Pathology of Perinatal Disorders.

Perinatal deaths including abortions, stillbirths, and neonatal deaths are an important economic and emotional challenge for companion animal owners, breeders, and veterinarians. A protocol for the investigation of perinatal deaths in dogs and cats is described, including examination of the placenta. Specific lesions of common infectious diseases leading to perinatal death are presented, as well as the more prevalent noninfectious causes. These include viruses, bacteria, protozoa, metabolic causes, "accidents of pregnancy," nutritional deficiencies, intoxications, hormonal causes, and heritable and nonheritable congenital defects.

Detection of Anaplasma phagocytophilum in fetal and placental tissue of bovine abortions and perinatal mortalities.

BACKGROUND: Anaplasma phagocytophilum is a tick-borne zoonotic bacterium that is the aetiologic pathogen of tick-borne fever (TBF) in ruminants. In clinical bovine cases of TBF, abortion and stillbirth may be observed. However, in this regard, the pathophysiology of TBF has not yet been completely elucidated, and no clear guidelines to diagnose A. phagocytophilum-related abortions and perinatal mortalities (APM) are available. METHODS: This exploratory study aimed to investigate the presence of A. phagocytophilum in bovine cases of APM and determine whether placental or fetal spleen tissue has the greatest sensitivity for A. phagocytophilum identification. The placenta and fetal spleen of 150 late-term bovine APM cases were analysed using real-time PCR to detect A. phagocytophilum. RESULTS: A total of 2.7% of sampled placentas were positive for A. phagocytophilum, while none of the fetal spleen samples was. LIMITATIONS: No histopathology to detect associated lesions was performed. Consequently, no evidence of causality between the detection of A. phagocytophilum and APM events could be achieved. CONCLUSION: The detection of A. phagocytophilum suggests a potential role of this pathogen in bovine APM, and placental tissue seems to be the most suitable tissue for its identification.

Infectious Causes of Equine Placentitis and Abortion.

A variety of infectious agents including viral, bacterial, and fungal organisms can cause equine abortion and placentitis. Knowledge of normal anatomy and the common pattern distribution of different infectious agents will assist the practitioner in evaluating the fetus and/or placenta, collecting appropriate samples for further testing, and in some cases, forming a presumptive diagnosis. In all cases, it is recommended to confirm the diagnosis with molecular, serologic, or microbiological testing. If a causative agent can be identified, then appropriate biosecurity and vaccination measures can be instituted on the farm.

ABORTION AND NEONATAL MORTALITY DUE TO TOXOPLASMA GONDII IN BIGHORN SHEEP (OVIS CANADENSIS).

Low lamb recruitment can be an obstacle to bighorn sheep (Ovis canadensis) conservation and restoration. Causes of abortion and neonate loss in bighorn sheep, which may affect recruitment, are poorly understood. Toxoplasma gondii is a major cause of abortion and stillbirth in domestic small ruminants worldwide, but no reports exist documenting abortion or neonatal death in bighorn sheep attributable to toxoplasmosis. Between March 2019 and May 2021, eight fetal and neonatal bighorn lamb cadavers from four western US states (Idaho, Montana, Nebraska, and Washington) were submitted to the Washington Animal Disease Diagnostic Laboratory for postmortem examination, histologic examination, and ancillary testing to determine the cause of abortion or neonatal death. Necrotizing encephalitis characteristic of toxoplasmosis was identified histologically in six of eight cases, and T. gondii infection was confirmed by PCR in five cases with characteristic lesions. Other lesions attributable to toxoplasmosis were pneumonia (3/5 cases) and myocarditis (2/5 cases). Protozoal cysts were identified histologically within brain, lung, heart, skeletal muscle, adipose tissue, or a combination of samples in all five sheep with PCR-confirmed T. gondii infections. Seroprevalence of T. gondii ranged from 40-81% of adult females sampled in the Washington population in October and November 2018-2021, confirming high rates of exposure before detection of Toxoplasma abortions in this study. Of 1,149 bighorn sheep postmortem samples submitted to Washington Animal Disease Diagnostic Laboratory between January 2000 and May 2021, 21 of which were from fetuses or neonates, a single case of chronic toxoplasmosis was diagnosed in one adult ewe. Recent identification of Toxoplasma abortions in bighorn sheep suggests that toxoplasmosis is an underappreciated cause of reproductive loss. Abortions and neonatal mortalities should be investigated through postmortem and histologic examination, particularly in herds that are chronically small, demographically stagnant, or exhibit reproductive rates lower than expected.

