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Background Brucellosis is listed as a priority disease in low-income countries like Guinea, facing challenges in logistics, equipment, competence, and cost limitations for diagnosis. Serological diagnosis is mainly performed by the Rose Bengal agglutination test (RBT) in the veterinary sector. We have compared its discriminative capacity with more sophisticated and expensive serological tests, such as multi-species or species-specific ELISA kits and Complement Fixation test (CFT). Methodology/principal findings A panel of 554 serum samples of pigs, goats, sheep, and cattle collected throughout Guinea from 2017 to 2019 where tested by RTB and ELISA tests in parallel at the Institut Pasteur de Guinée (Conakry) and the Brucellosis WOAH/EU Reference Laboratory of the French Agency for Food, Environmental and Occupational Health & Safety (Maisons-Alfort, France). ELISAs performed equally across laboratories (Kappa =0.867–0.958); RBT and ELISA showed 94–95% concordance. The CFT value of positive cattle samples also logically followed the RBT scores Conclusions/significance In low-income countries like Guinea, the less expensive RBT can be regarded as a convenient routine Brucella diagnosis tool, assuming a solid experience of the operator following standard operating protocols and regular proficiency tests. As WOAH recommends confirmatory methods, the multispecies ELISA kit appears as a good candidate for conveniently trained and equipped laboratories.

Brucellosis, a contagious zoonotic bacterial infection affecting livestock and wildlife, is primarily caused by Brucella abortus, globally. However, in South Africa, the true prevalence of bovine brucellosis remains unknown because of a lack of epidemiological data. Therefore, this study used diagnostic data to evaluate and determine the seropositivity of bovine brucellosis based on Rose Bengal test (RBT) screening and confirmation with complement fixation test (CFT) in Limpopo and Free State provinces between 2013 and 2022. The use and limitations of this data were also evaluated based on the bovine brucellosis scheme in South Africa. The study revealed the overall seropositivity of 4.2% (n = 8980/212 440) for bovine brucellosis based on RBT and CFT in series. In Limpopo province, the brucellosis seropositivity was slightly higher at 4.3% (n = 7488/173 011) compared to 3.8% (n = 1492/39 429) in Free State province. Analysis of brucellosis distribution over the study period indicated significant variation (p < 0.001) both between and within the provinces. Notably, the highest prevalence in Limpopo occurred during 2013–2017, whereas in Free State, peak prevalence was observed in 2013 and 2016. Challenges preventing an accurate reflection of the brucellosis seropositivity in these provinces for the period include a lack of data on vaccination history and herd status of the samples submitted, as well as the inability to match the CFT results from different laboratories, because some laboratories are only accredited to perform the RBT.ContributionInsights gained from retrospective studies such as this study can play crucial roles in shaping effective control and preventative measures against bovine brucellosis. Given the challenges in obtaining confirmatory test results, we suggest that brucellosis tests be conducted at a single central laboratory or that the government provides a central database where all laboratories can enter their data. Furthermore, information submitted to the laboratories must make herd and vaccine history compulsory for sample submission to ensure more accurate data.

Brucellosis is one of the most important zoonotic diseases in Greece, causing a significant burden on both human and animal vitality as well as economic loss. The present study was conducted from 2015 to 2022 on 711,415 serum samples by determining the seroepidemiology of Brucellosis among livestock in 24 geographical areas in Greece using the Rose Bengal Test (RBT) and the complement fixation test (CFT) and further performing genetic analysis of Brucella spp. by species-specific real-time PCR and MLVA Brucella analysis. A total of 3086 serum samples from goats, sheep, and cattle showed positive results using the RBT and CFT, and only strongly positive samples (n = 800) were preserved in the Βlood Bank of the Veterinary Laboratory of Brucellosis. From these, 212 sera samples were randomly selected for molecular and genetic analysis. The results indicated that the incidence rate of Brucellosis is higher in cattle herds in comparison with other animal species. Overall, 48 samples tested positive by real-time PCR, of which forty-seven of them were B. abortus and one was B. melitensis. Genetic analysis of two B. abortus samples revealed a common pattern, indicating two Bruce04, two Bruce18, four Bruce07, two Bruce09, three Bruce16, and four Bruce30 for both samples, which, interestingly, were not identical with the known genotypes in the public MLVA Brucella database. Our findings substantiate that animal Brucellosis remains a health issue in Greece, with a stable but apparent incidence rate, and further investigation is needed to fully characterize the newly identified Brucella strains in Greece.

The involvement of Brucella infection in causing abortion was investigated in a breeding female subpopulation of 283 cattle, 756 camels, and 757 goats. Serum samples were serially tested using the Rose Bengal test and complement fixation test. The study showed that anti-Brucella antibodies were prevalent in 10.6% (95% confidence interval (CI), 7.4, 14.9), 2.2% (95%CI, 1.4, 3.7), and 1.9% (95%CI, 1.1, 3.2) of cattle, camel, and goats, respectively. Abortion was more commonly reported in camels (23.4%) than cattle (13.8%) and goats (12.4%). The results of this study suggested that Brucella infections contribute significantly to abortion in cattle (odds ratio (OR), = 4.7; 95%CI, 2.0, 10.8) and goats (OR = 6.9; 95%CI, 2.2, 21.7) but not in camels. The number of young animals produced by breeding females seems to be apparently reduced in seropositive groups. Keeping more than two animal species at household level was found to be the risk factor for cattle (OR = 3.1; 95%CI, 1.2, 7.9) and camel (OR = 5.3; 95%CI, 1.2-23.5) seropositivity to Brucella infection when compared to those animals from households that keep only two animal species. This may suggest a possibility of cross species transmission of Brucella infection under such mixed herding. Wet season (OR = 4.8; 95%CI, 1.3, 18.1) was found to be associated with seropositivity in goats, linked to a coincidence of increased deliveries in flocks with possible excretion of Brucella organisms. The study results suggest that Brucella infection is the likely cause of abortion in cattle and goats while other causes largely outweigh brucellosis as a cause of abortion in camels in Borana, hence, contributing to reproductive loss.

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.

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.

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.

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. A retrospective study on brucellosis in South Africa.

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.

The sensitivity and specificity of three commercially available complement fixation test (CFT) antigens from c.c.pro (c.c.pro), Central Veterinary Institute of Wageningen UR (CIDC) and the United States Department of Agriculture (USDA) were comparatively evaluated by testing 410 sera collected from glanders-endemic and non-endemic areas (200 true-negative randomly collected sera and 210 sera collected from experimentally immunised animals (12 rabbits, 19 horses), clinically positive (135) and culture-positive (44) horses, donkeys and mules). Immunoblotting (IB) was used as the gold standard test. Highest sensitivity was shown for the CIDC antigen (100 per cent) followed by the c.c.pro antigen (99.39 per cent). However, the USDA antigen showed substantially less (p<0.05) sensitivity (62.19 per cent). Highest specificity was found for the USDA antigen (100 per cent) followed by the CIDC (97.5 per cent) and c.c.pro antigen (96.5 per cent). Positive and negative predictive values (assumed glanders prevalence of <0.1 per cent) for each antigen were calculated to be 95.88 and 99.48 (c.c.pro), 97.04 and 100 (CIDC), 100 and 76.33 per cent (USDA), respectively. Almost perfect agreement (0.96) was found between CFT using either c.c.pro or CIDC and IB.