Case-control study of late-term abortions associated with mare reproductive loss syndrome in central Kentucky

OBJECTIVE: To identify factors associated with abortions of mares during late gestation attributed to mare reproductive loss syndrome (MRLS). DESIGN: Case-control study. ANIMALS: 282 broodmares from 62 farms in central Kentucky, including 137 mares that had late-term abortions (LTAs) associated with MRLS, 98 mares from the same farms that did not abort, and 48 mares that aborted from causes other than MRLS. PROCEDURE: Farm managers were interviewed to obtain data on a wide range of management practices and environmental exposures for the mares. Data for case and control horses were compared to identify risk factors for a mare having a MRLS-associated LTA (MRLS-LTA). RESULTS: Several factors increased the risk of mares having MRLS-LTAs, including increased amount of time at pasture, less time in a stall, feeding concentrate on the ground, higher proportion of diet derived from grazing pasture, being fed in pasture exclusively during the 4-week period prior to abortion, access to pasture after midnight during the 4-week period prior to abortion, and drinking from a water trough or not having access to water buckets or automatic waterers. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis indicates that exposure to pasture predisposed mares to having MRLS-LTAs and stillborn foals. Methods for limiting exposure to pasture (keeping mares in stalls longer) during environmental conditions similar to those seen in 2001 should reduce the risk of mares having MRLS-LTAs.     

Case-control study of factors associated with excessive proportions of early fetal losses associated with mare reproductive loss syndrome in central Kentucky during 2001

OBJECTIVE: To identify factors associated with excessive proportions of early fetal losses associated with mare reproductive loss syndrome in central Kentucky during 2001. DESIGN: Case-control study. PROCEDURE: Questionnaires were used to collect information on farm-, pasture-, and individual animal-level factors purportedly associated with mare reproductive loss syndrome. Data were collected for 133 farms (97 with excessive proportions of early feta losses and 36 control farms) representing 6,576 mares. RESULTS: Factors significantly associated with an increased risk of excessive early fetal losses were exposure to moderate to high concentrations of Eastern tent caterpillars, exposure to cherry trees, farm size > or = 50 broodmares, being bred during February 2001, and frequent exposure to waterfowl. Feeding hay to mares outside was associated with a decreased risk of excessive proportions of early fetal losses. Pasture composition and management factors were not significantly different between affected and control pastures. Individual animal-level factors were investigated on 6 farms representing 340 mares, and age, parity, and pre- and postbreeding treatments were not significantly associated with risk of early fetal loss. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that limiting exposure to Eastern tent caterpillars and cherry trees and feeding hay to mares outside may help decrease the risk of excessive proportions of early fetal losses associated with mare reproductive loss syndrome.     

Temporality of early-term abortions associated with mare reproductive loss syndrome in horses

OBJECTIVE: To characterize the temporality of dates of breeding and abortion classified as mare reproductive loss syndrome (MRLS) among mares with abortions during early gestation. ANIMALS: 2,314 mares confirmed pregnant at approximately 28 days after breeding from 36 farms in central Kentucky, including 515 mares that had early-term abortions. PROCEDURE: Farm veterinarians and managers were interviewed to obtain data for each mare that was known to be pregnant to determine pregnancy status, breeding date, last date known to be pregnant, and date of abortion. RESULTS: Mares bred prior to April 1, 2001, appeared to be at greatest risk of early-term abortion, both among and within individual farms. Mares bred in mid-February appeared to be at greatest risk of abortion, with an estimated weekly incidence rate of abortion of 66% (95% CI, 52% to 80%). CONCLUSIONS AND CLINICAL RELEVANCE: Mares in central Kentucky bred between mid-February and early March were observed to be at greatest risk of early-term abortion, and risk gradually decreased to a background incidence of abortion of approximately 11%. Mares bred after April 1, 2001, appeared to be at markedly less risk, indicating that exposure to the cause of MRLS likely occurred prior to this date.     

Descriptive epidemiology of late-term abortions associated with the mare reproductive loss syndrome in central Kentucky.

