Congenital spirochetosis in calves: association with epizootic bovine abortion

Congenital spirochetosis was encountered as a newly recognized infection of cattle. The spirochete was seen in blood of fetuses with lesions of epizootic bovine abortion. A spirochete with morphologic features similar to those found in the fetuses was detected in Ornithodoros coriaceus ticks. Ticks collected from rangelands were allowed to feed on cows that then produced epizootic bovine abortion-affected fetuses, and the fetuses had spirochetosis. Inapparent spirochetosis also was found in fetuses in clinically normal cattle sent to slaughter. Only a few lesions were seen in abattoir-collected fetuses. Fetal spirochetosis was common in the bovine population studied, and it appeared that infection may be limited only by the availability of the tick vector.     

Histopathologic changes in bovine fetuses after repeated reintroduction of a spirochete-like agent into pregnant heifers: association with epizootic bovine abortion

A spirochete-like organism was found in the plasma of bovine fetuses affected with epizootic bovine abortion (EBA). The spirochete-like organism was frequently found in abattoir-collected fetuses as an inapparent infection, and EBA was found in cattle on foothill rangeland where the vector tick Ornithodorus coriaceus could repeatedly reintroduce the infectious agent into pregnant cattle (superinfection). Epizootic bovine abortion resembled a naturally acquired superinfection in circumstances where the agent was frequently present in the environment under conditions favoring transmission. Therefore, to determine whether fetal lesions could be experimentally induced in utero, spirochete-like organisms collected from clinically normal fetuses at an abattoir were inoculated IV and subcutaneously into 2 pregnant heifers 5 times over a 4-month period to mimic repeated tick transmission in the field. Macroscopic and microscopic examinations of tissues from 2 cesarean-collected fetuses and from 3 calves born at term with the naturally acquired spirochete infection indicated that the calves had evidence of an infection that caused morphologic changes compatible with immunologic stimulation and mild reticuloendothelial hyperplasia. Compared with findings in the calves, lesions in the superinfected fetuses were more severe, and the lesion distribution in various organs was more extensive. The spirochete-like organism appeared to be a mild pathogen because of its persistence in the host. Clinical disease from the infection may only develop with repeated superinfections. Therefore, a relationship between this microorganism and EBA is probable.     

Comparison of antigenic structure and pathogenicity of bovine intestinal Chlamydia isolate with an agent of epizootic bovine abortion.

A bovine fecal Chlamydia, isolated from a clinically normal cow, had pathogenic capabilities and antigenic structure similar to those of a chlamydial agent of epizootic bovine abortion (EBA). Intravenous inoculation of the fecal chlamydia into a pregnant heifer caused abortion and lesions in the fetus indistinguishable from those of experimentally induced EBA. In serotests with species-specific complement fixation antigens, the intestinal agent was similar to the EBA agent.     

Frequency and causes of infectious abortion in a dairy herd in Queretaro, Mexico

The objective of this study was to determine the frequency of infectious bovine abortion and to identify some of its causes, specifically brucellosis, leptospirosis, bovine viral diarrhea, infectious bovine rhinotracheitis, and neosporosis. The study was carried out in a dairy herd in the state of Queretaro, Mexico, between September 2002 and March 2003. At the beginning of the study, blood samples were taken from a random 33% of the 300 lactating or pregnant cows; antibodies against Leptospira interrogans were the most commonly identified, in 91% of the 99 samples. Blood samples were also taken 14 to 28 d after the 26 subsequent abortions in the herd in the 6-mo study period, as well as from 22 cows that had not aborted within 5 d after the abortions in the other group. Seroconversion was most frequent for L. hardjo, occurring in 8 (67%) of the 12 dams that aborted after the initial serologic sampling and for which paired serum samples were therefore available. Of the 16 collected fetuses, 10 had histologic lesions suggesting infection in various organs, the features correlating with the serologic results for the dams in 7 cases. Thus, the abortions may have been caused by more than 1 infectious agent.     

