Predicting the probability of abortion in dairy cows: a hierarchical Bayesian logistic-survival model using sequential pregnancy data

Although abortion contributes substantially to poor reproductive health of dairy herds, little is known about the predictability of abortion based on age, previous abortion or gravidity (number of previous pregnancies). A poor understanding of effects of maternal factors on abortion risk exists, in part, because of methodological difficulties related to non-independence of multiple pregnancies of the same cow in analysis of fetal survival data. We prospectively examined sequential pregnancies to investigate relationships between fetal survival and putative dam risk factors for 2991 abortions from 24,706 pregnancies of 13,145 cows in nine California dairy herds. Relative risks and predicted probabilities of abortion (PPA) were estimated using a previously described hierarchical Bayesian logistic-survival model generalized to incorporate longitudinal data of multiple pregnancies from a single cow. The PPA increased with increasing dam age at conception, with increasing number of previous abortions, and if the previous pregnancy was aborted >60 days in gestation. The PPA decreased with increasing gravidity and with increasing number of days open. For cows that aborted, the median time to fetal death decreased slightly as gravidity increased. The study considers several methodological issues faced in epidemiologic investigations of fetal health, including multi-modal hazard functions, extensive censoring and non-independence of multiple pregnancies. The model improves our ability to predict bovine abortion and to characterize fetal survival, which have important applications to herd health management.     

Chlamydial infection in aborted and stillborn lambs

Chlamydia psittaci is a major cause of ovine abortion in the fourth to fifth months of gestation. During the lambing seasons of 1986, 1987, and 1988, fetuses from 52 cases of ovine abortion, stillbirth, or perinatal death were submitted to the laboratory for necropsy examination. Placenta or fetal tissues from 34 cases were cultured on mouse L cells for C. psittaci. Chlamydia psittaci was identified by immunofluorescence on cultures in 20 of these cases. The major gross lesion consistently associated with chlamydial abortion was placentitis with multifocal cotyledonary necrosis and accumulation of red-brown exudate in the intercotyledonary placenta. Chlamydiae appeared as spherical organisms, less than 1 micron in diameter, in the cytoplasm of trophoblasts in impression smears of cotyledons. Histologically, placentitis was sometimes accompanied by pneumonia or encephalitis in the fetus. Chlamydia psittaci was considered the cause for fetal death when chlamydial isolation was associated with placentitis or inflammation of other fetal tissues and when other abortifacient agents were not detected.     

Bovine abortion caused by Bacillus cereus

Bacillus cereus was identified as an infrequent abortigenic agent in cattle. Necrotizing placentitis with no or sporadic lesions in fetal tissues was seen. Bacillus cereus was isolated in pure culture from fetal tissue and/or placenta. The recent identification of a bovine abortion caused by B cereus, prompted a retrospective survey of the pathology files. Eight of 947 bovine abortions were attributed to B cereus. Bacillus cereus is often mistaken as a contaminant in bovine abortion because of the failure to identify lesions in fetal tissues compatible with bacterial invasion. A necrotizing toxin may be responsible for the placentitis, with expulsion of the fetus before bacterial colonization.     

Salmonella enterica subsp. arizonae-associated abortion in a mare

Abortion in mares can be a financially devastating event to horse owners and the equine industry. In this case report, we describe the gross, histopathological and microbiological findings associated with an abortion caused by Salmonella enterica subsp. arizonae, which should be considered in cases of abortion and metritis. Gross and histopathological evaluation of fetal and placental tissue were consistent with ascending septic placentitis resulting in fetal septicaemia. S. enterica subsp. arizonae was isolated from the placenta, lung and fetal gastric contents. Diagnosis was confirmed by MALDI-TOF MS. To our knowledge, this subspecies of S. enterica has not been previously reported as an isolate in a case of late-term equine abortion. A retrospective search of abortion cases at our institution was done, and no similar cases were identified for comparison. This case demonstrates S. enterica subsp. arizonae, although not previously reported as a cause of abortion in pregnant mares and uncommonly isolated from equids in the literature, may cause late-term abortions in susceptible animals. This pathogen should be considered in cases of abortion outbreaks on a farm and merits investigation of the carrier status of mares within a herd.     

Field studies on enzootic abortion of ewes in south east Scotland

Statistical data were collected from 20 sheep flocks in which enzootic abortion of ewes (EAE) was diagnosed by examination of fetal material and maternal blood samples. The abortion rate in the 10,645 ewes in these flocks was 7.6 per cent. Thirteen of the flocks were fully vaccinated against EAE but the abortion rate in these was still in the order of 6.5 per cent. EAE was the major cause of the problem as ascertained from the laboratory examination of samples from a wide range of cases of abortion. There is therefore evidence that, in some flocks in south east Scotland, the efficacy of the vaccine against EAE is unsatisfactory. The reasons for this are discussed.     

