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.     

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.     

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.     

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.     

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 early-term abortions (early fetal losses) associated with mare reproductive loss syndrome in central Kentucky

OBJECTIVE: To identify factors associated with abortions during early gestation classified as mare reproductive loss syndrome (MRLS). DESIGN: Case-control study. ANIMALS: 324 broodmares from 43 farms in central Kentucky, including 121 mares from 25 farms that had early-term abortions (ETAs) associated with MRLS (case horses), 120 mares from the same farms but that did not abort, and 83 mares from 18 farms that were not severely Impacted by MRLS. PROCEDURE: Farm managers were interviewed to obtain data on various management practices and environmental exposures for the mares. Data for case and control horses were compared to identify risk factors for mares having MRLS-associated ETAs. RESULTS: Several factors increased the risk of MRLS-associated ETAs, including feeding hay in pasture, greater than usual amounts of white clover in pastures, more eastern tent caterpillars in pastures, abortion during a previous pregnancy, and sighting deer or elk on the premises. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis indicates that certain characteristics of pastures predisposed mares to MRLS-associated ETAs. Methods for limiting exposure to pasture (keeping mares in stalls longer) during environmental conditions similar to those of 2001 (ie, sudden freezing in mid-April following warmer-than-usual springtime temperatures and larger-than-usual numbers of eastern tent caterpillars in and around pastures) should reduce the risk of mares having MRLS-associated ETAs. It was not possible to determine whether exposure to white clover or caterpillars were causal factors for MRLS or were merely indicators of unusual environmental conditions that resulted in exposure of mares to a toxic or infectious agent.     

The effect of natural toxins on reproduction in livestock.

Reproductive efficiency is the most important economic factor in livestock production. Thus, the hypothalamo-pituitary-gonadal regulatory axis, accessory sexual organ functionality, and the complex events involved in fertilization, implantation, and embryonic and fetal development may be sensitive to therapeutic agents, environmental pollutants, and natural toxicants. There are many factors that adversely affect reproduction, one of which is toxic substances in the diets of animals. Toxic materials can affect reproductive success by causing abortions, interfering with libido, estrus, oogenesis, or spermatogenesis, causing emaciation and subsequent abnormal mating behavior, birth defects, and increasing the time between parturition and rebreeding. Examples of natural toxicants in poisonous plants interfering with reproduction are numerous. Abortion in livestock from locoweeds, ponderosa pine needles, broom snakeweeds, fescue, and others are reported in studies. Selenium and seleniferous forage inhibit estrus in cattle and swine. Emaciation and temporary illness from sneezeweeds, bitterweed, locoweed, larkspur, lupines, and others may interfere with mating. Embryonic loss and birth defects from Veratrum, lupines, locoweeds, poison hemlock, and so on, may occur. As suggested, toxins have many diverse and economically adverse effects on reproductive performance in livestock.     

苦马豆素对母畜繁殖机能的影响及防控

苦马豆素主要存在于豆科棘豆属和黄芪属植物如疯草中,动物采食大量疯草会发生神经中枢紊乱,苦马豆素中毒的母畜表现不孕、流产,产死胎、畸胎等,引起繁殖能力下降,妨碍家畜繁育和品种优化,给我国草原畜牧业造成严重的经济损失,严重制约了我国西部草原牧区养殖业的持续发展。现通过对疯草主要毒性成分苦马豆素对母畜的繁殖性能指标、生殖器官组织形态学病理变化和生殖激素的影响进行综述,旨在为母畜疯草中毒引起的繁殖机能紊乱疾病提供治疗及预防思路,降低牧区养殖业经济损失,提高我国畜牧业长远发展的能力。     

Developing vaccines to control protozoan parasites in ruminants: dead or alive?

Protozoan parasites are among some of the most successful organisms worldwide, being able to live and multiply within a very wide range of hosts. The diseases caused by these parasites cause significant production losses in the livestock sector involving reproductive failure, impaired weight gain, contaminated meat, reduced milk yields and in severe cases, loss of the animal. In addition, some protozoan parasites affecting livestock such as Toxoplasma gondii and Cryptosporidium parvum may also be transmitted to humans where they can cause serious disease. Data derived from experimental models of infection in ruminant species enables the study of the interactions between parasite and host. How the parasite initiates infection, becomes established and multiplies within the host and the critical pathways that may lead to a disease outcome are all important to enable the rational design of appropriate intervention strategies. Once the parasites invade the hosts they induce both innate and adaptive immune responses and the induction and function of these immune responses are critical in determining the outcome of the infection. Vaccines offer green solutions to control disease as they are sustainable, reducing reliance on pharmacological drugs and pesticides. The use of vaccines has multiple benefits such as improving animal health and welfare by controlling animal infections and infestations; improving public health by controlling zoonoses and food borne pathogens in animals; solving problems associated with resistance to acaricides, antibiotics and anthelmintics; keeping animals and the environment free of chemical residues and maintaining biodiversity. All of these attributes should lead to improved sustainability of animal production and economic benefit. Using different protozoan parasitic diseases as examples this paper will discuss various approaches used to develop vaccines to protect against disease in livestock and discuss the relative merits of using live versus killed vaccine preparations. A range of different vaccination targets and strategies will be discussed to help protect against: acute disease, congenital infection and abortion, persistence of zoonotic pathogens in tissues of food animals and passive transfer of immunity to neonates.     

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.     

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.     

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.     

