Association of farmers' knowledge, attitude and practices with bovine brucellosis seroprevalence in Myanmar.

OBJECTIVE: This study aimed to identify the relationship between bovine brucellosis prevalence, farmers' knowledge, attitude, practice (KAP), and social factors on migratory draft cattle and smallholder dairy farms in the central dry zone of Myanmar. METHODS: This cross-sectional study was conducted on 54 migratory and 38 dairy cattle farms between August 2020 and February 2021. A structured questionnaire was used to identify farmers' behaviors. Bulk milk was sampled and tested using indirect enzyme-linked immunosorbent assay (I-ELISA). STATA 17 was used for all the analyses. RESULTS: Migratory cattle farms had a higher farm level brucellosis prevalence (14.8%) than dairy farms (2.6%; χ2 = 3.75; df = 1; p = 0.05). Only 2.8% of the farmers had knowledge about brucellosis, while 39.1% and 41.6% had attitudes and farm practices with respect to brucellosis, respectively in the study area. Socio-economic attribute of training in animal husbandry (p<0.01), raising system (p<0.01), practice of separating the aborted cow (p<0.01) were negatively associated to brucellosis. The overall farm level brucellosis prevalence was strongly associated with cattle herd size (p = 0.01), free movement grazing practices (p<0.01), practice of self-removal of placental debris without using personal protective equipment (p<0.01) and farmers' attitudes towards eating cow placenta debris (p<0.01). CONCLUSION: Farmers had little knowledge of brucellosis. Attitudes and practices differed significantly between migratory and dairy farmers. Training and extension programs are necessary to make farmers aware of their KAP situation since livestock migration and the custom of eating cow placental debris contribute to the spread of brucellosis. Persistent efforts are required to reduce the adverse effects of brucellosis. Therefore, the study suggests that a feasible control intervention and public awareness campaigns need to be conducted regarding methods of preventing human exposure to brucellosis.

A novel Bayesian Latent Class Model (BLCM) evaluates multiple continuous and binary tests: A case study for Brucella abortus in dairy cattle.

Bovine brucellosis, primarily caused by Brucella abortus, severely affects both animal health and human well-being. Accurate diagnosis is crucial for designing informed control and prevention measures. Lacking a gold standard test makes it challenging to determine optimal cut-off values and evaluate the diagnostic performance of tests. In this study, we developed a novel Bayesian Latent Class Model that integrates both binary and continuous testing outcomes, incorporating additional fixed (parity) and random (farm) effects, to calibrate optimal cut-off values by maximizing Youden Index. We tested 651 serum samples collected from six dairy farms in two regions of Henan Province, China with four serological tests: Rose Bengal Test, Serum Agglutination Test, Fluorescence Polarization Assay, and Competitive Enzyme-Linked Immunosorbent Assay. Our analysis revealed that the optimal cut-off values for FPA and C-ELISA were 94.2 mP and 0.403 PI, respectively. Sensitivity estimates for the four tests ranged from 69.7% to 89.9%, while specificity estimates varied between 97.1% and 99.6%. The true prevalences in the two study regions in Henan province were 4.7% and 30.3%. Parity-specific odds ratios for positive serological status ranged from 1.2 to 2.2 for different parity groups compared to primiparous cows. This approach provides a robust framework for validating diagnostic tests for both continuous and discrete tests in the absence of a gold standard test. Our findings can enhance our ability to design targeted disease detection strategies and implement effective control measures for brucellosis in Chinese dairy farms.

Turning chaotic sample group clusterization into organized ones by feature selection: Application on photodiagnosis of Brucella abortus serological test.