Pathological and etiological characterization of cases of bovine abortion due to sporadic bacterial and mycotic infections.

Opportunistic bacteria and fungi are commonly reported causes of bovine abortion in a small percentage of fetal losses of infectious etiology in cattle. The objective of this study was to characterize the pathological and etiological findings in fetuses aborted due to secondary bacterial and fungal infections submitted for postmortem examination between 2004 and 2019 in the State of Rio Grande do Sul, Brazil. Nineteen cases of bacterial etiology and five cases of fungal etiology were assessed. In cases of bacterial etiology, gross changes were uncommon and two different microscopic patterns were observed: (1) primary bronchopneumonia with occasional dissemination in cases of Staphylococcus sp., Streptococcus sp., and Mannheimia haemolytica infections; and (2) systemic disease with sepsis in cases of Escherichia coli and Listeria sp. infections. Aspergillus sp. was the main fungal agent identified, and cases of mycotic abortion were characterized by placentitis, dermatitis, and pneumonia. Fetal membranes were available for examination in less than half of the submissions (11/24), and placental lesions were observed in all cases. This study reaffirms the importance of postmortem examinations in the determination of causes of fetal loss in cattle and highlights pathological findings commonly observed in fetuses aborted due to sporadic bacterial and fungal agents.

Investigation of the outbreaks of abortions and orchitis in livestock in Namibia during 2016-2018.

This study investigated outbreaks of seemingly related abortions and orchitis which occurred in the Khomas, Omaheke and Otjozondjupa regions of Namibia from 2016 to 2018, affecting cattle, sheep and goats. Fifty-nine questionnaires were administered, and 48 were completed giving an 81.4% return. The outbreaks were limited to Namibia's east and central regions, mainly on farms rearing cattle, sheep and goats and on farms with a mixture of these species. There was no significant difference between Khomas and other regions on abortion reporting at the farm level [X2 (1, N = 48) = 0.0002, p = 0.987851]. However, there was a significant difference in the abortions at the animal level among the three regions [X2 (2, N = 6246) = 239.8339, p = .00001]. In addition, the proportions of abortions calculated at the animal level at each farm were significantly different when the Khomas region was compared to the other regions. Seventeen cattle sera, 35 sheep sera, 52 caprine sera, 18 bovine liver samples, one caprine liver, five aborted cattle foetuses, two cattle placentas, 18 testes (one bull, eight bucks and nine rams) and ten bull sheath scrapings were collected and tested. Histopathology, microbiology, serology, immunohistochemistry, real-time PCR and mineral analytical techniques were used to establish the aetiology of the abortion and orchitis outbreaks. The gross and histopathological findings on the 18 testicles were characteristic of chronic orchitis. In aborted foetuses, significant histopathological findings included meconium aspiration, funisitis and cardiomyopathy. Placentitis and endometritis were the primary pathologies observed in cows. The bacteria isolated from microbiological samples included Enterococcus spp. (65.5% [19/29]), Enterobacter spp. (6.9% [2/29]) and Streptococcus spp. (10.3% [3/29]), Trueperella pyogenes (3.4% [1/29]), Stenotrophomonas maltophilia (3.4% [1/29]), Staphylococcus epidermidis (3.4% [1/29]), Providencia rettgeri (3.4% [1/29]) and Acinetobacter lwoffii (3.4% [1/29]), mostly opportunistic bacteria. On mineral analysis, 28%, 33%, 83%, 33% and 17% (n = 18) of cattle livers were low in copper, zinc, manganese, selenium and iron, respectively. Twenty-three percent (12/52) of the caprine sera were positive for Brucella melitensis on the Rose Bengal and complement fixation tests. Thirty-five ovine sera were tested for B. melitensis, B. ovis and Coxiella burnetii, and the prevalence for each was 2.9% (1/35). PCR tests on foetuses were all negative for Brucella spp., Coxiella burnetii, Chlamydia spp., Listeria monocytogenes, Salmonella spp., Campylobacter fetus spp., Leptospira pathogenic strains, bovine viral diarrhoea virus, Rift Valley fever virus, Anaplasma phagocytophilum and bovine herpes virus 4 Campylobacter fetus spp. and Trichomonas foetus spp. The authors concluded that Brucella spp., Enterococcus spp., Escherichia coli, Streptococcus spp., Trueperella pyogenes and Coxiella burnetii could have contributed to this outbreak. Micronutrient imbalances and pathogenic abiotic nanoparticles were also identified as possible contributors to the abortion outbreaks.

