testing and treatment were useful measures in the eradication of CEM from Japan.     

Horse species symposium: contagious equine metritis: an insidious threat to the horse breeding industry in the United States

Contagious equine metritis (CEM) has given rise to international concern since it was first recognized as a novel venereal disease of equids in 1977 and the etiologic agent was identified as a previously undescribed bacterium, Taylorella equigenitalis. Horse industry concerns over CEM centered on the ease with which this bacterium could be disseminated, the significance of T. equigenitalis as a cause of short-term infertility in the mare, and the existence of the carrier state in the stallion and the mare. The first known outbreak of CEM in the United States was in Kentucky in 1978. The economic impact on the Thoroughbred industry in the state was substantial. Before 2008, additional small-scale outbreaks occurred in Missouri in 1979, Kentucky in 1982, and Wisconsin in 2006, nearly all attributed to the importation of carrier animals. On each occasion, appropriate measures were taken to eliminate the infection, resulting in the United States regaining its CEM-free status. With the exception of the 1978 occurrence in Kentucky, none of the subsequent outbreaks significantly affected the horse industry. That changed dramatically in 2008, however, after the discovery of a Quarter horse stallion in Kentucky that cultured positive. Subsequent investigations turned up 23 carrier stallions and 5 carrier mares belonging to 11 breeds and located in 8 states. Shipment of infective semen and indirect venereal contact in stallion collection centers through the use of contaminated fomites were major factors in the spread of T. equigenitalis. Trace-back investigations of some 1,005 exposed and carrier stallions and mares in 48 states have failed to identify the origin of this latest CEM event. Neither clinical evidence of CEM nor decreased pregnancy rates were reportedly a feature in infected or exposed mares. In light of these findings, there was some question of whether or not the considerable expense incurred in investigating the latest CEM occurrence was warranted. Regaining CEM-free status for the United States will present considerable challenges.     

The occurrence of equine arteritis virus in Australia

This paper reports the first isolation of equine arteritis virus (EAV) in Australia and serological evidence of exposure to EAV in Australian horses. Twelve Standardbred stallions imported from North America were found to shed EAV in semen. One hundred and seven stallions were tested for serum antibodies to EAV and 73% of Standardbred stallions tested were seropositive as compared to 8% of Thoroughbred stallions. Serum antibody was detected in 71% of Standardbred mares, 6% of Standardbred racehorses and 1% of Thoroughbred mares and racehorses. Examination of stored serums demonstrated that EAV had been present in Australia since at least 1975.     

A survey of reproductive performance in Thoroughbred mares and morbidity, mortality and athletic potential of their foals

A survey was performed to evaluate the reproductive performance of Thoroughbred mares, estimate risks of dystocia and of morbidity and mortality in foals during the first year post partum and their physical acceptability at age one year. The study population consisted of registered Thoroughbred mares and their foals owned by residents of 4 Western Canadian provinces. Owners were identified using information obtained from the North American Jockey Club, and questionnaires were mailed regarding mares bred in 1988 and their foals born in 1989. Eighty-three per cent of mares were reported to be pregnant at some stage following breeding, and 80% of pregnant mares subsequently gave birth to live foals. Estimates of morbidity and mortality were greater than previously reported, 25% of foals had health problems and 5% died during the first 2 weeks postpartum. Twenty-seven per cent of foals surviving 2 weeks were reportedly affected by some health problem between age 15 days and one year, and 6% died during this period. The case fatality rates of horses with upper respiratory tract infections and diarrhoea were much lower than case fatality rates for infectious diseases occurring less frequently. The rate of death or euthanasia among horses with musculoskeletal problems was relatively high after age 2 weeks. Foals with health problems up to age 2 weeks, or between age 15 days and one year were 5 to 7 times more likely to be classified as physically unacceptable for athletic use. Angular limb deformity was the health problem most commonly reported in foals receiving unacceptable physical assessments, and assessments of longterm athletic potential were apparently not affected by the occurrence of infectious diseases.     