Bovine brucellosis diagnosis is a major problem to be solved; the disease has a tremendous economic impact with significant losses in meat and dairy products, besides the fact that it can be transmitted to humans. The sanitary measures instituted in Brazil are based on disease control through diagnosis, animal sacrifice, and vaccination. Although the currently available diagnostic tests show suitable quality parameters, they are time-consuming, and the incidence of false-positive and/or false-negative results is still observed, hindering effective disease control. The development of a low-cost, fast, and accurate brucellosis diagnosis test remains a need for proper sanitary measures at a large-scale analysis. In this context, spectroscopy techniques associated with machine learning tools have shown great potential for use in diagnostic tests. In this study, bovine blood serum was investigated by UV-vis spectroscopy and machine learning algorithms to build a prediction model for Brucella abortus diagnosis. Here we first pre-treated the UV raw data by using Standard Normal Deviate method to remove baseline deviation, then apply principal component analysis - a clustering method - to observe the group formation tendency; the first results showed no clustering tendency with a messy sample score distribution, then we properly select the main principal components to improve clusterization. Finally, by using machine learning algorithms (SVM and KNN), the predicting models achieved a 92.5% overall accuracy. The present methodology provides a test result in an average time of 5 min, while the standard diagnosis, with the screening and confirmatory tests, can take up to 48 h. The present result demonstrates the method's viability for diagnosing bovine brucellosis, which can significantly contribute to disease control programs in Brazil and other countries.

Retrospective benefit-cost analysis of bovine brucellosis control in the state of Mato Grosso, Brazil.

From 2001, due to the launching of the National Program for the Control and Eradication of Brucellosis and Tuberculosis, Brazil began a series of studies aimed at the epidemiological characterization of bovine brucellosis in its federative units. In the state of Mato Grosso, studies conducted in 2003 and 2014 showed a reduction in the prevalence of infected herds from 41.2 % to 24 % and in the prevalence of seropositive animals from 10.2 % to 5.1 %. These results stemmed from the efficient heifer vaccination program with S19 implemented by the state. From this scenario, the present study quantified and compared the benefits and costs of bovine brucellosis control in the state through a financial analysis. When considering private and public costs, the Net Present Value (NPV) was estimated at US$ 37.5 million, the Internal Rate of Return (IRR) was calculated at 31 %, and the Benefit-Cost Ratio (BCR) was 2.3. When considering only the private costs, the NPV was US$ 51.3 million, the IRR was 46 % and the BCR was 3.43, indicating that the cattle producer in Mato Grosso had a return of US$ 3.43 for each US$ 1.00 invested. Therefore, the bovine brucellosis control actions carried out in the state of Mato Grosso between 2003 and 2014 were highly advantageous from the economic point of view. The most rational way to continue decreasing bovine brucellosis prevalences at low cost in the state is to maintain the heifer vaccination program, using the RB51 vaccine in addition to S19.

Shedding rate of Brucella spp. in the milk of seropositive and seronegative dairy cattle.

Brucellosis in cattle herds has caused severe economic losses in many regions worldwide. A cross-sectional study was performed to investigate the presence of Brucella spp. in industrial dairy cattle farms in Iran. For this purpose, 935 blood and 935 milk samples were randomly collected from industrial dairy cattle farms in Iran's Alborz and Tehran provinces. Blood and milk samples were collected on the same day from each cow. Serological, bacteriological, and molecular characterization of Brucella isolates were performed using standard methods. Our results revealed the seroprevalence of brucellosis in dairy cattle farms in the Alborz and Tehran provinces, reaching 19.8%, 6.7%, 5.1%, 14.1%, and 13.1% using the Rose Bengal plate test (RBPT), serum agglutination test (SAT), 2-mercaptoethanol test (2-ME), indirect enzyme-linked immunosorbent assay (i-ELISA) and milk ring test (MRT), respectively. Furthermore, the results of bacterial culture and PCR analyses showed the presence of Brucella abortus among dairy cattle in the Alborz province and Brucella melitensis and B. abortus among dairy cattle in the Tehran province. Moreover, statistical analysis with Cohen's Kappa has highlighted the near-perfect agreement between RBPT and i-ELISA (k = 0.86). In contrast, substantial agreement was shown between RBPT and SAT performance (k = 0.70) and moderate agreement between RBPT and 2-ME (k = 0.67). The findings of this investigation showed shedding of Brucella in the milk of seropositive cows, which is a serious problem involving the maintenance and further spread of Brucella infection on the farm. Therefore, for brucellosis detection or eradication in dairy cattle farms, bacteriological and serological tests of milk samples should be performed along with blood analysis to inhibit the uncontrolled spread of the disease in animals and humans.

Economic analysis of bovine brucellosis control in the Rondônia state, Brazil.

