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Bovine abortion causes considerable economic losses to the livestock industry worldwide and is of concern for public health and food safety, given that many abortigenic infectious agents of cattle are zoonotic. Despite its importance, the etiological diagnosis of abortion in cattle is challenging both for veterinary practitioners and laboratory technicians, partly due to the difficulty in recovering aborted fetuses under extensive field conditions for pathological and microbiological diagnostic investigation, and in the early identification of aborted dams. Neospora caninum is a cosmopolitan protozoon identified as one of the main abortigenic agents in cattle worldwide. In this study we propose a comparative seroepidemiological approach for the diagnosis of abortion by N. caninum in dairy cattle. Samples from 12 to 93 cows/heifers with and without recent history of abortion (cases and controls) in four commercial dairy farms were tested. The ratio of controls to cases tested varied from 1:1 to 4.6:1. All samples (n=230) were analyzed by three commercial ELISA kits for the detection of anti-N. caninum antibodies. In all four dairy farms, the proportion of seropositive cows and/or heifers per kit was significantly higher in the cases than in the controls (Odds Ratios=5.13 to 36, p=0.0002 to 0.0485). The agreement among the three kits varied from weak to strong (Coheńs kappa coefficients=0.58 to 0.83). We conclude that, despite the imperfect agreement between these kits, all of them allowed to arrive at similar conclusions regarding the statistical association between N. caninum seropositivity and abortion, thus representing a useful tool for the diagnostic approach at the population level under field conditions., (Copyright © 2019 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.)

The objective of this study was to determine the association of beta-carotene concentration in plasma at the moment of artificial insemination (AI) on pregnancy/AI in lactating Holstein cows. A total of 399 events from 364 lactating Holstein cows were enrolled in the trial (143 primiparous and 221 multiparous). All cows were assigned to a timed AI protocol based on estradiol and progesterone. Blood samples were collected at the moment of AI and at 24 and 31d post-AI (samples on 31 d post-AI were collected only from cows that were diagnosed pregnant). The BCS were recorded at the time of AI. Plasma beta-carotene was quantified from blood samples taken at the time of AI using a single step denaturation and extraction into a solvent, followed by measurement using a portable spectrophotometer. Pregnancy-associated glycoproteins (PAG) were analyzed in blood samples taken at 24 and 31 d post-AI of pregnant cows. Milk production was collected for the entire experimental period. Pregnancy diagnosis was performed by ultrasound 31 and 60 d post-AI. Data was analyzed using the MIXED and GLIMMIX procedures of SAS. Cows classified as thin (<2.75) tended to have lower concentration of beta-carotene at AI when compared with those classified as Moderate (≥3.00; 3.8 ± 0.1 vs. 4.3 ± 0.1 μg/mL; P = 0.09). Concentration of beta-carotene were greater in multiparous compared with primiparous (P < 0.01). There was no correlation between concentration of beta-carotene and milk production (r = 0.04; P = 0.10). When plasma beta-carotene was categorized in quartiles, cows in the 1st quartile had lower pregnancy/AI and higher pregnancy losses when compared with cows that were in the 2nd, 3rd and 4th quartile (pregnancy/AI = 19.2 ± 4.5, 33.7 ± 4.7, 36.9 ± 5.0 and 39.8 ± 5.4%, respectively; P = 0.05; pregnancy losses = 41.9 ± 4.8, 20.4 ± 3.7, 22.1 ± 4.1, and 15.7 ± 4.2%, respectively; P < 0.05). There was no association between concentrations of beta-carotene at AI and PAG at 24 d post-AI (P = 0.60). Cows with greater concentrations of beta-carotene at AI were more likely to have greater concentrations of PAG at 31 d post-AI (P < 0.01). In conclusion, the concentration of beta-carotene at AI was affected by BCS and parity. Cows with higher concentrations of plasma beta-carotene at AI had greater pregnancy/AI, lower pregnancy losses and greater concentrations of PAG at d 31 post-AI, suggesting it may be associated with placental function in lactating dairy cows., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Double ovulation occurs more frequently in multiparous cows with high milk production than in primiparous cows and the rate of twin pregnancy/calving is increasing worldwide. Diagnosis of twin pregnancy is possible by ultrasound at the time of early pregnancy examination [28-34 days after artificial insemination (AI)]. Pregnancy proteins are also well-known indicators of gestation. The risk of pregnancy loss during the first trimester of gestation for cows carrying twins is three to nine times higher than for cows carrying singletons. Pregnancy-specific protein B (PSPB) is a good indicator not only of pregnancy but also of pregnancy loss. The aims of this study were (a) to collect calving data in some Hungarian Holstein-Friesian herds (n = 7,300) to compare PSPB serum concentrations (measured 29-35 days post insemination) in twin- and singleton-calving cows (Trial 1), and (b) to check the predictive value of PSPB serum concentration for twin pregnancy and pregnancy loss in high-producing Spanish Holstein-Friesian cows (n = 98; Trial 2). Our results showed almost 7% twin calving rate. Although hormonal treatments are commonly believed to be major causes of twin pregnancies, our data do not support this hypothesis. The only exception is the single PGF injection, which significantly increased twin calving. No effect of milk production on the risk of twin pregnancy was found, and twin pregnancy increased with parity. The AI bull, the bull's sire, the bull's grandfather and the cow's father also affected twin calving (P ≤ 0.02). We found much higher frequency of twin calving in cows diagnosed pregnant with higher than 3 ng/ml serum PSPB concentrations at 29-35 days after insemination. In Trial 2, non-significant but well-marked differences were found in PSPB serum concentration between singleton- and twin-pregnant cow samples (2.1 and 2.9 ng/ml) at different bleeding times. Probably the small size of the study population and the effects of milk production on PSPB values may explain this lack of significance.

Sixteen cases of spontaneous pregnancy loss (11 of singletons and five of pairs of twins) are described. The losses occurred between gestation Days 13 and 25 in 12 mares being monitored almost daily by transrectal ultrasonography (for measurement of conceptus growth) and blood sampling (for determination of maternal plasma progesterone concentrations as evidence of luteolysis) in experimental studies of early pregnancy. In 10 of the 16 cases the uterus was flushed and eight conceptuses were recovered for morphological assessment. Five of the 11 losses of singletons occurred before Day 16 and, with one exception, were preceded or accompanied by luteolysis. The remaining six singleton pregnancies failed after Day 16, with two cases evidencing luteolysis beforehand. Thus, overall, 6/11 singleton losses were associated with luteolysis while 5/11 were not. The five cases of simultaneous loss or degeneration of twin conceptuses all occurred on Day 19 or 20, preceded by luteolysis in only one case. These observations suggest that while the causes of spontaneous early pregnancy failure are multifactorial, luteolysis might contribute to the problem more often than has been previously contended.

This study was conducted to critically evaluate weekly and monthly circulating concentrations of immunoreactive relaxin throughout pregnancies that resulted in live births, stillbirths, and abortions in aquarium-based bottlenose dolphins. A relaxin RIA was used to analyze serum collected during 74 pregnancies involving 41 dolphins and 8 estrous cycles as well as 8 non-pregnant dolphins. Pregnancies resulted in live births (n=60), stillbirths (n=7), or abortions (n=7). Relative to parturition (Month 0), monthly changes (P<0.0001) in relaxin was indicated by relatively low concentrations during early pregnancy (Months -12 to -9) which subsequently increased (P<0.05) during mid- (Months -8 to -5) to late (Months -4 to -1) pregnancy; relaxin was highest (P<0.05) at the time of parturition. Post-parturition (Month 1), concentrations decreased (P<0.05). During the first 4weeks post-ovulation, relaxin concentrations were not different between pregnant and non-pregnant dolphins (status-by-week interaction, P=0.59). Status-by-month interaction (P<0.0002) involving different pregnancy outcomes was due, impart, to an increase in relaxin during early pregnancy (P<0.05) that was comparable among dolphins with live births, stillbirths, and abortions except concentrations were lower (P<0.05; 52%) at mid-pregnancy in association with pregnancy loss. Thereafter, concentrations increased (P<0.05) during late pregnancy in dolphins with stillbirths but not in dolphins with abortions. In conclusion, this study provided new information on the pregnancy-specific nature of relaxin, critical evaluation of the fundamental characteristics of relaxin during pregnancy and pregnancy loss, and clarification on the strengths and limitations of relaxin as a diagnostic aid to determine pregnancy status and assess maternal-fetal health in bottlenose dolphins., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Sera from 221 cattle were collected in 25 farms in Morocco to investigate the evidence and circulation of some of the main bovine abortive agents in the dairy cattle farming, where abortions are often reported. All sera were examined for brucellosis, 176 for neosporosis, 88 for leptospirosis, and 42 for Bovine viral diarrhoea (BVD/MD), Bovine Herpesvirus 1 (BHV-1) (Infectious bovine rhinotracheitis, IBR/IPV), and Bovine Herpesvirus 4 (BHV-4) infections (at least 1 sample per herd). Abortions were reported in 23 (10.4%) of the 221 tested cattle. Antibodies against the investigated pathogens were detected in all samples tested, with an overall seroprevalence of 33.48% for Brucella, 9.09% for Leptospira, 8.52% for Neospora, 37.71% for BVDV, 50% for BHV-1, 9.52% for BHV-4. As for Leptospira antibodies against serovars Hardjo, Pomona, and Tarassovi were identified. Mixed infections were common. The lack of evidence of non-infectious factors epidemiologically related to abortions suggested that the investigated agents are to be considered important risk factors in the dynamic of the abortion syndrome, even if further investigations are necessary to identify the abortion cause. Particular attention should be paid on brucellosis, considering the high seroprevalence and its zoonotic relevance.