Epidemiological and pathological findings of 433 late-term abortions associated with the mare reproductive loss syndrome (MRLS) in central Kentucky were identified by reviewing the records of the University of Kentucky Livestock Diseases Diagnostic Center. The distribution of dates of abortion was clustered during a brief period of time, presumably from a simultaneous environmental exposure. The most common pathological findings were microscopic pulmonary lesions consisting of squamous epithelial cells present in alveoli with or without concurrent infiltration of inflammatory cells (neutrophils, macrophages, or monocytes) in the interstitium or within alveoli. Isolation of a non-beta-hemolytic Streptococcus (52% of fetuses) or an Actinobacillus sp. (19% of fetuses) was common. Placentitis or funisitis was identified in 44% of fetuses. No single pathological finding, however, was pathognomonic for MRLS-associated late-term abortion. This report describes the pathological findings characterizing the MRLS-associated abortion. A cause of MRLS could not be determined from necropsy findings.     

An alternative model for fetal loss disorders associated with mare reproductive loss syndrome

Fertile chicken eggs were used as an alternative model for large animals to evaluate suspect toxic dietary ingredients for fetal loss disorders associated with mare reproductive loss syndrome (MRLS) and fetal losses in other livestock. Nitrate, ammonia, and sulfate may react with proteinaceous compounds to enable the formation of abiotic pathogenic nanoparticles which were constant findings in pathognomonic placental lesions associated with non-infectious fetal losses of previously unknown etiology in mares, chickens and other livestock. The pathogenic nanoparticles may be produced naturally by toxic elements associated with air pollution that affect pasture forages or crops, unintentionally by reactions of these elements in protein-mineral mixes in dietary rations, or endogenously within tissues of fetuses and adult animals. The nanoparticles may form niduses in small vessels and predispose animals to a host of secondary opportunistic diseases affecting the reproductive, respiratory and gastrointestinal tracts of animals. The newly recognized abiotic pathogenic micro and nanoparticles are associated with MRLS. The discovery of the pathogenic nanoparticles led to the identification of nitrate, ammonium, and sulfur, in the form of sulfate, that seemingly enable the formation of the pathogenic nanoparticles in embryonic and fetal tissues.     

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.     

Use of a Bayesian risk-mapping technique to estimate spatial risks for mare reproductive loss syndrome in Kentucky

OBJECTIVE: To estimate spatial risks associated with mare reproductive loss syndrome (MRLS) during 2001 among horses in a specific study population and partition the herd effects into those attributable to herd location and those that were spatially random and likely attributable to herd management. Animals-Pregnant broodmares from 62 farms in 7 counties in central Kentucky. PROCEDURE: Veterinarians provided the 2001 abortion incidence proportions for each farm included in the study. Farms were georeferenced and data were analyzed by use of a fully Bayesian risk-mapping technique. RESULTS: Large farm-to-farm variation in MRLS incidence proportions was identified. The farm-to-farm variation was largely attributed to spatial location rather than to spatially random herd effects. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that there are considerable data to support an ecologic cause and potential ecologic risk factors for MRLS. Veterinary practitioners with more detailed knowledge of the ecology in the 7 counties in Kentucky that were investigated may provide additional data that would assist in the deduction of the causal factor of MRLS via informal geographic information systems analyses and suggest factors for inclusion in further investigations.     

Case-control study of factors associated with fibrinous pericarditis among horses in central Kentucky during spring 2001

OBJECTIVE: To identify factors significantly associated with an epidemic of fibrinous pericarditis during spring 2001 among horses in central Kentucky. DESIGN: Case-control study. ANIMALS: 38 horses with fibrinous pericarditis and 30 control horses examined for other reasons. PROCEDURE: A questionnaire was developed to solicit information regarding a wide range of management practices and environmental exposures from farm owners or managers. RESULTS: The following factors were found in bivariate analyses to be significantly associated with an increased risk of pericarditis: being from a farm with mares and foals affected by mare reproductive loss syndrome, exposure to Eastern tent caterpillars in or around horse pastures, younger age, shorter duration of residence in Kentucky and at the farm of current residence, being fed hay grown outside Kentucky, a lack of access to pond water, access to orchard grass for grazing, and a lack of direct contact with cattle. In multivariate logistic regression analyses, only variables related to caterpillar exposure and age were significantly associated with fibrinous pericarditis. CONCLUSIONS AND CLINICAL RELVANCE: Results suggest that fibrinous pericarditis in horses may be associated with mare reproductive loss syndrome. Exposure to Eastern tent caterpillars was the greatest risk factor for development of fibrinous pericarditis. The distribution of times of diagnosis of fibrinous pericarditis was consistent with a point-source epidemic.     