Association between single nucleotide polymorphisms in p53 and abortion in Thoroughbred mares

Single nucleotide polymorphisms (SNPs) in the p53 gene have been studied extensively in humans. The aims of this study were to determine the frequency of the Arg/Pro SNP in p53 in Thoroughbred mares on one stud in Brazil and to correlate p53 genotypes with reproductive performance. SNPs were detected by PCR-restriction fragment length polymorphism in blood samples from 105 horses and confirmed by sequencing. The allele frequency in Thoroughbred mares at codon 72 in exon 4 was 73.3% Arg/Pro, 17.1% Arg/Arg and 9.6% Pro/Pro. The presence of Arg/Pro was significantly associated with abortion (P=0.02), while Pro/Pro mares had a lower probability of abortion (P<0.05). Using a logistic regression model, the dominance effect was significant (P=0.044; odds ratio 7.94) for abortion and additive effects were not significant (P=0.26). p53 may play a role in equine reproduction.     

Diagnostic and epidemiologic analysis of the 2008-2010 investigation of a multi-year outbreak of contagious equine metritis in the United States

Contagious equine metritis (CEM) is a highly contagious venereal disease of horses caused by Taylorella equigenitalis. During testing for semen export purposes, a stallion in Kentucky was found to be T. equigenitalis culture positive in December of 2008. This finding triggered an extensive regulatory investigation to search for additional positive horses, determine the extent of the outbreak, identify the potential source of the outbreak, and ultimately return the United States to CEM-free status. The investigation included over 1000 horses located in 48 states. Diagnostic testing found a total of 22 stallions, 1 gelding and 5 mares culture positive for T. equigenitalis. Epidemiologic analysis indicated that all of the positive horses were linked to a single common source, most likely a Fjord stallion imported into the United States in 2000. The T. equigenitalis strain subsequently spread to other stallions via undetermined indirect mechanisms at shared breeding facilities, and to mares via artificial insemination and live breeding. This CEM outbreak and investigation represent the largest ever in the United States based on the number of exposed horses tested and their geographic distribution.     

Occurrence of equine coital exanthema in pastured draft horses and isolation of equine herpesvirus 3 from progenital lesions

During the period from 2001 to the following year, progenital diseases had been epidemic among the draft stallions and mares pastured together in Iwate Prefecture, the northeastern district of Japan. A stallion and 8 of 31 mares were affected in 2001, and 1 of 2 stallions and 10 of 36 mares in 2002. The clinical symptoms consisted of the formation of papules, pustules, ulcers and scabs on the progenital skin and mucosa in stallions and mares. In 2002, Equine herpesvirus 3 (EHV3) was isolated from 2 mares and the glycoprotein G gene of the virus detected from a stallion and 4 mares by polymerase chain reaction. Serum neutralizing tests showed that 12 of 38 horses, 10 clinically and 2 subclinically affected, changed to be positive for the EHV3 antibody. The results suggest that the horses were affected with equine coital exanthema (ECE) through coitus. Five mares with the antibody at the pre-pastured period may have been the possible origins of EHV3 infection in 2002, although the exact origin in 2001 remains unknown. The artificial insemination was performed for the prevention of ECE spreading through coitus on the pasture in 2003. There was no epidemic of the disease in 31 mares, although 3 mares with the antibody at the pre-pastured period showed the significant increase in the titers during the pastured period.     

Factors Influencing the Frequency of Pregnancy Loss among Thoroughbred Mares in Hidaka,Japan

Pregnancy loss in mares is thought to be a main problem associated with reproductive efficiency. To clarify the situation of pregnancy loss in Thoroughbred mares in Japan, the occurrence of pregnancy loss before and after 35 days of gestation was investigated with 1,476 Thoroughbred mares in Hidaka, Japan, from 2007 to 2009. Pregnancy loss on days 17-35 was determined by ultrasound examination between 17 and 35 days after the last mating. Follow-up surveys were conducted between 35 days and foaling to determine pregnancy loss on day 35 until foaling in 843 of these mares. Using multiple logistic regression analysis, we assessed the influence of mare age, reproductive status, twin pregnancy reduction, body condition score (BCS), estrus type in foaling mares (foal heat or not), progesterone therapy, and endometrial cysts on pregnancy loss rates on days 17-35 and on day 35 until foaling in this population of mares. The pregnancy loss rates on days 17-35 and on day 35 until foaling were 5.8% and 8.7%, respectively. The overall pregnancy loss rate (day 17 until foaling, including parturient losses) was 14.7%. Risk factors for pregnancy loss included decrease in BCS between 17 and 35 days, <5 BCS at day 35, mating during foal heat, and endometrial cysts. In all, 14.7% of pregnancies were lost between day 17 and birth, contributing significantly to reduced reproductive efficiency in Thoroughbred mares in Japan. These observations indicate that mares should be maintained at high BCS and should be prevented from mating in foal heat to decrease the pregnancy loss rates.     

