Sero-prevalence and risk factors associated with occurrence of anti-Brucella antibodies among slaughterhouse workers in Uganda.

INTRODUCTION: Brucellosis is a febrile zoonosis occurring among high-risk groups such as livestock keepers and abattoir workers and is a public health priority in Uganda. The technical complexities of bacteriological and molecular methods make serological approaches the cornerstone of diagnosis of human brucellosis in resource limited settings. Therefore, proper application and interpretation of serological tests is central to achieve a correct diagnosis. MATERIALS AND METHODS: We conducted a cross-sectional study to estimate the seroprevalence and factors associated with anti-Brucella antibodies among slaughterhouse workers processing ruminants and pigs in three regions of the country with serial testing using a combination of the Rose Bengal Test (RBT) and the BrucellaCapt test. An authorized clinician collected 543 blood samples from consenting abattoir workers as well as attribute medical and social demographic data. Univariable and multivariable logistic regression were used to determine factors associated with anti-Brucella sero-positivity. RESULTS AND DISCUSSION: The sero-prevalence among ruminant slaughterhouse workers ranged from 7.3% (95% CI: 4.8-10.7) using BrucellaCapt to 9.0% (95% CI: 6.3-12.7) using RBT. Slaughterhouse workers from the Eastern regions (AOR = 9.84, 95%CI 2.27-69.2, p = 0.006) and those who graze animals for alternative income (AOR = 2.36, 95% CI: 1.91-6.63, p = 0.040) were at a higher risk of exposure to Brucella. Similarly, those who wore Personal Protective Equipment (AOR = 4.83, 95%CI:1.63-18.0, p = 0.009) and those who slaughter cattle (AOR = 2.12, 95%CI: 1.25-6.0, p = 0.006) were at a higher risk of exposure to Brucella. Those who slaughter small ruminants (AOR = 1.54, 95%CI: 1.32-4.01, p = 0.048) were also at a higher risk of exposure to Brucella. CONCLUSIONS AND RECOMMENDATIONS: Our study demonstrates the combined practical application of the RBT and BrucellaCapt in the diagnosis of human brucellosis in endemic settings. Both pharmaceutical (e.g., routine testing and timely therapeutic intervention), and non-pharmaceutical (e.g., higher index of suspicion of brucellosis when investigating fevers of unknown origin and observation of strict abattoir hygiene) countermeasures should be considered for control of the disease in high-risk groups.

Mapping the risk of Rift Valley fever in Uganda using national seroprevalence data from cattle, sheep and goats.

Uganda has had repeated outbreaks of Rift Valley fever (RVF) since March 2016 when human and livestock cases were reported in Kabale after a long interval. The disease has a complex and poorly described transmission patterns which involves several mosquito vectors and mammalian hosts (including humans). We conducted a national serosurvey in livestock to determine RVF virus (RVFV) seroprevalence, risk factors, and to develop a risk map that could be used to guide risk-based surveillance and control measures. A total of 3,253 animals from 175 herds were sampled. Serum samples collected were screened at the National Animal Disease Diagnostics and Epidemiology Centre (NADDEC) using a competition multispecies anti-RVF IgG ELISA kit. Data obtained were analyzed using a Bayesian model that utilizes integrated nested Laplace approximation (INLA) and stochastic partial differential equation (SPDE) approaches to estimate posterior distributions of model parameters, and account for spatial autocorrelation. Variables considered included animal level factors (age, sex, species) and multiple environmental data including meteorological factors, soil types, and altitude. A risk map was generated by projecting fitted (mean) values, from a final model that had environmental factors onto a spatial grid that covered the entire domain. The overall RVFV seroprevalence was 11.3% (95% confidence interval: 10.2-12.3%). Higher RVFV seroprevalences were observed in older animals compared to the young, and cattle compared to sheep and goats. RVFV seroprevalence was also higher in areas that had (i) lower precipitation seasonality, (ii) haplic planosols, and (iii) lower cattle density. The risk map generated demonstrated that RVF virus was endemic in several regions including those that have not reported clinical outbreaks in the northeastern part of the country. This work has improved our understanding on spatial distribution of RVFV risk in the country as well as the expected burden of the disease in livestock.

