Development and validation of a chemiluminescent western blot assay for glanders (Burkholderia mallei) serodetection.

Glanders, caused by Burkholderia mallei, is a zoonotic disease of equids. Serologic testing for glanders is required by disease-free countries before international movement of equids. The World Organisation for Animal Health Terrestrial Manual recommends the complement fixation test (CFT) for clearance of individual animals for movement, but the CFT is prone to false-positive results. A colorimetric western blot (WB) assay was developed and validated to resolve false-positive CFT results; however, that assay is relatively time-consuming, and the interpretation is subjective. We present here a procedurally similar chemiluminescent WB assay that performs comparably to the validated colorimetric WB assay and offers noticeable benefits of decreased time-to-result and greater ease of interpretation.

Comparison of coccidioidal complement fixation and quantitative immunodiffusion serology at a reference laboratory.

The incidence of coccidioidomycosis continues to increase. The diagnosis frequently relies on non-invasive diagnostic testing with immunodiffusion and complement fixation (CF) testing the current gold standard. A direct comparison of quantitative immunodiffusion and CF for IgG antibodies has not been previously reported. In a comparison of 368 samples, there was close concordance observed (360/368 = 97.8%) (P-value < .001). These tests can be considerably interchangeable in the reference laboratory setting.

There are several diagnostic methodologies available in coccidioidomycosis. Direct comparisons of these methods are limited. Prior studies have not compared quantitative immunodiffusion to complement fixation testing. Our results show these tests are highly concordant.

Comparative evaluation of RBPT, I-ELISA, and CFT for the diagnosis of brucellosis and PCR detection of Brucella species from Ethiopian sheep, goats, and cattle sera.

BACKGROUND: Brucellosis is an economically devastating animal disease and has public health concern. Serological methods such as Rose Bengal Plate Test (RBPT), Complement Fixation Test (CFT), and Indirect-Enzyme-Linked Immunosorbent Assay (I-ELISA) have been used to detect brucellosis. However, there is limited comparative evaluation studies and lack of molecular confirmation of the causative agents in the study areas. The study was aimed to compare RBPT, I-ELISA, CFT, and confirmation using Polymerase Chain Reaction (PCR). A total of 2317 sera samples were collected from brucellosis-affected areas of Ethiopia with no vaccination history. All sera were subjected to comparative serological assays. Post-cross tabulation, sensitivity, and specificity were determined using Receiver Operating Characteristics (ROC) curve analysis software. PCR was performed on 54 seropositive samples using genus- and species-specific primers. RESULTS: Among the 2317 sera tested for comparative serological assays, 189 (8.16%) were positive for RBPT, 191 (8.24%) for I-ELISA, and 48 (2.07%) for CFT. Sensitivity to RBPT was 100% (95%) in shoats and 74% (95%) in cattle. Specificity on RBPT was 98.69% (95%), 99.28% (95%), 100% (95%) in sheep, goats, and cattle, respectively. CFT sensitivity was 4 (95%) in sheep, 9.65 (95%) goats, and 72 (95%) cattle. Specificity on CFT was 100% (95%) for sheep, goats, and cattle. A 223bp Brucella genus-specific and 156bp B. abortus species-specific detected. However, B. melitensis not detected. CONCLUSION: In this study, I-ELISA was the most sensitive and specific test. RBPT detected all Brucellosis-infected sheep and goats; nevertheless, it showed false positive in sheep and goats and false negative in cattle. The presence of B. abortus in small and large ruminants was confirmed by PCR. This is the first report of B. abortus detection in small ruminant in Ethiopia. B.abortus detected in non-preferred hosts. The findings suggest further study on molecular epidemiology of Brucella species.

Designing of Western Blot Technique for Glanders Diagnosing in Iran.

