Mare Reproductive Loss Syndrome Research Articles

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.

Effects of Nitrate and Pathogenic Nanoparticles on Reproductive Losses, Congenital Hypothyroidism and Musculoskeletal Abnormalities in Mares and Other Livestock: New Hypotheses

Spontaneous abortions, congenital hypothyroidism and musculoskeletal abnormalities were attributed to high nitrate in the diet of pregnant mares on Thoroughbred farms in central Kentucky. These fetal losses, with an unknown etiology, and associated with the Mare Reproductive Loss Syndrome (MRLS) have plagued horse farms in central Kentucky for decades. Fetal losses occur in mares grazing spring pastures affected by climatic and environmental factors including droughts, cold-stress, nitrogenous fertilizers, and herbicides. These factors may cause nitrate to accumulate in pasture forages. On a Thoroughbred horse farm, mares affected with the MRLS, pregnant Boer goats grazing high nitrate pastures also were affected with fetal losses. When spring pastures were not fertilized with nitrogen, herbicides not applied, protein reduced in the ration and the diet increased in sodium, fetal losses did not occur and foals were normal at birth. Excessive nitrate, ammonia and sulfate in the diet were associated with the formation of toxic and pathogenic abiotic nanoparticles in the amnionic fluid and pathognomonic placental lesions consistent with the MRLS. Pathogenic nanoparticles were found in aborted fetuses of other livestock. The discovery of these toxic pathogenic abiotic micro and nanoparticles in developing fetuses is unique. This novel mechanism of action for the pathogenesis of fetal losses may be a predisposing factor for a host of opportunistic diseases in livestock. The pathogenic nanoparticles collect in the vessels of the placenta and other organs to form niduses that predispose fetuses to a host of opportunistic microorganisms.

Reproductive failure in horses: identifying the problem

REPRODUCTIVE efficiency has significant implications for the economic success of equine stud farms, especially large thoroughbred breeding farms. This is particularly evident if one looks at one episode of mare...

Invited Review: The role of caterpillars in mare reproductive loss syndrome: A model for environmental causes of abortion1

A new abortigenic disease, now known as mare reproductive loss syndrome (MRLS), significantly affected the horse industry in the Ohio River Valley of the United States in late April and early May of 2001 and 2002. In 2001, approximately 25% of all pregnant mares aborted within several weeks (over 3,000 mares lost pregnancies), and abortion rates exceeded 60% on some farms. Mare reproductive loss syndrome struck hard and without warning, it was caused by something in the environment, it was not transmitted between animals, and it was not associated with any known abortigenic agent or disease. These experiments demonstrated that horses will inadvertently consume Eastern tent caterpillars (ETC) when the insects are present in the pasture or other feedstuffs, and MRLS-type abortions were induced in experimental animals (mares and pigs) by mixing ETC with the feed of the animals. Eastern tent caterpillars are hirsute (hairy) caterpillars, and the only part of the caterpillar that caused MRLS abortions was the cuticle. The experiments revealed that the setae (hairs) embed into the submucosa of the alimentary tract creating microgranulomatous lesions. It is hypothesized that the alimentary tract lesions allow bacteria from the alimentary tract of the mare, principally streptococci, actinobacilli, and to a lesser extent enterococci, to invade the circulatory system of the mare. The bacteria then establish infections in tissues where the immune surveillance of the mare is reduced, such as the fetus and placenta. Fetal and placental fluid bacterial infections lead to fetal death and abortion characteristic of MRLS. Inadvertent ingestion of ETC by pregnant mares causes MRLS. Currently the only known means to prevent MRLS is to avoid exposure of horses, particularly pregnant mares, to ETC and probably most hirsute caterpillars.

