Scent detection of Brucella abortus by African giant pouched rats (Cricetomys ansorgei).

BACKGROUND: Brucellosis is a contagious zoonosis caused by bacteria of the genus Brucella. While the disease has been eradicated in most developed countries, it remains endemic in sub-Saharan Africa where access to reliable diagnostics is limited. African giant pouched rats (Cricetomys ansorgei) have been trained to detect the scent of Mycobacterium tuberculosis to increase case detection in sub-Saharan Africa. Given the similar diagnostic challenges facing brucellosis and tuberculosis, we explored the feasibility of training African giant pouched rats to detect Brucella. RESULTS: After 3 months of training, rats reliably identified cultured Brucella, achieving an average sensitivity of 93.56% (SD = 0.650) and specificity of 97.65% (SD = 0.016). Rats readily generalized to novel, younger Brucella cultures that presumably generated a weaker volatile signal and correctly identified at least one out of three fecal samples spiked with Brucella culture during a final test of feasibility. DISCUSSION: To our knowledge, these experiments are the first to demonstrate Brucella emits a unique odor profile that scent detection animals can be trained to identify. Importantly, cultured E. coli samples were included throughout training and test to ensure the rats learned to specifically identify Brucella bacteria rather than any bacteria in comparison to bacteria-free culture medium. E. coli controls therefore served a crucial function in determining to what extent Brucella abortus emits a unique odor signature. Further research is needed to determine if a Brucella-specific volatile signature is present within clinical samples. If confirmed, the present results suggest trained rats could serve as a valuable, novel method for the detection of Brucella infection.

Serological and molecular evidence of Brucella species in the rapidly growing pig sector in Kenya.

BACKGROUND: Brucellosis is an emerging yet neglected zoonosis that has been reported in Kenya. Epidemiological data on brucellosis in ruminants is readily accessible; however, reports on brucellosis in pigs remain limited. This study sought to detect Brucella infection in pig serum by both serological and molecular techniques. Serum from 700 pigs randomly collected at a centralized abattoir in Nairobi region, Kenya were screened in parallel, using both Rose Bengal Test (RBT) and competitive Enzyme-Linked Immuno-sorbent Assay (cELISA) for antibodies against Brucella spp. All sera positive by RBT and 16 randomly selected negative samples were further tested using conventional PCR targeting bcsp31 gene and real-time PCR (RT-PCR) assays targeting IS711 and bcsp31 genes. RESULTS: A prevalence of 0.57% (n = 4/700) was estimated using RBT; none of these samples was positive on cELISA. All RBT positive sera were also positive by both PCRs, while two sero-negative samples also tested positive on RT-PCR (n = 6/20). Brucella abortus was detected in four out of the six PCR positive samples through a real-time multiplex PCR. CONCLUSION: The detection of antibodies against Brucella spp. and DNA in serum from slaughterhouse pigs confirm the presence of Brucella in pigs. Therefore, investigation of the epidemiology and role of pigs in the transmission of brucellosis in Kenya is needed. Further targeted studies would be useful to systematically quantify and identify the spp. of Brucella in pigs.

Detection of serum neutralizing antibodies to Simbu sero-group viruses in cattle in Tanzania.

