The experimental infection of ponies with contagious equine metritis

Four pony mares were readily infected with the organism of contagious equine metritis by intracervical inoculation and one by coitus with an infected stallion. Infected mares developed an acute endometritis with local destruction of the endometrial epithelium. In 2 experimentally infected mares, infection appeared to have been spontaneously eliminated from the genital tract within 3 to 4 weeks. A third mare however remained persistently infected in the clitoral fossa over a long period and was a symptomless carrier. Four pony stallions were readily infected in the urethral fossa and the organism survived for varying periods without giving rise to any signs of infection. From 2 of these animals it appeared eventually to have been eliminated spontaneously. An experimentally infected stallion transmitted infection to a healthy mare by coitus. Bacteriological examination of infected pony stallions may occassionally give false negative results and fail to reveal the organism in the external genitalia. Repeated bacteriological examinations need to be undertaken before it can be concluded that a stallion is free of infection.     

Experimental reactivation of equine herpesvirus‐3 following corticosteroid treatment

State of latency, well known for several herpesviruses, has been proposed for equine herpesvirus‐3 (EHV‐3) and supported by epidemiological observations. No detailed assessment about reactivation, patterns of excretion and re‐excretion has been formally reported. An experimental reactivation study by corticosteroid treatment in previously naturally infected horses was therefore carried out. Two polo mares with clinical and virologically confirmed history of equine coital exanthema were injected with dexamethasone and prednisolone on 3 successive days. Clinical signs, body temperature and clinical samples for virological and serological studies were obtained daily. Mares did not show any systemic clinical signs or hyperthermia. EHV‐3 shedding, seroconversion and the presence of a small lesion were observed in one of the mares under study 2 weeks after corticosteroid treatment. The results demonstrate that this virus exhibits a latency‐reactivation behaviour similar to that of other alpha herpesviruses. Reactivation of latency may have an important bearing on the appearance of clinical signs in mares and/or stallions during the breeding season without the actual evidence of transfer from mare to stallion or vice versa.     

Status of equine viral arteritis in Kentucky, 1985

Clinical cases of equine arteritis virus infection have not been diagnosed in Kentucky since 1984, and there has been no indication that any of the horses involved in the 1984 epizootic have since been responsible for spread of the disease to horses in other states or other countries. Cases of abortion caused by naturally acquired infection with this virus have not been confirmed in 1984 or 1985. Neither field nor vaccine strains of equine arteritis virus have been shown to induce teratologic abnormalities or the carrier state in foals born to infected or vaccinated mares. The carrier stallion appears to have played a major epidemiologic role in the dissemination and perpetuation of the virus. A commercial modified live equine viral arteritis vaccine was found to be safe and efficacious for stallions and mares. The disease can be controlled by immunizing the stallion population and by restricting the breeding of equine arteritis virus-shedding stallions to vaccinated or seropositive mares, followed by an appropriate period of isolation from other nonvaccinated Equidae.     

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.     

Epizotiology and phylogeny of equine arteritis virus in hucul horses

The aim of the study was to determine the situation of equine arteritis virus (EAV) infections in hucul horses. A total of 176 horses (154 mares and 22 stallions) from the biggest hucul horse stud in Poland were tested. Antibodies against EAV were detected in 97 (55.1%) horses. The EAV seroprevalence among mares was 53.2% while in stallions - 68.2%. The percentage of positive mares increased with their age, thus amongst the mares of less than 2 years of age the percentage was 32.5%, while in the group of 3-5 years old increased to 59.4% and in the mares in the age of 6-10 years and older than 10 years 89.5% and 95% were seropositive, respectively. Among 11 seropositive stallions five were supposed to be shedders of EAV with their semen. It is likely that those persistently infected stallions were the reservoirs of the virus in the stud. Genetic studies using of ORF5 gene showed high homology between the viruses detected in the semen of those stallions what suggested lateral transmission between the stallions sharing the same stable. Persistent infection in an immature stallion, which has not yet been used for breeding, was established as a result of infection via respiratory route. Phylogenetic analysis confirmed that all hucul viruses shared the same ancestor and as most of EAV strains dominating in Polish horse population belonged to the European origin EAV subgroup (EU-1).     