Sero-prevalence and factors associated with anti-Brucella antibodies in slaughter livestock in Uganda.

Introduction: Brucellosis is endemic in Uganda and is a major cause of production losses in livestock. Early detection and quantification of the disease is vital for its control and eradication. The aim of this study was to assess the sero-prevalence and factors associated with anti-Brucella antibodies in slaughtered livestock. Materials and methods: Sera from 886 cattle, 925 small ruminants, and 900 pigs were collected from regional abattoirs in Northern, Eastern and Central Uganda. To estimate sero-prevalence, sera were serially tested using a combination of the Rose Bengal Test (RBT) and Native Hapten (NH) immunoprecipitation test. True sero-prevalence was estimated using the Rogan-Gladden estimator considering the sensitivity and specificity of the NH immunoprecipitation test. Multivariable logistic regression was used to assess factors associated with seropositivity for anti-Brucella antibodies. Results and discussion: Small ruminants showed the highest seroprevalence (6.7%, 95% CI = 4.2-7.1) followed by cattle (3.8%, 95% CI = 2.4-4.9) and pigs (2.8%, 95% CI = 1.1-2.9). Seropositivity for anti-Brucella antibodies was associated with region of origin (OR = 4.6,95%CI=1.49-17.75, p = 0.013) for cattle; sex (OR = 2.90, 95% C = 1.5-6.34, p = 0.004), age (OR=4.04, 95% CI = 1.07-8.52, p = 0.006) and species (OR = 2.53, 95% CI = 1.08-6.98, p = 0.048) for small ruminants; and finally sex for pigs (OR = 2.88, 95% CI = 1.07-8.52, p = 0.041). Progressive control interventions must include both cattle and small ruminants since they play a bigger role in the maintenance and dissemination of Brucella. The interventions should adopt a risk-based approach with regions at higher risk being given top priority. Bacteriological and molecular studies should be undertaken to clarify the role of pigs and the goat-cattle cross infections in the epidemiological cycle of brucellosis in Uganda.

Seroprevalence of brucellosis and risk factors associated with its seropositivity in cattle, goats and humans in Iganga District, Uganda.

INTRODUCTION: The burden of brucellosis among smallholder farmers is poorly-documented in Uganda. The disease burden is likely to be high, given the high levels of endemicity, lots of exposures and due to lack of control measures. In order to designate appropriate control measures, the magnitude and risk factors for brucellosis need to be known. We established the burden of and risk factors for Brucella seropositivity in cattle, goats, and humans in Iganga district, eastern Uganda. METHODS: A cross-sectional study was conducted in in Kigulamo Parish, Iganga District. We enrolled 226 households and administered a structured questionnaire to heads of households to capture data on socio-demographic characteristics, human brucellosis-related risk factors, and livestock farming practices. Human, cattle, and goat blood samples were collected and tested serologically using commercial indirect-ELISA kits manufactured by USDA, USA. RESULTS: Of 451 human blood samples, 20 (4.4%) were positive. Among 345 cattle blood samples, 4 (1.2%) were positive and among 351 goat blood samples, one (0.3%) was positive. Persons who reported consuming locally-made dairy products had 4 times higher odds of Brucella seropositivity (OR = 4.0, CI = 1.14-14.03, p = 0.031) than those who did not. None of the risk factors we asked about were significantly associated with seropositivity in cattle and goats. CONCLUSION: The seroprevalence of brucellosis in humans in smallholder households in Kigulamo was relatively low and associated with consumption of locally made dairy products. No risk factors were significantly associated with seropositivity in livestock, likely due to the small number of seropositive animals. We recommend a One Health approach to control brucellosis simultaneously in animals and humans needed to sustainably reduce the burden of brucellosis in Uganda and beyond.

Molecular detection and characterization of Brucella species in raw informally marketed milk from Uganda.

This study identified and characterized Brucella species in the informal milk chain in Uganda. A total of 324 cattle bulk milk samples were screened for the genus Brucella by real-time PCR with primers targeting the bcsp31 gene and further characterized by the omp25 gene. Of the samples tested, 6.5% were positive for Brucella species. In the omp25 phylogeny, the study sequences were found to form a separate clade within the branch containing B. abortus sequences. The study shows that informally marketed cattle milk in Uganda is a likely risk factor for human brucellosis and confirms that B. abortus is present in the cattle population. This information is important for potential future control measures, such as vaccination of cattle.