马eCG/LHβ亚基基因多态性研究

马绒毛膜促性腺激素(equine chorionic gonadotropin,eCG)是由妊娠期母马胎儿尿膜—绒毛膜细胞分泌的一种由α、β两个亚基组成的异源二聚体糖蛋白,具有FSH、LH的双重活性。采用PCR-SSCP和测序方法对28匹蒙古马和25匹纯血马的eCG/LHβ亚基基因的多态性进行研究,结果发现,在蒙古马eCG/LHβ亚基基因5′调控区存在一突变位点:G-415A分为AA、BB、AB 3种基因型,AA、BB、AB基因型频率分别为0.89、0.07、0.04;纯血马此基因中未检测到多态性。     

Relationship between stages of the estrous cycle and bone cell activity in Thoroughbreds

OBJECTIVE: To investigate the relationship between stage of estrous cycle and bone cell activity in Thoroughbreds. SAMPLE POPULATION: Blood samples collected from forty-seven 2-year-old Thoroughbred mares in training for racing. PROCEDURES: Blood samples were collected monthly (in April through September) from the mares. Stage of estrus was determined by assessing serum progesterone concentration. Bone cell activity was determined by measuring concentrations of 2 markers of bone formation (osteocalcin and the carboxy-terminal propeptide of type I collagen [PICP]) and a marker of bone resorption (the cross-linked carboxy-terminal telopeptide of type I collagen [ICTP]) in sera. RESULTS: When the relationship between stage of the estrous cycle and markers of bone cell activity was examined, serum concentrations of both osteocalcin and ICTP were significantly higher in mares that were in the luteal phase, compared with mares that were at other stages of the estrous cycle. Stage of estrus did not affect serum PICP concentration. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that bone cell activity in Thoroughbred mares fluctuates during the estrous cycle; serum concentrations of markers of bone formation and bone resorption are increased during the luteal phase. Further studies are required to determine whether these changes are of clinical importance and increase the risk of injury for mares in training during the breeding season. As in humans, stage of estrus must be considered as a source of uncontrollable variability in serum bone marker concentrations in horses.     

Factors influencing the development of stereotypic and redirected behaviours in young horses: findings of a four year prospective epidemiological study

Stereotypies are invariant and repetitive behaviour patterns that seemingly have no function, which tend to develop in captive animals faced with insoluble problems and may be indicative of reduced welfare. A 4 year prospective study of the factors influencing the development of stereotypic and redirected behaviours (abnormal behaviour) in a population of 225 young Thoroughbred and part-Thoroughbred horses was conducted between 1995 and 1999. Abnormal behaviour affected 34.7% of the population. Multivariable analysis showed that foals of low- or middle-ranking mares were less likely to develop abnormal behaviour than foals of dominant mares (rate ratio (RR) 0.23, P<0.01; RR 0.48, P<0.01, respectively). Weaning by confinement in a stable or barn was associated with an increased rate of development of abnormal behaviour, compared with paddock-weaning (RR 2.19, P<0.05), and housing in barns, rather than at grass after weaning, was associated with a further increase (RR 2.54, P<0.01). Specific stereotypic and redirected behaviours were then considered as separate outcomes. Crib-biting was initiated by 10.5% of horses at median age 20 weeks, weaving by 4.6% of horses at median age 60 weeks, box-walking by 2.3% of horses at median age 64 weeks and wood-chewing by 30.3% of horses at median age 30 weeks. Wood-chewing developed at a lower rate in horses born to subordinate or mid-ranking mares than in horses born to dominant mares (RR 0.29, P<0.01; RR 0.41, P<0.01, respectively), and at a higher rate in horses kept in barns or stables rather than at grass after weaning (RR 4.49, P<0.001; RR 1A6, P<0.001, respectively). Feeding concentrates after weaning was associated with a 4-fold increase in the rate of development of crib-biting (RR 4.12, P = 0.02). The results of this study support the idea that simple changes in feeding, housing and weaning practices could substantially lower the incidence of abnormal behaviour in young horses.     