Bovine abortion and necrotic placentitis by Aspergillus terreus.

A 31-month-old Japanese Black cow (Bos taurus) aborted at 5 months of gestation with no clinical symptoms. Histopathological examination of the placenta and fetus revealed severe necrotic placentitis associated with numerous irregular degenerative fungi and inflammatory cells. Regular filamentous fungi were also detected, without inflammatory response in the fetal digestive and respiratory organs. Both fungi had aleurioconidia and septa in the placenta and fetal organs and immunohistochemically stained with antibodies against Aspergillus spp. Aspergillus terreus was isolated from the fetal lung and abomasal contents as confirmed using mycological and molecular methods. This is the first immunohistochemical, morphological, and molecular identification of A. terreus in bovine placenta and aborted fetuses.

Equine abortion associated with placentitis caused by Pasteurella pneumotropica / Aborto equino associado a placentite por Pasteurella pneumotropica

Pasteurella pneumotropica is a bacterium that has so far not been described as a cause of placentitis in animals. Two cases of aborted equine fetuses were sent to the Department of Veterinary Pathology of the "Universidade Federal do Rio Grande do Sul" (SPV-UFRGS) for anatomopathological examination. Both cases presented suppurative placentitis associated with multiple basophilic bacterial cells. After bacterial isolation and biochemical analysis, P. pneumotropica was identified.

Pasteurella pneumotropica é uma bactéria que até o momento não foi descrita como causa de placentite em animais. Dois casos de fetos equinos abortados foram enviados ao Setor de Patologia Veterinária da Universidade Federal do Rio Grande do Sul (SPV-UFRGS) para exame anatomopatológico. Em ambos os casos se observou placentite supurativa associada a múltiplas colônias bacterianas basofílicas. Após o isolamento bacteriano e análise bioquímica, indentificou-se P. pneumotropica.

Equine abortion associated with placentitis caused by Pasteurella pneumotropica / Aborto equino associado a placentite por Pasteurella pneumotropica

Pasteurella pneumotropica is a bacterium that has so far not been described as a cause of placentitis in animals. Two cases of aborted equine fetuses were sent to the Department of Veterinary Pathology of the "Universidade Federal do Rio Grande do Sul" (SPV-UFRGS) for anatomopathological examination. Both cases presented suppurative placentitis associated with multiple basophilic bacterial cells. After bacterial isolation and biochemical analysis, P. pneumotropica was identified.(AU)

Pasteurella pneumotropica é uma bactéria que até o momento não foi descrita como causa de placentite em animais. Dois casos de fetos equinos abortados foram enviados ao Setor de Patologia Veterinária da Universidade Federal do Rio Grande do Sul (SPV-UFRGS) para exame anatomopatológico. Em ambos os casos se observou placentite supurativa associada a múltiplas colônias bacterianas basofílicas. Após o isolamento bacteriano e análise bioquímica, indentificou-se P. pneumotropica.(AU)

Coxiella burnetii abortion in a dairy farm selling artisanal cheese directly to consumers and review of Q fever as a bovine abortifacient in South America and a human milk-borne disease.

Coxiella burnetii is a highly transmissible intracellular bacterium with a low infective dose that causes Q fever (coxiellosis), a notifiable zoonotic disease distributed worldwide. Livestock are the main source of C. burnetii transmission to humans, which occurs mostly through the aerogenous route. Although C. burnetii is a major abortifacient in small ruminants, it is less frequently diagnosed in aborting cattle. We report a case of C. burnetii abortion in a lactating Holstein cow from a dairy farm producing and selling artisanal cheese directly to consumers in Uruguay, and review the literature on coxiellosis as a bovine abortifacient in South America and as a milk-borne disease. The aborted cow had severe necrotizing placentitis with abundant intratrophoblastic and intralesional C. burnetii confirmed by immunohistochemistry and PCR. After primo-infection in cattle, C. burnetii remains latent in the lymph nodes and mammary glands, with milk being a significant and persistent excretion route. Viable C. burnetii has been found in unpasteurized milk and cheeses after several months of maturing. The risk of coxiellosis after the consumption of unpasteurized dairy products, including cheese, is not negligible. This report raises awareness on bovine coxiellosis as a potential food safety problem in on-farm raw cheese manufacturing and sales. The scant publications on abortive coxiellosis in cattle in South America suggest that the condition has probably gone underreported in all countries of this subcontinent except for Uruguay. Therefore, we also discuss the diagnostic criteria for laboratory-based confirmation of C. burnetii abortion in ruminants as a guideline for veterinary diagnosticians.