BACKGROUND: Orthobunyaviruses belonging to the Simbu sero-group occur worldwide, including the newly recognized Schmallenberg virus (SBV) in Europe. These viruses cause congenital malformations and reproductive losses in ruminants. Information on the presence of these viruses in Africa is scarce and the origin of SBV is unknown. The aim of this study was to investigate the presence of antibodies against SBV and closely related viruses in cattle in Tanzania, and their possible association with reproductive disorders. RESULTS: In a cross-sectional study, serum from 659 cattle from 202 herds collected in 2012/2013 were analyzed using a commercial kit for SBV ELISA, and 61 % were positive. Univariable logistic regression revealed significant association between ELISA seropositivity and reproductive disorders (OR = 1.9). Sera from the same area collected in 2008/2009, before the SBV epidemic in Europe, were also tested and 71 (54.6 %) of 130 were positive. To interpret the ELISA results, SBV virus neutralization test (VNT) was performed on 110 sera collected in 2012/2013, of which 51 % were positive. Of 71 sera from 2008/2009, 21 % were positive. To investigate potential cross reactivity with related viruses, 45 sera from 2012/2013 that were positive in SBV ELISA were analyzed in VNTs for Aino, Akabane, Douglas, Peaton, Sabo, SBV, Sathuperi, Shamonda, Simbu and Tinaroo viruses. All 45 sera were positive for one or more of these viruses. Twenty-nine sera (64.4 %) were positive for SBV, and one had the highest titer for this virus. CONCLUSIONS: This is the first indication that Aino, Akabane, Douglas, Peaton, Sabo, SBV, Sathuperi, Shamonda and Tinaroo viruses circulate and cause negative effect on reproductive performance in cattle in Tanzania. SBV or a closely related virus was present before the European epidemic. However, potential cross reactivity complicates the interpretation of serological studies in areas where several related viruses may circulate. Virus isolation and molecular characterization in cattle and/or vectors is recommended to further identify the viruses circulating in this region. However, isolation in cattle is difficult due to short viremic period of 2 to 6 days, and isolation in vectors does not necessarily reflect the situation in cattle.

Effect of awareness training to frontline health workers and the use of e-based technology on reporting of brucellosis cases in selected pastoral communities, Tanzania: a quasi-experimental study.

INTRODUCTION: Brucellosis is a serious community health problem and endemic disease in Tanzania in both humans and animals. Frontline health workers (FHWs) play a vital role in reporting and hence prevent and control brucellosis in rural settings. This study aims to evaluate the effect of awareness training to frontline health workers and use of electronic technology (e- technology) on reporting of brucellosis cases. METHODS: A quasi-experimental design was implemented in two pastoral communities in eastern part of Tanzania with one as control and another as treatment involving 64 FHWs who were purposively selected from May 2020 to December 2020. A total of 32 FHWs from treatment pastoral community were purposively selected for awareness training, rapid diagnosis using Rose Bengal test (RBT) and use of electronic technology (AfyaData app) for brucellosis reporting while nothing was done in control community. Before and after training information about their knowledge, attitude and practices were collected from all participants using a structured questionnaires uploaded in the mobile phone powered by AfyaData application. Blood samples were collected from 141 febrile patients attending the selected facilities in treatment community. Serum obtained from collected blood were analyzed using RBT and Competitive Enzyme Linked Immunosorbent Assay (c-ELISA) for brucellosis screening and confirmatory, respectively. Results from this analysis were reported back to the health facility using AfyaData app. Chi-square was used to analyze categorical variables and t-test and/Anova test was used to assess the effectiveness of the intervention. RESULTS: Results revealed that before the training majority of the participants were ignorant about brucellosis, although they had good attitude towards brucellosis prevention. Participant's awareness, practice and attitude increased significantly (p = 0.003, p = 0.001, p = 0.032) respectively, after the intervention. Total of 17(12.1%) patients were positive on RBT and four (2.8%) were confirmed by c-ELISA. AfyaData app was proven to provide quick reports regarding brucellosis in the study area. CONCLUSION: The training program was effective in increasing the level of knowledge and practice about brucellosis. Electronic based technology (AfyaData app) improved the reporting of brucellosis cases. There is a need for the use of electronic based technology to improve timely management of brucellosis in pastoral communities. Also, continuous training on FHWs regarding the disease is needed to improved their awareness and practices.

Knowledge, attitude and practices of frontline health workers in relation to detection of brucellosis in rural settings of Tanzania: a cross-sectional study.