The Athletic Performance of Thoroughbred Racehorses out of Mares with Suspected Placentitis during Gestation

Thoroughbred mares treated for signs of placentitis are potentially costly to farms and owners. Although these treatments often are effective, the resulting foal may require expensive aftercare. The current study investigated the future racing performance of foals from mares treated for clinical and subclinical signs of placentitis compared with matched control mares that had no signs of placentitis producing foals on the same farms. In total, 190 horses born from mares with suspected placentitis met the criteria for analysis and were matched with 190 horses born on the same farm. Mare data and percentage of stakes winners in each group were analyzed with Student t test. Analysis between pairs of horses was done using a paired t test for number of starts, wins, places, shows, and amount earned. Significance was set at a P value of <.05. Mares with suspected placentitis were slightly older than control mares, had slightly shorter lengths of gestation, and had a longer interval to subsequent pregnancy than control mares (P < .05). Horses out of suspected placentitis mares that were racing as 2 year olds in the study had no difference in starts, wins, places, shows, and percentages of black-type race winners; however, this group earned more than their matched control pairs (P < .05). Older horses (3 year olds) out of suspected placentitis mares had no differences in number of starts, wins, places, shows or amount earned; however, the matched control older horses had more black-type race winners (P < .05). The information from this study may aid owners when making decisions about treatment options.     

Association between race history and risk of superficial digital flexor tendon injury in Thoroughbred racehorses

OBJECTIVE: To determine whether race history, including the number of races and total race distance, was associated with risk of superficial digital flexor tendon (SDFT) injury in Thoroughbred racehorses in Japan. DESIGN: Matched case-control study. ANIMALS: 515 Thoroughbred racehorses (case horses) that sustained an SDFT injury during training or racing in Japan during 2002 and 951 horses (control horses) without SDFT injury that were matched with case horses on the basis of age and month of the latest race. PROCEDURE: Variables related to race history were compared between case and control horses by means of conditional logistic regression. RESULTS: The odds of SDFT injury increased as mean race distance and mean body weight at race time increased. Compared with females that had never competed in steeplechase races, males regardless of steeplechase race history and females that had competed in steeplechase races had higher odds of SDFT injury. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that longer mean distance per race, heavier mean body weight at race time, steeplechase experience, and sex (male) increased the risk of SDFT injury in Thoroughbred racehorses.     

Epidemiology of EHV-1 and EHV-4 in the mare and foal populations on a Hunter Valley stud farm: are mares the source of EHV-1 for unweaned foals.

The prevalence of EHV-1 and EHV-4 antibody-positive horses was determined using a type specific ELISA on serum samples collected from 229 mares and their foals resident on a large Thoroughbred stud farm in the Hunter Valley of New South Wales in February 1995. More than 99% of all mares and foals tested were EHV-4 antibody positive, while the prevalence of EHV-1 antibody positive mares and foals were 26.2 and 11.4%, respectively. Examination of the ELISA absorbance data for the individual mares and foals suggested that the EHV-1 antibody positive foals had been infected recently with EHV-1 and that a sub-group of the mare population was the likely source of infectious virus for the unweaned foals.     

Evaluation of early fetal losses on four equine farms in central Kentucky: 73 cases (2001)

OBJECTIVE: To determine features of an early fetal loss (EFL) syndrome and evaluate potential risk factors for EFL in Thoroughbred broodmares on 4 farms in central Kentucky. DESIGN: Retrospective study. ANIMALS: 288 pregnant broodmares. PROCEDURE: Year-2001 breeding records for 288 Thoroughbred broodmares were examined. Early fetal loss was defined as loss of a fetus that was viable at > or = 40 days of gestation but was subsequently lost by 5 months of gestation. RESULTS: Overall 2001 EFL rate was 25% (73/288), median gestational age at time of fetal loss was 77 days, and median date of loss was May 7. Mares on 1 farm had significantly fewer fetal losses (5%) than mares on the other 3 farms (26 to 35%). Fetal losses were higher for maiden (42%) and barren (42%) mares than for foaling mares (18%). Fetal losses were greater in young than in older mares. Effects of broodmare farm, mare age, and reproductive status were all significant. Fetal losses were not associated with sire used for mating or stud farm. CONCLUSIONS AND CLINICAL RELEVANCE: Greatest risk for EFL occurred during the period from late April to May (ie, in mares bred during February through March). Higher incidence of EFL in maiden and barren mares and lower incidence of EFL on 1 farm suggest management or environmental influences may have affected outcome. Risk factors that should be investigated include environmental differences among farms and differences in management procedures used for lactating versus nonlactating mares.     