Equine coital exanthema and its potential economic implications for the equine industry

Equine coital exanthema (ECE) caused by equid herpesvirus 3 (EHV-3) is a contagious venereal disease characterised by the formation of painful papules, vesicles, pustules and ulcers on the external genitalia of both mares and stallions. EHV-3 is an alphaherpesvirus that is distinct from the other equine herpesviruses and endemic in most horse breeding populations worldwide. The negative impacts of ECE on equine breeding enterprises are the forced, temporary disruption of mating activities of mares and stallions, the additional care and supportive treatment that is required for affected horses, and the risk of virus spread by either fresh or frozen semen as well as by artificial insemination and embryo transfer. Because there are no effective surveillance systems to report ECE, its true prevalence and economic impact are difficult to assess and are probably underestimated. The purpose of this review is to describe the recent advances in understanding of EHV-3 infections and to consider the economic consequences of ECE within the current context of the equine industry.     

Contagious equine metritis eradicated from Japan

Contagious equine metritis (CEM), a contagious venereal disease of horses, invaded Japan in 1980 and spread in the Thoroughbred population of the Hidaka-Iburi district of Hokkaido. To eradicate CEM, we ran a program aimed at detecting Taylorella equigenitalis, the causal agent, in carrier horses by using the PCR test, followed by culling or treatment. In 2001, the first year of the program, 12,356 Thoroughbred racing stallions and mares were tested and 11 carriers were found. Four, two, one, and one carrier mares were detected in 2002, 2003, 2004, and 2005, respectively, by application of the program at the same scale as in 2001. No PCR-positive horses were found from 2006 to 2010. These results strongly suggest that CEM was eradicated from Japan by 2010.     

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.     

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.     

Three cases of virus isolation from horse fetuses diagnosed with equine arteritis virus (EAV) abortion from stud farms with different breeds]

Three cases of abortions were diagnosed as caused by Equine Arteritis Virus (EAV) by isolation and typing of this virus from the respective fetuses. All 3 abortions were single cases, one occurring on a stud with Iceland Ponies, one with Warmbloods, one with Lipizzaner horses. On each stud horses of the respective breed were kept exclusively, therefore there existed no epidemiologic link. By means of seroneutralization tests performed on in contact horses it could be shown, that EAV had only been introduced recently into the stud with the Iceland Ponies. An extraneous mare stabled temporarily for covering by the stud's stallion could be incriminated for introducing EAV. By means of post-abortion serology it could be demonstrated that the Warmblood stud had been harbouring EAV for a longer period of time. Likewise, the Lipizzaner stud could be shown to have been persistently infected, this time on pre-abortion serums stored frozen at our Institute. On both these studs preexisting neutralizing antibodies accounted for the single case of abortion and prevented serial abortions. By investigating frozen serums taken in earlier years we could show that the Lipizzaner stallions had reacted positively to EAV for several years already. However, the gestation period of the aborting mare allowed to exclude EAV-positive semen transmitted on copulation as cause of its abortion. Both the Iceland Pony stallion as well as the Warmblood stallion could be excluded as sources of infection for the respective aborting mares as both repeatedly were seronegative.(ABSTRACT TRUNCATED AT 250 WORDS)     

CASE STUDY: Use of Modified Phosphate-Buffered Saline as a Component of Semen Extenders Allows the Use of Transported Semen from Two Stallions

Conception rates for mares bred with transported-cooled and fresh stallion semen were collected over a 4-yr period (1998–2002) for two stallions. Both stallions stood at a commercial breeding farm. Semen from both stallions was used immediately after collection on the farm and after 24 to 48 h of cold storage when transported to locations in the U.S. and Canada. Semen for insemination of mares located on the farm was extended with a commercially available skim milk glucose extender (SKMG). Spermatozoal motility following cold storage for spermatozoa diluted in SKMG extender was unacceptable. Thus, semen from both stallions was centrifuged, and spermatozoa were resuspended in SKMG supplemented with modified PBS. In a previous study, the percentage of motile spermatozoa increased following centrifugation and reconstitution of the sperm pellet in SKMG-PBS as compared with semen dilution in SKMG (Stallion A: 15% vs 47%; Stallion B: 18% vs 43%). In the current study, 22 of 25 (88%) and 3 of 4 (75%) mares conceived with transported-cooled semen from Stallions A and B, respectively. Conception rates for mares inseminated with transported semen did not differ (P>0.05) from those inseminated on the farm with fresh semen. These data illustrate that stallion owners can modify standard cooled semen processing procedures and semen extender composition to improve post-storage spermatozoa motility and to obtain acceptable fertility.     

