Molecular and serological prevalence of Leptospira spp. among slaughtered cattle and associated risk factors in the Bahr El Ghazal region of South Sudan.

INTRODUCTION: Leptospirosis is a neglected emerging and zoonotic disease reported worldwide. This study sought to determine the molecular and serological prevalence of Leptospira spp. and the associated risk factors in slaughtered cattle from the Bahr El Ghazal region of South Sudan. MATERIALS AND METHODS: Between January 16th and February 25th, 2023, blood and urine samples were collected from 402 cattle at the Lokoloko Municipal Slaughterhouse in Western Bahr El-Ghazal State. Serum samples were tested using the microscopic agglutination test (MAT), with a panel of 12 serovars (sv) from 12 serogroups (sg) and 4 species (spp) of Leptospira spp. These serovars had been previously identified in Sudan and the East African region. Simultaneously, 400 corresponding urine samples were screened using qualitative real-time polymerase chain reaction (PCR) to detect the shedding of Leptospira spp. in urine. To identify the associated risk factors, the age, sex, breed and body condition score of each sampled cattle was noted at the time of sampling and subsequently analysed using logistic regression models. RESULTS: Among the 402 serum samples screened, a substantial 81.8% (329/402, 95% CI 77.9-85.3) displayed seropositivity for Leptospira spp. with a MAT titre ≥ 100. The prevalence of urine shedding determined by PCR was 6% (23/400, 95% CI 3.8-8.4), while probable recent leptospirosis with a MAT ≥ 1:800 was observed in 33.1% (133/402, 95% CI 28.6-37.8) of the cattle. Multiple reactions were detected in 34.8% (140/402, 95% CI 30.6-39.5) serum samples. The seropositivity was against L. borgpetersenii sg. Tarassovi (78.6%; 316/402, 95% CI 74.4-82.3), followed by L. borgpetersenii sg. Ballum at 20.4% (82/402, 95% CI, 16.7-24.4%), L. kirschneri sg. Autumnalis At 8.7% (35/402, 95% CI 5.7-11.7), L. interrogans sg. of Pomona at 7.0% (28/402, 95% CI 4.5-9.5), and L. interrogans sg. Hebdomadis was 5.0% (20/402, 95% CI 2.8-7.2). Several risk factors are associated with seropositivity. Older animals (≥ 2 years) had 2.0 times greater odds (95% CI 1.14-3.5) of being seropositive than younger animals (< 2 years), P-value = 0.016. Female animals demonstrated 2.1 times greater odds (95% CI 1.2-3.6) of seropositivity than males did (P-value = 0.008). Additionally, Felata/Mbororo cattle exhibited 2.4 times greater odds (95% CI 1.3-4.5) of being seropositive than did local Nilotic cattle (P-value = 0.005). The agreement between the MAT and PCR results was poor, as indicated by a kappa statistic value of 0.001 and a P-value of 0.913. But there was a moderate agreement between MAT high titres ≥ 800 and PCR positivity with a kappa statistic value = 0.501 and a P-value < 0.001. CONCLUSION: In addition to the high seroprevalence, Leptospira spp. were found in the urine of slaughtered cattle, suggesting that leptospirosis is endemic to the study area. This finding underscores the significance of cattle as potential sources of infection for slaughterhouse workers, the general public, and other animal species. To address this issue effectively in the Bahr El Ghazal Region and South Sudan, a comprehensive strategy involving a multidisciplinary approach is essential to minimize disease among animals, hence reducing potential zoonotic risks to humans.

Molecular characterization of Escherichia coli virulence markers in neonatal and postweaning piglets from major pig-producing districts of Uganda.

BACKGROUND: Piggery production is highly constrained by diseases, with diarrhoea in piglets being a major cause of economic losses to smallholder farmers in Uganda. Enterotoxigenic Escherichia coli (ETEC) is thought to be one of the major etiologies of this diarrhoea. A cross-sectional study was carried out in two high pig-producing districts of Uganda with the aim of determining the significance of piglet diarrhoea and the pathogenic determinants of causative E. coli. METHODOLOGY: A total of 40 households with piglets were visited in each district for a questionnaire survey and faecal sample collection. The questionnaire-based data collected included; demographic data and pig management practices. E. coli were isolated from diarrheic (43) and non-diarrheic (172) piglets and were subjected to antimicrobial susceptibility testing against nine commonly used antimicrobial agents. The E. coli isolates were further screened for the presence of 11 enterotoxin and fimbrial virulence gene markers using multiplex polymerase chain reaction. Data entry, cleaning, verification and descriptive statistics were performed using Microsoft Excel. Statistical analysis to determine any association between the presence of virulence markers and diarrhea in piglets was done using SPSS software (Version 23), with a p value of less than 0.05 taken as a statistically significant association. RESULTS: Escherichia coli were recovered from 81.4% (175/215) of the faecal samples. All the isolates were resistant to erythromycin, and most showed high resistance to tetracycline (71%), ampicillin (49%), and trimethoprim sulfamethoxazole (45%). More than half of the isolates (58.3%) carried at least one of the 11 virulence gene markers tested. EAST1 was the most prevalent virulence marker detected (35.4%), followed by STb (14.8%). Expression of more than one virulence gene marker was observed in 6.2% of the isolates, with the EAST1/STa combination being the most prevalent. Three adhesins; F17 (0.6%), F18 (6.3%) and AIDA-I (0.6%) were detected, with F18 being the most encountered. There was a statistically significant association between the occurrence of piglet diarrhoea and the presence of the AIDA-1 (p value = 0.037) or EAST1 (p value = 0.011) gene marker among the isolates. CONCLUSION AND RECOMMENDATION: The level of antimicrobial resistance among E. coli isolates expressing virulence markers were high in the sampled districts. The study established a significant association between presence of EAST1 and AIDA-I virulence markers and piglet diarrhea. Further studies should be carried out to elucidate the main adhesins borne by these organisms in Uganda and the actual role played by EAST1 in the pathogenesis of the infection since most isolates expressed this gene.