Bovine brucellosis, mainly caused by Brucella abortus, is a worldwide distribution anthropozoonosis that causes great economic losses. In 2001, Brazil launched the National Program for the Control and Eradication of Brucellosis and Tuberculosis (PNCEBT). Contemporaneously, a great effort to characterize the epidemiology of the disease in Brazilian states was started. In the state of Rondônia, a first epidemiological study was carried out in 2004, revealing a prevalence of 35.2% of infected herds and 6.22% of seropositive females. In 2014, after a successful heifer vaccination program with strain 19 (S19), a second study detected a reduction in the prevalence of infected herds to 12.3% and of seropositive females to 1.9%. The present study aimed to quantify and compare the costs and benefits related to the control of bovine brucellosis in the state through an accounting analysis. Vaccinating heifers and performing serological tests to move animals were computed as private costs. The expenditures of the state official veterinary service for brucellosis control were considered public cost. The considered benefits of lowering prevalence were decreased cow replacement, decreased abortions, decreased perinatal and cow mortality, and increased milk production. Considering private and public costs, the net present value (NPV) was estimated at US$ 18.3 million, the internal rate of return (IRR) was calculated at 23%, and the benefit-cost ratio (BCR) was 1.7. When considering only the private costs, the NPV was US$34.9 million, the IRR was 49%, and the BCR was 3.0, meaning that the bovine producer had a return of 3 for each unit of currency invested. The results showed that the bovine brucellosis control measures implemented in the state of Rondônia, which had as its main strategy the vaccination of heifers with S19, produced highly advantageous economic results. The state should continue with its vaccination program, stimulating the use of the RB51 vaccine in addition to S19, to achieve further reductions in prevalence at low cost.

Seroprevalence, associated risk factors, and molecular detection of bovine brucellosis in rural areas of Egypt.

The study was carried out on six villages in northern Egypt to evaluate the epidemiological situation of bovine brucellosis among 989 unvaccinated household cattle by Rose Bengal Plate Test (RBPT) and indirect ELISA (iELISA) and to investigate the existence of Brucella DNA using real-time PCR in 100 milk and 100 sera from seropositive cattle and 50 sera from seronegative cattle. The overall seroprevalence was 20.7% and 23.7% by RBPT and iELISA, respectively. Based on the iELISA results, the seroprevalence was significantly (P < 0.05) higher in the village II (34.7%) and cattle > 7 years (30.1%). More males than females were non-significant seropositive (P = 0.6). There was 95% agreement between RBPT and iELISA, although iELISA showed a higher positivity rate (23.7%, 95% CI: 0.21-0.26) than RBPT (20.7%, 95% CI: 0.18-0.24). DNA of Brucella was confirmed in 16 milk samples by IS711 qPCR from seropositive cattle, however, no Brucella DNA was detected in serum samples tested positive and negative. Brucella abortus was the only species detected based on the alkB gene. Prevalence is highly related to the sampling site and the age of the animals. In conclusion, although qPCR is more accurate and commonly used in the diagnosis of most infectious diseases but in this situation iELISA is preferred and recommended for continuous screening and animal movement restriction and vaccination protocols, especially in high-risk areas.

Seroprevalence of Bovine Brucellosis in Selected Sites of Central Highland of Ethiopia.

Background: Brucellosis is a contagious, economically significant bacterial disease that affects animals worldwide and is one of the most neglected zoonotic diseases in the world. The disease poses a barrier to the trade of animals and animal products, represents a public health hazard, and is an impediment to free animal movement. Methods: A cross-sectional study was carried out from December 2019 to May 2020 in order to determine seroprevalence and identify potential risk factors for brucellosis in dairy cows in the Central Highlands of Ethiopia with recent cases of abortion. Purposive sampling was carried out on the farms and kebeles in question to screen for recent cases of abortion in dairy cows. For the purpose of performing serological testing, 352 blood samples from dairy cattle were obtained. The Rose Bengal Plate test was used to initially screen the serum samples, and the Complement Fixation test was utilized as a confirmatory test. Results: Using combined RBPT and CFT tests, the overall seroprevalence of bovine brucellosis was 0.6% (95% CI: 0.16-2.09). Retained fetal membrane (OR = 32.74, p = 0.006), market-based stock replacement (OR = 16.55, p = 0.002), breeding method (OR = 7.58, p = 0.027), and late stage of abortion (OR = 14.74, p = 0.0002) are all significantly associated risk factors. Conclusion: The present seroprevalence study revealed that brucellosis is prevalent at a lower rate among dairy cattle in the study areas. However, there is a possible risk of brucellosis transmission in dairy cattle and the exposed human population in research locations because no control measures were put in place there. Implementing a test and slaughter method with compensation for farmers is advised due to the low prevalence of bovine brucellosis in government-owned and small-holder farms.