Method of predicting stallion to mare ratio for natural and artificial insemination programs

A method of predicting a stallion to mare ratio for horses entering either natural service or artificial insemination programs was discussed. Limiting factors for stallions in hand-breeding situations were age, libido and physical ability to cover mares presented for breeding. Other limiting factors for stallions involved in artificial breeding programs were scrotal width, daily sperm output, and the number of live spermatozoa inseminated per mare. Considerations were given for the seasonal effect upon pregnancy rate of the mare, but no considerations were made for seasonal effect upon the stallion. This method must only be applied to stallions that have undergone a thorough breeding soundness examination. It must be emphasized that it served only as a guide; the calculated values were merely useful estimates rather than firm numbers. The multitude of uncontrollable factors involved in breeding farm management, especially with regard to the mares that are presented for breeding were influences that could not be directly considered by this method.     

Lateral transmission of equine arteritis virus among Lipizzaner stallions in South Africa

REASONS FOR PERFORMING STUDY: A serological study conducted in 1995 revealed that 7 stallions at the Lipizzaner Centre, Gauteng, South Africa, were seropositive for antibody to equine arteritis virus (EAV). A Lipizzaner stallion imported into South Africa from Yugoslavia in 1981 had previously (1988) been confirmed to be an EAV carrier. Despite being placed under life-long breeding quarantine, EAV had been transmitted between stallions at the Lipizzaner Centre. OBJECTIVES: To investigate the phylogenetic relationships between the strain of EAV shed in the semen of the original carrier stallion and strains recovered from the semen of 5 other stallions; and to investigate the means whereby lateral transmission of EAV occurred among 7 in-contact, nonbreeding stallions at the Centre. METHODS: EAV was isolated from semen collected from the seropositive stallions using RK-13 cells. Viral RNA was reverse transcribed and amplified by polymerase chain reaction using ORF 5-specific primers, subjected to sequence and phylogenetic analysis. RESULTS: Phylogenetic analysis of strains of EAV recovered from the semen of 6 persistently infected stallions confirmed that all viruses were closely related and probably derived from a common ancestor, i.e. the stallion imported from Yugoslavia. Lateral transmission subsequently occurred among 7 in-contact, nonbreeding stallions at the Centre. It is speculated that these stallions may have been exposed to virus from bedding or fomites contaminated with semen. CONCLUSIONS: These data confirm that lateral transmission of EAV can occur from shedding stallions to susceptible, in-contact horses, including other stallions, which may become persistently infected with the virus. POTENTIAL RELEVANCE: The findings are consistent with lateral spread of a single, unique strain of EAV among a group; and suggest that transmission of EAV may be initiated by infection of one or more stallions with virus on bedding or other fomites contaminated with EAV- infected semen.     

Prevalence of equine herpesvirus-1 infection among Thoroughbreds residing on a farm on which the virus was endemic

OBJECTIVE: To determine the incidence of equine herpesvirus-1 (EHV-1) infection among Thoroughbreds residing on a farm on which the virus was known to be endemic. DESIGN: Prospective cohort study. ANIMALS: 10 nonpregnant mares, 8 stallions, 16 weanlings, 11 racehorses, and 30 pregnant mares and their foals born during the 2006 foaling season. PROCEDURES: Blood and nasopharygeal swab samples were collected every 3 to 5 weeks for 9 months, and placenta and colostrum samples were collected at foaling. All samples were submitted for testing for EHV-1 DNA with a PCR assay. A type-specific EHV-1 ELISA was used to determine antibody titers in mares and foals at birth, 12 to 24 hours after birth, and every 3 to 5 weeks thereafter. RESULTS: Results of the PCR assay were positive for only 4 of the 1,330 samples collected (590 blood samples, 590 nasopharyngeal swab samples, 30 placentas, and 30 colostrum samples), with EHV-1 DNA detected in nasal secretions from 3 horses (pregnant mare, stallion, and racehorse) and in the placenta from 1 mare. Seroconversion was detected in 3 of 27 foals during the first month of life. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that there was a low prevalence of EHV-1 infection among this population of Thoroughbreds even though the virus was known to be endemic on the farm and that pregnant mares could become infected without aborting. Analysis of nasopharyngeal swab samples appeared to be more sensitive than analysis of blood samples for detection of EHV-1 DNA.     

Serological surveillance of equine viral arteritis in the United Kingdom since the outbreak in 1993