Ultrasound studies of the effects of certain poisonous plants on uterine function and fetal development in livestock.

Ingestion of locoweed (Astragalus spp. and Oxytropis spp.) by pregnant livestock may result in fetal malformations, delayed placentation, reduced placental and uterine vascular development, hydrops amnii, hydrops allantois, abnormal cotyledonary development, interruption of fetal fluid balance, and abortion. Ultrasonography of pregnant sheep fed locoweed demonstrated that abortion was first preceded by changes in fetal heart rate and strength of contraction and structural changes of the cotyledons, followed by increased accumulation of fetal fluid within the placental membranes and death of the fetus. During pregnancy the toxic agent in locoweed (swainsonine) apparently passes through the placental barrier to the fetus and during lactation through the milk to the neonate. Poison-hemlock (Conium maculatum), wild tree tobacco (Nicotiana glauca), and lunara lupine (Lupinus formosus) all contain piperidine alkaloids and induce fetal malformations, including multiple congenital contractures and cleft palate in livestock. Ultrasonography studies of pregnant sheep and goats gavaged with these plants during 30 to 60 d of gestation suggests that the primary cause of multiple congenital contractures and cleft palate is the degree and the duration of the alkaloid-induced fetal immobilization.     

Preparation and use of a monoclonal antibody to detect Chlamydia psittaci antigen in paraffin-embedded tissue sections

A murine monoclonal antibody prepared against an ovine abortion isolate of Chlamydia psittaci (A22/Teramo) revealed specific binding to a 57 kDa chlamydial antigen in immunoblotting studies. The monoclonal antibody was able to detect intracytoplasmic chlamydial inclusions and scattered elementary bodies in infected McCoy cell culture, and on formalin-fixed paraffin-embedded tissue sections both from experimentally infected mice and from fetal membranes of cases of ovine enzootic abortion.     

Measurement of IgG concentration in ovine fetal fluids: a useful diagnostic test

The Veterinary Diagnostic Laboratory at Oregon State University received 172 aborted ovine fetuses during the 1985-1987 lambing seasons; from 120 of these, body fluids were evaluated for IgG levels. Fifty-two (43%) of the fetal fluids had immunoglobulin G (IgG) levels greater than 15 mg/dl. Forty-five (87%) of the fluids with elevated IgG levels were confirmed or presumed toxoplasma or chlamydia abortions. A mean fetal fluid IgG concentration of 111.5 +/- 78 mg/dl was found for the 26 toxoplasma abortions; for the 19 chlamydia abortions, a mean IgG concentration of 109 +/- 91 mg/dl was found. Antibody titers equal to or greater than 1:40 against Toxoplasma gondii were detected in 23 fetal fluids. Fetal fluid IgG concentration less than 15 mg/dl was found to be associated with bacterial organisms (i.e., Campylobacter sp.) as the confirmed or presumed cause of abortion. These results suggest that measurement of fetal fluid IgG concentration is a useful, supportive diagnostic test in determining the cause of ovine abortion, and should be included as a routine laboratory procedure for ovine abortion diagnosis.     

Chlamydia psittaci infection and associated infertility in sheep.

Nineteen ewes were injected subcutaneously with the agent of enzootic ovine abortion, Chlamydia psittaci serovar 1, at 50 days gestation. Placental and fetal tissues were examined at 15 days postinfection and thereafter at ten day intervals. Placental infection was detected at 15 days postinfection. Only postinoculation sera collected from postinfected ewes contained antibodies reactive to C. psittaci. Five (26%) chlamydial infected ewes experienced inapparent fetal loss before day 105 of gestation. This finding is significant since C. psittaci infection in sheep is commonly associated with abortion and not infertility.     

Pathology of maternal genital tract, placenta, and fetus in equine viral arteritis

Six pregnant mares were given equine viral arteritis virus intravenously. Tissues from genital tracts, placentae, and fetuses were examined by light and electron microscopy to study the mechanism of abortion. Four mares which died with acute disease had diffuse vacuolation of endometrial epithelium and systemic necrotizing vasculitis. Two of these mares had dead fetuses and two had live fetuses; virus was isolated from tissues of one live fetus. Placentae of mares dying from acute disease did not have lesions attributable to infection; virus was isolated from two of these placentae. One of the two mares which recovered from clinical disease aborted a dead fetus eight days after inoculation. The mare had severe necrotizing myometritis and virus was isolated from maternal ovaries and from fetal tissues. The fetus did not have lesions attributable to arteritis virus. These results suggest that although fetal death may occur in utero during acute equine viral arteritis, abortion probably is due to lesions in the uterus of the mare.     