Distribution of mecA-harboring staphylococci in healthy mares

The prevalence of staphylococci that harbor the mecA gene responsible for methicillin resistance was examined in healthy breeding mares. Staphylococci often cause diseases of horses such as metritis, keratitis, and abscess. Methicillin-resistant staphylococci would make antibiotic treatments ineffective, so it may be significant to know the distribution of mecA-harboring staphylococci in mares. Isolation of mecA-harboring staphylococci was achieved from nares and pasterns of 100 mares in Hokkaido, Japan. From 13% of the mares, mecA-harboring staphylococci, including 15 isolates of Staphylococcus sciuri and 3 of Staphylococcus lentus, were isolated. Isolates of S. sciuri were found to be genetically polyclonal by pulsed-field gel electrophoresis. These isolates produced no PCase and showed low or no resistance to beta-lactam and other classes of antibiotics. Distribution of staphylococcal species and levels of antibiotic resistance were found to be different between isolates from the present mares and those previously reported from riding-horses. Antibiotic pressure may lead to these differences. In addition, it appears that mecA-harboring S. sciuri may be native to horses.     

Helminths in horses: use of selective treatment for the control of strongyles

The current level of anthelmintic resistance in the horse-breeding industry is extremely high and therefore more emphasis is being placed on studies that focus on the judicious use of anthelmintic products. The aims of the study were to: 1) establish if there is variation in the egg excretion pattern of strongyles between the different age classes of Thoroughbred horses in the Western Cape Province (WCP), 2) test if a selective treatment approach successfully reduces the number of anthelmintic treatments and maintains acceptably low helminth burdens in adult Thoroughbred horses, and 3) evaluate the efficacy of subsampling large horse herds for faecal egg counts (FECs) to monitor the strongyle burden. In 2001 the FECs of 4 adult mare, 5 yearling and 3 weanling herds from 8 different farms were compared in the WCP Within the mare herds there were generally fewer egg-excreting individuals with lower mean FECs compared with the younger age classes. Individual faecal samples were collected every 3-4 weeks from 52 adult Thoroughbred mares from 1 farm in the WCP during a 12-month period (2002/2003). Animals with strongyle FECs > or =100 eggs per gram (epg ) were treated with an ivermectin-praziquantel combination drug (Equimax oral paste, Virbac). The mean monthly strongyle FEC for the entire group was <300 epg throughout the study and the number of treatments was reduced by 50 %. Resampling methods showed that an asymptote to mean FEC was reached at 55 animals for each of the pooled weanling, yearling and mare egg counts. Resampling within 4 different mare herds recorded asymptotes of between 24 and 28 animals. Subsampling entire herds for FECs therefore provided an effective approach to treatment management. This study demonstrates that selective treatment is both a practical and an effective approach to the management of anthelmintic resistance.     

Characteristic of Inflammatory Airway Disease in Japanese Thoroughbred Racehorses

Inflammatory airway disease (IAD) is a common cause of poor performance, interruption of training and premature retirement in racehorses. It is also reported that up to 80% of horses are affected at some point in the first years of training in UK and Australia. However, no studies with regard to the information on occurrence of IAD in Japanese Thoroughbred racehorses have been reported. To investigate the occurrence and the characteristics of IAD, epidemic research including endoscopic examination of the airway tract and trachea wash was conducted for Thoroughbred racehorses presenting coughs or poor performance which airway tract disease was suspected stalled in training facility managed by Japan Racing Association. Fifty-six out of 76 Thoroughbred racehorses (73.7%) presenting coughing or poor performance were diagnosed as IAD. Mean incidence rate of IAD was 0.3% and it has been confirmed that constant number of IAD exists in Japan. Up to 35.7% of IAD horses showed upper airway abnormalities in some extent. There was a trend for IAD horses to use wood shavings for bedding and fed hay from the ground compared with the control group. Therefore, improvement of stabling environment may aid in preventing IAD. This study demonstrated that Japanese Thoroughbred racehorses are affected by IAD likewise other countries as well as demonstrated the characteristics of IAD which may contribute to the clarification of the pathogenesis of IAD.