Rapid, ultrasensitive, and highly specific identification of Brucella abortus utilizing multiple cross displacement amplification combined with a gold nanoparticles-based lateral flow biosensor.

Brucella abortus (B. abortus) as an important infectious agent of bovine brucellosis cannot be ignored, especially in countries/regions dominated by animal husbandry. Thus, the development of an ultrasensitive and highly specific identification technique is an ideal strategy to control the transmission of bovine brucellosis. In this report, a novel detection protocol, which utilizes multiple cross displacement amplification (MCDA) combined with a gold nanoparticles-based lateral flow biosensor (AuNPs-LFB) targeting the BruAb2_0168 gene was successfully devised and established for the identification of B. abortus (termed B. abortus-MCDA-LFB). Ten specific primers containing engineered C1-FAM (carboxyfluorescein) and D1-biotin primers were designed according to the MCDA reaction mechanism. These genomic DNA extracted from various bacterial strains and whole blood samples were used to optimize and evaluate the B. abortus-MCDA-LFB assay. As a result, the optimal reaction conditions for the B. abortus-MCDA-LFB assay were 66°C for 40 min. The limit of detection of the B. abortus-MCDA-LFB was 10 fg/µl (~3 copies/µl) for genomic DNA extracted from pure cultures of B. abortus isolate. Meanwhile, the B. abortus-MCDA-LFB assay accurately identified all tested B. abortus strains, and there was no cross-reaction with non-B. abortus pathogens. Moreover, the detection workflow of the B. abortus-MCDA-LFB assay for whole blood samples can be completed within 70 min, and the cost of a single test is approximately 5.0 USD. Taken together, the B. abortus-MCDA-LFB assay is a visual, fast, ultrasensitive, low-cost, easy-to-operate, and highly specific detection method, which can be used as a rapid identification tool for B. abortus infections.

Erratum to "Evolution of animal and human brucellosis in Algeria: a mini narrative review: Anerni n tawla n Malta n yiɣersiwen d yimdanen di Lezzayer: tasɣunt tanalasant tilemzit" [New Microbes and New Infections March 46C, (2022) 100975].

[This corrects the article DOI: 10.1016/j.nmni.2022.100975.].

Accuracy of molecular diagnostic methods for the detection of bovine brucellosis: A systematic review and meta-analysis.

Background and Aim: Bovine brucellosis is a disease of global socio-economic importance caused by Brucella abortus. Diagnosis is mainly based on bacterial culture and serology. However, these methods often lack sensitivity and specificity. A range of molecular diagnostic methods has been developed to address these challenges. Therefore, this study aims to investigate the diagnostic accuracy of molecular tools, in comparison to gold standard bacterial isolation and serological assays for the diagnosis of bovine brucellosis. Materials and Methods: The systematic review and meta-analysis were conducted based on analyses of peer-reviewed journal articles published between January 1, 1990, and June 6, 2020, in the PubMed, Science Direct, Scopus, and Springer Link databases. Data were extracted from studies reporting the use of molecular diagnostic methods for the detection of B. abortus infections in animals according to Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines. The quality of included journal articles was assessed using the quality assessment of diagnostic-accuracy studies assessment tool and meta-analysis was carried out using Review Manager. Results: From a total of 177 studies, only 26 articles met the inclusion criteria based on PRISMA guidelines. Data from 35 complete studies were included in the meta-analysis and used to construct 2 × 2 contingency tables. Improved diagnostic performance was observed when tissue (sensitivity 92.7% [95% confidence interval (CI) 82.0-98.0%]) and serum samples (sensitivity 91.3% [95% CI 86.0-95.0%]) were used, while the BruAb2_0168 locus was the gene of preference for optimal assay performance (sensitivity 92.3% [95% CI 87.0-96.0%] and specificity 99.3% [95% CI 98.0-100.0%]). Loop-mediated isothermal amplification (LAMP) had a higher diagnostic accuracy than polymerase chain reaction (PCR) and real-time quantitative PCR with sensitivity of 92.0% (95% CI 78.0-98.0%) and specificity of 100.0% (95% CI 97.0-100.0%). Conclusion: The findings of this study assign superior diagnostic performance in the detection of B. abortus to LAMP. However, due to limitations associated with decreased specificity and a limited number of published articles on LAMP, the alternative use of PCR-based assays including those reported in literature is recommended while the use of LAMP for the detection of bovine brucellosis gains traction and should be evaluated more comprehensively in future.