Burkholderia mallei is the etiologic agent of glanders. It is difficult to diagnose this zoonotic disease in its early stages. Some methods such as the complement fixation test (CFT) cause some problems for veterinary authorities and financial losses to animal owners due to false-positive results. The mallein test requires appropriate laboratory equipment and skilled personnel. To quickly and accurately diagnose the disease, especially in areas where animals cannot be kept, new methods (such as the Western blot test [WBT]) should be used to identify the disease. This study designed and optimized the Western blot (immunoblot) test using sera from 84 glanderous equids, and the sensitivity and specificity of ELISA and CFT were compared with the WBT. ELISA tests are based on B. mallei antigens whereas a purified lipopolysaccharide-containing B. mallei antigen is used in the WBT. The sensitivity and specificity of the tests were estimated using the cut-off values recommended by the test developers. The WBT and ELISA were significantly more specific than the CFT. The ELISA based on B. mallei antigens was significantly less sensitive than the CFT. Given their comparable sensitivities and specificities, the CFT (95.7%, 98.5%), the WBT (95%, 100%) and the ELISA (85%, 100%) should be further developed. The CFT is still the prescribed technique for serological investigation of equids for trade purposes to certify individual animals without glanders. Therefore, more efforts should be made to further improve and optimize the WBT and ELISA tests.

A retrospective study (2007-2015) on brucellosis seropositivity in livestock in South Africa.

In South Africa, brucellosis testing and record-keeping are done by several laboratories, thus it is difficult to access any organized data to assess the status of the disease. This study evaluated the seropositivity for brucellosis using Rose Bengal test and complement fixation test in suspect cattle, sheep, goats and pigs sera submitted to Bacterial Serology Laboratory, Agricultural Research Council-Onderstepoort Veterinary Research (ARC-OVR) from nine provinces in the country during the period 2007-2015. This retrospective data analysis was conducted to estimate the occurrence of brucellosis in the country from the submitted samples, identify variables that affected seropositivity for brucellosis, investigate existing gaps in data recording and make recommendations on important variables to facilitate better data capture and inferences on brucellosis. Nine years of data were collated and analysed to detect association (seropositivity over time regarding animal species and location). Of the 764,276 animals tested, the distribution of samples was 90.50% (691,539/764,276), 5.19% (39,672/764,276), 3.92% (29,967/764,276) and 0.41% (3,098/764,276) for cattle, sheep, goats and pigs, respectively. The seropositivity for brucellosis by animal species was 6.31% (43,666/691,539, 95% CI: 6.26-6.37), 2.09% (828/39,672, 95% CI: 1.95-2.23), 0.63% (189/29,967, 95% CI: 0.55-0.73) and 0.13% (4/3,098, 95% CI: 0.05-0.33) in cattle, sheep, goats and pigs respectively. The data available did not capture information on the age, sex, breed and other host risk factors that would have been related to seropositivity for brucellosis. The data provide an understanding of the disease occurrence and confirm that brucellosis is enzootic in South Africa. Improved and standardized data collection can be used to pro-actively drive, monitor, change or formulate policies to mitigate the challenges brought about by brucellosis in the livestock sector in South Africa.

Serological screening for Coxiella burnetii in the context of early pregnancy loss in dairy cows.

Q fever is one of the commonest infectious diseases worldwide. A Coxiella burnetii prevalence of 97.6% has been found by ELISA and PCR tests of the bulk tank milk in dairy cattle farms of Hungary. The herd- and individual-level seroprevalence rates of C. burnetii in the examined dairy cows and farms have dramatically increased over the past ten years. Three high-producing industrial dairy farms were studied which had previously been found ELISA and PCR positive for C. burnetii by bulk tank milk testing. Coxiella burnetii was detected in 52% of the 321 cows tested by ELISA. Pregnancy loss was detected in 18% of the cows between days 29-35 and days 60-70 of gestation. The study found a higher seropositivity rate (80.5%) in the cows that had lost their pregnancy and a seropositivity of 94.4% in the first-bred cows that had lost their pregnancy at an early stage. The ELISA-positive pregnant and aborted cows were further investigated by the complement fixation test (CFT). In dairy herds an average of 66.6% individual seropositivity was detected by the CFT (Phase II) in previously ELISA-positive animals that had lost their pregnancy and 64.5% in the pregnant animals. A higher (Phase I) seropositivity rate (50.0%) was found in the cows with pregnancy loss than in the pregnant animals (38.5%). The high prevalence of C. burnetii in dairy farms is a major risk factor related to pregnancy loss.

Evaluation of the sensitivity and specificity of the lateral flow assay, Rose Bengal test and the complement fixation test for the diagnosis of brucellosis in cattle using Bayesian latent class analysis.

