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Introduction: Human brucellosis is one of the most common zoonosis infections, with an important impact on the health and economy worldwide. This study aimed to update and provide epidemiological information on this infection and evaluate Rose Bengal Test, which is used as an essential diagnostic test for brucellosis in Erbil. Methodology: A total of 325 participants seeking care and reporting fever at Rizgary Teaching Hospital were enrolled. Blood samples were tested for Brucella spp. antibodies using Rose Bengal Test and blood culture followed by species identification. A questionnaire was administered to detect the risk factors. Results: The prevalence of probable and confirmed brucellosis was 12.3% (95% CI 9.2–16.3) and 9.5% (95% CI 6.8–13.2) respectively. The majority of cases were in the age group of 18-39 years. Brucellosis was significantly associated with raw milk consumption (OR = 10.3 95% CI 5-22.4) and contact with livestock (OR = 11.5 95% CI 5.6-23.9). Brucella melitensis (58.1%) and Brucella abortus (41.9%) are the dominant species in the area. The sensitivity, specificity, positive predictive value, and negative predictive value of the Rose Bengal Test in comparison to the blood culture were 100%, 96.9%, 77.5 %, and 100% respectively. Conclusions: Brucellosis is a significant cause of fever in Erbil and could be diagnosed by the Rose Bengal Test taking into account the compatibility of clinical features with the positive result. The vaccination of livestock and boiling or pasteurization of milk are essential procedures to reduce the frequency of human brucellosis.

Brucellosis is an important bacterial zoonosis responsible for considerable economic losses in livestock and health-related burden worldwide. The objective of this study was to determine the seroprevalence of brucellosis in communal and smallholder cattle farming in four districts of the North West province of South Africa (Dr Ruth Segomotsi Mompati, Ngaka Modiri Molema, Bojanala platinum and Dr Kenneth Kaunda districts). Seven hundred and seventy blood samples from farmed animals (n = 378) and abattoir-slaughtered animals (n = 392) were collected. In addition, milk samples (n = 22) were collected from lactating farmed cows. Rose Bengal test (RBT), complement fixation test (CFT) and milk ring test (MRT) were used to detect antibodies against Brucella species. The RBT showed a seroprevalence of 2% at 95% confidence interval (CI: 1.35-3.35), CFT confirmed an overall seroprevalence of 1.95% (95% CI: 1.14-3.12) for all four districts sampled. Although the seroprevalence of brucellosis was found to be low, the possibility of undetected cases of the disease could not be ruled out. Overall, the findings of this study confirmed that brucellosis is endemic in the surveyed areas of the North West province of South Africa.Contribution: The outcome of this study will contribute to the National Brucellosis Project organised by the Department of Agriculture, Land Reform and Rural Development (2016-2026) to assist in the effective implementation of the disease control measures with a view to prevent further outbreaks in the country's cattle population.

Background Brucellosis in livestock is endemic in Mongolia, and efficient monitoring is required for clarifying its prevalence. Milk can be obtained noninvasively and is useful for monitoring brucellosis in livestock. However, the usefulness of milk in monitoring brucellosis should be clarified. Materials and methods Serum and milk samples were obtained from 326 cows from six farms near Ulaanbaatar City between 2020 and 2022. Serum and milk were assessed using the Rose Bengal Test (RBT) and Milk Ring Test (MRT), respectively. All milk samples were further subjected to bacterial isolation and DNA extraction. DNA samples were analyzed with quantitative PCR (qPCR) targeting Brucella genus-specific IS 711 insertion sequence to detect and estimate Brucella spp. levels in milk. qPCR-positive samples were further subjected to single nucleotide polymorphism (SNP) assay to discriminate B. abortus field strains from the S19 vaccine strain. Results Of the 326 milk samples, 108 (33.1%) were revealed to be positive for Brucella spp. by qPCR, whereas only 5 samples (1.5%) were deemed positive via the bacterial isolation. No S19 vaccine strain was identified in the IS 711 qPCR-positive milk samples by the SNP assay. Although qPCR detected Brucella spp. from milk, which was obtained from cows in three lactation stages, the detection ratio was significantly higher in the early lactation stage than in the middle lactation stage. Additionally, the five milk samples from which Brucella spp, were isolated exhibited the top 5 estimated colony forming units among the IS 711 qPCR-positive samples, indicating that detection sensitivity of the IS 711 qPCR is extremely higher than that of bacterial culture. There was a tendency that milk samples from RBT- and MRT-positive cows are more likely to be positive by IS 711 qPCR. Conclusion The results revealed that analysis of milk with qPCR is easy and sensitive monitoring method for detecting Brucella infection in livestock.