Classification of Actinobacillus spp isolates from horses involved in mare reproductive loss syndrome

OBJECTIVE: To identify Actinobacillus spp isolates recovered from fetuses and pericardial fluid from horses affected with mare reproductive loss syndrome (MRLS) and determine whether these bacterial species are the same as those isolated from clinically normal horses. SAMPLE POPULATION: Isolates of actinobacilli recovered from 18 horses with pericarditis and 109 fetuses aborted by mares affected by MRLS. Procedures-Actinobacillus spp isolates were identified to the level of species or subspecies by use of conventional phenotypic tests and biochemical and enzyme test kits. The 16S rRNA gene from selected isolates was amplified, purified, and sequenced. Sequence data were compared with sequence data for actinobacilli in GenBank. RESULTS: Of the 109 isolates obtained from fetuses, 14 were Actinobacillus equuli subsp equuli, 65 were A equuli subsp haemolyticus, 28 were Bisgaard taxon 10-like bacterium, and 2 were Actinobacillus genomospecies 1. Of the 18 isolates from horses with pericarditis, 4 were A equuli subsp equuli, 13 were A equuli subsp haemolyticus, and 1 was Bisgaard taxon 10-like bacterium. Comparisons with published data and GenBank data revealed that the isolates recovered from horses with MRLS were the same as those isolated from the oral cavity or alimentary tract of healthy horses. CONCLUSIONS AND CLINICAL RELEVANCE: Actinobacillus spp isolates recovered from fetuses and pericardial fluid samples of horses affected by MRLS in 2001 to 2003 were identical to Actinobacillus spp found in the oral cavity and alimentary tracts of healthy horses.     

Impact of reproductive efficiency over time and mare financial value on economic returns among Thoroughbred mares in central Kentucky

Reason for performing study: There have been no studies reporting the impact of reproductive efficiency and mare financial value on economic returns. Objective: To explore the economic consequences of differences in reproductive efficiency over time in the Thoroughbred mare. Methods: Complete production records for 1176 mares were obtained. Production history and drift in foaling date were calculated. Multiple logistic regression was used to identify factors influencing the probability of producing a registered foal in 2005. The ‘net present value’ and ‘internal rate of return’ were calculated for economic scenarios involving different initial mare financial values, levels of reproductive efficiency, and durations of investment. Results: Among mares that did not produce a foal every year (63%), the mean time before failing to produce a registered foal was 3.4 years. The majority of mares drifted later in their foaling dates in subsequent foaling seasons. Increasing mare age, foaling after 1st April, needing to be mated multiple times during the season, and producing a lower number of foals in continuous sequence during previous years decreased the probability of producing a registered foal. Over a 7 year investment period, live foals must be produced in all but one year to yield a positive financial return. Profitability was highest among mares of greatest financial value. Conclusions: Mares are long‐term investments due to the extended period before there is a return on the investment. Improving our understanding of mare, stallion and management factors that affect the likelihood of producing a live foal are critical to ensuring a positive financial return. Additional work is needed to test the robustness of the study's conclusions when the cost and revenue assumptions are varied. Potential relevance: This information can assist in assessing mare profitability and developing management strategies to maximise profitability.     

Theory of ammonia toxicity as the mechanism of abortion in the mare reproductive loss syndrome

In late April/early May of 2001, for the equine industry in Kentucky, a previously unknown factor led to the loss of up to 40% of the new pregnancies (20-60 days) for that year, with a following significant number of late abortions and ill new born foals. In addition, encephalopathy, ocular lesions, pericarditis, laminitis, and other diseases were suspected to be associated with the syndrome. This occurrence was known as the mare reproductive loss syndrome (MRLS). On examination of the weather data, this period was seen to have an unusual pattern of severe late spring frost. Acute changes in the pasture diet affected the horses gut microflora, allowing the overgrowth of opportunistic pathogens and resulting in the excessive absorption of nitrogen and ammonia from the large bowel. Clinical blood chemistries from the horses on pasture at this time showed abnormally high levels of blood urea nitrogen (BUN). Elevated blood ammonia levels were found in mares and newborn foals affected with MRLS. Ammonia is metabolically one of the most toxic substances to the body. It is present naturally as a breakdown product of nitrogen turnover. If the body nitrogen balance is disturbed for any reason, with nitrogen accumulation exceeding the body's ability to excrete it as urea (BUN), ammonia will accumulate in toxic levels. Ammonia toxicity can cause all the symptoms that were observed in the horses exhibiting MRLS.     