霉菌毒素对畜禽的危害及削减方法研究进展

霉菌毒素是由不同种属真菌产生的次级代谢物,其对多种动物（包括人在内）具有显著毒性效果,如降低畜禽生产性能、损害脏器功能、影响机体免疫功能、造成细胞和DNA氧化损伤,甚至导致动物死亡等。饲料及其原料霉菌毒素污染是一个广泛的、全球性的问题,既可在作物田间生长时产生,也可在仓储期间产生,给畜禽养殖业造成了巨大的经济损失,防控霉菌生长及去除饲料中霉菌毒素已成为国内外研究的热点。实际上,不同霉菌毒素对不同动物的损害具有差异性,而在畜禽养殖过程中添加适当物质（如蒙脱石和酶制剂等）可有效抑制霉菌毒素活性,缓解霉菌毒素的毒性。作者详细论述了玉米赤霉烯酮、烟曲霉毒素、呕吐毒素、T-2毒素、黄曲霉毒素和烟曲霉毒素对畜禽的危害,并从物理、化学、生物角度介绍了削减饲料及其原料中霉菌毒素的方法,旨在为合理有效防控霉菌毒素毒害及保障畜禽健康养殖提供参考。     

T-2毒素研究进展

T-2毒素是污染我国饲料的主要霉菌毒素,主要危害动物的造血组织和免疫器官,引起出血性综合征,出现白细胞减少、贫血,使胃肠道功能受损等,对畜禽健康和畜牧业生产的危害很大.本文主要从T-2毒素的一般性质、产毒菌株、毒性效应、毒性机制及预防控制等方面作了论述.     

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.     

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.     

布鲁菌病及其研究进展

布鲁菌病常发生于牛、羊、猪等家畜,且可传染给人和其他家畜,主要特征是生殖器官和胎膜发炎,从而导致繁殖障碍,给畜牧业和人类健康带来严重危害。主要结合文献对布鲁菌病相关知识进行介绍,以期为有效防控该病提供参考。     

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.     

Transgenesis may affect farm animal welfare: a case for systematic risk assessment

This paper considers (potentially) harmful consequences of transgenesis for farm animal welfare and examines the strategy of studying health and welfare of transgenic farm animals. Evidence is discussed showing that treatments imposed in the context of farm animal transgenesis are by no means biologically neutral and may compromise animal health and welfare. Factors posing a risk for the welfare of transgenic farm animals include integration of a transgene within an endogenous gene with possible loss of host gene function (insertional mutations), inappropriate transgene expression and exposure of the host to biologically active transgene-derived proteins, and in vitro reproductive technologies employed in the process of generating transgenic farm animals that may result in an increased incidence of difficult parturition and fetal and neonatal losses and the development of unusually large or otherwise abnormal offspring (large offspring syndrome). Critical components of a scheme for evaluating welfare of transgenic farm animals are identified, related to specific characteristics of transgenic animals and to factors that may interact with the effects of transgenesis. The feasibility of an evaluation of welfare of transgenic farm animals in practice is addressed against the background of the objectives and conditions of three successive stages in a long-term transgenic program. Concrete steps with regard to breeding and testing of transgenic farm animals are presented, considering three technologies to generate transgenic founders: microinjection, electroporation and nuclear transfer, and gene targeting including gene knockout. The proposed steps allow for unbiased estimations of the essential treatment effects, including hemi- and homozygous transgene effects as well as effects of in vitro reproductive technologies. It is suggested that the implementation of appropriate breeding and testing procedures should be accompanied by the use of a comprehensive welfare protocol, specifying which parameters to monitor, at which stages of the life of a farm animal, and in how many animals. Some prerequisites and ideas for such a protocol are given. It is anticipated that systematic research into the welfare of farm animals involved in transgenesis will facilitate the use of the safest experimental protocols as well as the selection and propagation of the healthiest animals and, thereby, enable technological progress that could be ethically justified.     

Economic aspects of reproductive failure in dairy cattle. I. Financial loss at farm level

Some years ago, systematic research was initiated to improve quantitative insight into the economic impact of diseases and disease control in livestock. This paper deals with the financial loss at farm level caused by reproductive failure in dairy cattle. Economic calculations concerning the calving interval of cows which differ in age, relative production level and persistence of milk production during lactation were made. On average, an optimal interval of 1 year or less was established, while the loss per day lengthening of the interval (norm-loss) amounted to 1–2 Dutch guilders (Dfl.). Also, 676 norm-losses due to forced replacement because of reproductive failure were determined, dependent on age and relative production level of the culled cow and average herd life of the farm. On average, this norm-loss was determined to be almost Dfl. 500 per culled cow. Based on 71 farms in an investigation in the province of Overijssel, total loss per farm was determined, using the calving interval data, forced replacement data and norm-loss rate described above. On average, the calculated loss amounted to Dfl. 63 per cow per year, of which Dfl. 35.50 resulted from sub-optimal calving interval and Dfl. 27.50 from forced replacement due to reproductive failure. Costs for veterinary treatment and drugs were not included in these elements. Total loss due to reproductive failure was estimated to average about Dfl. 80 per cow per year, which equals about 2% of the gross production value or 10% of an average farmer's income.Finally, considerable differences in loss between farms have been determined. The difference between the 20% of farms with the highest and the 20% of farms with the lowest calculated loss is even more than the average loss. Thus improvment may be achieved on many farms, e.g., with the help of a herd health programme.