Nocardioform placentitis in Australia with implications for Equine Amnionitis and Foetal Loss (EAFL) and Mare Reproductive Loss Syndrome (MRLS)

Nocardioform placentitis in Australia with implications for Equine Amnionitis and Foetal Loss (EAFL) and Mare Reproductive Loss Syndrome (MRLS)

An unidentified toxin associated with Eastern Tent Caterpillars causes Mare Reproductive Loss Syndrome (MRLS)

An unidentified toxin associated with Eastern Tent Caterpillars causes Mare Reproductive Loss Syndrome (MRLS)

STILLBORN INFANT WITH CALCIFIED CHORIONIC EPITHELIUM, CORNEAL SCARRING, AND PERICARDITIS

This autopsy of a stillborn term infant revealed a constellation of unusual features including calcification of the chorion membrane and portions of the umbilical vascular media, extensive white matter gliosis, arthrogryposis multiplex congenita, adhesions of one eyelid to the globe, pericarditis, a miniature left foot, and a cleft palate. We hypothesized that the membrane and umbilical cord lesions resulted from an episode of resolved chorioamnionitis earlier in the pregnancy. Mare reproductive loss syndrome (MRLS) demonstrates a bacteremic infection of the amniotic cavity, pericarditis, and uniocular endophthalmitis in the mare. On the basis of analogy, we speculated that this infant suffered an intrauterine bacteremia with tissue predilection similar to that of MRLS.

Hormone Profiles of Mares Affected by the Mare Reproductive Loss Syndrome

While searching for the cause of the Mare Reproductive Loss syndrome (MRLS), we postulated that 1 of 3 tissues in 40-120 D pregnant mares was the likely primary target of the noxious factor that caused early abortions: The corpora lutea (CL), the endometrium or the fetus and/or its membranes. At this stage of gestation, progesterone (P4) is solely produced by luteal tissue, eCG by endometrial cups in the endometrium and oestrogens by the feto-placental unit. We determined whether concentrations of P4, eCG and/or total conjugated oestrogens (CE) would indicate which tissue was targeted during the MRLS. P4, eCG and CE were measured in single serum samples collected from 216 mares, 60-110 D after ovulation during the 2001 MRLS outbreak. All mares had previously been confirmed pregnant by ultrasonography. The following data was obtained from each mare: Interval from ovulation, pregnancy status and normalcy of fetal fluids at the time of sampling, and pregnancy status 3 weeks after sampling and at term. There were no meaningful differences in hormone concentrations between pregnant mares that had normal and excessively echogenic fetal fluids at the time of sampling. CE were lower (p < 0.05) in mares that aborted after sample collection than in mares the carried to term. In 8 mares from which multiple samples were obtained, CE consistently decreased prior to any decreases in P4 or eCG. Arguments are presented that lead to the hypothesis that the fetal trophoblast was the primary target of the MRLS agent.

Review Paper: Mare Reproductive Loss Syndrome

An epidemic of early fetal loss (EFL), late fetal loss (LFL), fibrinous pericarditis, and unilateral uveitis which occurred during the spring of 2001, are together now known as the mare reproductive loss syndrome (MRLS). A similar epidemic with less intensity was reported during the same period of time from southern Ohio, West Virginia, and Tennessee. The same syndrome with lesser intensity recurred in 2002. The estimated economic loss from the syndrome in 2001 and 2002 together was approximately $500 million. Both EFL and LFL were characterized by the absence of specific clinical signs in aborting mares. Nonhemolytic Streptococcus spp. and Actinobacillus spp. accounted for 65% of the organisms isolated from fetuses submitted for a postmortem during the MRLS period in 2001 and 2002. The pathologic findings in fetoplacental units of LFL included bronchopneumonia and funisitis, and there were no findings in EFL. Epidemiologic studies conducted in 2001 suggested an association between the presences of eastern tent caterpillars (ETC) in pastures with MRLS. Experimental studies in pregnant mares by exposure to ETC, or administration by stomach tube or with feed material, reproduced EFL and LFL. Similar experimental studies in mouse, rats, and goats with ETC were unsuccessful. Currently, 2 hypotheses are proposed for MRLS. One hypothesis proposes that an ETC-related toxin with secondary opportunistic bacterial invasion of the fetus leads to MRLS. The second hypothesis suggests that a breach of gastrointestinal mucosal integrity by hairs of ETC leads to a bacteremia and results in MRLS. In 2004, a similar equine abortion storm was reported from Australia and caterpillar exposure was identified as a risk factor for the abortion. In 2006, the syndrome was observed in Florida and New Jersey.