Alpha-fetoprotein has proved to be a good indicator of fetal well-being in human medicine for decades. Although this molecule is present in most of the mammalian species including horses, reference values in healthy and high-risk pregnant mares have not yet been published. The aim of the present study was to determine whether equine alpha-fetoprotein (eqAFP) is a good indicator of complicated pregnancies in Lipizzaner mares. A total of 111 serum samples from 30 mares have been analyzed for eqAFP levels throughout gestation (Days 60-325). After the pregnancy was confirmed, 23 mares had normal pregnancies with viable foals, six had late embryonic loss, and one of the mares aborted in the ninth gestational month. Equine alpha-fetoprotein concentrations significantly differed in the normal group (72.93 ± 49.25 pg/mL; mean ± standard deviation) and in the complicated pregnancy loss group (152 ± 36.48 pg/mL; mean ± standard deviation). The mares' age, gestational age, and the conception rate significantly affected the alpha-fetoprotein concentrations in the normal group. Furthermore, notable individual differences occurred in eqAFP concentrations between mares. Equine alpha-fetoprotein seems to be an important indicator of fetal well-being in horses, but there are still some unanswered questions (levels in foals of different age, ponies, and draft horses) regarding this serum protein. Large-scale studies are needed to assess the specificity, sensitivity, and reliability of this test as a possible future diagnostic tool for fetal well-being in horses., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Rift Valley fever is a mosquito-borne zoonotic disease of domestic ruminants. This disease causes abortions in pregnant animals, and it has a high mortality rate in newborn animals. Recently, a Rift Valley fever virus (RVFV) outbreak in the Arabian Peninsula increased its potential spread to new regions worldwide. In non-endemic or disease-free countries, early detection and surveillance are important for preventing the introduction of RVFV. In this study, a serological surveillance was conducted to detect antibodies against RVFV. A total of 2382 serum samples from goats and cattle were randomly collected from nine areas in South Korea from 2011 to 2013. These samples were tested for antibodies against RVFV, using commercial ELISA kits. None of the goats and cattle were positive for antibodies against RVFV. This finding suggests that this disease is not present in South Korea, and furthermore presents the evidence of the RVFV-free status of this country.

In a recent lambing season (2012/2013), the seroprevalence of ovine chlamydiosis was monitored in small ruminant abortion cases in Croatia. Blood samples of 93 sheep and 69 goats were examined. In addition, 50 sheep and 61 goat samples were tested using molecular methods. Furthermore, 14 sheep blood samples, one goat blood sample and one sheep placenta sample from Bosnia and Herzegovina (BIH) were also tested as a part of inter-laboratory cooperation. Overall high seroprevalence was detected in sheep, 19.6% with the ELISA IDEXX kit and 20.5% with the ClVTEST kit. Seroprevalence in goats was 11.4%. In BIH, four sheep and one goat blood sample were seropositive for chlamydiosis. The disease causing agent, Chlamydia abortus (C. abortus) was confirmed using molecular methods in two sheep flocks in continental Croatia and in one sheep flock in BIH. In this study, C. abortus infection in sheep was identified for the first time in Croatia using species specific molecular methods. Ovine chlamydiosis is present in national sheep and goat flocks in Croatia and BIH. Thus should be subject to ongoing controls in the case of abortion. A combination of serological and molecular methods should be used for optimal laboratory diagnostics of C. abortus.

The use of live vaccine strain RB51 for vaccination of domestic water buffaloes (Bubalus bubalis) at risk of infection with Brucella abortus is permitted notwithstanding the plans for the eradication and only under strict veterinary control. The antibodies induced by RB51 vaccination are not detectable using conventional diagnostic techniques; therefore, it is necessary to have a specific diagnostic tool able to discriminate vaccinated from unvaccinated animals. The combination of a complement fixation test (CFT) with specific RB51 antigen (RB51-CFT) and a brucellin skin test has been demonstrated to be a reliable diagnostic system to identify single cattle (Bos taurus) vaccinated with RB51. So far, no data are available in the international scientific literature regarding the use of this test association in water buffalo. For this reason the suitability of this test combination has been evaluated in a water buffalo herd. One hundred twenty-seven animals farmed in a herd of Salerno province (Campania, Southern Italy), in the context of a presumptive unauthorized use of RB51 vaccine were chosen for this study. All tested animals resulted negative to Rose Bengal test (RBT) and complement fixation test (CFT) used for the detection of specific antibodies against Brucella field strains. Seventy-one animals (56%) developed RB51 antigen-specific CFT (RB51-CFT) antibodies against RB51 vaccine in a first sampling, while 104 animals (82%) gave positive result to a second serum sampling conducted 11 days after the intradermal inoculation of the RB51 brucellin. One hundred and seven animals (84%) showed a positive reaction to the RB51-CFT in at least 1 sampling, while 111 animals (87%) resulted positive to the RB51 brucellin skin test. Thus, analysing the results of the 3 testing in parallel, 119 animals (94%) were positive to at least 1 of the performed tests. The results suggest that the use in parallel of the RB51 brucellin skin test with RB51-CFT may represent a reliable diagnostic system to identify water buffaloes vaccinated with RB51 vaccine.

A serological survey was conducted to estimate the seroprevalence of 5 major abortive infections in 13 sheep flocks and 10 goat herds in 2 regions of Morocco. A total of 308 from aborted females (202 ewes and 106 does) and 197 sera (97 ewes and 99 does), were tested for brucellosis, chlamydiosis, Q fever, toxoplasmosis, and for 9 major serovars of Leptospira. An average abortion rate of 12.10% was found in ewes and 10.26% in does. The serological analyses revealed the presence of all 5 abortive infections, both in sheep and in goats. Ten (43%) herds/flocks were positive to brucellosis, 21 (91%) to chlamydiosis, 17 (74%) to toxoplasmosis, 13 (57%) to Q fever, and 5 (22%) to leptospirosis. Leptospira spp. serovars Copenhageni and Grypothyphosa were found in a single sheep flock, while Tarassovi and Copenhageni were detected in 4 goat herds. Of the 23 investigated herds/flocks, 22 (96%) showed mixed infections. The findings of this study confirmed the possible involvement of the 5 selected abortive infections in abortion outbreaks occurring in the investigated regions. Further investigations are needed to better understand the aetiology of infectious abortions in herds and flocks within investigated regions.

Neospora caninum is an important cause of abortion in cattle worldwide, but the isolation of a viable parasite from an abortus is difficult, and viable N. caninum has not been isolated from any host in China. In the present study, peripheral blood samples were collected from a jugular vein of an adult dairy cow that had aborted; the cow was seropositive to N. caninum antibodies by ELISA. White blood cells were separated and seeded onto Vero cell monolayer cultures for parasite isolation. Tachyzoites were first observed in cell culture on day 84 after initial inoculation. The parasite was confirmed to be N. caninum by gene sequencing and immunofluorescence, and by bioassays in BALB/c mice. The new N. caninum isolate (NC-Bj) has a unique pattern on microsatellite Cont-14. To our knowledge, this is the first successful isolation of N. caninum in China from any host.