This study was conducted to evaluate the sensitivity (Se) and specificity (Sp) of the Rose Bengal test (RBT), complement fixation test (CFT), the serum lateral flow assay (LFAserum) and the blood lateral flow assay (LFAblood) for the detection of antibodies to Brucella spp. using Bayesian latent class models (BLCMs). Sera and whole blood were collected from naturally infected cattle reared in smallholder, small-scale commercial and large-scale commercial farms in Zimbabwe (n = 1022) and Botswana (n = 770). The BLCMs were fitted under the assumption that conditional dependences existed between the tests. Based on the conditional dependence model, the RBT had the highest Se of 0.897 (95 % Probability Intervals: 0.854; 0.932) compared to 0.827 (0.773; 0.872), 0.812 (0.76; 0.858) and 0.809 (0.785; 0.832) for the LFAserum, LFAblood and CFT, respectively. The CFT recorded a higher Sp of 0.999 (0.995; 1.000) than the LFAserum 0.996 (0.99; 1.000), the LFAblood 0.984 (0.976; 0.991) and the RBT 0.969 (0.959; 0.978). The data indicated that both the Se and Sp of RBT and CFT and the Sp of LFAserum and LFAblood were conditionally independent, while the Se appeared to be conditionally dependent. These results indicated that none of the evaluated tests had perfect Se and Sp and consequently could not be used alone for the diagnosis of brucellosis in cattle from the studied farming sectors. Thus, based on high Se and Sp, respectively, a brucellosis testing regimen using the RBT (screening) and the LFA (confirmatory) may be considered.

Molecular detection of 7SL-derived small RNA is a promising alternative for trypanosomosis diagnosis.

Equine trypanosomosis comprises different parasitic diseases caused by protozoa of the subgenus Trypanozoon: Trypanosoma equiperdum (causative agent of dourine), Trypanosoma brucei (nagana) and Trypanosoma evansi (surra). Due to the absence of a vaccine and the lack of efficacy of the few available drugs, these diseases represent a major health and economic problem for international equine trade. Development of affordable, sensitive and specific diagnostic tests is therefore crucial to ensure the control of these diseases. Recently, it has been shown that a small RNA derived from the 7SL gene (7SL-sRNA) is produced in high concentrations in sera of cattle infected with Trypanosoma congolense, Trypanosoma vivax and Trypanosoma brucei. Our objective was to determine whether 7SL-sRNA could serve as a marker of active infection in equids experimentally infected with Trypanosoma equiperdum by analysing the sensitivity, specificity and stability of the 7SL-sRNA. Using a two-step RT-qPCR, we were able to detect the presence of 7SL-sRNA between 2 and 7 days post-infection, whereas seroconversion was detected by complement fixation test between 5 and 14 days post-infection. There was a rapid loss of 7SL-sRNA signal from the blood of infected animals one day post-trypanocide treatment. The 7SL-sRNA RT-qPCR allowed an early detection of a treatment failure revealed by glucocorticoid-induced immunosuppression. In addition, the 7SL-sRNA remains detectable in positive sera after 7 days of storage at either 4°C, room temperature or 30°C, suggesting that there is no need to refrigerate serum samples before analysis. Our findings demonstrate continual detection of 7SL-sRNA over an extended period of experimental infection, with signals detected more than six weeks after inoculation. The detection of a strong and consistent 7SL-sRNA signal even during subpatent parasitemia and the early detection of treatment failure highlight the very promising nature of this new diagnostic method.

Factors associated with seroprevalence of bovine anaplasmosis in Mississippi, USA.