Introduction: Brucellosis is a zoonotic endemic disease in Lebanon. It is caused by direct transmission of Brucella from contaminated animal products to humans. If left untreated brucellosis might lead to several complications and a chronic disease state. The prompt diagnosis of brucellosis has the ability to limit the progression of the disease, especially if the correct treatment is administered for the adequate amount of time. The aim of this study is to determine the optimal diagnostic tool and to assess Brucella burden in Lebanon. Methodology: This retrospective study was performed by reviewing the medical charts of 46 brucellosis patients from three Lebanese hospitals. Brucellosis diagnostic tests were compared and sensitivity of each test was calculated, as well as, the level of agreement with other standard diagnostic tools. Data retrieved were analyzed for relevance and statistical significance using the statistical package for social sciences version 23. Results: Sensitivity results of the diagnostic tests were: Rose Bengal test (RBT) 94.7%, blood culture 65.6%, standard agglutination test (SAT) melitensis 95.1% and SAT abortus 97.6%. The level of agreement between RBT and SAT melitensis as well as abortus is 98% and 90.18%, respectively. While the level of agreement between Blood culture and SAT melitensis as well as abortus is 66.88% and 64.5%, respectively. Discussion: Culture techniques require further optimization in order to find the best diagnostic tool for brucellosis. Meanwhile, Blood Rose Bengal test held a significant potential for identifying Brucella infection in a highly sensitive, cost effective and time saving manner.

Brucellosis is caused by Gram-negative coccobacilli of the Brucella genus, with cattle mainly infected with Brucella abortus. The disease burden is a threat to socioeconomic development (agriculture/tourism) as well as to animal health, biodiversity and to human health due to the zoonotic nature of this pathogen. In South Africa (S.A), the prevalence of the disease in cattle and livestock in general is mostly unknown in communal farms. A cross-sectional study with a multistage sampling strategy was applied in communal areas from three district municipalities, i.e., Mopani, Capricorn and Sekhukhune of Limpopo province, South Africa. Sera (n = 1133) were collected and screened for antibodies against the Brucella species using the Rose Bengal Test (RBT) and confirmation of positive reactors with a Complement Fixation Test (CFT). The brucellosis seroprevalence was found to be 0.79% (95% CI: 0.38–1.45) by a CFT. Univariate analysis indicated that only the frequency of birth was significantly associated with CFT positivity (OR = 20; 95% Cl: 1.61–247.99; p = 0.039). The multivariable logistic regression model revealed that the frequency of birth, age, breed, gender, municipality and district were not statistically significant predictors at 0.05 level. However, some variables like cattle aged more than five years, had higher odds of CFT positivity compared to those younger than five years (OR = 5.66; 95% CI: 0.36–87.97), although the association was not statistically significant (p = 0.215). All positive reactors detected originated from the Mopani district municipality. Overall, the findings reveal a much lower seroprevalence of brucellosis in the communal farms of Limpopo province than previously assumed. We are of the opinion that the low seroprevalence is attributed to effective control strategies implemented by the Limpopo provincial veterinary services and hence provide important information to assist the regulatory bodies in the control and eradication of the disease.