In this cross-sectional study, we assessed and mapped the seroprevalence of brucellosis in small-scale dairy farming in an urban and peri-urban area of Tajikistan and investigated factors associated with seropositivity. As urban and peri-urban farming is both an opportunity to improve the livelihood for small-scale farmers and a potential public health hazard, studies are warranted to reveal possible peculiarities in the epidemiology of brucellosis in this type of dairy farming. In total, 904 cows of breeding age belonging to 443 herds in 32 villages were serologically tested with indirect enzyme-linked immunosorbent assay (ELISA) and positive samples confirmed with competitive ELISA. Two logistic regression models were used to investigate an association between seropositivity and risk factors at herd and individual level. The herd and individual seroprevalences were 4.1 and 2.0 %, respectively. Herds with a history of abortions were found to be associated with seropositivity [odds ratio (OR) = 5.3; 95 % confidence interval (CI), 1.3-21.3]. Large herds with more than eight cattle were more likely to be seropositive compared to smaller herds with one to two cattle (OR = 13.9; 95 % CI, 1.6-119). The number of calves produced per cow (indicating age) was found to be associated with seropositivity. Younger cows with one to two produced calves were less likely to be seropositive compared to older cows with more than six produced calves (OR = 0.24; 95 % CI, 0.06-1.0). Neither introduction of new cattle to the herd nor communal grazing was associated with seropositivity. This study shows that infection with Brucella (1) is present in small-scale urban and peri-urban dairy farming in Tajikistan and (2) has significant negative effects on reproductive performance in this farming system and (3) that some previously known risk factors for seropositivity in rural farming system were absent here.

Chlamydophila abortus is an important amphixenosis which in a wide range of animals, associated with reproductive disorders in yaks. In order to assess the prevalence of this infection in yaks in Qinghai, China, a cross-sectional study was carried out, and a total of 674 serum samples were collected from June to October 2012 in six counties, and antibodies to C. abortus were examined by indirect hemagglutination (IHA) test. The overall seroprevalence of C. abortus in yaks was 17.66 % (119/674), and the seroprevalence of antibodies to C. abortus in yaks ranged from 11.82 to 28.43 % among the six different areas, and the difference was statistically significant (P < 0.05). The seropositivity of C. abortus infection in different age groups varied from 16.33 to 18.49 %, and prevalence in yaks of ≥3 year (18.49 %) was slightly higher than that in yaks of <3 year, but the differences among the age groups were not statistically significant (P > 0.05). The seroprevalence of C. abortus infection in male yak (16.8 %) was slightly lower than that in females (17.85 %), and the difference was not statistically significant (P > 0.05). So far, this is the first systematic and comprehensive investigation of C. abortus infectionin in yaks in this area.

Reasons for Performing Study: Placentitis is a prevalent cause of abortion, premature delivery and neonatal death in mares. Early diagnosis is paramount for the survival of the fetus and delivery of a live foal., Objectives: To determine: 1) Serum amyloid A (SAA) profile in healthy mares during late gestation; 2) if placentitis affects SAA concentrations and 3) the effects of therapy on SAA concentrations and pregnancy outcome in mares with placentitis., Methods: In Experiment I, 15 healthy pregnant mares were evaluated from 280 days of gestation to 60 h post partum. In Experiment II, pregnant mares were inoculated intra-cervically with Streptococcus zooepidemicus (Day 280-295) and assigned to control (n = 5) and treatment (n = 9) groups. Treatment was initiated at the onset of clinical signs. Serum amyloid A concentrations were determined prior to inoculation and then weekly until abortion or delivery., Results: Serum amyloid A remained at low concentrations (95% confidence interval [CI]: 3.2-8.1 mg/l) during late gestation followed by a significant increase within 36 h post partum; SAA returned to basal concentrations by 60 h post partum. In Experiment II, SAA significantly increased within 96 ± 56 h of inoculation in control mares followed by abortion. Therapy was effective (P<0.05) in preventing the rise in SAA in 66% (6/9) of mares and only one out of 3 mares with increased SAA aborted. Overall, the incidence of abortion was higher in mares with increased SAA concentrations (75%; 6/8) compared with mares in which SAA remained at baseline concentrations (0/6)., Conclusions: Mares with placentitis had significant increased SAA within 96 h post inoculation and concentrations remained increased until abortion in untreated mares. Successful treatment either prevented the rise of SAA concentration or decreased its concentration to baseline concentrations, followed by delivery of a live foal., Potential Relevance: Serum amyloid A may be used as a prognostic indicator in cases of ascending placentitis in the mare., (© 2012 EVJ Ltd.)

The present study sought to determine: (i) the effects of Neospora caninum infection and twin pregnancy on plasma pregnancy-associated glycoprotein-2 (PAG-2) concentrations throughout pregnancy and (ii) whether plasma PAG-2 concentrations could predict abortion in N. caninum-infected cows. The study was performed on a commercial Holstein-Friesian dairy herd in northeastern Spain and the final data included those recorded in 53 non-aborting and 19 aborting animals. Blood samples were collected immediately before pregnancy diagnosis (on Days 40, 90, 120, 150, 180 and 210 post-insemination) in non-aborting cows or until the time of abortion detection in aborting cows. General lineal models (GLM) repeated measures anova revealed the different behaviour of PAG-1 and PAG-2, and significant effects of Neospora seropositivity, cool season and twin pregnancy on plasma PAG-2 concentrations throughout gestation (between-subject effects). In addition, based on the odds ratios, the likelihood of abortion increased in Neospora-seropositive cows (by a factor of 7.0) compared to seronegative animals and decreased in cows with a high plasma PAG-2 concentration (>4.5 ng/ml) on Day 120 of pregnancy (by a factor of 0.24), compared to the remaining cows. In conclusion, there is a relationship between plasma PAG-2 concentrations and the risk of abortion in Neospora-infected dairy cows. Thus, plasma PAG concentrations measured using anti-boPAG-2 antiserum on Day 120 of gestation could serve as an indicator of the abortion risk in N. caninum infected animals; values <4.5 ng/ml indicating a high risk of abortion in chronically infected animals., (© 2012 Blackwell Verlag GmbH.)

Background: Latency is a key feature of the animal pathogen Chlamydia abortus, where infection remains inapparent in the non-pregnant animal and only becomes evident during a subsequent pregnancy. Often the first sign that an animal is infected is abortion occurring late in gestation. Despite this, little is understood of the underlying mechanisms that control latency or the recrudescence of infection that occurs during subsequent pregnancy. The aim of this study was to develop an experimental model of latency by mimicking the natural route of infection through the intranasal inoculation of non-pregnant sheep with C. abortus., Methodology/principal Findings: Three groups of sheep (groups 1, 2 and 3) were experimentally infected with different doses of C. abortus (5×10(3), 5×10(5) and 5×10(7) inclusion forming units (IFU), respectively) prior to mating and monitored over 2 breeding cycles for clinical, microbiological, pathological, immunological and serological outcomes. Two further groups received either negative control inoculum (group 4a,b) or were inoculated subcutaneously on day 70 of gestation with 2×10(6) IFU C. abortus (group 5). Animals in groups 1, 2 and 5 experienced an abortion rate of 50-67%, while only one animal aborted in group 3 and none in group 4a,b. Pathological, microbiological, immunological and serological analyses support the view that the maternal protective immune response is influenced by initial exposure to the bacterium., Conclusions/significance: The results show that intranasal administration of non-pregnant sheep with a low/medium dose of C. abortus results in a latent infection that leads in a subsequent pregnancy to infection of the placenta and abortion. In contrast a high dose stimulates protective immunity, resulting in a much lower abortion rate. This model will be useful in understanding the mechanisms of infection underlying latency and onset of disease, as well as in the development of novel therapeutics and vaccines for controlling infection.