Anecdotally, Veterinary Feed Directive prescriptions in the southeastern United States (U.S.) are written most often for treatment and prevention of bovine anaplasmosis (BA) but seroprevalence estimates and factors associated with this disease are currently unavailable in Mississippi (MS). Bovine anaplasmosis, a tick-borne disease of cattle caused by Anaplasma marginale, remains an economically important disease in U.S. The lack of recent seroprevalence of BA throughout the U.S. makes accurate assessment of production losses incurred by the cattle industry in the U.S. difficult, if not impossible to estimate. This study was aimed at determining the seroprevalence of and factors associated with BA in MS. Data were obtained from an active survey of 207 beef cows slaughtered between May 2013 and December, 2014 as well as from reviewing 5182 Veterinary Diagnostic Laboratories (VDLs) records of specimens from MS submitted for BA testing between 2002 and 2018. From the active surveillance, the overall observed apparent seroprevalence of BA in MS with cELISA was 28.99% (95% CI: 23.23 - 35.50%) while the estimated true seroprevalence was 29.02% (22.74 - 36.07%). However, from the laboratory records, the overall apparent period seroprevalence of BA in MS between 2002 and 2018 irrespective of diagnostic assay used was 16.72% (15.73 - 17.76%) and yearly increase in the diagnosis of BA followed a significant trend (P < 0.0001). With cELISA, the apparent seroprevalence of BA was 22.11% (20.78 - 23.49%) and the estimated true seroprevalence was 21.62% (20.18 - 23.11%). However, with CFT, the apparent seroprevalence of BA was 13.50% (10.75 - 16.81%) and the estimated true seroprevalence was 47.90% (36.30 - 61.87%). Factors associated with positive BA results were age, cattle type, and quarter of the year the specimens were submitted. The odds of the outcome were 22 as high in adults, 27 times as high in beef cattle, and 2 times as high between October to December in comparisons to juveniles, dairy cattle, and between April to June, respectively. Cattle population in the counties was not associated with positive BA results. Current records from the VDLs appear to accurately estimate the seroprevalence of BA in MS and thus serves as a reliable surveillance tool BA in the state. Because the burden of BA appears to be distributed throughout the state, future prevention and control measures for BA should focus on the identified putative risk factors and be intensified throughout MS.

Seroprevalence of Brucella suis in eastern Latvian wild boars (Sus scrofa).

Brucellosis due to Brucella suis biovar 2 is one of the most important endemic diseases in wild boar (Sus scrofa) populations in Europe. The aim of the present study was to determine the seroprevalence of brucellosis in wild boars in the eastern part of Latvia. Wild boars killed by hunters in the period from January to April 2015 (n = 877) and from March to April in 2016 (n = 167) were examined for antibodies against B. suis by the Rose Bengal test (RBT), a complement fixation test (CFT), and by enzyme-linked immunosorbent assays. In 2015, 199 samples (22.7%) were positive by RBT and/or CFT while 36 samples (21.6%) were seropositive in 2016. Of the Brucella seropositive samples from 2015 and 2016 (n = 235), 162 (68.9%) were also seropositive to Yersinia enterocolitica. Considering cross-reactivity of serological tests, the seroprevalence of B. suis biovar 2 exposure in wild boars in the eastern part of Latvia was calculated to 14.0% in 2015 and 9.6% in 2016. From selected seropositive samples (42 in 2015 and 36 in 2016) total DNA was extracted and analyzed with an IS711-based nested polymerase chain reaction (PCR) assay. Species and biovar identification was conducted for bacteria isolated in monoculture from PCR positive samples by species specific primers and Bruce-ladder multiplex PCR. Brucella suis biovar 2 was isolated from 12/20 samples in 2015 and 9/9 samples in 2016. The average seroprevalence was relatively low compared to that found in certain other European countries. Males and females had an equal level of seropositivity, but a positive age-trend was observed for both males and females.

Simultaneous detection of antibodies to five Actinobacillus pleuropneumoniae serovars using bead-based multiplex analysis.

We developed and made a preliminary validation of a bead-based multiplexed immunoassay for simultaneous detection of porcine serum antibodies to Actinobacillus pleuropneumoniae serovars 1, 2, 6, 7, and 12. Magnetic fluorescent beads were coupled with A. pleuropneumoniae antigens and tested with a panel of serum samples from experimentally infected pigs and with serum samples from uninfected and naturally infected pigs. The multiplex assay was compared to in-house ELISAs and complement fixation (CF) tests, which have been used for decades as tools for herd classification in the Danish Specific Pathogen Free system. Assay specificities and sensitivities as well as the corresponding cutoff values were determined using receiver operating characteristic (ROC) curve analysis, and the A. pleuropneumoniae multiplex assay showed good correlation with the in-house ELISAs and CF tests with areas under ROC curves ≥ 0.988. Benefits of multiplexed assays compared to ELISAs and CF tests include reduced serum sample volumes needed for analysis, less labor, and shorter assay time.

An evaluation of serological tests in the diagnosis of bovine brucellosis in naturally infected cattle in KwaZulu-Natal province in South Africa.