Identification and distribution of Rhipicephalus microplus in selected high-cattle density districts in Uganda: signaling future demand for novel tick control approaches.

BACKGROUND: Rhipicephalus (Boophilus) microplus (Canestrini, 1888), the Asian blue tick, is a highly invasive and adaptable ectoparasite. This tick species has successfully established itself in most regions of the world, with movement of cattle being a major driver for its spread. In the recent past, R. microplus ticks have been reported in three districts of Uganda. Information on its spread and distribution are vital in deepening our understanding of the ecological scenarios that lead to tick persistence and in the formulation of control strategies. This is especially important in the cattle-dense districts. METHODS: We randomly collected tick specimens from 1,461cattle spread across seven cattle dense districts located in the Central, Karamoja and West Nile regions of Uganda from January to September 2020. The ticks were identified using standard morpho-taxonomic keys and the R. microplus tick species identities were confirmed by sequencing of the ITS2 region, 12S rRNA and 16S rRNA genes and phylogenetic analyses. RESULTS: Adult ticks (n = 13,019) were collected from 1,461 cattle. Seventeen tick species were identified based on morpho-taxonomic keys and the majority (47.4%; n=6184) of these were R. appendiculatus. In total, 257 R. microplus ticks were found infesting cattle in 18 study sites in the districts of Amudat, Kaabong, Napak (Karamoja region) and Arua (West Nile region). The identity of R. microplus was confirmed using molecular technics. No R. microplus tick was recorded in the districts of Lyantonde and Nakaseke (Central region). Arua district accounted for 82.1% (n=211) of the R. microplus ticks recorded followed by Napak district at 16.3% (n=42), while Amudat and Kaabong districts accounted for 1.5% (n=4). Rhipicephalus microplus and R. decoloratus co-existed in 6 of the 13 study sites in Arua district, while in another 6 study sites, no R. decoloratus was recorded. In the Karamoja region districts R. decoloratus co-existed with R.microplus. Of the total 618 ticks belonging to four species of the subgenus Boophilus recorded in this study, R. decoloratus accounted for 50.04% (n=334), followed by R. microplus at 41.58% (n=257), R. geigyi at 2.75% (n=17) and R. annulatus at 1.61% (n=10). In the districts of Amudat, Kaabong and Napak, R. decoloratus was more dominant (76.1%; n=179) of the three Rhipicephalus (Boophilus) tick species recorded, followed by R. microplus (19.5%; n=46) and R. geigyi (4.2%; n=10). Contrariwise, R. microplus was more dominant (84%; n=211) in Arua district followed by R. decoloratus (10.7%; n=27), R. annulatus (3.9%; n=10) and R. geigyi (1.1%; n=3). Phylogenetic analyses of the ITS2 region, 12S rRNA and 16S rRNA genes revealed subgrouping of the obtained sequences with the previously published R. microplus sequences from other parts of the world. CONCLUSION: Rhipicephalus microplus ticks were found infesting cattle in four districts of Uganda. The inability to find R. decoloratus, an indigenous tick, from six sites in the district of Arua is suggestive of its replacement by R. microplus. Rhipicephalus microplus negatively affects livestock production, and therefore, there is a need to determine its distribution and to deepen the understanding of the ecological factors that lead to its spread and persistence in an area.

Prevalence of cystic echinococcosis among livestock in pastoral and agro-pastoral areas in Uganda.