Serological analysis of blood samples submitted to the Animal Health Trust showed that during 1995, 185 of 9203 unvaccinated horses (2.0 per cent) tested positive for antibodies to equine arteritis virus (EAV), and that during 1996, 46 of 8851 unvaccinated horses (0.52 per cent) tested positive. During both years thoroughbreds were the predominant breed tested and only a small proportion of these (<0.3 per cent), consisting predominantly of imported mares, were seropositive. In contrast, among standardbred horses, from which samples were actively solicited in 1995, 84 of 454 (18.5 per cent) were seropositive. Among standardbreds there was a difference in prevalence between types of horses, with 3.7 per cent of racing horses, 25 per cent of non-racing horses and 41 per cent of stallions testing seropositive. Investigations of seropositive stallions identified during 1994 and 1995 demonstrated that clinically inapparent equine viral arteritis (EVA) had occurred previously in the UK. Of 50 seropositive stallions, nearly half were standardbreds and nearly all had been imported from either North America or the European Union. Whether 34 seropositive stallions were shedding virus in their semen was established either by test mating, by the serology of the covered mares, or by investigation by MAFF following the introduction of the Equine Viral Arteritis Order 1995. Nine of the stallions (26.5 per cent) were identified as presumptive shedders of EAV in semen and among specific breeds, viral shedding was identified in six of 15 (40 per cent) standardbreds and three of nine (33 per cent) warmbloods. In contrast with the outbreak of EVA in the UK in 1993, no signs of disease typical of EAV infection were reported during these investigations, even in mares test mated to stallions shedding the virus.     

Genetic studies of neuraxonal dystrophy in the Morgan

A naturally occurring disease condition, neuraxonal dystrophy, was identified in 27 male and 15 female Morgans, many of which were closely related. Five-generation pedigrees were constructed for 33 of these horses and were compared with those for 29 randomly selected age- and sex-matched Morgans. Their mean coefficient of inbreeding was not statistically different (P greater than 0.1, 1 tailed t test; P less than 0.001, median test). Breeding of 2 severely affected and 2 normal stallions to 2 severely affected, 3 mildly affected, and 6 normal mares produced 10 mildly affected, 8 suspect affected, and 6 normal offspring. Breeding 4 normal stallions to 17 normal mares produced 22 normal offspring. Neuraxonal dystrophy seems to have a familial component, although neither a simple dominant not recessive mode of inheritance could be demonstrated. The condition could be inherited in a polygenic mode or as a dominant disorder with variable expression.     

Review on Effects of Fescue Grass Ergot Alkaloids in the Horse and Preliminary Study on Effect of Fescue Grass Ergot Alkaloid in the Stallion

Feed with ergot alkaloids ingested by horses has a deleterious clinical and economic impact on the industry. The clinical manifestation of the effects in mares is early embryonic mortality, abortions, prolonged gestation, dystocia, thickened (edematous) or retained placental membranes, agalactia, and increased rates of newborn mortality. For the stallion, very little is known, although ergot alkaloids decrease the ejaculate volume. However, a large number of breeding stallions graze endophyte infected (E+) pastures. This is especially true in the southeastern United States, and clinically we do not perceive that any stallions have any defined problems that could be attributed to ergot alkaloids. However, the number of spermatozoa produced by any stallion might mask the effects. The effects on fertility may be more subtle and only evident with sperm cell manipulation such as chilling or freezing. A preliminary study was performed on six breeding stallions fed feed containing infected fescue seed. We were not able to determine any significant (P < .05) detrimental effects on sperm motility, number morphology, and sperm morphology when compared with controls.     

Effect of book size and age of mare and stallion on foaling rates in thoroughbred horses

The entire population data for Thoroughbred horses in the United States, Canada and Puerto Rico registered with the Jockey Club of America were utilized to study the effect of stallion book size, age of mare and age of stallion on live foal percentage for the 1987 and 1988 breeding seasons. There were 17,260 stallions bred to 179,009 mares, with a resulting overall live foaling rate of 58.1%. There was an increase in live foal percentage as book size increased (P<0.001) and a decrease in live foal percentage as mare age increased (P<0.0001). Stallion age had no effect on live foal percentage.     

Long-Term Follow-up of Partial Limb Amputation in 13 Horses

Thirteen cases of partial limb amputation in horses were reviewed. Nine horses (69%) survived 18 to 111 months (mean, 41 months). Four unsuccessfully treated horses were euthanatized within 21 days of surgery. The horses' ages at amputation were 4 to 13 years (mean, 7.3 years). There was no predilection for the left or right limb nor for fore or hind limb involvement. Osteomyelitis in six horses (46%) was the most common cause for amputation. Severe, open, comminuted fractures of the third metacarpal bone that were impossible to stabilize by any currently available technique in four horses (31%) were the second most common cause for amputation. One gelding was salvaged because of his sentimental value, two horses were used as breeding stallions, and six were broodmares. One stallion lived 30 months, completed two breeding seasons, and started a third. The other stallion began breeding, but became sterile. Three of the mares have produced five foals. One mare died while in foal. One mare aborted near-term twin fetuses and died of uterine hemorrhage. One mare aborted twice before the femoral head ligament on the contralateral limb ruptured, and she was euthanatized.