The diagnostic sensitivity (DSe) of the Rose Bengal test (RBT), the complement fixation test (CFT), the serum agglutination test (SAT), the competitive enzyme-linked immunosorbent assay (cELISA) and the indirect ELISA (iELISA) were determined in naturally infected cattle in KwaZulu-Natal province of South Africa with known infectious status from culture (gold standard). Natural brucellosis infection status of animals was determined by culturing and identification of Brucella abortus biovar 1 from abomasal fluid, milk, hygroma fluid, lymph nodes or uterine discharges samples. The diagnostic specificity (DSp) of the tests mentioned above was determined using samples from known negative herds. There was no statistically significant difference between the tests in their ability to diagnose brucellosis. The RBT and iELISA had the highest DSe of 95.8%, whereas RBT and CFT had the highest DSp of 100%. In South African laboratories, the RBT and CFT serological tests are used, because of the cost efficacy of CFT when compared to the less labour intensive but more expensive iELISA.

Evaluation of three competitive ELISAs and a fluorescence polarisation assay for the diagnosis of bovine brucellosis.

Bovine brucellosis is an infectious disease of worldwide public health and economic importance. The usual tests for the diagnosis of this disease include the Rose-Bengal test (RBT), complement fixation test (CFT), serum agglutination test (SAT) and indirect ELISA. New tests such as competitive ELISAs (C-ELISA) and fluorescence polarisation assay (FPA) have been developed. However, C-ELISA may correspond to different protocols and a wide variation may exist in their diagnostic performance. The aim of this study was to evaluate three commercially available C-ELISA kits (C-ELISA1-3) and FPA for the diagnosis of bovine brucellosis and compare test performance with RBT, CFT, indirect ELISA and FPA. Sera submitted to EU laboratories in 2011 from 5111 adult cattle were tested. Individual test sensitivities (Se) and specificities (Sp) were estimated. Threshold assessment using the receiver operating characteristic method was also performed. The most sensitive tests were FPA (99.0%; 95% confidence interval [CI], 97.9-100%), C-ELISA1 (98.4%; 95% CI, 97.0-99.8%) and RBT (97.7%; 95% CI, 95.9-99.3%). The most specific tests were CFT (99.98%; 95% CI, 99.93-100%), SAT (99.98%; 95% CI, 99.93-100%) and RBT (99.89%; 95% CI, 99.79-99.99%). Among the new tests, none of the three C-ELISA kits studied could be recommended as a single screening test because of their low specificity, especially when used in a herd. C-ELISA3 could not be recommended as confirmatory test on individual animals to determine whether false positive serological test results had occurred.

Detection of Yersinia enterocolitica serotype O:9 in the faeces of cattle with false positive reactions in serological tests for brucellosis in Ireland.

Intestinal infection by Yersinia enterocolitica serotype O:9 (YeO9) in cattle has been linked to false positive serological reactivity (FPSR) in diagnostic tests for brucellosis. Although eradicated in Ireland, brucellosis monitoring still identifies seropositive animals, usually one or two (termed singletons) per herd, which are classed as FPSR. To investigate a link between FPSR and YeO9, faeces and blood were collected from singleton FPSR cattle, and from companion animals, in eight selected herds with more than one FPSR animal, for YeO9 culture and Brucella serology. YeO9 was isolated from 76/474 (16%) FPSR singletons in 309 herds, but not from any of 621 animals in 122 control non-FPSR herds. In the FPSR herds 52/187 (27.8%) animals were culture positive, and 17% of the isolates were from seronegative animals. Seropositive animals were more likely to have a rising antibody titre when culture positive.

Interlaboratory ring trial to evaluate CFT proficiency of European laboratories for diagnosis of glanders in equids.

To evaluate the routine complement fixation test (CFT) used to detect Burkholderia mallei antibodies in equine sera, an interlaboratory proficiency test was held with 24 European laboratories, including 22 National Reference Laboratories for glanders. The panels sent to participants were composed of sera with or without B mallei antibodies. This study confirmed the reliability of CFT and highlighted its intralaboratory reproducibility. However, the sensitivity of glanders serodiagnosis and laboratory proficiency may be improved by standardising critical reagents, including antigens, and by developing a standard B mallei serum.

Seroprevalence and "Knowledge, Attitudes and Practices" (KAPs) survey of endemic ovine brucellosis in Egypt.