Efficacy of calfhood vaccination with Brucella abortus strain RB51 in protecting bison against brucellosis

In the studies reported here, protection induced by calfhood vaccination of bison with 1.2-6.1 x 10(10)CFU of Brucella abortus strain RB51 (SRB51) against a virulent strain of B. abortus was evaluated. Non-vaccinated and SRB51-vaccinated bison were intraconjunctivally challenged during midgestation with 3 x 10(7)CFU of virulent B. abortus strain 2308 (S2308). Maternal and fetal tissues were obtained within 24hour after abortion or parturition. Incidence of abortion was greater (P<0.05) in non-vaccinated as compared to SRB51-vaccinated bison (62% and 15%, respectively), with abortions occurring between 5 and 8 weeks after experimental challenge. Calves from bison vaccinated with SRB51 had a reduced (P<0.05) prevalence of fetal infection with S2308 as compared to calves from non-vaccinated bison (19% and 62%, respectively). Although the ability to recover the 2308 challenge strain from maternal tissues did not differ (P>0.05) between nonvaccinates and vaccinates (100% and 78%, respectively), calfhood vaccination with SRB51 reduced (P<0.05) recovery of S2308 from uterine or mammary gland tissues. In bison which did not abort, S2308 was routinely recovered in low numbers from maternal lymphatic tissues; particularly the parotid, bronchial, supramammary, and mandibular lymph nodes. The RB51 vaccine strain was not recovered at any time from maternal or fetal samples obtained at necropsy. Histological lesions associated with Brucella-induced abortions were suppurative placentitis, fetal broncho-interstitial pneumonia, and fetal histiocytic splenitis. The results of our studies suggest that calfhood vaccination of bison with SRB51 is efficacious in protecting against intramammary, intrauterine, and fetal infection following exposure to a virulent strain of B. abortus during pregnancy. As brucellosis is transmitted horizontally through fluids associated with the birth or abortion of an infected fetus, or vertically to the calf through the ingestion of milk containing B. abortus, our data suggest that calfhood vaccination with SRB51 will be beneficial in preventing transmission of brucellosis in bison.     

Equine abortion and premature birth associated with Cellulosimicrobium cellulans infection.

During the 2002 and 2003 foaling seasons, Cellulosimicrobium (Cellumonas) cellulans (formerly Oerskovia xanthineolytica) was the principal microorganism isolated from fetal tissues or placentas from cases of equine abortion, premature birth, and term pregnancies. Significant pathologic findings included chronic placentitis and pyogranulomatous pneumonia. In addition, microscopic and macroscopic alterations in the allantochorion from 4 of 7 cases of placentitis were similar to those caused by Crossiella equi and other nocardioform bacteria. This report confirms a causative role of C. cellulans infection in equine abortion.     

Single tube nested PCR for the detection of Toxoplasma gondii in fetal tissues from naturally aborted ewes.

A single tube nested polymerase chain reaction (PCR) assay targeting the multicopy 18S-5.8S rRNA internal transcribed spacer (ITS1) region has been developed for the diagnosis of Toxoplasma gondii-induced abortion in ovine fetal tissues. In all, 145 ovine fetal samples including brain, spleen, lung, liver, kidney, placenta and fetal fluids from 53 fetuses and stillborns of 32 farms in Northern Spain were analyzed. Thirty-six samples belonging to nine fetuses and one stillborn lamb were T. gondii PCR-positive. Although T. gondii DNA was amplified from different types of tissues, brain was the tissue with the highest detection rate. All animals that had histopathological lesions associated to T. gondii infection were positive by PCR. In addition, four fetuses whose histological examination was hindered by autolysis were PCR-positive. Results obtained by PCR and indirect fluorescent antibody test (IFAT) showed good correspondence, demonstrating the diagnostic value of the two techniques. However, PCR has the advantage over serology in its ability to diagnose T. gondii infection at earlier stages of gestation when the fetus is not yet immunocompetent and in lambs that have taken colostrum. Once other abortifacient agents are ruled out, PCR detection of the ITS1 region in fetal tissues is a valuable and relatively rapid technique for the diagnosis of ovine abortion caused by T. gondii.     

Detection of chlamydial antibody by fetal serology--an aid to the diagnosis of ovine abortion.