BACKGROUND: Brucellosis an important zoonotic disease worldwide, which frequently presents as an undifferentiated febrile illness with otherwise varied and non-specific clinical manifestations. Despite its importance, there are few reports on its awareness among frontline health workers. This study aimed at assessing the baseline knowledge, attitude and practice (KAP) related to detection and management of brucellosis among frontline health workers (FHWs) namely; healthcare workers (HWs) and community health workers (CHWs). METHODS: A cross-sectional study was conducted from December 2019 to January 2020 in Kilosa and Chalinze districts of Tanzania. Data on demographic characteristics, knowledge, attitude and practices regarding brucellosis were collected from the study participants using a structured questionnaire. Interviews were conducted with 32 HWs and 32 CHWs who were systematically selected in study districts. Chi square/fisher Exact was used to assess the association between sociodemographic variables and those related to knowledge, attitude and practices. RESULTS: Overall, a total of 30 (93.8%) HWs and nine (28.1%) CHWs from the study districts heard about brucellosis, with (34.4%) of HWs having knowledge about the causative organism. Overall, knowledge showed almost half (46.9%) HWs and (28.1%) CHWs were aware of the symptoms, clinical signs, diagnosis and control regarding brucellosis. Knowledge difference was statistically significant with HWs' age (p = 0.016). Almost half (46.9%) HWs and less than quarter (12.5%) CHWs had good practices regarding brucellosis control. Almost three quarters (71.9%) of HWs and (21.9%) CHWs had positive attitude regarding brucellosis control; overall attitude was statistically significant with CHWs age (p = 0.028) and education level (p = 0.024). Lack of awareness and unavailability of diagnostic tools were the main challenges faced by FHWs in the two districts. CONCLUSION: The majority of participants were not aware of human brucellosis. Moreover, their overall knowledge was inadequate and the common practices were diagnostic tools, and adequate knowledge to manage brucellosis cases. These findings highlight the need to strengthen frontline health workers knowledge, practices and diagnostic capacities related to brucellosis.

Gendered asymmetry of access to knowledge for brucellosis control among pastoral communities in north-west Côte d'Ivoire.

Brucellosis is an infectious zoonotic disease considered as a threat to public health and pastoralist livelihoods. Symptoms of the disease can lead to gender-specific ailments such as abortions in women and orchitis in men. Pastoralists and their families are at high risk of contracting the disease. Access to health information reinforces existing knowledge and contributes to disease prevention. However, in developing countries, interventions for knowledge sharing on zoonotic diseases predominantly target men. This study aimed to describe mechanisms of knowledge production and transfer on brucellosis according to gender, by assessing the way knowledge affects behaviours of pastoral communities. A community-based cross-sectional survey was conducted among a pastoral community (PC) of the Folon region in north-west Côte d'Ivoire. The study included transhumant pastoralists, sedentary livestock owners, shepherds and their wives. By using mixed methods, 26 semi-structured interviews were conducted, and 320 questionnaires were completed. Statistical analysis with chi-square (χ 2) comparison tests was performed to compare variables between men and women. Findings were interpreted through the concept of specialisation of the social exclusion theory. We found that gender influences access to information on brucellosis and transfer of knowledge on brucellosis appeared gender-biased, especially from veterinarians towards men in the community. The social labour division and interventions of veterinarians through awareness reinforce the knowledge gap on brucellosis between men and women. Men and women consume raw milk, whilst only men in general handle animal discharges with bare hands. To improve the control of brucellosis, knowledge on best practice should be shared with pastoral communities using the One Health approach that encourages mutual learning. Innovative strategies based on gender daily tasks such as safe dairy processing by women and safe animal husbandry to expand their herd for men can be the entry point for the prevention of brucellosis.

Crossing the Line: Seroprevalence and Risk Factors for Transboundary Animal Diseases Along the Tanzania-Zambia Border.