Quantitative evaluation of the infection dynamics of bovine brucellosis in Tanzania.

Brucellosis is endemic in Tanzania. A cross-sectional study was conducted at 17 cattle farms in agro-pastoral areas in Tanzania to identify risk factors associated with the within-farm prevalence of bovine brucellosis and to quantitatively assess the infection dynamics through disease modelling. Cattle blood sampling and interviews with farmers using a structured questionnaire were conducted. A total of 673 serum samples were screened using the Rose-Bengal plate test (RBPT), and sero-positivity of RBPT-positive samples was confirmed using a competitive enzyme-linked immunosorbent assay. Zero-inflated binomial regression was performed for univariable and multivariable risk factor analyses of within-farm prevalence. Several susceptible-infectious (SI) models were compared based on deviance information criteria, and age-dependent force of infection (FOI) was measured using age-specific prevalence data for the 10 infection-positive farms. Using the diagnoses of cows on the 17 farms, the basic reproduction number, R0, was also calculated. The farm-level prevalence and animal-level adjusted prevalence were 58.8 % (10/17, 95 % confidence interval: 33.5-80.6 %) and 7.0 % (28/673, 95 % credible interval: 5.7-8.4 %), respectively. The risk factor for high within-farm prevalence was introduction of cattle from other herds. A mathematical model with constant FOI showed the annual probability of infection as 1.4 % (95 % credible interval: 1.0 %-2.0 %). The R0 was 1.07. The constant FOI could have been due to the predominant mode of infection being transmission of Brucella from contaminated aborted materials during grazing. Direct purchase of infected cattle could facilitate efficient transmission between susceptible animals through abortion.

Epidemiology of Chlamydia psittaci infections in pregnant Thoroughbred mares and foals.

Late-term foal loss due to the traditional avian pathogen Chlamydia psittaci recently emerged as a threat to the Australian Thoroughbred industry. A longitudinal study of 14 stud farms was undertaken to better understand C. psittaci infection in pregnant mares and their foals by evaluating C. psittaci prevalence, equine herpesvirus-1 (EHV-1) co-infection, avian reservoirs, and potential risk factors. Mucosal swabs taken from 228 healthy pregnant mares and their foals were tested for C. psittaci and EHV-1 using species-specific qPCR assays. No foal loss was recorded due to either pathogen, and no mare tested positive to either C. psittaci or EHV-1. However, healthy newborn foals tested positive to both pathogens, at low levels, with 13.2% (n = 30/228) and 14.5% (n = 33/228) prevalence for C. psittaci and EHV-1, respectively. Co-infection occurred in 1.3% (n = 3/228) of foals. In avian environmental faecal samples collected from the same studs, C. psittaci was detected at 5.3% (n = 5/94). Multiple logistic regression modelling found that foals born in winter were more likely to be infected with C. psittaci (adjusted odds ratio = 15.83; P < 0.001; Confidence Interval 5.12-48.49). Being a maiden mare, absence of prophylactic vaginal suture, interventions in the last trimester and residing on a farm with prior history of C. psittaci abortion posed no higher risk to infection in the newborn. Analysis of all reported C. psittaci abortion cases (Hunter Valley, 2016-2019) revealed a dominant C. psittaci sequence type (denoted ST24) and a significant correlation with frost events (Spearmans' rho = 0.44; P = 0.002).

Chlamydia pecorum detection in aborted and stillborn lambs from Western Australia.

Lamb survival is an important welfare and productivity issue for sheep industries worldwide. Lower lamb survival has been reported for primiparous ewes, but the causes of this are not well studied. The aim of this study was to determine causes of perinatal deaths for lambs born to primiparous ewes in Western Australia, and identify if infectious diseases are implicated. Lamb mortality from birth to marking were determined for 11 primiparous ewe flocks on 10 farms in Western Australia. Lamb mortality from birth to marking averaged 14% for single-born and 26% for multiple-born lambs. Lamb necropsies (n = 298) identified starvation-mismosthering-exposure (34%), dystocia (24%) and stillbirth (15%) as the most common causes of perinatal lamb death. There was no evidence of exotic abortigenic pathogens in aborted and stillborn lambs (n = 35). Chlamydia pecorum was detected by qPCR in 15/35 aborted and stillborn lambs on 5/6 farms. Preliminary molecular characterisation of C. pecorum detected in samples from aborted and stillborn lambs (n = 8) using both Multilocus Sequence Typing and ompA genotyping indicated all strains were genetically identical to previously described pathogenic livestock strains, denoted ST23, and dissimilar to gastrointestinal strains. High frequency of detection of a pathogenic C. pecorum strains ST23 associated with ovine abortion and stillbirth on multiple farms located across a wide geographic area has not been previously reported. Chlamydia pecorum may contribute to reproductive wastage for primiparous sheep in Western Australia. Further investigation to understand C. pecorum epidemiology and impact on sheep reproduction is warranted.