母马生殖损失综合征研究现状及其在中国流行的可能性

母马生殖损失综合征(MRLS)是指由天幕毛虫相关因素引起的孕马早期胎儿流产(EFL)、晚期胎儿流产(LFL)、纤维素性心包炎(FP)和单侧葡萄膜炎(UU)等综合症状的疾病。2001年,美国首次报道该病,该病的暴发对美国养马业造成了巨大的经济损失。流行病学调查研究表明,母马生殖损失综合征在牧场间的传播与东方天幕毛虫(ETC)有关。其原因可能为:天幕毛虫毒素中毒并发细菌二次感染或天幕毛虫通过胃肠道黏膜进入血液导致菌血症,从而引发母马生殖损失综合征。在中国,天幕毛虫分布广泛,尤其是每年的春夏季节是天幕毛虫大量繁殖的季节。据大量报道表明,中国母马流产多发生在4~6月份天幕毛虫活动频繁的季节,而这些马流产病例中是否存在由天幕毛虫引起的母马生殖损失综合征尚未知。因此,我们对母马生殖流产综合征的流行现状以及临床症状的研究现状做了首次中文综述,希望对国内马流产的临床诊断、流行病学研究有所借鉴。     

Premature centromere division (PCD) identified in a hucul mare with reproductive difficulties

A hucul mare with reproductive abnormalities was examined during karyotype analysis. The karyotype was analysed based on evaluation of 860 metaphase plates in chromosome preparations. The use of fluorescence in situ hybridization (FISH) with an X chromosome painting probe showed premature X chromosome separation in 9.5% cases of examined chromosome spreads. In this report, we present the first identify premature centromere division (PCD) as a possible cause of abnormal X chromosome morphology in horses and as a probable cause of reproductive difficulties.     

Equine amnionitis and fetal loss: mare abortion following experimental exposure to Processionary caterpillars (Ochrogaster lunifer)

Reasons for performing study: In Australia, there have been recent reports of unusual abortions in mid‐ to late‐gestation mares. These were clinically distinct from other recognised causes of pregnancy loss and the term ‘equine amnionitis and fetal loss’ (EAFL) was adopted to describe this syndrome. Initial investigations concluded that possible causal factors included the presence on affected stud farms of Processionary caterpillars (Ochrogaster lunifer). Objectives: To determine if exposure of pregnant mares to Processionary caterpillars or their shed exoskeletons can induce EAFL. Methods: Processionary caterpillars and their shed exoskeletons were collected and stored frozen. Mid‐gestation mares were dosed with a slurry of caterpillars or shed exoskeleton by nasogastric intubation. Their clinical responses and times to abortion were recorded. All aborted fetuses were autopsied and samples taken for bacteriological and virological culture and histopathology. Results: Intubating mares in mid‐pregnancy with preparations of either whole Processionary caterpillars or shed caterpillar exoskeletons induced abortion with few impending clinical signs. The gross pathological and bacteriological findings of the aborted fetuses were similar to those observed in field cases of EAFL. Potential relevance: Possible exposure to Processionary caterpillars should be considered when examining cases of fetal loss in the mare. The present results provide a starting point to further explore the aetiology and pathogenesis of EAFL.     

Insulin-dependent diabetes mellitus associated with presumed autoimmune polyendocrine syndrome in a mare

A 5-year-old Thoroughbred-cross mare was diagnosed with insulin-dependent diabetes mellitus. Partial glycemic control and clinical improvement were achieved with daily insulin administration for 18 mo. The mare subsequently developed evidence of hypoadrenocorticism and died. Necropsy findings included lymphocytic infiltration of the pancreas, adrenal cortex, adrenal medulla, and thyroid glands, suggestive of an immune-mediated polyendocrinopathy.     

Prevalence and serovars of leptospira involved in equine abortions in central Kentucky during the 1990 foaling season.