Background: Epizootic bovine abortion (EBA) is a tick-transmitted abortive disease of beef cattle in the western United States. Infected cattle do not have clinical signs until abortion occurs, usually within the last trimester of gestation. There is little information on the hematologic response of the dam following infection., Objective: The purpose of this study was to determine if changes in blood leukocytes and serum IgG concentrations could be detected following experimental infection of pregnant heifers with the etiologic agent of EBA (aoEBA)., Methods: Twelve Angus heifers were infected during gestation with the aoEBA using an inoculum prepared from the thymus of an infected fetus. Five pregnant heifer controls were given an inoculum prepared from the thymus of an aoEBA-negative calf. PCVs, total and differential leukocyte counts, and serum IgG concentrations were measured weekly following administration of the inocula until abortion or calving. Gross and microscopic examinations were performed on all aborted fetuses to confirm infection., Results: Eleven of 12 heifers in the treatment group aborted, and significant findings were decreased lymphocyte counts at weeks 1 and 14 postinoculation and increased monocyte counts at week 4 compared with control animals. Serum IgG concentrations were significantly increased at weeks 6-8 and 11 in the treatment group., Conclusions: Leukogram changes are subtle in infected cattle. Future research efforts should be aimed at development of an antibody test specific for detection of previously infected animals, which could graze safely on EBA-endemic pastures., (© 2012 American Society for Veterinary Clinical Pathology.)

Prenatal death can occur due to several genetic and environmental factors which alter normal embryo development, maternal environment to support normal fertilisation, development of embryos, placenta and foetus, or affect the necessary relationship between embryo and endometrium. The aim of this work was to study gestational losses and progesterone, 17 β-estradiol and IGF I serum levels in a rabbit line selected for growth rate (paternal line). In this study, a maternal line well characterised in previous studies was used as a reference line. A total of 211 laparoscopies were carried out, and the number of corpora lutea and implanted embryos at 12th days, total born and live born were recorded per female. To analyse the endocrine levels, blood serum was collected from 54 females with implanted embryos at 12th and 24th day of gestation (27 from each line). The paternal line showed the lowest ovulation frequency, number of implanted embryos, total born and live born (0.70, 11.3, 7.4, and 6.4 vs 0.86, 12.8, 11.1 and 10.6 for maternal line, respectively) and consequently, the highest implantation, gestational, foetal and perinatal losses (0.31, 0.60, 0.40, and 0.15, respectively). Progesterone serum levels at 12th days of gestation were similar between lines; however, progesterone serum level at 24th day of gestation was significantly lower in the paternal line (4.8 vs 8.2 ng/mL). Serum levels of 17β-estradiol and IGF-I at 12th days of gestation were different between lines (14.6 vs 26.5 pg/mL, 237 vs 149 ng/mL for paternal and maternal lines respectively). These higher gestational losses of the paternal line could be explained by differences in 17 β-estradiol level at 12th days of gestation and the possible effect on low progesterone serum levels at 24th days of gestation. Further studies in steroid production and bioavailability have to be done during oestrus and pregnancy related with metabolic activity of this line., (Copyright © 2012 Elsevier Inc. All rights reserved.)

The study involved 46 healthy purebred Arabian mares exhibiting regular oestrous cycles that underwent artificial insemination (AI). Pregnancy was detected ultrasonographically (US) in 40 mares. In 15 mares in foal, early embryonic death (EED) was observed during the pregnancy days 14-21. Blood for determinations of serum acute phase proteins (SAA and Hp) and progesterone (P4) was sampled 12-24 h before ovulation and the first insemination, at 12, 24, 72, 96 h and on day 7, 10, 14, 21, 35 and 55 after ovulation. The results revealed that in 25 mares without EED, the serum levels of P4, SAA and Hp were within physiological limits; in 15 mares with EED, the levels of SAA and Hp were significantly increased. In seven mares with EED, high levels of SAA and Hp were already found before ovulation and at 12, 24, 72, 96 h as well as on day 7 and 10 post-ovulation, whereas the level of P4 was normal for early pregnancy. In the remaining eight mares with EED, increased levels of SAA and Hp were found at 72 h after ovulation and maintained until day 55. In this group, the level of P4 decreased since 96 h after ovulation. Determinations of SAA, Hp and P4 in mares in early pregnancy (EP) are useful for monitoring normal development of pregnancy and for diagnosis of subclinical genital inflammations, which may lead to EED., (© 2011 Blackwell Verlag GmbH.)

Thirteen female Bedouin goats living in arid land of Algeria Sahara desert were used in this study. These goats were pregnant but they sustained an abortion because of unidentified causes. None of the goats showed any signs of general disease. Plasma concentrations of caprine pregnancy-associated glycoproteins (cPAGs) and progesterone (P4) were determined during pregnancy using radioimmunoassay. The cPAGs concentration was undetectable (<0.8 ng/ml) throughout the first 2 weeks of gestation. From week 3 after mating, cPAGs concentration was detectable with significant individual variations (p < 0.05) reaching a maximum secretion (436.1 ng/ml). Throughout gestation, cPAGs concentration remained relatively constant but decreased few days before abortion, on an average of 9.2 ± 1.2 days (n = 11), except for two females where the concentrations decreased later (1-2 days before abortion). One or two peaks of cPAGs concentrations (in 4/13 and in 9/13 females, respectively) have been measured few weeks before abortion (77-124 days after mating), when a decline of cPAGs was detected. The P4 concentration increased after mating, and was high from the first week till the end of pregnancy. The P4 concentration (9.1 ± 0.9 ng/ml) decreased rapidly (<0.5 ng/ml) after 4 ± 0.7 days (n = 6) or 9.4 ± 1.6 days (n = 7) before abortion. A positive relationship (p < 0.01) was found between P4 and cPAGs concentrations during gestation. Results indicate that cPAGs and P4 measurements can be used for monitoring gestation and for abortion prediction., (© 2009 Blackwell Verlag GmbH.)

The objective of the present study was to analyse the immune status of mares in the early stages of pregnancy and mares which experienced early embryonic death (EED). Forty-eight fertile purebred Arabian mares were used for the study. The mares were divided into two groups. Group I was the control group composed of 32 mares in the early stages of a normal pregnancy. Group II was composed of 12 mares which had experienced EED. In both groups, subpopulations of T lymphocytes containing CD2(+), CD4(+) and CD8(+) were characterized using flow cytometry. The percentages of CD3(+) lymphocytes (adding up the cells with CD4(+) and CD8(+) phenotypes) and of NK cells (subtracting CD3(+) cells from CD2(+) lymphocytes) calculated. Due to its suppressive effects on T lymphocytes during pregnancy, TGF-beta1 was determined using ELISA. The findings revealed that in mares with EED, the percentages of lymphocytes TCD2(+), TCD4(+), NK cells at EED, i.e. between days 14 and 21, were significantly higher compared to mares without EED. The changes in the CD4(+)/CD8(+) ratio were similar. In the EED group, this ratio significantly increased between the 96th hour and the 30th day after ovulation compared to mares without EED. Otherwise, serum levels of TGF-beta1 in the EED group were significantly lower than in the control group. The results show that mares which experienced an EED develop inflammatory reactions which activate Th1 cell immune responses., (Copyright 2010 Elsevier B.V. All rights reserved.)

An indirect enzyme-linked immunosorbent assay (ELISA) was modified and validated to detect antibodies against Salmonella Abortusovis in naturally infected sheep. The ELISA was validated with 44 positive and 45 negative control serum samples. Compared with the immunoblot, the sensitivity and specificity of the assay were 98% and 100%, respectively. To follow antibody levels over time, samples from 12 infected ewes were collected at 1, 3, and 10 months after abortion. All animals showed antibody levels above the cutoff value throughout the observation period. One and 3 months after abortion, high antibody levels could be detected in all but one animal, whereas after 10 months, 9 animals had markedly lower but still positive antibody levels. The test characteristics and evidence for the persistence of detectable antibody levels in all infected animals for up to 10 months indicates that the ELISA can be used for herd surveillance testing.

The aim of this study was to determine seroprevalence of Neospora caninum in dairy cattle ranches with higher abortion rates and co-existence of Neospora caninum, Toxoplasma gondii, Brucella abortus, and Listeria monocytogenes antibodies. The blood samples were collected from dairy cows with history of abortion (n=234) as well as from pregnant cows that were (n=323) kept in the same ranches. N. caninum seroprevalence was 10.77% (60/557). The co-existence rate of N. caninum seropositivity with T. gondii, B. abortus and L. monocytogenes was 24.77% (138/557), 13.82% (77/557) and 42.85% (162/378), respectively. Only one animal had significant antibody titers for all analyzed infectious agents. The seroprevalence of N. caninum and T. gondii were significantly higher in pregnant cows than aborted cows (p<0.05 and p<0.001, respectively). As a result, the comprehensive data generated through this study can significantly contribute to understanding of serologic association of N. caninum with T. gondii, B. abortus and L. monocytogenes in pregnant and aborted dairy cows.

