Dataset Information

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

ABSTRACT:

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.

SUBMITTER: Akoko J

PROVIDER: S-EPMC7216537 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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Publications

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

Akoko James J Akoko James J Pelle Roger R Kivali Velma V Schelling Esther E Shirima Gabriel G Machuka Eunice M EM Mathew Coletha C Fèvre Eric M EM Kyallo Victoria V Falzon Laura C LC Lukambagire AbdulHamid S AS Halliday Jo E B JEB Bonfoh Bassirou B Kazwala Rudovick R Ouma Collins C

BMC veterinary research 20200511 1

<h4>Background</h4>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 competiti ...[more]

PMID: 32393374

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Project description: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.

Project description:Brucellosis, one of the most common zoonotic diseases and has significant public health and economic importance worldwide. Few studies and reports have been performed to estimate the true prevalence of animal brucellosis in the Sultanate of Oman; however, no incidence of the disease was previously reported in Al Jabal Al Akhdar. The purpose of this study was to investigate the prevalence of brucellosis in goats in eight villages in Al Jebal Al Akhdar, Sultanate of Oman, namely: Al Aqaieb, Al Helailat, Al Ghilayil, Hail Al Hedap, Da'an Al Hamra, Shnoot, Al Qasha'e and Al Sarah, Al Jabal Al Akhdar in the Sultanate of Oman. In this study we used different diagnostic serological tests, namely, RBT, I-ELISA and CFT to study the prevalence of Brucella infection in goats in Al Jabal Al Akhdar. Statistical analysis using Kappa statistics was used to compare the performance of the serological tests. Biochemical tests and species-specific Multiplex PCR were used to identify the brucella species involved in the infection. A structured questionnaire and Chi-square (x2 ) statistical analysis was used to identify related brucellosis risk factors. This study is the first to reveal brucellosis infection in goats in eight villages in Al Jebal Al Akhdar, Sultanate of Oman, namely: Al Aqaieb, Al Helailat, Al Ghilayil, Hail Al Hedap, Da'an Al Hamra, Shnoot, Al Qasha'e and Al Sarah, with an overall seroprevalence of 11.1%. The study also compared the performance of three different serological tests, namely, RBT, I-ELISA and CFT. Statistical analysis using Kappa statistics showed that the degree of agreement was best seen between RBT and CFT (96%), followed by RBT, I- ELISA (91.4%) and CFT and I- ELISA (89.2%). Biochemical tests and species-specific Multiplex PCR showed the typical profile for B. melitensis. A structured questionnaire and Chi-square (x2 ) statistical analysis indicated that the presence of abortion is the major risk factor for the prevalence of brucellosis, whereas age and sex were not significant factors in the tested animals. Besides, poor knowledge about brucellosis, consumption of unpasteurized milk or milk products, free trade of animals and the introduction of new animal breeds to herds were all contributing risk factors to the prevalence of brucellosis. The prevalence of human brucellosis obtained verbally from pastoralists gave an insight that brucellosis could pose a public health hazard, especially in those high-risk groups, mainly the pastoralists in the study area. Because of their constant and increasing interaction with their animals, pastoralists could be at a high risk of occupational infection.

Project description:Middle East Respiratory Syndrome Coronavirus (MERS-CoV) is an emerging viral disease and dromedary camels are known to be the source of human spill over events. A cross-sectional epidemiological surveillance study was carried out in Kenya in 2017 to, 1) estimate MERS-CoV antibody seropositivity in the camel-dense counties of Turkana, Marsabit, Isiolo, Laikipia and Nakuru to identify, and 2) determine the risk factors associated with seropositivity in camels. Blood samples were collected from a total of 1421 camels selected using a multi-stage sampling method. Data were also collected from camel owners or herders using a pre-tested structured questionnaire. The sera from camel samples were tested for the presence of circulating antibodies to MERS-CoV using the anti-MERS-CoV IgG ELISA test. Univariate and multivariable statistical analysis were used to investigate factors potentially associated with MERS-CoV seropositivity in camels. The overall seropositivity in camel sera was 62.9 %, with the highest seropositivity recorded in Isiolo County (77.7 %), and the lowest seropositivity recorded in Nakuru County (14.0 %). When risk factors for seropositivity were assessed, the "Type of camel production system" {(aOR = 5.40(95 %CI: 1.67-17.49)}, "Age between 1-2 years, 2-3 years and above 3 years" {(aOR = 1.64 (95 %CI: 1.04-2.59}", {(aOR = 3.27 (95 %CI: 3.66-5.61)}" and {(aOR = 6.12 (95 %CI: 4.04-9.30)} respectively and "Sex of camels" {(aOR = 1.75 (95 %CI: 1.27-2.41)} were identified as significant predictors of MERS-CoV seropositivity. Our studies indicate a high level of seropositivity to MERS-CoV in camels in the counties surveyed, and highlights the important risk factors associated with MERS-CoV seropositivity in camels. Given that MERS-CoV is a zoonosis, and Kenya possesses the fourth largest camel population in Africa, these findings are important to inform the development of efficient and risk-based prevention and mitigation strategies against MERS-CoV transmission to humans.

Project description:A Brucella melitensis 16M DNA fragment of 17,119 bp, which contains a large region deleted in B. abortus strains and DNA flanking one side of the deletion, has been characterized. In addition to the previously identified omp31 gene, 14 hypothetical genes have been identified in the B. melitensis fragment, most of them showing homology to genes involved in the synthesis of a polysaccharide. Considering that 10 of the 15 genes are missing in B. abortus and that all the polysaccharides described in the Brucella genus (lipopolysaccharide, native hapten, and polysaccharide B) have been detected in all the species, it seems likely that the genes described here might be part of a cluster for the synthesis of a novel Brucella polysaccharide. Several polysaccharides have been identified as important virulence factors, and the discovery of a novel polysaccharide in the brucellae which is probably not synthesized in B. abortus might be interesting for a better understanding of the pathogenicity and host preference differences observed between the Brucella species. However, the possibility that the genes described in this paper no longer encode the synthesis of a polysaccharide cannot be excluded. Brucellae belong to the alpha-2 subdivision of the class Proteobacteria, which includes other microorganisms living in association with eucaryotic cells, some of them synthesizing extracellular polysaccharides involved in the interaction with the host cell. The genes described in this paper might be a remnant of the common ancestor of the alpha-2 subdivision of the class Proteobacteria, and the brucellae might have lost such extracellular polysaccharide during evolution if it was not necessary for survival or for establishment of the infectious process. Nevertheless, further studies are necessary to identify the entire DNA fragment missing in B. abortus strains and to elucidate the mechanism responsible for such deletion, since only 9,948 bp of the deletion was present in the sequenced B. melitensis DNA fragment.

Dataset Information

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

Background

Results

Conclusion

Publications

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

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