Cystic echinococcosis (CE) remains a significant challenge in Uganda with precise status largely undocumented in most communities. To determine CE prevalence, post-mortem examination was done on 14 937 livestock (5873 goats, 1377 sheep, 3726 zebu cattle and 3054 Ankole cattle) slaughtered in abattoirs in the districts of Moroto in Karamoja region, Kumi in Teso region and Nakasongola and Luwero in Buganda region. The overall CE prevalence was 21.9% in sheep, 15.2% in zebu cattle, 5.5% in goats and 2.1% in Ankole cattle. Moroto district had a higher prevalence of CE than other districts with 31.3% in zebu cattle, sheep 28%, goats 29.1% and (0%) in Ankole cattle. On organ locations, the lungs were the most affected in all livestock in all the study areas. Considering cyst fertility, 33.9, 1.7 and 6.4% of Ankole cattle, sheep and zebu cattle respectively had fertile cysts in the liver while 4.5% of goats and 4% Ankole cattle had fertile cysts in the lungs. In conclusion, CE is widespread and occurs among cattle, sheep and goats in pastoral and agro-pastoral areas in Uganda. Therefore, there is an urgent need to create awareness among the communities on role of livestock in CE epidemiology and transmission.

Spatio-temporal analysis of sheep and goat pox outbreaks in Uganda during 2011-2022.

BACKGROUND: Sheep and goat pox (SGP) caused by sheep poxvirus (SPV) and goat poxvirus (GPV) respectively; are transboundary and World Organisation for Animal Health (WOAH)-notifiable viral diseases. There is barely any coherent information about the distribution and prevalence of SGP for Uganda. We therefore conducted this study to describe the temporal and spatial distribution of SGP suspected outbreaks in Uganda for the period 2011-2020 as well as serologically confirm presence of SGP antibodies in suspected SGP outbreaks reported in 2021-2022. RESULTS: Thirty-seven [37] SGP outbreaks were reported across the country during the study period. North-eastern region [that comprises of Karamoja region] had the highest number of outbreaks [n = 17, 45%]; followed by Central [n = 9, 2.4%], Northern [n = 8, 2.2%] and Western region [n = 3, 0.08%]. Reports from district veterinary personnel indicate that the prevalence of; and mortality rate and case fatality rate associated with SGP were 0.06%, 0.02% and 32% respectively. There was a steady increase in the number of reported SGP outbreaks [x̄ = 4] over the study period. Seropositivity of SGPV antibodies in outbreak sheep and goats that were investigated during the study period [2021-2022] was [n = 41, 27%, 95 CI;] CONCLUSION: Our analyses of SGPV passive and active reports indicate that SGP is present in Uganda with a decade long average of four outbreaks per annum. During this period, about a third of all SGPV-clinically infected animals died. SPG is therefore a major constraint to small ruminant health and productivity in Uganda. Introduction of animals from infected herds and breach in farm biosecurity were the most important predictors of SGP outbreaks. In addition to the already existing SGP commercial vaccines, small ruminant screening for SGPV before introducing them to naïve herds and ensuring on farm biosecurity should be part of the SGP control tool pack for Ugandan small ruminant farmers.

Economic losses associated with respiratory and helminth infections in domestic pigs in Lira district, Northern Uganda.

This study sought to quantify direct economic losses due to respiratory and gastrointestinal (GI) helminth infections in domestic pigs in Uganda. In a longitudinal study design with repeated measures, farm visits were made at 2 month intervals from October 2018 to September 2019. Weaner and grower pigs (n = 288) aged 2-6 months from 94 farms were sampled. The pigs were monitored for growth and screened for exposure to four important respiratory pathogens: porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSv), Mycoplasma hyopneumoniae (M. hyo), Actinobacillus pleuropneumoniae (App) using ELISA tests. Farm management practices were recorded and used to generate management level scores. Treatment expenses incurred were recorded throughout the study. A mixed effects model was fitted to quantify effects of respiratory and helminth infections on average daily weight gains (ADGs), with farm and pig as random effects. Analysis of variance (ANOVA) was used to determine differences in mean treatment costs by farm management standard. Financial losses were estimated from average carcass dressing percentage, ADG reductions during fattening (200 days). Results showed a grower pig in a given farm exposed to PRRSv and Ascaris spp. had significantly lower ADG by 17.10 gr/day and 16.80 grams/day respectively, compared to a similar unexposed pig (p < 0.05). Mean treatment costs per pig declined significantly with increase in management standard scores (MSS), from USD 1.13 per pig in MSS 1 (poor management) farms to USD 0.95 for MSS 3 (better management) farms (p < 0.05). We show that monetary losses due to PRRSv and Ascaris spp. infection amounted to USD 6.6 ± 2.7 and 6.50 ± 3.2 (Mean ± SEM) per pig, respectively during 200 days of fattening. This study strengthens evidence that improving management practices to reduce infections mitigates economic losses. To guide interventions, further studies are required to unravel the full extent of indirect economic losses.

Co-infections of respiratory pathogens and gastrointestinal parasites in smallholder pig production systems in Uganda.