Genetic features of Salmonella enterica subspecies diarizonae serovar 61:k:1,5 isolated from abortion cases in sheep, United States, 2020.

Salmonella enterica subspecies diarizonae serovar 61:(k):1, 5, (7) (sheep associated S. diarizonae, SASd) is the most common Salmonella serotype identified in sheep flocks. Despite the involvement with animal and human infections, there is limited information regarding virulence profiles of SASds and their antibiotic resistance gene complement, particularly for those circulating in the U.S. In this study, we genetically characterized three SASds, 20-265, 20-269, and 20-312, isolated from sheep placental tissues during an abortion storm affecting a flock in Connecticut during 2020. SASds were the only bacteria isolated from analyzed sheep tissues. The isolates were sensitive to all the antibiotics tested, but all these SASd isolates carry the aminoglycoside resistance gene, aac(6')-Iaa, and a chromosomal substitution in the parC gene. The proportion of pseudogenes (5.3-5.5%) was similar among the isolates, and these SASds carry IncX1 type plasmids. Comparing with the SASds isolates from Enterobase, the three isolates showed an identical genomic virulence profile carrying virulence genes in the conserved set of other SASd isolates except for steC, iagB, iacP, sseI, and slrP genes. In the SNP-based phylogenetic analysis, SASd sequences were grouped into group A-C, and the group C was further subdivided into subgroup C1-C6. The three isolates clustered with other SASd isolates from the U.S. and Canada in subgroup C6. SASd isolates in the identical phylogenetic groups tended to have similar geographical origin. The results of our study did not provide conclusive evidence about which are the genetic traits that trigger SASds to become virulent in sheep, but our data will provide a point for comparative studies of this Salmonella serovar.

Lay attitudes and misconceptions and their implications for the control of brucellosis in an agro-pastoral community in Kilombero district, Tanzania.

Brucellosis is a priority zoonotic disease in Tanzania that causes ill-health in people and affects livestock productivity. Inadequate awareness and behavior risking transmission can impede control efforts. We conducted a cross-sectional survey of 333 livestock owners in three villages in the Kilombero district, Tanzania, to understand their awareness, knowledge and behavior associated with brucellosis. Six Focus Group Discussions (FGDs), two in each village, were conducted, as well as an additional FGD with male herders from one of the villages. Factors associated with knowledge on brucellosis, food consumption and animal husbandry behavior risking transmission of this disease, were identified using generalized linear models. Predictors for knowledge of brucellosis were being male and having a higher educational level, while age was positively associated with a higher level of knowledge. Faith and ethnicity were associated with the performance of practices risking transmission. Following traditional religion and belonging to the Wamaasai ethnicity significantly increased the odds of carrying out these practices. Qualitative analysis gave insight into risk practices and reasoning. Of the 333 respondents, 29% reported that they had experienced abortions in their herds, 14% witnessed retained placentas, and 8% had seen still-births in their cattle within the previous year. However, survey results also showed that only 7.2% of participants had heard about brucellosis as a disease in livestock. Of those who had heard about brucellosis in livestock, 91% associated abortions with it and 71% knew that humans can get infected through raw milk consumption. People overwhelmingly attributed symptoms and transmission of brucellosis in livestock to infection with trypanosomiasis and to supernatural reasons instead. In the community, consumption of raw milk was valued and handling of aborted material was not considered a risk for infection. This agro-pastoralist community holds on to long-held beliefs and practices and lacks understanding of the biomedical concept of brucellosis. Transmission routes and symptoms of brucellosis in humans and livestock are completely unknown. The disparity between risk perception and actual transmission risk related to animal handling and consumption of animal products presents a challenge for disease awareness communication. This study recommends focused community engagement and sensitization to address the limited awareness and misconceptions among agro-pastoralists.

Epidemiologic case investigation on the zoonotic transmission of Staphylococcus aureus infection from goat to veterinarians.