Transboundary pathogens pose a threat to livelihood security in countries such as Zambia and Tanzania. This study aimed to investigate the seroprevalence of peste des petits ruminants virus (PPRV), foot and mouth disease virus (FMDV), sheep and goat pox virus (SGPV), Rift Valley fever virus (RVFV) and Brucella spp. in sheep and goats along the Tanzania-Zambia border. Another aim was to assess the association between certain predictor variables and seroprevalence, focusing on trade and proximity to an international border, to a town and to the Tanzania-Zambia highway. During September-October 2018, 486 serum samples from small ruminants in Zambia and 491 in Tanzania were collected and analyzed using enzyme-linked immunosorbent assays (ELISA). A questionnaire focused on management strategies was administered to each household. The animal-level seroprevalence in Zambia was 0.21% [95% confidence interval (CI) (0.01-1.14) for PPRV, 1.03% (95% CI 0.33-2.39) for FMDV, 0% (95% CI 0-0.76) for SGPV, 2.26% (95% CI 1.14-4.01) for RVFV and 1.65% (95% CI 0.71-3.22) for Brucella spp.]. In Tanzania, animal-level seroprevalence was 2.85% (95% CI 1.57-4.74) for PPRV, 16.9% (95% CI 13.7-20.5) for FMDV, 0.20% (95% CI 0.01-1.13) for SGPV, 3.26% (95% CI 1.87-5.24) for RVFV and 20.0% (95% CI 14.5-26.5) for Brucella spp. For PPRV (OR 6.83, 95% CI 1.37-34.0, p = 0.019) and FMDV (OR 5.68, 95% CI 1.58-20.3, p = 0.008), herds situated more than 30 km from an international border were more likely to be seropositive, while being located 10-30 km (OR 4.43, 95% CI 1.22-16.1 p = 0.024) from a border was identified as a risk factor for Brucella spp. For FMDV (OR 79.2, 95% CI 4.52-1388.9, p = 0.003), being situated within 30 km from a town was associated with seropositivity. Furthermore, contact with wild ruminants (OR 18.2, 95% CI 1.36-244), and the presence of sheep in the household (OR 5.20, 95% CI 1.00-26.9, p = 0.049), was associated with seropositivity for PPRV, and FMDV. No significant associations between trade or distance to the Tan-Zam highway and seroprevalence were found. We recommend that the impact of trade and proximity to borders, towns and roads should be further evaluated in larger studies, ideally incorporating aspects such as temporal trade fluctuations.

Brucellosis testing patterns at health facilities in Arusha region, northern Tanzania.

BACKGROUND: Brucellosis is listed as one of six priority zoonoses in Tanzania's One Health strategic plan which highlights gaps in data needed for the surveillance and estimation of human brucellosis burdens. This study collected data on current testing practices and test results for human brucellosis in Arusha region, northern Tanzania. METHODS: Retrospective data were extracted from records at 24 health facilities in Arusha region for the period January 2012 to May 2018. Data were captured on: the test reagents used for brucellosis, procurement and testing protocols, the monthly number of patients tested for brucellosis and the monthly number testing positive. Generalised linear mixed models were used to evaluate relationships between health facility characteristics and the probability that brucellosis testing was conducted in a given month, and the proportion of individuals testing positive. RESULTS: Four febrile Brucella agglutination tests were used widely. The probability of testing for brucellosis in a given month was significantly associated with an interaction between year of testing and facility ownership. Test probability increased over time with more pronounced increases in privately owned as compared to government facilities. The proportion of individuals testing positive for brucellosis was significantly associated with facility type and district, with individuals tested in hospitals in Meru, Monduli and Ngorongoro districts more likely to test positive. CONCLUSIONS: Febrile Brucella agglutination tests, known for their poor performance, were the mainstay of brucellosis testing at health facilities in northern Tanzania. The study indicates that historical data on human brucellosis in Arusha and other regions are likely to provide an inaccurate measure of true disease burden due to poor performance of the tests used and variation in testing practices. Measures to address these identified shortcomings could greatly improve quality of testing and surveillance data on brucellosis and ultimately inform prevention and control of this priority disease.

Seroprevalence of brucellosis in small ruminants and related risk behaviours among humans in different husbandry systems in Mali.

Mali has a high pastoral potential with diverse coexisting production systems ranging from traditional (nomadic, transhumant, sedentary) to commercial (fattening and dairy production) production systems. Each of those systems is characterised by close interactions between animals and humans, increasing the potential risk of transmission of zoonotic diseases. The nature of contact network suggests that the risks may vary according to species, production systems and behaviors. However, the study of the link between small ruminants and zoonotic diseases has received limited attention in Mali. The objective of this study was to assess brucellosis seroprevalence and determine how the husbandry systems and human behaviour expose animal and human to infection risk. A cross-sectional study using cluster sampling was conducted in three regions in Mali. Blood was collected from 860 small ruminants. The sera obtained were analysed using both Rose Bengal and cELISA tests. In addition, 119 farmers were interviewed using a structured questionnaire in order to identify the characteristics of farms as well as the risk behaviors of respondents. Husbandry systems were dominated by agro-pastoral systems followed by pastoral systems. The commercial farms (peri-urban and urban) represent a small proportion. Small ruminant individual seroprevalence was 4.1% [2.8-5.6% (95% CI)]. Herd seroprevalence was estimated at 25.2% [17.7-33.9% (95% CI)]. Peri-urban farming system was more affected with seroprevalence of 38.1% [18.1-61.5 (95% CI)], followed by pastoral farming system (24.3% [11.7-41.2 (95% CI)]). Identified risk behaviors of brucellosis transmission to animals were: exchange of reproductive males (30.2%); improper disposal of placentas in the farms (31.1%); and keeping aborted females in the herd (69.7%). For humans, risk factors were: close and prolonged contact with animals (51.2%); consumption of unpasteurized dairy products (26.9%); and assisting female animals during delivery without any protection (40.3%). This study observed a high seroprevalence of brucellosis in small ruminants and also identified risky practices that allow cross transmission between the two populations. This calls for control strategy using a multi-sectoral and multidimensional approach.