A cross-sectional study was conducted to identify factors for infections of pigs with key respiratory pathogens: porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus (PPRSv), Mycoplasma hyopneumoniae (M. hyo), Actinobacillus pleuropneumoniae (App), and gastrointestinal (GI) parasites in Uganda. A structured questionnaire was used to collect data on management practices associated with infections. Ninety (90) farms and 259 pigs were sampled. Sera were screened against 4 pathogens using commercial ELISA tests. The Baerman's method was used to identify parasite species in faecal samples. Logistic regression was done to identify risk factors for infections. Results showed individual animal seroprevalence of PCV2 was 6.9% (95% CI 3.7-11.1), PRRSv 13.8% (95% CI 8.8-19.6), M. hyo 6.4% (95% CI 3.5-10.5), and App 30.4% (95% CI 24.8-36.5). The prevalence of Ascaris spp. was 12.7% (95% CI 8.6-16.8), Strongyles spp was 16.2% (95% CI 11.7-20.7), and Eimeria spp. was 56.4% (95% CI 50.3-62.4). Pigs infested with Ascaris spp. were more likely to test positive to PCV2, odds ratio (OR) 1.86 (CI 1.31-2.60; p = 0.0002). For M. hyo, infection with Strongyles spp. was a risk factor (OR 12.9, p < 0.001). Pigs that had Strongyles and Ascaris spp. Infections (ORs 3.5 and 3.4, p < 0.001 respectively) were likely to have co-infections. The model showed that use of cement, elevated floor, and limiting contacts with outside pigs were protective while using mud and helminth infestations increased risks of co-infections. This study provided evidence that improved housing and biosecurity are critical in reducing pathogen incidence in herds.

Seroprevalence and spatial distribution of livestock brucellosis using three serological tests in Kajiado County, Kenya.

Background: Brucellosis is a serious zoonotic infection with a global socioeconomic impact on both the livestock industry and human health. In Kenya, brucellosis is endemic but there is limited information on the true burden of the disease due to weak or peace-meal surveillance. The true burden and spread of animal brucellosis in Kajiado County is not known. Aim: The aim of the study was to determine the current seroprevalence and spatial distribution of livestock brucellosis in Kajiado County and also to compare the three serological tests, namely; Rose Bengal plate test (RBPT), indirect enzyme-linked immunosorbent assay (i-ELISA), and competitive ELISA (c-ELISA) in the detection of seropositive animals. Methods: A cross-sectional study was undertaken in 5 sub-counties and 13 wards, where a total of 782 serum samples from unvaccinated bovine (n = 278; 34 herds), ovine (n = 256; 25 flocks), and caprine (n = 248; 28 flocks), were screened for anti-Brucella antibodies using RBPT, i-ELISA, and c-ELISA tests, in parallel. Results: Overall animal seroprevalence was 6.91% (54/782); while that for bovine, ovine, and caprine was 18.35% (51/278), 0.78% (2/256), and 0.4% (1/248), respectively. Bovine seroprevalence was 2.2% (6/278), 14.4% (40/278), and 4.7% (13/278) in RBPT, i-ELISA, and c-ELISA tests, respectively; while ovine 0.78% (2/256) and caprine 0.4% (1/248) were positive only in c-ELISA. Bovine herd seropositivity was 67.65% (23/34), whereas ovine and caprine flock seropositivities were 8% (2/25) and 3.6% (1/28), respectively. Conclusion: The findings indicate a moderate seroprevalence of brucellosis in bovine, while that of ovine and caprine was low in Kajiado County. Indirect ELISA was found superior to both c-ELISA and RBPT in detecting bovine seropositive animals, while c-ELISA was superior to both RBPT and i-ELISA in detecting seropositive ovines and caprines. These results will contribute to baseline data for further study of Brucella infection and a starting point for the formulation of a strategy for the control of brucellosis in Kajiado County.

Perceptions and practices of farmers of indigenous poultry towards Salmonella infections in North-Central Nigeria.

Background: Salmonella infections continue to be of global concern to poultry health, productivity, and public health. About 44% of the poultry in Nigeria are indigenous and kept in close interaction with farmers who are mostly rural dwellers and have limited access to veterinary and extension services. Aim: The perceptions and practices of farmers of indigenous poultry toward Salmonella infections were assessed to obtain and document baseline data that can be used to create awareness among farmers about these infections and their attendant public health implications. Methods: A cross-sectional approach using a multistage sampling method was used in this survey. A total of 419 farmers keeping indigenous poultry were interviewed using a pre-tested electronic questionnaire in three randomly selected states within North-Central Nigeria. Data were analyzed using descriptive and regression analysis. Results: Out of the 419 respondents, 138 (32.9%), 141 (33.7%), and 140 (33.4%) were from Benue, Kwara, and Plateau States, respectively. Of the 419, 55.4% were females, 40.8% were above 40 years, and 35.8% have over 10 years of farming experience. The majority of the poultry are not housed (58.5%) and farmers predominantly rear chickens (51.8%). Also, 49.9% of the birds were 1-6 months with 41.5% of the flock sizes being 11-20. Respondents had a poor level of perception toward Salmonella infection as the majority did not know that Salmonella affects poultry (89.3%) and that Salmonella infections are zoonotic (94.5%). Significant (p = 0.000) associations existed between categorized perception score and age, educational status, family size, and farming experience of farmers. There were significant (p = 0.000) associations of categorized practice scores with gender, age, education status, family size, and farming experience of farmers. Conclusion: This study has revealed the poor perception of farmers on Salmonella infections and has highlighted their practices. There is a need to raise awareness about these infections to improve indigenous poultry health and productivity as well as public health.

