The seroprevalence and risk factors of bovine brucellosis were studied at animal and herd level using a combination of culture, serological and molecular methods. The study was conducted in 253 randomly selected cattle herds of the Potohar plateau, Pakistan from which a total of 2709 serum (1462 cattle and 1247 buffaloes) and 2330 milk (1168 cattle and 1162 buffaloes) samples were collected. Data on risk factors associated with seroprevalence of brucellosis were collected through interviews using questionnaires. Univariable and multivariable random effects logistic regression models were used for identifying important risk factors at animal and herd levels. One hundred and seventy (6.3%) samples and 47 (18.6%) herds were seropositive for brucellosis by Rose Bengal Plate test. Variations in seroprevalence were observed across the different sampling sites. At animal level, sex, species and stock replacement were found to be potential risk factors for brucellosis. At herd level, herd size (≥9 animals) and insemination method used were important risk factors. The presence of Brucella DNA was confirmed with a real-time polymerase chain reaction assay (qRT-PCR) in 52.4% out of 170 serological positive samples. In total, 156 (6.7%) milk samples were positive by milk ring test. B. abortus biovar 1 was cultured from 5 positive milk samples. This study shows that the seroprevalence of bovine brucellosis is high in some regions in Pakistan. Prevalence was associated with herd size, abortion history, insemination methods used, age, sex and stock replacement methods. The infected animal may act as source of infection for other animals and for humans. The development of control strategies for bovine brucellosis through implementation of continuous surveillance and education programs in Pakistan is warranted.

Brucella species occasionally cause spontaneous human abortion. Brucella can be transmitted commonly through the ingestion of raw milk or milk products. The objective of this study was to determine the sero-prevalence of and to identify potential risk factors for brucellosis in pregnant women from Rawalpindi, Pakistan. We conducted a cross-sectional study at the Gynecology Outdoor Patient department of the Benazir Bhutto Hospital, Rawalpindi, Pakistan from March to June 2013. Data related to potential risk factors and clinical history was collected by individual interviews on the blood sampling day. The 429 serum samples collected were initially screened by Rose Bengal Plate Agglutination test for the detection of Brucella antibodies. We applied standard descriptive statistics and logistic regression analyses. Twenty five (5.8 %; 95 % confidence interval (CI): 3.8 % -8.5 %) serum samples were found to be seropositive. Brucellosis-related clinical symptoms were recorded in various seropositive cases. Animal contact, raw milk consumption, having an abortion history and the experience of an intrauterine fetal death were associated with seropositivity for brucellosis in univariate analyses (all p <0.05). In multiple logistic regression models only the contact with animals remained as independent and robust risk factor (odds ratio 5.21; 95 % CI: 1.88-13.75; p = 0.001) for seropositivity. Brucellosis is a serious threat for pregnant women and their unborn children in Pakistan. Pregnant women having brucellosis-related symptoms or previous history of abortions, miscarriages, intrauterine fetal death and other brucellosis-related manifestations should be screened for brucellosis – especially those exposed to animals given the increased risk – and medication should be administered according to state of the art.

Brucellosis is a zoonotic disease which affects humans, farm animals and wildlife as well. In Pakistan, the status of human and farm animal brucellosis is documented but the information about brucellosis in wildlife is lacking. To access the seroprevalence of Brucella antibodies in avian (turkeys, peafowl, guineafowl, mallard ducks and Indian blue rock pigeon) and selected wild species (reptiles and amphibians), a total of 117 serum samples (reptile = 34, amphibian = 04, avian n = 79) were collected from Karachi and Pattoki regions of Pakistan. Serum samples were screened for Brucella antibodies using Rose Bengal plate test (RBPT) and 11.1% serum samples were found positive. Overall, 2.5% birds, 29.4% reptiles and 25% amphibians were seropositive for Brucella antibodies. This is first report of detection of Brucella antibodies in reptiles and amphibians in Pakistan.

Molecular Identification of Bovine Brucellosis Causing Organisms at Selected Private Farms in Pothohar Plateau, Pakistan