A Seroprevalence Study of Brucellosis in Boran (Zebu) Breeds of Pastoral Area.

Purpose: The economic and health implications of brucellosis are of particular concern in developing countries, primarily in the vulnerable sector of rural herders. A cross-sectional study was done in Boran breeds to estimate the seroprevalence of bovine brucellosis, identify risk variables and assess public health implications in Borena zone, Oromia region, Ethiopia in the questionnaire survey. Methods: The sampling animals were chosen from smallholders using a simple random sampling procedure. The study involved a total of 788 animals. Animals of both sex and different age groups with the age of 6 months or above found during the study interval were included. The Rose Bengal plate test was used to screen sera, and positive samples were subsequently retested using a Direct Enzyme-Linked Immunosorbent Assay for confirmation. Results: The overall seroprevalence of bovine brucellosis was 7.6% (60/788) in the Direct Enzyme-Linked Immunosorbent Assay test. Herd size, age, history of abortion, testicular hygroma, and retained fetal membrane were statistical significance for the Brucella seropositivity (P<0.05). Whereas, district, sex, body condition score, and management did not influence the disease occurrence (P>0.05). The majority of the participants, 91.7% (55/60) did not aware of the zoonotic implications of brucellosis. Only 10% (6/60) of interviewed respondents disposed of aborted fetuses and retained fetal membrane properly and the rest 90% (54/60) left in the environment. Ninety percent and 83.3% of the respondents revealed that they consumed raw milk and meat, respectively. Conclusion: The presence of Brucella infection is highly correlated with age, history of abortion, and testicular hygroma. According to the collected data: sex, body condition score, district, and management had no statistically significant effect on Brucella occurrence. The majority of respondents were unaware of the disease's zoonotic consequences. Finally, creating community awareness about its transmission, zoonotic significance, and hygienic practices were recommended.

Development of a novel glycoprotein-based immunochromatographic test for the rapid serodiagnosis of bovine brucellosis.

AIMS: Bovine brucellosis is a worldwide zoonotic disease that causes important economic losses and public health concerns. Because control of the disease depends on vaccination, serodiagnosis and isolation of the infected animals, affordable, rapid and accurate point of care (POC) tests are needed. METHODS AND RESULTS: We developed and evaluated a novel glycoprotein-based immunochromatographic test for the detection of IgG antibodies against the O-polysaccharide of Brucella in bovine serum samples. Brucella GlycoStrip combines the power of immunochromatographic and bacterial glycoengineering technologies for the diagnosis of bovine brucellosis. The analysis of positive and negative reference samples indicated that the test has a diagnostic sensitivity and specificity of 96.9% (95% CI: 92.7%-100.0%) and 100%, respectively. CONCLUSIONS: Due to the recombinant glycoprotein-based antigen OAg-AcrA, which consists of the O-side chain of Brucella smooth lipopolysaccharide (sLPS) covalently linked to the carrier protein AcrA, the test is highly accurate, allows the differentiation of infected animals from those vaccinated with a rough strain or with a single dose of a smooth strain and fulfil the minimum diagnostic requirements established by the national and international regulations. SIGNIFICANCE AND IMPACT OF STUDY: This strip test could provide a rapid (10 min) and accurate diagnosis of bovine brucellosis in the field contributing to the control of the disease.

Efficacy of Brucella abortus S19 and RB51 vaccine strains: A systematic review and meta-analysis.