Identification of the etiologic agent of epizootic bovine abortion in field-collected Ornithodoros coriaceus Koch ticks

Epizootic bovine abortion (EBA), or foothill abortion as it has often been termed, is a tick-borne disease of pregnant cattle recognized in California, Nevada and Oregon. The primary objective of this study was to better define the relationship of a novel deltaproteobacterium, the putative etiological agent of EBA (aoEBA), with the Pajaroello tick (Ornithodoros coriaceus Koch), the recognized vector of EBA. Three developmental stages of O. coriaceus (larva, nymph, and adult) were collected from five locations in California, Nevada and Oregon. A polymerase chain reaction (PCR), developed for detection of aoEBA, was applied to DNA extracted from ticks. Southern blotting of the PCR products increased the number of ticks determined to be carrying the bacteria by seven-fold, suggesting the majority of infected ticks carry relatively low numbers of the pathogen. An effort was made to determine if an artificial blood meal would stimulate replication of the bacterial pathogen, thereby increasing the frequency in which aoEBA could be identified; no statistically significant effect was evident. The number of ticks determined to be carrying aoEBA varied with geographic location and ranged from 5 to 20%. aoEBA was found in both adults (12% of the males and 12% of the females) and nymphs (13%) but not larvae. Comparative analysis of dissected ticks provided strong evidence that the salivary gland was the most common location of aoEBA in field-collected ticks. No significant correlations were identified between the frequency of infection and tick weight, suggesting that increasing tick age and increased number of blood meals did not increase infectivity.     

Adult Amblyomma hebraeum burdens and heartwater endemic stability in cattle.

Several conclusions of importance to studies on the epidemiology of heartwater were drawn from an investigation in which the numbers of adult Amblyomma hebraeum ticks, to which a closed herd of Hereford cattle were exposed over a period of 6 1/2 years, were manipulated. With a tick Cowdria ruminantium infection rate of 3-5%, an endemically stable situation was created by dipping the herd only when an average of 10 adult male and female A. hebraeum ticks were counted on 10 animals. When the average was increased to 15 during the calving period, 97% of calves acquired a tick-mediated immunity at the age of 6 months. Because only adult ticks confined to the hindquarters are counted, this procedure is recommended as a feasible and practical guideline to stock owners wishing to determine a dipping programme that would ensure endemic stability. The indirect fluorescent antibody test gave a true reflection of the infection rate through ticks in calves 3-6 months old, but not in older animals that had been re-infected more than once. This is because on one hand antibody may persist for 2 years after withdrawal from tick exposure and on the other the artificial re-infection of cattle with a tick acquired immunity is not always followed by a rise in antibody titres and may even result in seronegativity. Four cows infected and re-infected through ticks, remained immune to challenge for 2 years after withdrawal from tick exposure. Within the confines of one farm 3 isolates of C. ruminantium that differed in pathogenicity and immunogenicity were recovered from ticks. One of these isolates was almost non-pathogenic to cattle.     

Prospective serology and analysis in diagnosis of dairy cow abortion

A prospective serologic investigation was undertaken on 3 California dairies (herds 1, 3, and 4, as previously designated in a report on abortion surveillance) to determine if fetal loss was associated with infectious disease agents in cows. The diagnostic problem in these herds was typical of many dairies in that abortions were not discovered for several months and aborted fetuses were seldom recovered. Blood from approximately 100 pregnant cows in each herd was sampled at monthly intervals, beginning when the cows were palpated at approximately 40 days gestation. Sera were tested for antibodies to infectious bovine rhinotracheitis virus and bovine viral diarrhea virus (BVDV) and to the Leptospira serovars pomona, hardjo, grippotyphosa, icterohemorrhagiae, canicola, and szwajizak. Logn antibody titers were examined for an association with fetal loss, using multivariate methods of logistic regression and survival analysis. Of the 325 cows followed, 37 aborted, and no fetuses were recovered. Statistical analyses indicated that significant fetal loss was associated with high titers to L. hardjo and with low titers to L. szwajizak (herds 1 and 4) and BVDV (herd 1). Results for herd 3 revealed a connection between abortion and L. icterohemorrhagiae (P = 0.036) and L. canicola (P = 0.050) and possible vaccinal protection against abortion caused by L. grippotyphosa (P = 0.027 and 0.015). For herd 4, there was a marginally significant tendency for the first vaccination of the gestation against leptospirosis to have protected against fetal death (P = 0.077). Advantages of the diagnostic design were that it permitted comparison of titers of aborted cows with those of nonaborted cows and it considered vaccination-induced titers in analyses.(ABSTRACT TRUNCATED AT 250 WORDS)     

Bovine virus diarrhoea-mucosal disease virus: pathogenicity for the fetal calf following maternal infection