Two serological tests (indirect immunofluorescence and enzyme-linked immunosorbent assay) were developed for the detection of fetal antibody to Chlamydia psittaci. Fetal blood and thoracic fluid from 126 field cases of suspected ovine chlamydial abortion were examined using both tests. Placenta and fetal tissues (lung, liver, and kidney) from the same animals were also examined by the following conventional diagnostic methods: isolation in McCoy cells, detection of chlamydial lipopolysaccharide (LPS), modified Ziehl-Nielsen staining, and direct fluorescent antibody staining of chlamydia in frozen cryostat sections. Seventy cases were positive by fetal serology, and of these, 68 were also positive by isolation and/or LPS detection. The remaining 56 cases had negative fetal serology, and of these, 39 were positive by isolation and/or LPS detection. Results indicate that fetal serology, although less sensitive than either isolation in McCoy cells or detection of chlamydial LPS antigen, may be of particular use when placenta is not available.     

The live attenuated bovine viral diarrhea virus components of a multi-valent vaccine confer protection against fetal infection

Fetal infection with bovine virus diarrhea virus (BVDV) causes severe economic loss and virus spread in cattle. This study investigated the ability of modified live BVDV I and II components of a commercially available modified live virus (MLV) vaccine (Breed-Back FP 10, Boehringer Ingelheim Vetmedica Inc.) to prevent fetal infection and abortion, and therefore the birth of persistently infected animals. Heifers immunized with vaccine 4-8 weeks before insemination showed no adverse effects. All vaccinated animals had seroconverted to BVDV 4 weeks after immunization. Pregnant heifers were divided into two vaccination and two control groups and challenged with type I or II BVDV on days 60-90 of gestation. Seroconversion, clinical signs, immunosuppression, viremia, mortality, abortion rate, and fetal infection were studied. Post-challenge, 6/11 (type I challenged) and 8/11 (type II challenged) vaccinated heifers were free from clinical signs of BVD. Post-challenge clinical signs noted in the vaccinated groups were mild to moderate, while all unvaccinated controls had clinical signs ranging from moderate to severe. Viremia was not detected post-challenge in any of the vaccinated heifers. However, 100% of the controls were BVDV viremic on at least 1 day post-challenge. One of 22 vaccinated heifers had transient leukopenia, whereas 2/8 and 6/7 unvaccinated heifers in control groups I and II, respectively, had transient leukopenia. Type II BVDV infection led to abortion or death in 86% of unvaccinated heifers. The corresponding vaccinated group showed no deaths or abortions. All control group fetuses were infected with BVDV. The test vaccine gave 91% (type I BVDV challenged) and 100% (type II BVDV challenged) protection from fetal infection. This vaccine is safe and effective against fetal infection, abortion (type II BVDV) and the birth of persistently infected animals.     

Involvement of Parachlamydia in bovine abortions in Scotland

Bovine abortion represents a major animal welfare issue and a cause of substantial economic loss yet the rate of successful diagnosis remains low. Chlamydia-related organisms including Parachlamydia have recently emerged as putative cattle abortifacients. Placental tissue samples and fetal lung from bovine abortion submissions across Scotland in Spring 2011 were investigated by histopathology for the presence of suspect Chlamydia-related organisms. Evidence of Chlamydia-related organisms was observed in 21/113 (18.6%) placenta samples. Thirteen of the suspect cases and 18 histopathology negative cases were analysed by molecular and immunohistochemical methods. All samples were PCR positive for Parachlamydia but sequencing revealed high homology between identified environmental 16S sequences in all but three cases. Parachlamydial antigen was detected in 10/31 placental samples (32.2%) with pathology consistent with chlamydial infection. This work supports the need for further surveillance investigations and experimental studies to determine the role of Parachlamydia in bovine abortion.     

Prepartum changes of plasma concentrations of prostaglandin F and 13,14-dihydro-15-ketoprostaglandin metabolites in pregnant animals exposed to Sarcocystis cruzi or Campylobacter fetus

Pregnant cows at 4- to 5-months' of gestation were exposed to Sarcocystis cruzi or Campylobacter fetus. Plasma prostaglandin F (PGF) and 13,14-dihydro-15-ketoprostaglandin metabolite (PGM) concentrations were determined at intervals from before exposure until abortion or parturition. The plasma PGF concentration of pregnant infected cattle remained at 0.02 +/- 0.04 ng/ml until 24 to 48 hours before abortion or parturition when it increased 5-fold to 0.11 +/- 0.12 ng/ml. The plasma PGM concentration of these cattle remained at 0.10 +/- 0.07 ng/ml until 24 to 48 hours before abortion or parturition when it increased over 10-fold to 1.36 +/- 0.60 ng/ml. This change in PGF and PGM was similar to that of cattle exposed to each of the infective agents and to that of normal cows at parturition. Thus, changes in PGF and PGM concentrations in bovine plasma cannot be used as a diagnostic tool to determine fetal distress or fetal death for these infections.