This report deals with the acute onset of an abortion outbreak and high sow mortality in one pig herd consisted of 1,200 pigs and 120 sows on Jeju Island, Korea. Affected pregnant sows showed clinical signs, including high fever, gradual anorexia, vomiting, depression, recumbency, prostration, abortion, and a few deaths. Four dead sows, five aborted fetuses from the same litter, and 17 sera collected from sows infected or normal were submitted to the Pathology Division of the National Veterinary Research and Quarantine Service for diagnostic investigation. Grossly, hepatomegaly and splenomegaly were observed in sows. Multiple necrotic foci were scattered in the lungs, liver, spleen, and lymph nodes. Microscopically, multifocal necrotizing lesions and protozoan tachyzoites were present in the lesions. Tachyzoites of Toxoplasma (T.) gondii were detected immunohistochemically. Latex agglutination showed that the sera of 7 of 17 (41.2%) sows were positive for antibody to T. gondii. The disease outbreak in this herd was diagnosed as epizootic toxoplasmosis. To our knowledge, this is the first report of porcine toxoplasmosis with a high abortion rate and sow mortality in Korea.

Chlamydophila abortus is one of the principal causes of late-term abortion (enzootic abortion of ewes or EAE) in sheep across Europe. Serological diagnosis of EAE is routinely carried out by the complement fixation test, although the interpretation of results can often be difficult because of cross reaction with Chlamydophila pecorum, which also commonly infects sheep. The purpose of this study was to evaluate and compare four ELISAs developed at Moredun Research Institute and based on whole C. abortus elementary bodies (EBs), an outer membrane preparation of the whole organism (SolPr) and two recombinant polymorphic outer membrane protein fragments (rOMP90-3 and rOMP90-4), with 3 commercial tests, the CHEKIT Chlamydophila Abortus, Pourquier ELISA Chlamydophila abortus and ImmunoComb Ovine Chlamydophila Antibody tests. The tests were evaluated using a panel of 202 sera from experimentally and naturally infected animals, as well as from EAE-free flocks. The EB, SolPr and CHEKIT ELISAs performed similarly to the CFT, all lacking in specificity by cross reacting with sera from C. pecorum infected animals. The ImmunoComb also lacked specificity with C. pecorum sera, but also badly cross reacted with sera from EAE-free flocks. The rOMP90-3, rOMP90-4 and Pourquier ELISAs were the most specific, although the Pourquier test appeared less sensitive with sera from naturally infected animals. Overall, the rOMP90-3 ELISA performed the best, with high sensitivity (96.8%) and no cross reaction with sera from C. pecorum infected animals or from EAE-free flocks (100% specificity) and so would be a suitable alternative to the CFT for the serological diagnosis of EAE.

The effects of restricted feeding (20 g/day from gestational day (GD) 6 to 28) on pregnancy outcome and blood parameters were examined in pregnant rabbits. As compared with the group which was allowed free access to diet throughout the gestational period (NT group), the group subjected to restricted feeding (R group) showed significantly lower values in many parameters such as total protein, albumin and triglyceride on GDs 22 and 28, reflecting low nutritive conditions. In addition, there were significant changes in blood coagulation-related parameters, suggesting an imbalance between coagulation and anti-coagulation factors. Moreover, abortions occurred in about half of the animals of the R group between GDs 23 and 27. When aborted rabbits were compared with those which could maintain pregnancy under restricted feeding, total protein, albumin, platelets and antithrombin III values and especially blood progesterone concentration were significantly lower in aborted rabbits on GD 22, prior to occurrence of abortion. These results suggested that abortions due to restricted feeding might be brought about by lower nutritive conditions, an imbalance of blood coagulation-related factors and lower blood progesterone concentrations.

The objective was to validate a high-sensitivity chemiluminescent assay of serum progesterone concentrations for pregnancy diagnosis in manatees. Assay analytical sensitivity was 0.1 ng/mL, with mean intra- and inter-assay coefficients of variation of 9.7 and 9.2%, respectively, and accuracy had a mean adjusted R(2) of 0.98. Methods comparison (relative to Siemen's Coat-A-Count RIA) demonstrated r=0.98, Deming regression slope of 0.95, and an intercept of 0.01. Based on ROC analysis, a progesterone concentration >or=0.4 ng/mL was indicative of pregnancy. Assay results were not significantly altered by two freeze-thaw cycles of samples. Characteristic progesterone concentrations during pregnancy were Months 1-4 (1.7-4.7 ng/mL), 5-8 ( approximately 1.0 ng/mL), and 10 and 11 (0.3-0.5 ng/mL), whereas two late-pregnant females with impending abortion had progesterone concentrations of 0.1 ng/mL. Among pregnant females, maximum progesterone concentrations occurred in autumn (3.9+/-1.8 ng/mL), and were greater during all seasons than concentrations in non-pregnant females (0.1-0.2 ng/mL). Progesterone concentrations were also significantly higher in pregnant females than in non-pregnant females and males. This highly sensitive, specific, and diagnostic assay will be valuable for monitoring pregnancy and abortion in manatees.

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.

Ovine toxoplasmosis is a major cause of abortion and placentitis. Lambs with congenital infection are born with encephalitis. Abortions occurred between May and August 2006 in an estate in Três Palmeiras, southern Brazil. This study aimed to assess the frequency of abortion associated with toxoplasmosis among ewes in southern Brazil using indirect hemagglutination and pathoanatomical and immunohistochemical analyses. Blood samples were collected from nine animals for indirect hemagglutination. One fetus underwent necropsy and histopathological and immunohistochemical analyses. Three samples, one of which belonged to the necropsied animals mother, were positive on the serological test. The necropsy revealed pale foci in the liver, lungs with a marbled appearance, heart with severe pallor and brain and cerebellar congestion. Fragments of all organs were collected, fixed in 10% buffered formalin, processed, and stained with hematoxylin-eosin. Microscopically, there were cysts and tachyzoites in the brain in areas of malacia with microgliosis and lymphoplasmacytic infiltration, suggesting toxoplasmic encephalitis. There was also interstitial pneumonia, centrolobular necrosis with structures compatible with tachyzoites, focal lymphocytic myocarditis and acute tubular nephrosis. The immunohistochemical test was positive for Toxoplasma gondii. The results allowed diagnosing the ovine toxoplasmosis.

Water buffalo (Bubalus bubalis) are affected by high rates of embryonic mortality and abortion related to infectious diseases and non-infectious factors. A number of viral and bacterial infections have been associated with reproductive failure, but there is limited information on the role of chlamydial infections. In order to investigate the presence and the role of Chlamydiaceae in water buffalo a retrospective study was performed in a herd with a history of reproductive failure. During an 11-month period, the pregnant heifers suffered an abortion rate of 36.8% between the 3rd and 7th month of pregnancy. Antibodies to Chlamydiaceae were detected in 57% of the aborted cows, and in 0% of the overtly healthy cows used as control. By a nested-PCR assay, three of 14 vaginal swabs from aborted animals tested positive for Chlamydophila agents and, additionally, three out of seven aborted fetuses tested positive for Chlamydophila spp., with two being co-infections by Cp. abortus and Cp. pecorum and one being characterised as Cp. abortus. Sequence analysis of the amplicons confirmed the results of the nested-PCR. The presence of anti-Chlamydiaceae antibodies in more than half of the aborting animals (P<0.002) and the detection of Chlamydophila agents in several fetal organs and in the vaginal swabs are consistent with the history of abortions observed in the herd and suggest an abortifacient role by Chlamydophila spp. in water buffalo (B. Bubalis) herds.