This systematic review and meta-analysis aimed to recalculate the efficacy of Brucella abortus S19 and RB51 vaccine strains and discuss the main variables associated with controlled trials to evaluate bovine brucellosis vaccine efficacy (VE). The most commonly used vaccine strain was S19, at a dose of 1010 colony forming units (CFU), followed by RB51 at 1010 CFU. The most commonly used challenge strain was B. abortus 2308, at a dose of 107 CFU, by the intraconjunctival route. Regarding the meta-analysis, trials were grouped according to the vaccine strain and dose to recalculate protection against abortion (four groups) or infection (five groups) using pooled risk ratio (RR) and VE. Regarding protection against abortion (n = 15 trials), the S19 vaccine at 109 CFU exhibited the highest protection rate (RR = 0.25, 95% confidence interval (CI) : 0.12-0.52; VE = 75.09%, 95% CI: 48.08-88.05), followed by RB51 at 1010 CFU (RR = 0.31, 95% CI: 0.16-0.61; VE = 69.25%, 95% CI: 39.48-84.38). Regarding protection against infection (n = 23 trials), only two subgroups exhibited significant protection: S19 at 109 CFU (RR = 0.28, 95% CI: 0.14-0.55; VE = 72.03%, 95% CI: 57.70- 81.50) and RB51 at 1010 CFU (RR = 0.43, 95% CI: 0.22-0.84; VE = 57.05%, 95% CI: 30.90-73.30). In conclusion, our results suggest that a dose of 109 CFU for S19 and 1010 CFU for RB51 are the most suitable for the prevention of abortion and infection caused by B. abortus.

Seropositividad de Brucella abortus en ganado bovino de Aguazul- Casanare / Seropositivity of Brucella abortus in cattle from Aguazul- Casanare

Resumen Introducción. La Brucelosis bovina, es una enfermedad bacteriana e infecto contagiosa, causada por Brucella abortus. Se transmite a través de la ingestión de forrajes y aguas contaminadas con descargas vaginales infectadas, que conlleva a una patología del sistema reproductor en bovinos, que impactan la sanidad pecuaria y la economía de la agroindustria. Objetivo. Evaluar el comportamiento de la brucelosis bovina en el municipio de Aguazul, Casanare (Colombia) y los factores asociados al desarrollo de esta enfermedad. Metodología. Estudio descritptivo de 26.187 muestras de suero sanguíneo de ganado bovino evaluadas, que corresponden a 260 predios del municipio de Aguazul. Se empleó técnicas serológicascomo rosa de bengala, fluorescencia polarizada y ELISA competitiva para la evaluación de positividad a Brucella abortus y se evaluaron las pérdidas económicas asociadas a positividad en los ensayos de laboratorio. Resultados y conclusiones. La positividad a brucelosis bovina correspondió al 1%, que corresponde a hembras menores de 24 meses de edad y entre 37 a 48 meses y, machos entre los 57 a 68 meses de edad. Se sugiere consolidar esfuerzos en investigación para evaluar los factores que contribuyen a la seropositividad en el ganado y el riesgo para la propagación y mantenimiento de la enfermedad.

Abstract Introduction. Bovine brucellosis is a contagious bacterial and infectious disease caused by Brucella abortus. It is transmitted through the ingestion of forage and water contaminatedwith infected vaginal discharges, which leads to a pathology of the reproductive system inbovines, which impacts livestock health and the economy of agribusiness. Aim. To evaluatethe behavior of bovine brucellosis in the municipality of Aguazul, Casanare (Colombia) and the factors associated with the development of this disease. Methodology. Descriptive study of 26,187 bovine blood serum samples evaluated, corresponding to 260 farms in the municipality of Aguazul. Serological techniques such as rose bengal, polarized fluorescence, and competitive ELISA were used to evaluate positivity to Brucella abortus and the economic losses associated with positivity in laboratory tests were evaluated. Results and conclusions. The positivity to bovine brucellosis corresponded to 1%, which corresponds to females under 24 months of age and between 37 to 48 months and males between 57 to 68 months of age. It is suggested to consolidate research efforts to evaluate the factors that contribute to seropositivity in cattle and the risk for the spread and maintenance of the disease

Seroprevalence of Bovine Brucellosis and Associated Risk Factors in Western Ethiopia.