Molecular characterization of porcine reproductive and respiratory syndrome virus (PRRSv) identified from slaughtered pigs in northern Uganda.

BACKGROUND: A cross sectional study was conducted to detect and characterize species of porcine reproductive and respiratory syndrome virus (PRRSv) identified from slaughtered pigs in Lira district, northern Uganda. The study was conducted from March to September 2019 in three selected slaughter slabs. Pigs brought for slaughter were randomly sampled. At necropsy, lungs were extracted from the thoracic cavity and examined for pneumonic lesions. Seventy-three (73) pigs with gross lung lesions were sampled, from which one hundred and one (101) tissue samples were taken. A real-time reverse transcriptase PCR (RT-qPCR) was used to characterize PRRSv species. RESULTS: A total of 20 samples tested positive for PRRSv. The respective prevalence of PRRSv type 1 and type 2 were 24.65% (n = 18) and 2.73% (n = 2) respectively. Of the pigs sampled (n = 73), only two pigs, 2.73% (n = 2) tested positive to both species. The likelihood of PRRSv detection decreased with pig age, but increased with gross pneumonic pathology. CONCLUSIONS: This study demonstrated dual circulation of both species in northern Uganda. The association between PRRSv and lung pathology suggests that it may be an important cause of lung disease in pigs in Uganda and hence loss of production. This calls for further investigations on potential economic impacts of PRRSv on pig productivity. These findings contribute to discussions about the need of surveillance and possible vaccination strategies against PRRSv in Uganda.

Mycobacterium avium subsp. paratuberculosis Virulence: A Review.

To propose a solution for control of Mycobacterium avium subsp. paratuberculosis (MAP) infections in animals as well as in humans, and develop effective prevention, diagnostic and treatment strategies, it is essential to understand the molecular mechanisms of MAP pathogenesis. In the present review, we discuss the mechanisms utilised by MAP to overcome the host defense system to achieve the virulence status. Putative MAP virulence genes are mentioned and their probable roles in view of other mycobacteria are discussed. This review provides information on MAP strain diversity, putative MAP virulence factors and highlights the knowledge gaps regarding MAP virulence mechanisms that may be important in control and prevention of paratuberculosis.

Correlations between lung pneumonic lesions and serologic status for key respiratory pathogens in slaughtered pigs in northern Uganda.

BACKGROUND: A cross-sectional study of slaughtered pigs was conducted in Lira district, Uganda, to (1) determine the prevalence and severity of pneumonia and (2) establish relationships between pneumonia types and the serological status for key respiratory pathogens. Using enzyme-linked immunosorbent assays (ELISAs), sera were screened for antibodies against Mycoplasma hyopneumoniae (M. hyo), Actinobacillus pleuropneumoniae (App), porcine reproductive and respiratory syndrome virus (PRRSv) and porcine circovirus type 2 (PCV2). Postmortem, lungs were grossly scored for pneumonia types and pneumonic lesions. Pneumonia types were characterized as catarrhal purulent bronchopneumonia (CPBP), pleuropneumonia (PLP) and pleuritis. The percent of lung surface affected by pneumonia was determined by estimating the affected surface area of each lung lobe. Each lobe was assigned scores based on the approximate volume represented and the total percentage of lung surface affected obtained as a sum of individual lobe scores. Metastrongylus spp. helminth infection was determined by examining lungs for gross presence or absence. RStudio was used for data analysis and presentation. Wilcoxon rank sum tests were used to compare median pneumonia lesion scores and serostatus for each studied pathogen. An ordinal logistic regression model was fitted to evaluate the odds of multiple pneumonia, with pathogen serostatus and Metastrongylus spp. infection as predictors. RESULTS: One hundred sixty-seven (n = 167) lungs were examined for pneumonic lesions. The prevalences of CPBP, PLP and pleuritis were 29.9% (95% CI 22.9-36.9), 74.2% (95% CI 67.5-80.9) and 17.3% (95% CI 22.4-36.3), respectively. The true prevalence of PCV2 was 9.7% (95% CI 4.5-16.8), that of PRRSv was 7.5% (95% CI 2.7-14.2), that of M. hyo was 11.5% (95% CI 7.2-18.0), that of App was 25.1% (95% CI 18.5-38.0), and that of Metastrongylus spp. was 29.3% (95% CI 22.9-36.6). The odds of multiple pneumonia forms increased in pigs with multiple pathogens (ORs 2.6, p = 0.01) and Metastrongylus spp. infestation (OR 2.5, p = 0.003), suggesting synergistic effects of coinfections in the induction of lesions. CONCLUSIONS: This study revealed a high prevalence and severity of pneumonic lesions in slaughtered pigs. It provides baseline information and evidence for the magnitude of pneumonia associated with the studied pathogens and justifies future studies on their potential economic impacts on Ugandan pigs.