Fifteen pregnant, bovine virus diarrhoea-mucosal disease (BVD-MD) antibody-free Jersey heifers were infected experimentally with a mixture of 10 cytopathic strains of BVD-MD virus isolated from cattle in Britain. Each cow was inoculated intramuscularly on gestation day 100 with a high or a low dose of virus grown in primary calf testis tissue cultures. None of the cows showed clinical signs of illness following exposure, but all had seroconverted within six weeks. Six fetuses, including one set of twins, died in utero following infection. Of these five were aborted between days 136 and 154; the sixth one was mummified and still retained at day 300. The remaining 10 fetuses survived to term, but all showed evidence of intrauterine growth retardation with or without gross malformation and/or dysmyelination of the central nervous system. Three were clinically affected with congenital nervous disease. Of the 10 liveborn fetuses, two had specific serum antibodies to BVD-MD. Non-cytopathic BVD-MD virus was recovered from all of the remaining eight. When non-immune cows become infected with BVD-MD virus in mid gestation: transplacental infection of the fetus will probably result; apart from the risk of fetal death, with or without abortion, there is a high probability of fetal mal-development which may not always be clinically obvious; the immunological competence of the fetus may be impaired; congenital infection is likely in a substantial proportion of liveborn calves. About one in 16 bovine fetuses in British herds are estimated to be at risk from BVD-MD virus infection.     

Experimental transmission of epizootic bovine abortion (foothill abortion)

Advances in defining the biology of epizootic bovine abortion (EBA), including identification of the etiologic agent, have been hampered by the inability to reproduce the disease with confidence. Experimental reproduction of EBA, by feeding the tick vector Ornithodoros coriaceus on susceptible pregnant heifers, is not reliable. The primary objectives of this study were to identify specific tissue(s) obtained from EBA-infected fetuses that could transmit the disease, and then utilize such an infectious challenge system to better define the pathogen, host immunity and geographic distribution of the agent. Described here is the ability to routinely reproduce EBA following inoculation of cryopreserved suspensions of homogenized thymus into susceptible pregnant heifers. This challenge system permitted experiments demonstrating the agent was non-filterable, inactivated upon sonication and susceptible to antibiotics. These findings suggest a prokaryotic microbe and represent a major advance in EBA research. Additional experiments demonstrated that inoculation of the cryopreserved EBA-infectious tissue into heifers, prior to breeding, conferred immunity. Furthermore, such immunized heifers were resistant to challenge with heterologous sources of infectious tissue, suggesting monovalent vaccine development might be feasible. Lastly, challenge studies employing animals from Central Nevada, an area considered free of EBA, demonstrated partial immunity, suggesting the pathogen, and possibly the disease, enjoy a broader distribution than previously thought.     

Preliminary observations on rinderpest in pregnant cattle

A Kabete 'O' strain of rinderpest virus enhanced in virulence was inoculated subcutaneously into four cows which were between six and eight months pregnant. All the cows developed clinical signs of rinderpest from the third day after inoculation and shed high titres of virus in their ocular and vaginal secretions during the course of the clinical disease. Three of the cows died of rinderpest on the third day after the onset of fever but no virus was isolated from their fetuses which were examined post mortem. The fourth cow showed complete clinical and virological recovery by the eighth day after the onset of fever and aborted an eight-and-a-half-month-old fetus on the 12th day after it recovered. Rinderpest virus was demonstrated in a wide range of the aborted fetal tissues. Virus was also detected in the maternal vaginal discharges up to 24 hours after abortion. The only gross pathological change observed was a severe necrotising placentitis.     

In utero infection of swine fetuses with infectious bovine rhinotracheitis virus (bovine herpesvirus-1)