This trial was conducted on three Hungarian dairy farms between March 2003 and April 2004. The aims of this study were to examine the effect of milk production and milk contents, the day after calving (DAC) and the sire for the pregnancy rate (PR) and late embryonic loss (LEL) in dairies. Five thousand three hundred and eighty insemination was carried out in this period. Pregnancies (n = 1969) were detected by BioPryn test from 3919 blood samples between 30-36 days post insemination (PI). Retention of pregnancy was determined by rectal palpation on Day 60. LEL has been determined by the optical density (OD) of blood samples/cutoff ratio and the serum progesterone (P4) concentration 30-36 days PI. According to serum progesterone concentration the authors predicted a presumed or possible embryonic loss or maintenance of the pregnancy. The efficiency of the artificial inseminations (AI) was significantly lower 60 days within post-partum. Significant positive correlation was found between the result of early and late PR and DAC and the milk protein/fat ratio. Significant negative correlation was detected between the early and late PR the milk protein and fat content. A later stage (60 days PI) of pregnancy the milk production has a negative effect for retaining of pregnancy. More data would be necessary to state that sires can influence the reproduction ability of their progenies. This is a really important issue, because the AI bulls have a great impact on the herds and the one-way selection may cause decreased fertility on breed level as well.

The aim of present experiment was to evaluate the plasma concentrations of estrone sulfate (E(1)S) and progesterone (P(4)) during late gestation in recipient cows transferred embryos produced by nuclear transfer (NT) and in vitro fertilization (IVF). Blood samples were collected from recipients transferred embryos produced by NT (n=9) and IVF (n=13) at 160, 220, 240, 260 and 270 d of gestation and then at 5 d intervals until parturition. Plasma samples were analyzed for E(1)S and P(4) by ELISA. One NT and three IVF cows aborted between days 220 and 260 of gestation. Two NT and one IVF cow had prolonged gestation (over 290 d). One IVF cow had an overweight fetus (50 kg) after abortion (257 d). The patterns of changes in the concentrations of E(1)S during late gestation in the NT and IVF cows were almost identical. The NT and IVF cows that aborted had prolonged gestation and much higher E(1)S concentrations than the average. One NT cow aborted after 220 d of gestation and had a sudden high increase in its E(1)S concentration from 160 d to 220 d of gestation. The NT and IVF cows that had prolonged gestation also had significantly higher (P<0.05) P(4) concentrations than the average. These results raise the possibility that the E(1)S and P(4) profiles can be used to monitor some late gestational problems, such as higher birth weight, abortion and prolonged gestation.

Over a 25-month period 8118 blood samples were assayed for the presence of the serum pregnancy specific-protein B (PSPB) and progesterone (P4) concentrations on three Hungarian large-scale dairy farms. Pregnancy (n = 4085) was checked by BioPRYN assay at 30-36 days post-insemination (PI). Samples from all cows that tested not pregnant and from cows with an optical density (OD) reading in the BioPRYN test that was between 0% and 30% above the cutoff OD value were tested for serum P4 concentration. According to serum P4 concentration, cows were assigned to three categories: high (>4 ng/ml), medium (2-4 ng/ml) and low (<2 ng/ml) serum progesterone. The authors predicted a presumed (low) or possible (medium) late embryonic loss (LEL) or maintenance of the pregnancy (high). A total of 710 LELs were detected (17.4%) and 31.8% of them were predicted because of a low OD value at 30-36 days after insemination. Lower PSPB serum level significantly refers for LEL (p < 0.0001). The prediction rate for the true embryonic loss was 31.8% when OD cutoff from 0% to + 30% of cutoff was examined while it was 62.5% when the threshold was OD cutoff of 0% to 10% of cutoff. The authors conclude that BioPRYN was useful for prediction of a part of LEL in dairy cows and serum P4 concentration in these cows related to the rate of LEL.

The aim of the present study was to investigate the effects of two medications on two subsequent abortions and plasma hormone concentrations of dogs. For this purpose, two groups of bitches (n=5 each), received the antiprogesterone aglepristone (Alizine) at 10mg/kg body weight on two subsequent days around day 30 after mating. In group II, the antiprolactin cabergoline (Galastop) was additionally administered po at 5 microg/kg body weight until the start of abortion. The plasma concentrations of relaxin, progesterone (P4) and estradiol-17beta (E2) were measured before, during and after each abortion. During the next cycle after the abortion, the same bitches were mated again and in pregnant animals, induction of abortion was performed as before. During the third cycle, pregnant bitches were allowed to whelp. Termination of first pregnancy occurred significantly earlier after the combined treatment (6.8 versus 10.6 days, p<0.05). In both groups and during both abortions, relaxin varied between individuals; however, there was a continuous decrease after the abortions and no significant differences between groups (p>0.05). In one bitch with high relaxin concentrations before treatment (11.6 ng/ml), a cystic endometrial hyperplasia was diagnosed. In the aglepristone only group, P4 concentrations increased significantly after the first application (p<0.05), then decreased continuously until day 45 after the beginning of abortion. In the combined group, there was a continuous decrease until day 45 (p>0.05). At this time, P4 concentrations between 0.47 and 84.9 nmol/l were measured in both groups. The level of E2 over time was not influenced by any medication. We therefore note that the two medications mainly influenced plasma concentrations of P4 in different ways, probably due to specific treatment-hormone interactions. However, all measurements fell within the range considered normal.

A 1-year-old, 5- to 6-week pregnant cat was presented with a history of aborting 3 kittens the previous night. During the examination, the cat began to seizure and died. At necropsy, formalin-fixed tissues and blood smears prepared from an EDTA blood sample collected via cardiac puncture were submitted to a diagnostic laboratory. Examination of the blood smears revealed numerous large (15-75 microm) clumped macrophages containing dark blue intracytoplasmic organisms consistent with Cytauxzoon merozoites. Scattered erythrophagocytic macrophages were also observed. Within several erythrocytes, 1-2 small, round, ring-like piroplasms consistent with Cytauxzoon were observed. Histologic examination revealed numerous large, schizont-laden macrophages filling the blood vessels of multiple organs. The cytologic and histologic findings were diagnostic for Cytaxuzoon felis infection. This case was of particular interest because of the unusually large number of organism-laden macrophages in the cardiac blood sample, an uncommon finding in peripheral blood. Although the cat was presented for complications of abortion, it remains uncertain whether C felis organisms crossed the placenta and infected the fetuses or lead to the abortions.

The present study was designed to establish whether plasma pregnancy-associated glycoprotein-1 (PAG-1) measurements during the early fetal period can be associated with early fetal loss. Blood samples were obtained and ultrasound controls performed on days 35, 42, 49, 56, and 63 of gestation or until pregnancy loss from 98 lactating dairy cows. Radioimmunoassay systems were used to determine PAG-1 and progesterone concentrations. Of the 98 pregnancies investigated 18 (18.4%) suffered early fetal loss: 15 (18.5%) in cows carrying singletons, and 3 (16.7%) in twin pregnancies. In cows suffering pregnancy loss, all living embryos registered on day 35 seemed normal in size and development in all weekly ultrasound controls before fetal expulsion. Using analysis of variance, plasma PAG-1 and progesterone values were not different between no loss and fetal loss groups for every gestation period. Based on the odds ratio, and considering only PAG-1 values obtained on day 35 of gestation, the risk of fetal loss was 10 and 6.8 times more likely in cows with low (<2.5 ng/ml) and high (>4 ng/ml) PAG-1 values, respectively, than in cows with medium PAG-1 values, used as reference. Of the 10 inseminating bulls included in the study, one was related to increased fetal loss by odds ratio of 21.7, whereas one bull was attributed fetal loss rate reduced by odds ratio of 12.5 (1/0.08) These findings can have a clear clinical application: PAG-1 measurements from one single sample taken on day 35 of gestation provided more useful information than a series of values obtained from day 35 to 63 of gestation, and can be indicators of subsequent fetal loss.