Background Brucellosis is an emerging disease that causes a significant impact on productive and reproductive performance in dairy cattle. Though Brucella is a pivotal microorganism for dairy cattle, the scenario of brucellosis in Sylhet District is unknown. Objectives A cross‐sectional study was carried out to assess the prevalence and determinants associated with brucellosis in dairy cattle of Sylhet District. Methods A total of 386 sera and data on determinants from 63 dairy herds were collected from 12 sub‐districts using simple random sampling. The sera were tested with Rose Bengal Brucella antigen test, Brucella abortus plate agglutination test and serum agglutination test to find out the sero‐positivity. Results Overall, 17.09% (95% CI: 13.67–21.18) prevalence in cows were calculated. Relatively higher prevalence (56.08%; 95% CI: 42.23–70.32) was recorded in cows having parity ≥4 and were at higher risk (OR = 7.28) than the other cows with parity 0–3. Prevalence was significantly higher in cows with history of abortion 90.63% (95% CI: 75.79–96.76), repeat breeding 79.17% (95% CI: 65.74–88.27) and reproductive abnormalities 48.54% (95% CI: 39.12–58.07). Farm‐level prevalence was high in farms with the previous history of abortion 95.45% (95% CI: 78.20–99.19) and repeat breeding 90.00% (95% CI: 74.38–96.54). Conclusions The prevalence was high in Sylhet district, which might be a public health concern. Therefore, this study would represent the baseline information for guiding brucellosis control and prevention. Brucellosis is an emerging disease, which is highly prevalent in Sylhet district of Bangladesh. Prevalence was significantly higher in cows with history of abortion, repeat breeding and reproductive abnormalities, which might be a public health concern.

Brucellosis is one of the most important bacterial zoonotic diseases worldwide, characterized in domestic animals by long-term reproductive disorders. As known, wild boars (Sus scrofa) are natural hosts for Brucella suis biovar 2, in which the infection passes in inapparent form, increasing the pathogen transmission risk to domestic pigs, other domestic animals and humans. So far, no studies regarding brucellosis in wild boars in Serbia have been published. During the hunting season 2020/2021, 480 sera of wild boars living in Serbia were collected and tested for the presence of anti-Brucella antibodies. For the serological survey, the Rose Bengal Test (RBT) and competitive enzyme-linked immunosorbent assay (c-ELISA) were used. Of the 480 sera, 45 sera tested positive, indicating the acquired Brucella seroprevalence in wild boars of 9.4%. The greatest numbers of Brucella seropositive animals were detected in the eastern parts of the country and in one of the central districts, i.e., Pomoravski, Branicevski, Borski and Juznobanatski. This study provides the first data regarding brucellosis in the wild boar population in Serbia, revealing the seroprevalence of Brucella, thus indicating that wild boars as natural hosts and/or vectors of Brucella likely present a risk for the infection of other animals.

Rose Bengal antigen and smooth lipopolysaccharide (s-LPS) were produced from a field strain of Brucella ceti (“homologous” antigens) and from the reference strain B. abortus S99 (“heterologous” antigens); they are currently used for the diagnosis of brucellosis in cattle, water buffaloes, sheep, goats, and pigs, as recommended in the Manual of Diagnostic Tests and Vaccines for Terrestrial Animals of the World Organization for Animal Health (WOAH). “Homologous” and “heterologous” antigens were used in a rapid serum agglutination test (Rose Bengal test, RBT) and a competitive ELISA assay (c-ELISA) to test a panel of sera, blood, and other body fluids (cerebrospinal fluid, pericardial fluid, tracheal fluid, and aqueous humor) collected from 71 individuals belonging to five cetacean species (Stenella coeruleoalba; Tursiops truncatus; Grampus griseus; Globicephala melas; and Ziphius cavirostris), which were found stranded on the Italian coastline. Six animals were positive for Brucella spp. for bacterial isolation and/or PCR, and 55 animals were negative; for the remaining 10 animals, no PCR/isolation data were available. A total of 90 samples were tested. Results obtained from the two tests were compared in order to identify the most suitable antigen for the serological diagnosis of Brucella infection in cetaceans. The RBT performed with the “homologous” antigen showed the best results in comparison with the “heterologous” antigen: diagnostic sensitivity, specificity, and accuracy were 80.0%, 44.1%, and 46.9% for the “homologous” antigen and 80.0%, 17.0%, and 21.9% for the “heterologous” antigen. For the c-ELISA, “homologous” and “heterologous” s-LPS showed similar results (diagnostic sensitivity 66.7%, diagnostic specificity 97.3%, and diagnostic accuracy 95.0%). Therefore, the RBT using the “homologous” antigen and c-ELISA with “homologous” or “heterologous” s-LPS could be used in parallel for the detection of antibodies against Brucella spp. in cetaceans.