Pheromone Trap for the Eastern Tent Caterpillar Moth

The discovery that the eastern tent caterpillar Malacosoma americanum (F.) causes mare reproductive loss syndrome (MRLS), and thus has the potential to continue to result in major economic losses to the equine industry of Kentucky, has resulted in an intensive effort to identify practical means to monitor and control this defoliator, including these experiments to optimize a sex pheromone trap for this pest. A pheromone-baited delta trap with a large opening, such as InterceptST Delta, was more effective than other tested traps. Orange delta traps caught more moths than other tested colors. ETC males are caught at all tested heights within the tree canopy. For monitoring flights, setting traps at 1.5 m would allow easy counting of moths. A 9:1 blend of (E,Z)-5,7-dodecadienal (ETC-Ald) and (E,Z)-5,7-dodecadienol (ETC-OH) was most effective in capturing males. Increasing loading doses of a 3:1 blend (Ald:OH) resulted in the capture of increasing numbers of moths, but a 9:1 blend was more effective than 3:1 blend even at a nine-fold lower loading rate. Pheromone-impregnated white septa caught more moths than gray septa at the same loading dose. The advantages and limitations of using pheromone traps for monitoring M. americanum are discussed.

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

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. 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. 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. 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.

A Murine model of Mare Reproductive Loss Syndrome

Background Mare reproductive loss syndrome (MRLS), first reported in 2001 from Kentucky, has caused thousands of equine abortions and estimated $1/2 billion in economic loss. MRLS affects mare after ingesting caterpillars. This may result in irritation, lesions and increased gut permeability. The á hemolytics Strep. sp and Actinobacillus sp, present in normal digestive tract, have been isolated in field and experimental aborted cases. Bacterial replication in the fetal fluids is likely to compromise the fetus and causes abortion. A murine model would facilitate the study of MRLS. Methods Seventy programmed pregnant ICR mice were injected via tail vein or IP with 1X104–8 CFU Strep. cricetus and/or Actinobacillus sp isolated from aborted fetus. Control animals received saline. Results Injected dams did not develop any clinical symptoms. There were sporadic cases of aborted fetuses that correlated with the amount of bacteria. No significant difference was noted between the number of delivered pups and their weight between groups. Day old pups when injected with bacteria developed focal abscess, lost weight but recovered after one week. However, SRA/CD36 deficient pregnant mice developed 100% fetal death and abortion 24–74h after bacterial injection. We conclude that deficient mice may serve as a pivotal model to study MRLS.

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

To characterize the temporality of dates of breeding and abortion classified as mare reproductive loss syndrome (MRLS) among mares with abortions during early gestation. 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. 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. 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%). 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.

Orientation and Dispersal Patterns of the Eastern Tent Caterpillar, Malacosoma americanum F. (Lepidoptera: Lasiocampidae)

Larvae of the eastern tent caterpillar, Malacosoma americanum, undergo density-dependent dispersal in response to depleted resources. Because these caterpillars have recently been implicated in abortions of pregnant mares (equine Mare Reproductive Loss Syndrome, or MRLS), there is increased interest in managing caterpillar populations, potentially through manipulation of caterpillar dispersal behavior. Consequently, we investigated dispersal patterns of food-deprived eastern tent caterpillars in artificial arenas with respect to distance, direction, and response to visual stimuli. Distance traveled is influenced by time of day, and is strongly correlated with time elapsed. Movement is non-random, and correlates closely with the position of the sun. The pattern is more pronounced with foraging third instars than with penultimate fifth instars. Visual cues appear important in caterpillar orientation, and caterpillars are responsive to vertically oriented, black objects.

Microbiologic and Pathologic Findings in an Epidemic of Equine Pericarditis

During the spring and summer of 2001 and in association with the mare reproductive loss syndrome, 22 terminal and 12 clinical cases of equine pericarditis were diagnosed in central Kentucky. Actinobacillus species were the principal isolates from 8 of 10 nontreated, terminally affected and 3 of 10 clinically affected horses. Enterococcus faecalis and Streptococcus zooepidemicus were cultured from the remaining 2 nontreated terminal cases. No viruses were isolated in tissue culture. Nucleic acid of equine herpesvirus-2 was detected in pericardial and tracheal wash fluids of 3 and 1 individuals, respectively. Microscopic alterations in sections of heart and parietal pericardium were consistent with chronic fibrinous bacterial pericarditis. This report confirms a significant role of Actinobacillus species in equine pericarditis and describes an epidemic of this infrequently observed syndrome in the horse.