The pathogenesis of infectious bovine rhinotracheitis (IBR) virus (bovine herpesvirus-1) was studied in porcine fetuses after in utero inoculation. Laparotomies were performed on 8 seronegative pregnant sows at 34 to 86 days of gestation, and all fetuses in 1 uterine horn of each sow were exposed to IBR virus via inoculation into the amniotic sacs. Fetuses in the other horn served as controls. Clinical signs of infection were not observed in the sows, except for 2 sows that aborted at postinoculation days (PID) 11 and 15. Fetuses of the remaining 6 sows were collected at slaughter on PID 15 to 28. Fetuses were examined for gross abnormalities, presence of IBR virus in tissues, and the formation of neutralizing antibodies to IBR virus. Of 33 inoculated fetuses from 6 sows, 10 were mummified, 11 were hemorrhagic and/or edematous, and 12 were alive. Necrotic lesions were observed on the skin and in the liver of dead and live fetuses. Virus was recovered from 29 of 33 inoculated fetuses. Infectious bovine rhinotracheitis virus was isolated from fetal skin, liver, lungs, kidney, spleen, stomach contents, brain, amniotic fluid, and placenta. Virus was isolated from 4 of 11 fetuses recovered from 1 aborting sow. Antibodies to IBR virus were not detected in sera from the sows. However, antibodies were detected in 6 of 15 fetuses inoculated at 63 to 86 days of gestation and collected at slaughter at 86 to 112 days of gestation. The youngest fetus with detectable IBR antibody was estimated to be 74 days of gestation by measuring crown-rump length of the fetus.     

Borrelia theileri: isolation from ticks (Boophilus microplus) and tick-borne transmission between splenectomized calves

The bovine spirochete, Borrelia theileri, was detected in Giemsastained blood smears from a splenectomized calf 17 days after exposure to a laboratory colony of the tropical cattle tick, Boophilus microplus. Spirochetes were detected in the hemolymph and ovary of all engorged female ticks examined, indicating a high infection rate in this tick colony. Spirochetes were detected in a 2nd splenectomized calf 15 days after exposure to the larval offspring of ticks from the 1st calf. The only observable effect of infection in the 2 calves was a maximum rectal temperature increase to 40.2 C, which coincided with the first detectable parasitemia. The tick colony did not have any adverse effects, despite extensive multiplication of spirochetes in their tissues.     

Seroconversion and abortion in cattle experimentally infected with Brucella sp. isolated from a Pacific harbor seal (Phoca vitulina richardsi).

Previously unrecognized Brucella species have been isolated from a number of marine mammals, including harbor seals (Phoca vitulina richardsi) in the Puget Sound area of the state of Washington. Because of the presence of dairy herds in proximity to the harbor seal populations, a study was conducted to determine the effects of the harbor seal Brucella isolate in experimentally inoculated cattle. Six pregnant cattle were exposed by intravenous injection (n = 3) or intraconjunctival inoculation (n = 3). Two pregnant cows were intravenously injected with saline and served as controls. All of the cows receiving the Brucella seroconverted on 1 or more tests commonly used for the detection of Brucella abortus infection. Two of the cattle receiving the intravenous inoculation aborted, and brucellae were demonstrated in the fetuses and dams immediately following abortion. The remaining 4 Brucella-inoculated animals and their fetuses were culture negative for the organism at 14 weeks postinoculation. Results of this study indicate the marine mammal Brucella is capable of producing seroconversion and abortion in cattle but is less pathogenic in that species than B. abortus.     

Plasma alpha-fetoprotein concentrations in pregnant cows exposed to Sarcocystis cruzi, Campylobacter fetus, or Aspergillus fumigatus

Bovine alpha-fetoprotein (AFP) was determined in maternal plasma, using radioimmunoassay in an attempt to detect and monitor fetal distress in pregnant cows. Plasma from pregnant cows in the 4th to 5th month of the gestation which had been exposed to Sarcocystis cruzi, Campylobacter fetus, or Aspergillus fumigatus was used. Plasma AFP concentrations were determined at intervals from before the cows were exposed until they had aborted or calved. The plasma AFP concentration of the exposed pregnant cattle remained at 6.5 +/- 5.0 mg/ml until 24 to 48 hours before abortion or parturition, when the value increased to 25.0 +/- 8.0 ng/ml. This pattern was similar for cattle exposed to each of the infective agents. Unlike in persons, rats, or monkeys, fetal-maternal transfer of AFP seems to be minimal in cows even with inflammation or necrosis of the placentome, Thus, changes in AFP concentrations in bovine plasma cannot be used as a diagnostic tool for fetal distress or fetal death.     