Age-dependent carriage of alleles and haplotypes of Plasmodium falciparum sera5, eba-175, and csp in a region of intense malaria transmission in Uganda.

BACKGROUND: The development of malaria vaccines is constrained by genetic polymorphisms exhibited by Plasmodium falciparum antigens. The project the age-dependent distribution of alleles or haplotypes of three P. falciparum malaria vaccine candidates, Circumsporozoite Protein (csp), Erythrocyte Binding Antigen 175 (eba-175) and Serine Repeat Antigen 5 (sera5) in a region of intense malaria transmission in Uganda. METHODS: A cross-sectional study was carried out between August and November 2009 in which 250 study participants were selected from a population of 600. Finger prick blood samples were collected after informed consent from participants below 5 years, 5-10 years, and above 10 years of age. Blood was used for microscopy, RDT and dried blood spots. Plasmodium falciparum DNA was extracted by chelex method. Alleles of sera5 and eba-175 were determined by polymerase chain reaction (PCR) amplification followed by resolution of products by agarose gel electrophoresis. Allele calling was done using gel photographs from ethiduim bromide stained gels. Haplotypes of csp were identified by sequencing 63 PCR products using the P. falciparum 7G8 laboratory strain sequence as a reference. The data were analysed using SPSS 16, EQX for windows and Chi-square test was used to calculate associations (P-values), Excel was used to generate graphs. The BioEdit and NCBI blast software programs were used to analyse the sequences from which csp haplotypes map was constructed. RESULTS: Eba-175 FCR3 (48/178) and CAMP (16/178) alleles were observed, the FCR3 (24/67) allele being predominant among children aged below 5 years old while the CAMP (12/67) allele was predominant among older participants. Sera5 alleles ORI (6/204) and ORII (103/204) were observed in the population, ORII was more prevalent and was significantly associated with age (P values < 0.0001), parasite density (P-value < 0.0001) and clinical outcomes (P value = 0.018). There was marked csp diversity in the Th2/Th3 region. Out of 63 sequences, 16 conformed to the reference strain and one (1/16) was similar to a West African haplotype and the majority (14/16) of the haplotypes were unique to this study region. There was an age-dependent distribution of csp haplotypes with more haplotypes being harbored by children < 5-year of age, (10/16) compared to adults (2/16). Interestingly, the csp haplotype corresponding to 3D7 whose prototypical sequence is identical to the sequence of the leading malaria vaccine candidate RTS, S was not observed. CONCLUSION: This data suggest that the eba-175 FCR3 allele, sera5 ORII allele, and csp haplotypes are targets of host immunity and under immune selection pressure in Apac District. These molecules could provide alternative malaria vaccine candidates as sub-unit vaccines.

Knowledge, attitude and practices about rabies management among human and animal health professionals in Mbale District, Uganda.

OBJECTIVE: To assess the knowledge, attitude and practices (KAP) of animal and human health professionals towards rabies management and also to establish the level of relationship between KAP. METHODS: A cross-sectional study was conducted between December 2012 and March 2013 among 147 randomly selected animal and human health professionals in Mbale District. Of these, only 16 were animal health professionals. Quantitative data was obtained using a semi-structured questionnaire while qualitative data was obtained from 4 Focus Group Discussions (FGDs) and 2 Key Informant (KI) interviews. Quantitative data was entered into EpiInfo version 3.5.1 and proportions computed while qualitative data was summarised into themes and sub-themes resulting from content analysis of interview scripts. FINDINGS: Of all the respondents, only 44% (65/147) had sufficient knowledge about rabies while 25% (37/147) had positive attitude towards rabies management. A half of the respondents (50%, 73/147) had limited good practices. Respondents knowledgeable about rabies were more likely to have positive attitude towards rabies management (OR = 3.65; 95% CI: 1.60-8.3) while respondents with positive attitudes, were more likely to have good practices towards rabies management (OR: 2.22; 95% CI: 1.01-4.86). CONCLUSION: Respondents had low knowledge, negative attitude and limited good practices of rabies management. Regular refresher trainings about rabies to broaden staff knowledge and improve their attitudes and hence practices of rabies management should be conducted by the District leaders. Harnessing multi-sectoral and multi-disciplinary collaborative efforts ("One Health" approach) for rabies control should be instituted to reduce the incidence of the disease in the District. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s42522-020-00031-6.

Prevalence and risk factors for brucellosis in prolonged fever patients in post-conflict Northern Uganda.