The milk delivery chain and presence of Brucella spp. antibodies in bulk milk in Uganda.

This study examined the influence of informal milk delivery chains on the risk of human exposure to Brucella spp. through milk consumption in two regions of Uganda (Gulu and Soroti Districts). The work involved describing milk delivery chains, investigating brucellosis awareness amongst milk deliverers and determining the presence of Brucella spp. antibodies in cattle milk on delivery to primary collection points (boiling points and dairies). Milk samples (n = 331) were collected from deliverers at primary collection points and from street vendors at point of sale and analysed using indirect enzyme-linked immunosorbent assay (I-ELISA). A written questionnaire was used to collect data from deliverers (n = 279) on their milk delivery chains and their brucellosis awareness. The most common delivery points in Gulu District were small dairies and in Soroti District boiling points. The presence of Brucella spp. antibodies in milk samples was higher in Soroti (40 %) than in Gulu (11 %) (P < 0.0001). There are possible public health risk consequences of this finding as 42 % of deliverers in Soroti District reported drinking raw milk, compared with 15 % in Gulu District (P < 0.0001). Awareness of brucellosis was low, with 70 % of all milk deliverers reporting not having heard of the disease or the bacterium. Application of quality controls for milk (colour and odour) along the delivery chain varied depending upon supply and demand. This study provides evidence of the diversity of informal milk markets in low-income countries and of the potential public health risks of consuming unpasteurised milk. These results can be useful to those planning interventions to reduce brucellosis.

Isolation and molecular characterization of Brucella isolates in cattle milk in Uganda.

Brucellosis is endemic in livestock and humans in Uganda and its transmission involves a multitude of risk factors like consumption of milk from infected cattle. To shed new light on the epidemiology of brucellosis in Uganda the present study used phenotypic and molecular approaches to delineate the Brucella species, biovars, and genotypes shed in cattle milk. Brucella abortus without a biovar designation was isolated from eleven out of 207 milk samples from cattle in Uganda. These isolates had a genomic monomorphism at 16 variable number tandem repeat (VNTR) loci and showed in turn high levels of genetic variation when compared with other African strains or other B. abortus biovars from other parts of the world. This study further highlights the usefulness of MLVA as an epidemiological tool for investigation of Brucella infections.

Prevalence of and factors associated with Brucella sero-positivity in cattle in urban and peri-urban Gulu and Soroti towns of Uganda.

Brucellosis is a key zoonosis of major public health, animal welfare and economic significance, and is endemic in livestock in Uganda. A cross-sectional epidemiological study was carried out to estimate the sero-prevalence of brucellosis and identify factors associated with sero-positivity in cattle in urban and peri-urban Gulu and Soroti towns of Northern and Eastern Uganda, respectively. A total of 1007 sera and data on biologically plausible risk factors from 166 herds and their spatial locations, were collected from cattle reared in urban and peri-urban Gulu and Soroti towns of Uganda. The sera were analyzed using indirect ELISA and sero-positive reactors confirmed by competitive ELISA. Multivariable models were used to investigate for risk factors. The overall animal-level and herd-level sero-prevalence was 7.5% (76/1007, 95% Confidence Interval (CI): 6.15-9.4%) and 27.1% (45/166, 95% CI: 20.9-34.3%), respectively. Herd-level sero-prevalence was significantly (P<0.001) higher in Soroti than Gulu. In Gulu town, sero-positivity increased with an increase in herd size (P=0.03) and age (P=0.002), and was higher in cattle brought in from western Uganda (P<0.0001). In Soroti town, introduction of new cattle into a herd was significantly (P=0.027) associated with herd sero-positivity. There was a geographically differential risk (clustering) of Brucella sero- positivity in herds in Soroti, while sero-positivity was homogeneously distributed in Gulu. The data highlight brucellosis occurrence and major risk factors for its transmission in cattle in urban and peri-urban areas.