Histopathologic findings in Brucella abortus-infected, pregnant goats

Twenty-eight pregnant goats in midgestation were exposed to a bovine pathogenic strain of Brucella abortus to determine the histologic changes associated with infection. Does were necropsied 0 to 7 days or 4 to 6 weeks after delivery of aborted, stillborn, or live, full-term fetuses. Aborted and stillborn fetuses were necropsied within 16 hours of delivery. Selected, paired tissue specimens were collected for histologic and bacteriologic examination. An avidin-biotin-peroxidase complex immunostaining procedure was used to detect Brucella antigen in tissue section. Histologic changes were evident in specimens from infected does and aborted fetuses. Postpartum does had endometritis, lymphoid hyperplasia in lymph nodes and spleen, and lymphocytic mastitis. The most prominent finding in aborted fetuses was an interstitial pneumonia. Lesions in does and fetuses were similar to those described in Brucella-infected cows and fetuses; however, lesions were less consistently observed in goat fetuses than has been observed in bovine fetuses. Brucella antigen was detected by immunoperoxidase staining within the cytoplasm of placental chorioallantoic trophoblastic cells, macrophages, neutrophils, and uterine epithelial cells. Also, stained brucellae were free in placental and fetal vascular lumens and in the interstitium of autolyzed fetal tissues.     

Early abortion in cattle induced by experimental intrauterine infection with pure cultures of Actinomyces pyogenes

Actinomyces pyogenes from a case of endometritis was used to study the effects of infection of the bovine embryo between days 27 and 41 of pregnancy. From 10(9) to 10(10) washed organisms were introduced into the uterine lumen of four pregnant cows. Two pregnant cows were inoculated with sterile saline and four pregnant cows were treated with cloprostenol. Embryonic death and abortion followed 29 to 144 hours after the inoculation of the live bacteria. The aborted embryos were macerated or clearly degenerating and yielded profuse pure cultures of A pyogenes. Abortion was accompanied by a sustained increase in uterine tone, opening of the cervix, presence of vaginal pus and a vulval discharge and the persistence of the corpus luteum for at least eight days after abortion. Intrauterine inoculation with saline did not affect pregnancy, but embryonic death, abortion and regression of the corpus luteum occurred 66 to 72 hours after the treatment with cloprostenol. The results suggest that A pyogenes is a primary pathogen and is capable of causing embryonic death and abortion.     

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.     

Some immune responses in cattle exposed to Mycobacterium paratuberculosis after injection with modified-live bovine viral diarrhea virus vaccine

Delayed-type hypersensitivity responses to Mycobacterium paratuberculosis purified protein derivative (PPD) were decreased in cows experimentally exposed to M. paratuberculosis 7 days after exposure to a modified-live bovine viral diarrhea virus (ML-BVDV) vaccine. In vitro lymphocyte blastogenic responses to phytohemagglutinin were decreased in each of 3 cows 7 days after exposure to ML-BVDV vaccine. Also, decreased lymphocyte blastogenic responses to M. paratuberculosis PPD were observed in cultures of 2 of 3 cows 7 days after exposure to ML-BVDV vaccine. No significant differences in enzyme-linked immunosorbent assay reactions were detected in sera of M. paratuberculosis-infected cattle collected before and at 4 and 12 weeks after exposure to ML-BVDV vaccine.     

Risk of abortion associated with Neospora caninum during different lactations and evidence of congenital transmission in dairy cows

OBJECTIVE: To examine the relationship between exposure to Neospora caninum and abortion in dairy cows during their first, second, third, and fourth or later lactations and to establish the main mode of transmission in female calves from birth until their first pregnancy was terminated by abortion or parturition. DESIGN: Prospective observational study. ANIMALS: 460 Holstein cows and 79 female calves. PROCEDURE: Cows were classified as seropositive or seronegative to N caninum within 7 days after calving; incidence of abortion was compared between groups during different lactations. Blood samples were collected from female calves before ingestion of colostrum and every 6 months until their first pregnancy was terminated by abortion or parturition; number of seropositive calves was compared between seropositive and seronegative dams. RESULTS: During the first pregnancy of their second lactation, risk of abortion for seropositive cows was 2.8 times that of seronegative cows. Among 10 calves born to seropositive cows, 4 were classified as seropositive at birth and thereafter. Among 69 calves born to seronegative cows, all were classified as seronegative at birth; 67 calves remained seronegative thereafter. CONCLUSIONS AND CLINICAL RELEVANCE: Exposure to N caninum alone was not significantly associated with abortion in cows during the first, third, and fourth or later lactations. Seropositive cows that have aborted previously may have subsequent abortions attributable to N caninum. Congenital infection was the main mode of N caninum transmission in a cohort of female calves.