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

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. 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. 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. 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.

Managing Eastern tent caterpillars Malacosoma americanum (F) on horse farms to reduce risk of mare reproductive loss syndrome

An equine disease now known as mare reproductive loss syndrome (MRLS) struck the Ohio Valley in 2001-2002 causing thousands of foal abortions and enormous economic loss. Evidence that pregnant mares' exposure to Eastern tent caterpillars Malacosoma americanum (F) induces MRLS created an urgent call for control measures suitable for use on horse farms. We surveyed egg mass distribution and monitored emergence in wild cherry trees, and evaluated reduced-risk treatment strategies including foliage sprays, trunk injections, winter egg mass treatments and barrier sprays to intercept larvae entering pastures. Egg masses were concentrated in the lower canopy, on exposed sides of trees. Larval emergence began in mid-March. Emergence was prolonged (3-4 weeks) in 2002, a typically cool spring, but more synchronized in warmer 2003. Winter treatment of egg masses with bifenthrin or permethrin in a penetrating solvent prevented emergence, but 3% horticultural oil was ineffective for that purpose. Insecticidal soap or oil sprayed directly on neonates gave relatively poor control. Bifenthrin and spinosad were effective as foliage sprays against all instars, their field-weathered residues active for at least 7 days. Bacillus thuringiensis Berliner var kurstaki controlled neonates within 3 days but was less active against late instars, with shorter residual action than bifenthrin or spinosad. Larvae did not avoid insecticide-treated leaves. Trunk microinjection of cherry trees with dicrotophos was effective against all instars, whereas microinjection with milbemectin or abamectin gave poor or inconsistent control. Trunk injection with emamectin benzoate also was effective. Dry permethrin residues controlled late instars crawling in pasture grass for at least 7 days. Factors complicating M americanum management on horse farms are discussed.

Eastern tent caterpillars ( Malacosoma americanum) cause mare reproductive loss syndrome

A new equine abortigenic disease, mare reproductive loss syndrome (MRLS), was recognized and significantly impacted the Ohio Valley in the springs of 2001 and 2002. MRLS caused ~$ 330 million in losses in 2001. An epidemiological investigation of MRLS associated occurrence of the disease with exposure to eastern tent caterpillars ( M. americanum). This work investigates the epidemiological association between M. americanum and MRLS to determine if this association was correlative or causative. A pilot study and Experiment 1 simulated exposure to M. americanum and their excreta on pasture grasses. The pilot study advanced exposure of pregnant mares to M. americanum materials and 18 of the 29 mares in the study aborted with symptoms of MRLS before other cases were reported in the region. In Experiment 1, three of seven mares exposed to M. americanum aborted, while mares in control ( n=6) and M. americanum frass ( n=7) treatments had no losses. In Experiment 2, mares were fed frozen insect larvae in feed buckets mixed with oats. Abortions occurred in three of five mares receiving frozen M. americanum, while mares that were fed autoclaved M. americanum ( n=5) or frozen gypsy moth larvae ( n=4) had no abortions due to MRLS. In Experiment 3, M. americanum larvae were dissected and fractionated. Statistically significant numbers of abortions occurred only in the positive control group and in association with the M. americanum exoskeleton. All abortions induced by exposure to M. americanum exhibited changes in echogenicity of fetal fluids and bacteriological findings post abortion that were consistent with MRLS. These studies support the hypothesis that ingestion of M. americanum larvae induces the MRLS-type equine abortions, and provide experimental evidence that this lepidopteran larva can cause an abortigenic disease in a vertebrate host.

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

To identify factors significantly associated with an epidemic of fibrinous pericarditis during spring 2001 among horses in central Kentucky. Case-control study. 38 horses with fibrinous pericarditis and 30 control horses examined for other reasons. A questionnaire was developed to solicit information regarding a wide range of management practices and environmental exposures from farm owners or managers. 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.

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.