BACKGROUND: Brucellosis is a disease with significant public and economic implications but strategies for controlling this disease remain problematic. OBJECTIVES: This study sought to determine the sero-prevalence of brucellosis in prolonged fever patients and to identify modifiable risk factors for the infection in humans in post conflict Northern Uganda. METHODS: The study employed a cross-sectional method among prolonged fever patients who had visited selected health facilities in the study districts in Northern Uganda. Sero-prevalence of brucellosis was calculated for i-ELISA IgG/IgM. A structured questionnaire was used to obtain data on possible risk factors for brucellosis. Associations between sero-prevalence and risk factors were measured using the Odds Ratio. RESULTS: Brucellosis was confirmed in 18.7% of the 251 patients that tested positive for the disease, with the rapid Brucella Plate Agglutination Test, and ages 10-84 years (median age 47+0.86). Sex (p = 0.001; OR 3.79; 95% CI 1.75 - 8.24), rearing livestock (p < 0.005; OR 8.44; 95% CI 2.84-25.03) and consumption of unpasteurised milk (p = 0.023; OR 2.57; 95% CI 1.14-5.80) were factors associated with brucellosis. CONCLUSION: Control of brucellosis in animals, training and sensitisation of the community on brucellosis is needed to stimulate action on human brucellosis control.

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.

Molecular Detection of Torque Teno Sus Virus and Coinfection with African Swine Fever Virus in Blood Samples of Pigs from Some Slaughterhouses in Nigeria.

Torque teno sus virus 1 (TTSuV1a/TTSuV1b) infection is present in pig herds worldwide. This study investigated the prevalence of TTSuV1a/TTSuV1b infections in domestic pigs from some slaughterhouses in Nigeria as well as coinfection with African swine fever virus (ASFV) and described the phylogeny in relation to global strains. One hundred and eighty-one (181) blood samples from four slaughterhouses were used for the study and viral nucleic acid detection was carried out by PCR. Comparative sequence analysis was carried out to infer phylogeny. The overall prevalence of TTSuV1a/b was 17.7%. Prevalence of individual genotypes was 10.5% and 7.2% for TTSuV1a and TTSuV1b, respectively. Coinfection of ASFV/TTSuV1a/b was 7.7% while that of TTSuV1a and TTSuV1b was 1.7%. ASFV alone was detected in 11.91% of the total samples. The Nigerian TTSuV1a and TTSuV1b shared a sequence identity of 91-100% and 95-100%, respectively, among each other. The ASFV sequences were 100% identical to members of genotype 1. This is the first report on the presence of TTSuV1a/b in domestic pigs in Nigeria and coinfection with ASFV. Although the prevalence of TTSuV1a/b in Nigeria was low, we recommend further studies to establish the trend and possible role in the pathogenesis of ASFV.

The frequency distribution of vitamin D Receptor fok I gene polymorphism among Ugandan pulmonary TB patients.

Background:Mycobacterium tuberculosis (TB) is still a major problem globally and especially in Africa. Vitamin D deficiency has been linked to TB in the past and studies have found vitamin D deficiency to be common among Ugandan TB patients. The functional activity of vitamin D is dependent on the genotype of the vitamin D receptor (VDR) polymorphic genes. Recent findings have indicated that VDR polymorphisms may cause increased resistance or susceptibility to TB. The vitamin D ligand and its receptor play a pivotal role in innate immunity by eliciting antimicrobial activity, which is important in prevention of TB. The fok I vitamin D receptor gene has extensively been examined in TB patients but findings so far have been inconclusive. Objectives: This study sought to investigate the frequency distribution of the VDR fok I gene polymorphisms in pulmonary TB patients and controls. Methods: A pilot case control study of 41 newly diagnosed TB patients and 41 healthy workers was set up. Vitamin D receptor fok I gene was genotyped. Results: The frequency distribution of fok I genotype in Ugandan TB patients was 87.8% homozygous-dominant (FF), 7.3% (Ff) heterozygous and 4.8% (ff) homozygous recessive. For normal healthy subjects the frequencies were (FF) 92.6%, (Ff) 2.4% and (ff) 4.8%. No significant difference was observed in the FF and ff genotypes among TB patients and controls. The Ff heterozygous genotype distribution appeared more in TB patients than in controls. A significant difference was observed in the fok I genotype among gender p value 0.02. No significant difference was observed in ethnicity, p value 0.30. Conclusions: The heterozygous Ff fok I genotype may be associated with TB in the Ugandan population.

Carbapenem resistant Pseudomonas aeruginosa and Acinetobacter baumannii at Mulago Hospital in Kampala, Uganda (2007-2009).