BACKGROUND: Between February and July 2014, a cross-sectional study to estimate the seroprevalence of brucellosis in sheep in the Kafrelsheikh district of Egypt was carried out, together with a survey of knowledge, attitudes and practices (KAPs) among local shepherds. A total of 273 serum samples were collected from 28 sheep flocks in 10 villages within the study area. These samples were analysed by the Rose Bengal Plate test (RBPT) test, with all positive samples being confirmed by complement fixation test (CFT). RESULTS: True seroprevalence was 20 % (95 % CI 15.3-24.7 %) with the prevalence of villages with at least one seropositive sheep estimated at 95.5 % (95 % CI 92.2-100 %); village flock seroprevalence ranged from 0 to 46.8 %. Results of the KAPs survey demonstrated that despite good knowledge regarding brucellosis being potentially present within their flocks, shepherds lacked knowledge regarding routes of livestock to humans disease transmission and the symptoms of brucellosis in humans. This lack of knowledge regarding disease transmission resulted in high-risk practices being widespread-practices such as assisting parturition without protective measures, throwing aborted material into water canals and a reluctance to remove animals that had aborted from the flock. CONCLUSIONS: This study proposes potential measures to reduce seroprevalence of brucellosis in sheep and reduce public health risks from brucellosis such as culling aborted livestock and educational campaigns among shepherds regarding disease risks and modes of transmission.

Seroprevalence and risk factors for brucellosis in cattle in selected districts of Jimma zone, Ethiopia.

A cross-sectional study was carried out in Jimma town and Chora Botor district of Jimma zone from February 2014 to May 2014 to determine seroprevalence and risk factors of brucellosis in cattle. A total of 348 blood samples (174 each from zebu and crossbreed) were collected. The sera were separated and screened by Rose Bengal plate test (RBPT), and positive sera were retested by complement fixation test (CFT) for confirmation. The overall seroprevalence of bovine brucellosis was 1.4 and 0.3 % as tested by RBPT and CFT, respectively. The seroprevalence of bovine brucellosis in indigenous and crossbreed cattle was 1.1 and 0.6 % and 1.7 and 0 % using RBPT and CFT, respectively. Retained fetal membrane was the only risk factor found to be significantly associated with seropositivity of brucellosis in this study (p = 0.019). The overall seroprevalence of brucellosis was very low. However, due to the zoonotic and economic importance of the disease, prevention and control measures are required to stop further spread of the disease. To effectively implement this, the One Health (OH) is the most constructive approach we recommend.

Retrospective analysis of the prevalence of Brucella antibodies in sheep in the Karas Region of Namibia.

In this retrospective study, complete Brucella serology data from the annual national brucellosis testing program and disease investigation for the years 2008-2010 was collated and analyzed to estimate the prevalence of brucellosis in sheep in the Karas Region of Namibia. A total of 22,994 serological results from 762 flocks screened using the Rose Bengal test (RBT) and confirmed using the complement fixation test (CFT) were analyzed. An overall prevalence of 0.14% was recorded over the 3 years. Yearly prevalence was 0.19% (2008), 0.05% (2009), and 0.18% (2010). At district level, brucellosis prevalence was estimated to be between 0 and 0.49%. On positive farms (n = 32), prevalence was between 2.25 and 30%. True prevalence was zero at district level and in all the three study years. We concluded that the prevalence of Brucella antibodies in sheep was low taking into account that some farmers may have vaccinated against the disease. The low prevalence confirms the effectiveness of existing brucellosis control measures implemented by the official veterinary services.

Brucellosis in nomadic pastoralists and their goats in two provinces of the eastern Algerian high plateaus.

A 31-months study was conducted to elucidate the prevalence of brucellosis in nomadic pastoralists and their goats in two provinces of the eastern Algerian high plateaus. Five hundred eight human and 4955 animal sera were screened with the Rose Bengal plate test and the complement fixation test for confirmation. Uterine fluids from aborting goats were subjected to microbiological analyses to determine the biovars responsible for abortions. The overall seroprevalence was 0.98% among animals and 15.84% among herds. A significant correlation was recorded between occurrence of brucellosis and herd size (r = 0.4046, P < 0.0001) as well as age (χ(2) = 5.809, P = 0.0159) and sex of animals (χ(2) = 20.09, P < 0.0001); 89.65% of human cases were related to positive herds and the infection rate was higher in men (7.6%) than in women (6%) and children (0.92%). Brucella melitensis biovar 3 was the only aetiology of brucellosis-associated abortion in goats of the studied region.