A study to determine the prevalence of leptospira-induced abortions in the central Kentucky equine population during the 1990 foaling season and to determine the leptospira serovars responsible was conducted. From July 1, 1989 through June 30, 1990, 32 (4.4%) of 726 submissions (fetuses, stillborn foals, and/or placentas) were diagnosed as leptospirosis by the fluorescent antibody test and/or microscopic agglutination test. Attempts were made to isolate leptospires from the fetal tissues and/or the dam's urine in 31 of these cases. Leptospira interrogans serovar kennewicki was isolated from 11 (35.5%) and serovar grippotyphosa from 2 (6.5%) of the 31 cases. Of 12 cases that were culture negative with serologically positive fetal fluids, 8 had titers against serovar pomona, 1 against bratislava, 1 against grippotyphosa, 1 against hardjo, and 1 against both bratislava and pomona.     

Factors associated with early and mid-to-late fetal loss in lactating and nonlactating Holstein cattle in a hot climate

The purpose of this study was to evaluate associations of lactation, somatic cell count score (SCCS) at breeding, milk yield, lactation number, interval from calving to breeding (days open), number of times inseminated, and season of breeding on fetal loss for lactating Holstein females (both first-parity and multiparous cows) and nonlactating Holstein heifers in a hot climate. Females were palpated between d 40 and 50 of gestation and again at d 70 to 80 to determine pregnancy status. Early fetal loss was defined as a loss that occurred after d 40 to 50 but before d 70 to 80. Mid-to-late fetal loss represented losses after d 70 to 80 but before expected calving. Lactating females had higher early (P = 0.055) and mid-to-late fetal loss (P < 0.05) than nonlactating heifers. Those lactating females with increased days open experienced greater early (P < 0.05) and mid-to-late fetal loss (P = 0.055), whereas lactating females with an elevated SCCS encountered greater mid-to-late fetal loss (P < 0.01). Milk yield, lactation number, number of times inseminated, and season were not associated with early or mid-to-late fetal loss. For nonlactating heifers, there were no associations between number of times inseminated, season, or age at breeding on early or mid-to-late fetal loss. In conclusion, lactating females were more likely to suffer early and mid-to-late fetal loss than nonlactating heifers. Also, days open and SCCS at breeding were related to ability of lactating females to maintain pregnancy, but there were no relationships between fetal loss and milk yield, lactation number, number of times inseminated, or season.     

The mare reproductive loss syndrome and the eastern tent caterpillar: a toxicokinetic/statistical analysis with clinical, epidemiologic, and mechanistic implications

During 2001, central Kentucky experienced acute transient epidemics of early and late fetal losses, pericarditis, and unilateral endophthalmitis, collectively referred to as mare reproductive loss syndrome (MRLS). A toxicokinetic/statistical analysis of experimental and field MRLS data was conducted using accelerated failure time (AFT) analysis of abortions following administration of Eastern tent caterpillars (ETCs; 100 or 50 g/day or 100 g of irradiated caterpillars/day) to late-term pregnant mares. In addition, 2001 late-term fetal loss field data were used in the analysis. Experimental data were fitted by AFT analysis at a high (P <.0001) significance. Times to first abortion ("lag time") and abortion rates were dose dependent. Lag times decreased and abortion rates increased exponentially with dose. Calculated dose x response data curves allow interpretation of abortion data in terms of "intubated ETC equivalents." Analysis suggested that field exposure to ETCs in 2001 in central Kentucky commenced on approximately April 27, was initially equivalent to approximately 5 g of intubated ETCs/day, and increased to approximately 30 g/day at the outbreak peak. This analysis accounts for many aspects of the epidemiology, clinical presentations, and manifestations of MRLS. It allows quantitative interpretation of experimental and field MRLS data and has implications for the basic mechanisms underlying MRLS. The results support suggestions that MRLS is caused by exposure to or ingestion of ETCs. The results also show that high levels of ETC exposure produce intense, focused outbreaks of MRLS, closely linked in time and place to dispersing ETCs, as occurred in central Kentucky in 2001. With less intense exposure, lag time is longer and abortions tend to spread out over time and may occur out of phase with ETC exposure, obscuring both diagnosis of this syndrome and the role of the caterpillars.