BACKGROUND: Brucellosis is a contagious bacterial disease, which affects domestic animals, humans, and wildlife in Ethiopia and other countries. METHODS: A cross-sectional study design using a simple random sampling technique was conducted in Diga, Guto, Gida, and Sibu Sire districts of East Wollega Zone, Western Oromia, from November 2019 to July 2020 to determine the prevalence of bovine brucellosis and identify associated risk factors. A lottery method was used during serum collection and risk factors identified during interview. RESULTS: A total of 1152 serum samples were collected from cattle of 6 months age and above. Serum samples were tested by enzyme linked immunosorbent assay (I-ELISA). An overall seroprevalence of brucellosis was 1.82% (21/1152). Statically significant variation (P<0.05) in seroprevalence of bovine brucellosis was recorded among potential risk factors such as breed [P≤0.05; OR:8.905; CI:1.568-50.573], parity [P≤0.05; OR:0.017; CI:0.042-5.195], retained fetal membrane [P≤0.001; OR:0.018; CI:0.00-2.169], and abortion history [P≤0.001; OR:0.030; CI:0.004-0.212]. Conversely, seroprevalence variations in sex, district, villages, body condition score, and age were statistically insignificant (P>0.05). CONCLUSION: The current study revealed bovine brucellosis is posing a threat to livestock with no strict control and prevention scheme in place. Therefore, public awareness creation about the zoonotic importance of the disease is crucial and the Government should establish legislation, support, control, and prevention of the disease recommended.

Effect of co-positivity for brucellosis and tuberculosis on milk yield and fertility of Holstein cows.

This study aimed to determine whether cows detected as tuberculosis (bTB) reactors and seropositive to brucellosis (bBR), as well as co-positive to bBR and bTB (bBR-bTB) and with a complete lactation before slaughter, were associated with reduced milk production and fertility. A total of 8068 productive and reproductive records of high-yielding Holstein cows from a single large dairy herd with a high prevalence of bTB and bBR were collected from 2012 to 2015. Lactation derived either from calving (n = 6019) or hormonally induced lactation (n = 2049), and all cows received growth hormone throughout lactation. For cows not induced into lactation, pregnancy rate to first service for healthy cows (C; 26.6%) was higher (P < 0.01) than bBR (15.2%), bTB (15.8%), and bBR-bTB (1.3%) cows. For induced cows, pregnancy rate to first service did not differ significantly among C, bBR, and bTB (14.5-17.3%) cows, but the percentage success of first service was extremely low (1.3%; P < 0.01) in bBR-bTB cows. Services per pregnancy (only pregnant cows) were lowest for C (3.3 ± 2.9; P < 0.01) and highest (6.4 ± 3.4) for bBR-bTB non-induced cows. This variable was lowest for C (2.9 ± 2.5; P < 0.01) and highest for bBR-bTB non-induced cows (6.3 ± 3.1). Pregnancy rate to all services did not differed for C (79.5%), bBR (76.7%), and bTB (75.9%) but was lower (58.9%; P < 0.01) for bBR-bTB non-induced cows. For induced cows this variable was highest for bBR (53.3%) and lowest for bBR-bTB (34.1%; P < 0.01) non-induced cows. 305-d milk production was increased by 4%, and total milk yield by 7% in TB-positive cows compared to that of the negative cows non-induced hormonally into lactation. This study showed the negative impact of the co-positivity for bTB and bBR on the reproductive efficiency of Holstein cows, although positive bTB and bBR tests enhanced milk yield.

Epidemiology of bovine brucellosis in Hisar, India: identification of risk factors and assessment of knowledge, attitudes, and practices among livestock owners.