Staphylococcus aureus infection led to a case of goat abortion, and four veterinarians contracted S. aureus infection from the goat during and after the abortion. Three veterinarians assisted a doe during the dystocic delivery of a dead foetus. Seventy-two hours after the dystocia, which ended with the goat's death, the veterinarians who assisted during the kidding and the veterinarian who performed the necropsy showed the presence of multiple, isolated, painful pustules 1-5 mm in diameter located along their forearms and knees. S. aureus was isolated from the pustules of the veterinarians, the placenta and uterus of the goat, the organs (brain, thymus gland, abomasum, liver and spleen) of the foetus, the scrotum and eye swabs of the buck, and mammary pustules of another goat from the same herd. Histological analysis revealed purulent metritis and inflammation of the placental cotyledons. Additional investigations eliminated the chances of other infections. S. aureus isolates recovered from the veterinarians, goats, foetus and buck were sensitive to the tested anti-microbials and did not encode staphylococcal enterotoxin genes (sea, ser, sep, see, seg and sei). The isolates were closely related, as indicated by the results of Fourier-transform infrared spectroscopy and comparative whole-genome sequencing analysis. The results of this study clearly support the hypothesis that an episode of professional zoonosis was caused by S. aureus infection during the abortion and also highlight the need for bacterial subtyping in epidemiological surveys.

Molecular detection of Coxiella burnetii infection in aborted samples of domestic ruminants in Iran.

BACKGROUND: Coxiella burnetii is the causative agent of Q fever which is a highly infectious zoonotic disease. C. burnetii has become one of the most important causes of abortion in livestock, which can lead to widespread abortions in these animals. There are very limited studies on the prevalence of C. burnetii infection in cases of animal abortion in Iran. The aim of this study was to investigate the occurrence of C. burnetii in ruminant abortion samples in Iran. METHODS: Abortion samples from cattle, sheep and goats were collected from different parts of Iran and were tested using Real-time PCR targeting the IS1111 element of C. burnetii. RESULTS: In this study, 36 samples (24.7%) of the 146 collected samples were positive for C. burnetii. The prevalence of C. burnetii was 21.3% (20 of 94 samples) in sheep samples. Also, 10 of 46 cattle samples (21.7%) were positive. All six goat abortion samples were positive for C. burnetii. CONCLUSIONS: The findings of the study demonstrate that C. burnetii plays an important role in domestic ruminant abortions in Iran, suggesting that more attention should be paid to the role of C. burnetii in domestic animal abortions by veterinary organizations. The risk of transmitting the infection to humans due to abortion of animals should also be considered.

Infectious bovine abortions: observations from an organized dairy herd.

Abortions in dairy animals can be caused by several infectious agents. Identification of the actual causal agent(s) is important for formulating suitable control strategies. A 3-year (2016-2018) longitudinal study was conducted in a dairy farm following an abortion storm in the mid- to late gestations. The investigation focused on the seven major infectious abortifacient in cattle, viz. bovine alphaherpesvirus-1 (BoHV-1), bovine viral diarrhoea virus (BVDV), Neospora caninum, Brucella abortus, Coxiella burnetii, Leptospira Hardjo, and Listeria monocytogenes. High seroprevalence was observed for BVDV (79.4%), Leptospira (70.5%), BoHV-1 (53.5%), and Brucella (45.0%) at the beginning of the investigation (August 2016). The incidence proportion increased for BVDV, Leptospira, and Brucella in the following years of the investigation. A strong association of Brucella seropositivity with history of abortion (OR = 3.27) was recorded. Incidence of BoHV-1 reduced during the period of study coincident with systematic IBR inactivated marker vaccination of the herd. Sixty-four abortion cases were investigated for the identification of causative agent(s) by microbial culture, serological (ELISA), and molecular detection (PCR/ real-time PCR). Antibodies to BVDV, Brucella, BoHV-1, Leptospira, Neospora, and Coxiella were detected in 63, 61, 56, 35, 5, and 6 aborting cattle, respectively. Real-time PCR/PCR of clinical specimens detected DNA of Brucella, BoHV-1, Coxiella, Leptospira, and Listeria in 34, 13, 12, 9, and 4 abortion cases, respectively. BVDV and Neospora were not detected in any specimen samples. Brucella abortus isolated from the farm was determined as ST1 by multi-locus sequence typing (MLST). DNA of multiple agents were detected in 21 of the 64 cases (43.75%). Overall, the data suggests, Brucella was the major causative agent, although multiple causative agents circulated in the farm.