Molecular epidemiology of Brucella species in mixed livestock-human ecosystems in Kenya.

Brucellosis, caused by several species of the genus Brucella, is a zoonotic disease that affects humans and animal species worldwide. Information on the Brucella species circulating in different hosts in Kenya is largely unknown, thus limiting the adoption of targeted control strategies. This study was conducted in multi-host livestock populations in Kenya to detect the circulating Brucella species and assess evidence of host-pathogen associations. Serum samples were collected from 228 cattle, 162 goats, 158 sheep, 49 camels, and 257 humans from Narok and Marsabit counties in Kenya. Information on age, location and history of abortion or retained placenta were obtained for sampled livestock. Data on age, gender and location of residence were also collected for human participants. All samples were tested using genus level real-time PCR assays with primers specific for IS711 and bcsp31 targets for the detection of Brucella. All genus positive samples (positive for both targets) were further tested with a speciation assay for AlkB and BMEI1162 targets, specific for B. abortus and B. melitensis, respectively. Samples with adequate quantities aggregating to 577 were also tested with the Rose Bengal Test (RBT). A total of 199 (33.3%) livestock and 99 (38.5%) human samples tested positive for genus Brucella. Animal Brucella PCR positive status was positively predicted by RBT positive results (OR = 8.3, 95% CI 4.0-17.1). Humans aged 21-40 years had higher odds (OR = 2.8, 95% CI 1.2-6.6) of being Brucella PCR positive compared to the other age categories. The data on detection of different Brucella species indicates that B. abortus was detected more often in cattle (OR = 2.3, 95% CI 1.1-4.6) and camels (OR = 2.9, 95% CI 1.3-6.3), while B. melitensis was detected more in sheep (OR = 3.6, 95% CI 2.0-6.7) and goats (OR = 1.7, 95% CI 1.0-3.1). Both B. abortus and B. melitensis DNA were detected in humans and in multiple livestock host species, suggesting cross-transmission of these species among the different hosts. The detection of these two zoonotic Brucella species in humans further underpins the importance of One Health prevention strategies that target multiple host species, especially in the multi-host livestock populations.

Performance characteristics and costs of serological tests for brucellosis in a pastoralist community of northern Tanzania.

The control of brucellosis across sub-Saharan Africa is hampered by the lack of standardized testing and the use of tests with poor performance. This study evaluated the performance and costs of serological assays for human brucellosis in a pastoralist community in northern Tanzania. Serum collected from 218 febrile hospital patients was used to evaluate the performance of seven index tests, selected based on international recommendation or current use. We evaluated the Rose Bengal test (RBT) using two protocols, four commercial agglutination tests and a competitive enzyme-linked immunosorbent assay (cELISA). The sensitivity, specificity, positive predictive value, negative predictive value, Youden's index, diagnostic accuracy, and per-sample cost of each index test were estimated. The diagnostic accuracy estimates ranged from 95.9 to 97.7% for the RBT, 55.0 to 72.0% for the commercial plate tests, and 89.4% for the cELISA. The per-sample cost range was $0.69-$0.79 for the RBT, $1.03-$1.14 for the commercial plate tests, and $2.51 for the cELISA. The widely used commercial plate tests performed poorly and cost more than the RBT. These findings provide evidence for the public health value of discontinuing the use of commercial agglutination tests for human brucellosis in Tanzania.