Embryonic and fetal mortality reduce lambing rates and litter sizes, thus contributing to economic losses in the sheep industry. In the current study, the timing of late embryonic and fetal loss in ewes and the factors with which these losses were associated were examined. Ewes lambing and lambs born were compared with pregnancy diagnosis and counts of embryos by ultrasonography near d 25, 45, 65, or 85 of gestation. Approximately 19.9% of the ewes experienced late embryonic loss, fetal loss, or both; and 21.2% of the embryos or fetuses were lost from d 25 to term. Potential offspring were lost throughout gestation; 3.7% of embryos from d 25 to 45, 4.3% of fetuses from d 45 to 65, 3.3% from d 65 to 85, and 11.5% from d 85 to parturition; thus, approximately 3 to 4% of the potential offspring were lost for each 20-d period of pregnancy beyond d 25. A greater proportion of ewes lost one (36.7%) rather than all (20.5% single; 3.8% multiple) embryos or fetuses. The patterns of loss were similar in ewes mated during the anestrous season and the transitional period and did not vary with service period within breeding season or method of synchronization of estrus. Late embryonic or fetal losses were not related to the temperature-humidity index. Maternal serum collected near d 25, 45, 65, or 85 of gestation was assayed for concentrations of progesterone, estradiol-17beta , and vascular endothelial growth factor (VEGF). The proportions of embryos or fetuses lost were associated with breed type (P < 0.05), as were concentrations of progesterone (P < 0.01), estradiol (P < 0.05), and VEGF (P < 0.01). The relationships of loss or retention of pregnancy to hormonal variables at the 4 stages studied were limited. Complete and partial losses increased rapidly as maternal progesterone at d 25 decreased below 2 ng/mL (P < 0.05). Survival of fetuses within a litter from d 25 to 65 was greater for ewes with medium concentrations of VEGF near d 25 and from d 65 to parturition was greater for ewes with high concentrations of VEGF near d 45 (P < 0.05). In summary, late embryonic or fetal losses occurred from d 25 throughout gestation and varied with breed type and with concentrations of progesterone in maternal serum on d 25.

In order to evaluate the efficacy, the safety and the variation in plasma concentrations of estrogens, progesterone, PGFM, oxytocin, cortisol and prolactin after mid-pregnancy termination induced by aglepristone, 61 pregnant queens (33.3 + 4.2 days), were injected subcutaneously with 15 [corrected] mg/kg aglepristone, (Alizine) [corrected] repeated once 24 h later. Five queens served as control and received a placebo. The efficacy of aglepristone was 88.5% and termination of pregnancy was achieved in 50% of the queens within 3 days. Brief periods of depression and anorexia were noted in 9.3% of the queens before fetal expulsion (these symptoms were attributed to the phenomenon of fetal expulsions). Not one of the queens that aborted developed uterine disease. There were no changes in plasma concentrations of estrogen, prostaglandin, prolactin or oxytocin following aglepristone administration. However, there were significant increases in plasma concentrations of progesterone and cortisol 60 and 30 h, respectively, after aglepristone administration. Termination of pregnancy occurred with high plasma progesterone concentrations. Fetal expulsion was characterised by an increase in estrogen, PGFM and oxytocin concentrations, whereas prolactin and cortisol levels remained at a basal level.

The aim of this study was to investigate the plasma concentrations of folic acid, vitamin B12 and progesterone at different stages of the sexual cycle and pregnancy, during induced abortion and in bitches with pyometra. Bitches (n = 97) were assigned to groups as follows: a) oestrous cycle (n = 42) b) pregnancy (n = 25) c) induction of abortion (n = 10 and d) pyometra (n = 20). Oestrous cycle stages were determined by vaginal inspection and cytology. Pregnancies were estimated by ultrasound (5.0 Mhz; linear transducer; Schimadzu) at days 15-25, 35-45 and 46-63 of pregnancy. Treatments for the induction of abortion were started between days 25 and 35 after mating (5 microg/kg cabergoline daily, Galastop; 5-10 microg/kg Alfaprostol every other day, Gabbrostim). Diagnosis of pyometra was confirmed by ultrasound and vaginoscopy. Folic acid and vitamin B12 concentrations did not differ among different stages of the oestrous cycle. The mean concentration of folic acid during early pregnancy (days 15-25) exceeded levels of later stages (days 46-63): 9.4 +/- 3.7 microg/ml and 4.7 +/- 1.8 microg/ml, respectively (p < 0.01). A positive correlation between folic acid and vitamin B12 was determined in pregnant dogs ( r = 0.925; p < 0.02). Before the induction of abortion, the concentration of folic acid was 9.6 +/- 5.2 microg/ml; during abortion it decreased to 5.0 +/- 3.2 microg/ml (p < 0.01). A significant correlation (r = 0.925; p < 0.02) between progesterone and folic acid was obtained in bitches with abortion. The mean concentration of folic acid in bitches with pyometra significantly differed from that of bitches at different stages of the oestrous cycle (p < 0.05). The mean concentration of folic acid was significantly lower in metoestrous bitches when compared to bitches with pyometra (p < 0.05). The decrease of serum concentrations of folic acid during pregnancy and induced abortion show that fetal growth and abortion caused higher consumption of folic acid. Concerning bitches did not show any deficiency symptoms, which is why it can be concluded that this decrease is physiological.

Objective: To test horses for serologic evidence of an association between vesiviral antibodies and abortion., Sample Population: Sera from 141 horses., Procedures: 2 experiments were conducted. Experiment 1 comprised sera obtained in 2001 and 2002 from 3 groups of horses (58 mares from farms with a history of abortion problems, 25 mares between 3 and 13 years of age with unknown reproductive histories that were sold at auction [breeding-age control mares], and 29 mixed-age males and yearling females sold at auction [negative control population]). Experiment 2 comprised sera from 3 groups of pregnant mares (10 pregnant mares fed Eastern tent caterpillars [ETCs], 9 pregnant mares fed ETC frass only, and 10 pregnant control mares). Sera were analyzed for antibodies against vesivirus by use of a validated recombinant vesivirusspecific peptide antigen in an indirect ELISA., Results: For experiment 1, 37 of 58 (63.8%) mares from farms with abortion problems were seropositive for vesivirus antibodies, whereas 10 of 25 (40%) breeding-age control mares were seropositive. All 29 mixed-age males and yearling females were seronegative for vesivirus antibodies. For experiment 2, 17 of 29 mares aborted (some from each group). Seropositive status for vesivirus antibodies increased from 47.1% (8/17) to 88.2% (15/17) for the pregnant mares that aborted during the experiment., Conclusion and Clinical Relevance: Significant association was detected between seropositive status for vesivirus and abortion in mares; consequently, vesivirus appears to be a pathogenic virus associated with abortion in mares. These data support adding vesivirus antibody testing into diagnostic screening to determine the cause for abortion in mares.

An immunochromatographic test (ICT) with recombinant surface antigen 1 of Neospora caninum (NcSAG1) was developed for the rapid detection of antibodies to N. caninum in cattle. The ICT was used to clearly discriminate between immunofluorescent-antibody test (IFAT)-positive bovine sera and IFAT-negative bovine sera. Serum samples collected from cattle in Yanbian, China, were examined by the ICT. Of the 96 serum samples, 23 (24.0%) were positive by the ICT, and 19 (19.8%) samples were positive by a previously developed enzyme-linked immunosorbent assay (ELISA). Eighteen of 19 ELISA-positive samples were positive according to the ICT. A good agreement was found between the results of the ICT and the ELISA. The results presented here suggest that the ICT with recombinant truncated NcSAG1 fused to glutathione S-transferase is a useful and reliable method for the detection of antibodies to N. caninum in cattle.

Body temperature responses and the timing of abortions were evaluated in pregnant bitches with the anti-progestin aglepristone. Fifteen purebred and crossbred, 25-45 days pregnant, were included in this study and seven untreated bitches at the same stage of pregnancy served as controls. Treated bitches were administered two applications of aglepristone (10 mg/kg SC) 24 h apart for pregnancy termination. Pregnancy termination was confirmed by ultrasonographic assessment. Body temperature was rectally measured three times a day for 6 days beginning 24 h before treatment or pregnancy diagnosis in the treated and control bitches, respectively. Additionally, serum progesterone concentrations were assessed at time points during the study in the treated bitches. Pregnancy was terminated in 14 treated bitches in a mean+/-S.E.M. of 4.3+/-0.7 days after treatment. Control bitches remained pregnant. In the treated bitches, but not in the controls, body temperature significantly decreased 24 h after the beginning of the treatments (P < 0.01) and then gradually returned to pre-treatment values. Correlation between the day of mean minimum body temperature and the day of pregnancy termination was low (0.07; > 0.05). Progesterone did not show significant change throughout the study. Body temperature does not seem to be a suitable variable to clinically monitor the aborting effect of aglepristone. Decrease of body temperature after aglepristone treatment could represent further evidence of its hypothalamic effects.