Abstract Brucellosis is known as one of the most common zoonotic diseases worldwide affecting both livestock and humans. It causes abortions, reduces milk production, and infertility in infected animals. The disease is routinely diagnosed through three serological techniques, such as rose bengal plate test (RBPT), standard agglutination test (SAT), and indirect enzyme-linked immunosorbent assay (I-ELISA). The aim of this study was to identify and compare the brucellosis seroprevalence among dairy cattle farms through these different serological tests. From 2112 sampled dairy cattle in different parts of Iran, RBPT, SAT, and I-ELISA led to 296 (14.02%), 215 (10.18%), and 297 (14.06%) positive results, respectively. Brucella abortus biovar 3 (62 cases) was identified as the most common cause of brucellosis in tested animals. Our results showed that the specificity and sensitivity of I-ELISA were higher than those obtained by RBPT and SAT. In this study, the overall agreement of RBPT and SAT with I-ELISA reached 95.21% and 94.12% in dairy cattle farms, respectively. Furthermore, Cohen’s kappa statistical analysis revealed that the best degree of agreement was seen between RBPT and I-ELISA (0.80), followed by RBPT and SAT (0.78) and finally SAT and I-ELISA (0.72), thereby indicating a strong agreement between RBPT and I-ELISA methods and good agreement between SAT and I-ELISA methods. The McNemar analysis also showed that a significant difference exists between positive and negative results determined by SAT and I-ELISA methods (p < 0.0001). However, the positive and negative results determined by I-ELISA and RBPT did not show a significant difference (p = 0.9207). Therefore, I-ELISA was a more specific and sensitive serological test when compared to RBPT and SAT and could remarkably decrease non-specific reaction by improving the serological screening specificity for an accurate brucellosis diagnosis in endemic areas.

Brucellosis is a zoonotic disease caused by Brucella spp., with the highest prevalence found in the northern cities of China. In this case report, we present an occurrence of spinal infection caused by B. melitensis in a 67-year-old man residing in a non-endemic area of southern China. The patient initially presented with chest and back pain, which was not accurately diagnosed and treated at a local hospital. Subsequently, due to worsening pain, he was admitted to our hospital. To determine the cause of the infection, we performed CT-guided aspiration biopsy and collected biopsy tissue for metagenomic next-generation sequencing (mNGS) on the second day of hospitalization. Imaging investigations revealed involvement of the thoracic vertebrae, specifically thoracic 4-7 with the main focus on 5-6, accompanied by stenosis of the intervertebral space. The mNGS results indicated that the spine infection was caused by B. melitensis. The patient's history as a shepherd and a positive Rose Bengal plate test (RBPT) further supported the diagnosis of brucella spondylitis. In order to alleviate pain and restore spinal function, the patient underwent posterior internal fixation of the thoracic spine. Treatment was initiated with cefoperazone/sulbactam, followed by doxycycline. Subsequently, the patient was switched to a combination therapy of rifampicin and doxycycline for a duration of six weeks. The patient responded well to treatment, and his condition remained stable. In conclusion, brucellosis is a common disease that can be easily misdiagnosed. This case report highlights the potential value of mNGS in early and rapid diagnosis. We believe that mNGS can serve as an effective tool to improve the diagnosis of spine infections caused by this pathogen. Keywords: spine infection, Brucella melitensis, metagenomic next-generation sequencing, Rose Bengal plate test