BACKGROUND: Multidrug resistant Pseudomonas aeruginosa and Acinetobacter baumannii are common causes of health care associated infections worldwide. Carbapenems are effective against infections caused by multidrug resistant Gram-negative bacteria including Pseudomonas and Acinetobacter species. However, their use is threatened by the emergence of carbapenemase-producing strains. The aim of this study was to determine the prevalence of carbapenem-resistant P. aeruginosa and A. baumannii at Mulago Hospital in Kampala Uganda, and to establish whether the hospital environment harbors carbapenem-resistant Gram-negative rods. RESULTS: Between February 2007 and September 2009, a total of 869 clinical specimens were processed for culture and sensitivity testing yielding 42 (5 %) P. aeruginosa and 29 (3 %) A. baumannii isolates, of which 24 % (10/42) P. aeruginosa and 31 % (9/29) A. baumannii were carbapenem-resistant. Additionally, 80 samples from the hospital environment were randomly collected and similarly processed yielding 58 % (46/80) P. aeruginosa and 14 % (11/80) A. baumannii, of which 33 % (15/46) P. aeruginosa and 55 % (6/11) A. baumannii were carbapenem-resistant. The total number of isolates studied was 128. Carbapenemase genes detected were bla IMP-like (36 %, 9/25), bla VIM-like (32 %, 8/25), bla SPM-like (16 %, 4/25); bla NDM-1-like (4 %, 1/25) in carbapenem-resistant P. aeruginosa, and bla OXA-23-like (60 %, 9/15), bla OXA-24-like (7 %, 1/15), bla OXA-58-like (13 %, 2/15), and bla VIM-like (13 %, 2/15) in carbapenem-resistant A. baumannii. Furthermore, class 1 integrons were detected in 38 % (48/128) of P. aeruginosa and Acinetobacter, 37 % (26/71) of which were in clinical isolates and 39 % (22/57) in environment isolates. Gene cassettes were found in 25 % (12/48) of integron-positive isolates. These were aminoglycoside adenylyltransferase ant(4')-IIb (3 isolates); trimethoprim-resistant dihydrofolate reductase dfrA (2 isolates); adenyltransferase aadAB (3 isolates); QacE delta1 multidrug exporter (2 isolates); quinolone resistance pentapeptide repeat protein qnr (1 isolate); and metallo-ß-lactamase genes bla VIM-4-like, bla IMP-19-like, and bla IMP-26-like (1 isolate each). Gene cassettes were missing in 75 % (36/48) of the integron-positive isolates. CONCLUSIONS: The prevalence of carbapenem-resistant P. aeruginosa and Acinetobacter among hospitalized patients at Mulago Hospital is low compared to rates from South-East Asia. However, it is high among isolates and in the environment, which is of concern given that the hospital environment is a potential source of infection for hospitalized patients and health care workers.

Prevalence of adhesin and toxin genes in E. coli strains isolated from diarrheic and non-diarrheic pigs from smallholder herds in northern and eastern Uganda.

BACKGROUND: Enterotoxigenic E. coli (ETEC) significantly contribute to diarrhea in piglets and weaners. The smallholder pig producers in Uganda identified diarrhea as one of the major problems especially in piglets. The aim of this study was to; i) characterize the virulence factors of E. coli strains isolated from diarrheic and non-diarrheic suckling piglets and weaners from smallholder herds in northern and eastern Uganda and ii) identify and describe the post-mortem picture of ETEC infection in severely diarrheic piglets. Rectal swab samples were collected from 83 piglets and weaners in 20 herds and isolated E. coli were characterized by PCR, serotyping and hemolysis. RESULTS: The E. coli strains carried genes for the heat stable toxins STa, STb and EAST1 and adhesins F4 and AIDA-I. The genes for the heat labile toxin LT and adhesins F5, F6, F18 and F41 were not detected in any of the E. coli isolates. Where the serogroup could be identified, E. coli isolates from the same diarrheic pig belonged to the same serogroup. The prevalence of EAST1, STb, Stx2e, STa, AIDA-I, and F4 in the E. coli isolates from suckling piglets and weaners (diarrheic and non-diarrheic combined) was 29, 26.5, 2.4, 1.2, 16, and 8.4 %, respectively. However the prevalence of F4 and AIDA-I in E. coli from diarrheic suckling piglets alone was 22.2 and 20 %, respectively. There was no significant difference in the prevalence of the individual virulence factors in E. coli from the diarrheic and non-diarrheic pigs (p > 0.05). The main ETEC strains isolated from diarrheic and non-diarrheic pigs included F4/STb/EAST1 (7.2 %), F4/STb (1.2 %), AIDA/STb/EAST1 (8 %) and AIDA/STb (8 %). At post-mortem, two diarrheic suckling piglets carrying ETEC showed intact intestinal villi, enterocytes and brush border but with a layer of cells attached to the brush border, suggestive of ETEC infections. CONCLUSION: This study has shown that the F4 fimbriae is the most predominant in E. coli from diarrheic piglets in the study area and therefore an F4-based vaccine should be considered one of the preventive measures for controlling ETEC infections in the piglets in northern and eastern Uganda.