Progesterone (P4), pregnenolone (P5) and their metabolites are present in maternal plasma in pregnant mares. It is believed that one of these progestagens may maintain myometrial quiescence. The aims of this study were to identify specific progestagens in pregnant mares' plasma and determine whether these differed between mares with healthy or compromised pregnancies. Jugular blood samples were collected between 243 and 351 days gestation from 19 healthy Thoroughbred mares and 14 mares with placental pathology, including placentitis, and other clinical problems (uterine torsion/rupture, colic, laminitis). Ten progestagens were identified using gas chromatography-mass spectrometry, of which seven increased significantly with gestational age in healthy mares while P4 was undetectable. Mares with placentitis had increased concentrations of either P5 and/or P4 and several metabolites (5alpha-DHP, P5betabeta, betabeta-diol, betaalpha-diol, 20alpha-5P) suggesting increased fetal production of P5 and/or P4 and increased metabolism in the utero-placental tissues in response to chronic stress. Mares with other placental pathology had raised P4 concentrations while 5alpha-DHP and 3beta-5P were low possibly due to reduced placental function. In mares with problems unrelated to the placenta, most progestagens were substantially lower than control values. Although progestagen profiles differed between normal and abnormal pregnancies, no clear link was demonstrated between maternal plasma concentrations of P4, 5alpha-DHP or any other progestagen and the maintenance of pregnancy.

Our aim was to assess the seroprevalence of Chlamydophila (Cd) abortus (Chlamydia psittaci serotype 1), denoted ovine enzootic abortion (OEA), in the Swiss sheep population. A competitive enzyme-linked immunosorbent assay (cELISA) was adapted for the investigation of pooled serum samples (pool approach) and receiver-operator characteristic (ROC) analysis was applied to define the cut-off of the pool approach. At a cut-off value of 30% inhibition, the flock-level pooled sensitivity and specificity were 92.9% and 97.6% when compared to classifying the flock based on individual-animal samples. Subsequently, sera from 775 randomly selected flocks out of 11 cantons of Switzerland were investigated using the pool approach. The cantons included in the study represented 72% of the Swiss sheep flocks and 76% of Swiss sheep population. Antibodies against Cd. abortus were found in almost 19% (144) of the 775 examined sheep flocks. Test prevalences were adjusted for the imperfect test characteristics using the Rogan-Gladen estimator and Bayesian inference. Seroprevalence was highest (43%) in the canton Graubunden. In the remaining 10 cantons the seroprevalence ranged from 2 to 29%. The cELISA in combination with testing pooled sera and statistical methods for true prevalence estimation provided a good survey tool at lower costs and time when compared to other approaches.

This report describes a retrospective study based on serological screening, performed on a Neospora-associated abortion episode over a 12-month period in a dairy herd in north-east Spain. During the 1-year study period, the overall abortion rate for the herd was 23.2% (38 abortions of 164 diagnosed pregnancies). The data analysed were those derived from blood samples collected from the whole herd (n = 237) and from diagnosed pregnancies. Antibodies to Neospora caninum were found in 35.4% of the cattle with 44% of seropositive pregnant animals aborting over 1-year period. Based on the odds ratio, the risk of abortion was 12.2 times higher (P < 0.0001) in the Neospora-seropositive animals than in seronegative animals and significantly higher during the second term of gestation (P < 0.01) than during the first and third terms. Abortions were not found to be associated with parity status or season of pregnancy, and the common risk factors associated with pregnancy loss in the geographical area of the study. Age-related differences in N. caninum seroprevalence were not statistically significant, indicating vertical transmission as the main route of infection. Indeed, a high percentage of congenitally infected offspring was observed (90.6%) and the farm had been free of dogs for the last 7 years. Our results suggest that, when a dairy herd shows an increased incidence of abortions due to Neosporosis, maternal serology can be a good indicator of the abortion risk in individual cows, and that the effects of factors normally related to abortion, such as parity and pregnancy season, may be masked.

A pilot study of 10 Dutch dairy herds was performed to investigate possible causes of stillbirth/perinatal weak calf syndrome in heifers. Possible causes and advice for further investigations on farms affected by this syndrome are discussed based on the results of investigations of blood and urine from pregnant young stock, postmortem examination of still-born calves, and a questionnaire held among farmers. Infections with Neospora caninum, Salmonella spp. or Leptospira hardjo, or a deficiency of iodine could be excluded.

We studied the frequency and timing of abortion and the serum levels of 17beta-estradiol and progesterone in Korean native cows fed pine needles during pregnancy. Fifteen pregnant cows were randomly assigned to groups of 5. The control group was fed a concentrate and rice straw, and the other 2 groups were fed, in addition, either 1.3 or 2.7 kg (dry weight) of Korean pine needles daily, starting at an average of 91 d of gestation and continuing until 245 d of gestation. The health status of the dams and the viability of the fetuses were ascertained by rectal palpation and ultrasound scanning during pregnancy. Simultaneously, blood samples were collected for analysis of serum 17beta-estradiol and progesterone. Two abortions in mid-pregnancy (at 126 and 150 d of gestation) occurred in the group ingesting the higher daily amount of pine needles. Premature parturition occurred at 259 d of gestation in this group and at 241 and 252 d of gestation in the group ingesting the lower daily amount of pine needles. The serum 17beta-estradiol concentration was significantly higher (P < 0.05) at 4 mo of gestation and the serum progesterone level significantly lower (P < 0.05) at 8 mo of gestation in the group ingesting more pine needles daily than in either of the other 2 groups. These results suggest that the ingestion of pine needles may play a role in abortion in Korean native cows by increasing the serum 17beta-estradiol concentration and decreasing the serum progesterone concentration.

An enzyme-linked immunosorbent assay (ELISA) was established to measure the avidity of bovine IgG directed against p38, a surface antigen (NCSRS2) of Neospora caninum tachyzoites. In the sera of dams intravenously infected with N. caninum NC-1, the p38-specific avidity increased from initially below 40% up to avidity indices between 50 and 80% after days 23 and 91 p.i. The p38-avidity-ELISA was used to examine various herds that had experienced endemic and epidemic N. caninum-associated bovine abortions. In herds with endemic abortion, generally high avidity indices of N. caninum-specific IgG were detected. This finding and the observation of an association between the seropositivity of dams and that of their offspring suggested a predominantly vertical transmission of the parasite among the animals of these herds, thus indicating chronic infection of these cattle. In contrast, for herds experiencing epidemic abortion: (i) an association regarding seropositivity of dams and their daughters could not be shown and (ii) a generally low avidity of p38-specific IgG in the sera from aborting dams was determined. This indicates recent postnatal transmission of N. caninum in these herds. A linear regression model explaining the avidity of p38-specific IgG was significantly influenced by the time span between sampling and the occurrence of the first abortion of the epidemic in herds with an N. caninum-associated abortion storm. Another factor significantly contributing to the model was the proportion of dams at risk that had aborted in the herds (i.e. the severity of the abortion epidemic). A possible explanation for this observation is that herds experiencing heavy abortion are sampled earlier after the onset of the epidemic than others that have a less severe abortion storm.

Objective: To develop a method of probability diagnostic assignment (PDA) that uses continuous serologic measures and infection prevalence to estimate the probability of an animal being infected, using Neospora caninum as an example., Animals: 196 N caninum-infected beef and dairy cattle and 553 cattle not infected with N caninum; 50 dairy cows that aborted and 50 herdmates that did not abort., Procedure: Probability density functions corresponding to distributions of N caninum kinetic ELISA results from infected and uninfected cattle were estimated by maximum likelihood methods. Maximum likelihood methods also were used to estimate N caninum infection prevalence in a herd that had an excessive number of abortions. Density functions and the prevalence estimate were incorporated into Bayes formula to calculate the conditional probability that a cow with a particular ELISA value was infected with N caninum., Results: Probability functions identified for infected and uninfected cattle were Weibull and inverse gamma functions, respectively. Herd prevalence was estimated, and probabilities of N caninum infection were determined for cows with various ELISA values., Conclusions and Clinical Relevance: Use of PDA offers an advantage to clinicians and diagnosticians over traditional seronegative or seropositive classifications used as a proxy for infection status by providing an assessment of the actual probability of infection. The PDA permits use of all diagnostic information inherent in an assay, thereby eliminating a need for estimates of sensitivity and specificity. The PDA also would have general utility in interpreting results of any diagnostic assay measured on a continuous or discrete scale.