Brucellosis caused by facultative intracellular bacteria, Brucella, remains a global threat to both animal and human health. In this study we aimed to identify potential risk factors of bovine brucellosis and to assess the knowledge, attitudes, and practices (KAPs) of livestock keepers in Hisar, India. A standardized questionnaire was used to collate information regarding potential risk factors of bovine brucellosis and livestock owners' KAPs. A total of 127 livestock keepers were involved. Serum samples from their animals (n = 635) were tested for the presence of antibodies against Brucella by Rose Bengal Plate Test (RBPT) and indirect enzyme-linked immunosorbent assay (iELISA). Out of these, 78 (61.4%) of the herds had at least one seropositive animal, and 302 (47.6%) of the cattle were seropositive. Univariate and multivariate analysis revealed significant associations between intensive farm type (OR = 4.6; 95% CI, 1.6-16.7; P = 0.009), hygienic disposal of aborted fetuses (OR = 0.3; 95% CI, 0.08-0.9; P = 0.04) and herd seropositivity for brucellosis. The majority, 96 (75.6%) of the respondents, were males aged 18-50, and 82 (64.6%) owned a small-backyard farm. Only 51 (40.2%) of the participants knew about brucellosis; out of them, 54.9% (28/51) could not identify clinical signs of brucellosis. Six (11.8%) participants indicated abortion as the most noticeable clinical sign, and 45.1% indicated that consumption of raw milk is associated with high risk of contracting brucellosis. A large proportion of respondents confirmed that milk from their animals was regularly consumed (86.6%) and sold (59.8%) to other people. These results suggest that bovine brucellosis is endemic in Haryana, where Brucella-contaminated milk is likely being regularly sold. Brucellosis control efforts in Haryana should include education programs to raise awareness of the disease and means to control it in cattle and to prevent zoonotic transmission.

Documenting the absence of bovine brucellosis in dairy cattle herds in the southern region of Malawi and the associated knowledge, attitudes and practices of farmers.

There is paucity of Brucella prevalence data in Malawi. For this reason, a cross-sectional study was conducted, from 06 January 2020 to 27 February 2020, to estimate the seroprevalence of brucellosis in dairy cattle herds amongst smallholder farmers, government and private dairy farms in the southern region. A total of 529 serum samples were screened for anti-Brucella antibodies using the Rose Bengal test (RBT) and a competitive enzyme-linked immunosorbent assay (cELISA). A pre-tested electronic (Epicollect tool, Wellcome Sanger Institute, United Kingdom) questionnaire was administered to 378 smallholder farmers to assess their knowledge, attitudes and practices towards brucellosis. Descriptive statistics were used to analyse the data in Microsoft Excel® and Statistical Package for Social Sciences (SPSS®) version 21. No animal tested positive for presence of anti-Brucella antibodies, indicating 0% prevalence (individual and herd levels). The majority (94.2%; 95% confidence interval [CI]: 91.8-96.5) of smallholder farmers had never heard about brucellosis. Furthermore, assisting during parturition without protective equipment (41.3%; 95% CI: 36.3-46.2) and using bulls for breeding (75%; 95% CI: 70.2-78.9) were amongst the common risk practices that were identified. We could not detect brucellosis in this study that indicates the disease could be very rare or even absent in the dairy cattle herds of the southern region of Malawi. However, further Brucella studies need to be conducted in cattle, small livestock, wildlife and humans to document the true status of brucellosis in the country. Brucellosis surveillance, monitoring, awareness and preventive measures are required to maintain this favourable situation.

Development of BruAb2_0168 based isothermal polymerase spiral reaction assay for specific detection of Brucella abortus in clinical samples.

Bovine brucellosis, predominantly caused by Brucella abortus is one of the most neglected zoonotic diseases causing severe economic losses in the dairy industry. The early and precise diagnosis of the disease is required to reduce the transmission of infection in humans as well as animals. In the current study, a rapid and novel isothermal amplification-based polymerase spiral reaction (PSR) was developed for the specific detection of Brucella abortus by targeting the BruAb2_0168 gene. The assay could be conducted at 65 °C in a water bath and results can be obtained after 60 min. The detection limit of the PSR assay was found to be 1.33 fg. The sensitivity of the assay was found to be 104 fold higher than conventional PCR and equivalent to real-time PCR (RT-PCR). The assay didn't exhibit cross-reaction with selected pathogenic non-Brucella bacteria and Brucella spp. other than B. abortus. Forty clinical samples were also tested using this novel assay and it was able to detect 25 samples as positive, however, conventional PCR could detect the targeted organism in 22 samples only. To the extent of our knowledge, this is the first report towards the development of a PSR assay for specific detection of B. abortus. The assay can be used as a quick, sensitive and accurate test for the diagnosis of bovine brucellosis in the field setting. Relatively one of the paradigm-shifting aspects of this assay would be it does not require any expensive equipment and the results can be easily visualized by the unaided eye, therefore making PSR a valuable diagnostic tool in field conditions.