Isolation And Molecular Characterization Of Shiga Toxin Producing Escherichia coli In Cattle, Water And Diarrhoeal Children From The Pastoral Systems Of Southwestern Uganda

This study describes the molecular characteristics of STEC isolated from the pastoralist community of Nyabushozi in Southwestern Uganda. Faecal samples or rectal swabs of children with diarrhoea obtained in phases 1 and 2 were investigated for the presence of STEC by PCR detection of stx genes. During phase 1, cattle reared on range which were associated with households of sick children were investigated in parallel to the children for STEC excretion. STEC was isolated from E. coli in 7 of 80 (8.8%) children and in 15 of 216 (6.9%) bovines in phase 1. Similarly, STEC was isolated from 11 of 142 (7.7%) E. coli carrying children and 3 of 45 (6.7%) water samples in phase 2. Molecular characterization further ascertained the genetic relatedness of STEC. PFGE pro les of up to 10 colonies obtained from an individual source (child, bovine or water) and in total 185 STEC colonies were analysed. Nine pro les from 43 colonies (phase 1) and 15 pro les from 38 colonies (phase 2) obtained from children were not or were distally related, indicating the genetic diversity of clinical STEC. The intra-host analysis of STEC pro les revealed that strains from 11 of the 13 children exhibited multiple clonal subgroups. The 101 colonies from 15 bovines clustered in 18 di erent pro les. Clonal subgroups were observed in multiple STEC colonies from 11 of 12 bovines. Closely related pro les indicated that STEC isolated from two children (Hh2 and Hh4) was acquired from bovines or their environment. While none of the clinical or bovine STEC were related to 5 genetically diverse water strains. A single isolate of STEC representing each PFGE pro le in association with stx gene content was serotyped for the O antigen. Twenty four bovine STEC were typed into 10 O serogroups including O8, O76, O111 and O113, which were also identi ed among the clinical STEC. The 25 clinical STEC belonged to 15 serogroups of which O29, O149 and O176 are being reported for the rst time as clinical STEC. STEC xxi Abstract O166 was isolated from a child and water during the same sampling, indicating the potential health hazard of drinking STEC-contaminated water. The production of Shiga toxin (Stx) investigated using Duopath Verotoxin detection kits showed that a majority of STEC from di erent sources produced Stx1 or Stx2 or both Stx. Using PCR or PCR-RFLP assays, stx2 and eae gene types were analysed. Variant stx2 vhc was most prevalent and closely associated with stx2d2 in clinical and bovine STEC. The frequency of eae-positive STEC among clinical and bovine STEC was 15 of 25 (60%) and 14 of 24, (58.3%), respectively. eae- 2/ was predominant among the bovine STEC, eae- / in clinical STEC, while eae- 1 was associated with STEC from di erent sources including water. Previously undescribed eae-positive serogroups O28ac, O113, O142 and O158 were identi ed. Studies of the genetic background showed that both clinical and bovine STEC obtained in phase 1 predominantly belonged to phylogenetic group A and B1, while phase 2 clinical and water STEC belonged to group D and A, respectively. Seropathotype classi cation of clinical STEC, separated most strains (20 of 24 strains) into seropathotype D. These STEC belonged to phylogenetic groups A, B1 and D. Thus, the characterised genetic attributes of STEC from Nyabushozi suggests that the pathogens have the potential to cause a wide spectrum of childhood illnesses ranging from mild to bloody diarrhoea and haemolytic uraemic syndrome. xxi

The potential for the double risk of rabies and antimicrobial resistance in a high rabies endemic setting:Detection of antibiotic resistance in bacterial isolates from infected dog bite wounds in Uganda

BACKGROUND: Post-exposure treatment for dog bites in humans aims at alleviating the risk of rabies and promoting wound healing. Wound healing may be complicated by bacteria. This study identified the different bacteria and their antibiotic susceptibilities in infected dog bite wounds (DBWs) in Uganda. METHODS: A cross-sectional study was conducted among 376 dog bite patients. Wound swabs from patients with infected DBWs were collected and inoculated into recommended media. They were cultured for both aerobic and anaerobic bacteria. All isolated bacteria were identified based on colony characteristics, gram stain, and standard biochemical tests. Molecular identification was performed for strains that were resistant to three or more antibiotics. Antibiotic susceptibility testing was conducted using the disc diffusion method following the modified Kirby-Bauer method. The data were analysed using Stata version 15 software. RESULTS: Approximately half of the patients (52.9%, 199/376) presented with infected wounds. Majority of the swabs (84.4%, 168/199) were culture positive, and yielded a total of 768 isolates where about half (52.9%, 406/768) were gram positive bacteria, and about two-thirds (64.6%, 496/768) were recovered from category II wounds. Among the gram positive bacteria, 339 (83.5%) were aerobes where Staphylococcus aureus (103, 30.4%), Coagulase-negative staphylococci (68, 20.1%), and Corynebacterium spp (33, 9.7%) had the highest prevalence. For the 362 Gram negative isolates, 217 (59.9%) were aerobes and the commonest isolates were P. maltocida (64, 29.5%), Capnocytophaga canimorsus (36, 16.6%) and P. canis (26, 12.0%). Gram-positive isolates were resistant to metronidazole (93.6%), oxacillin (68.5%), ceftriaxone (14.6%) and amoxicillin/clavulanic acid (14.0%). Gram negative isolates were resistant to metronidazole (100%), ampicillin (30.7%), oxacillin (29.3%), and doxycycline (22.9%). Multidrug resistance was in 105 (29.0%) and 121/406 (29.8%) of the gram-negative and gram-positive isolates, respectively. All gram-positive isolates were susceptible to vancomycin and ciprofloxacin. CONCLUSIONS: Infection rates of DBWs in Uganda are high and the dominant bacterial isolates are Staphylococcus aureus, Pasteurella spps, and Capnocytophaga canimorsus. Multidrug resistance to commonly used antibiotics is high. The recommendation in the Uganda Clinical Guidelines to use metronidazole in the management of DBWs should be reviewed. DBWs should be enlisted for routine antimicrobial resistance surveillance and rational use of antimicrobial agents should be promoted. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13756-022-01181-0

Sero-prevalence and associated risk factors of Brucellosis among Malaria negative febrile out-patients in Wakiso district, Central Uganda

Abstract Objective Brucellosis is a zoonotic disease usually acquired through direct contact with the infected animals and consumption of contaminated milk and meat products. In humans Brucellosis presents similar signs with other febrile diseases like Malaria, typhoid and other febrile conditions. This study was carried out to determine the prevalence of Brucella abortus among patients with fever but were negative for Malaria. Results A cross-sectional study was carried out in Namayumba Health Centre IV, Wakiso district involving 200 participants. Blood samples was screened for B. abortus using Serum Agglutination Test and confirmed with Tube Agglutination test. A questionnaire was used to collect data on socio-demographic characteristics and human Brucellosis related risk factors. Human B. abortus sero-prevalence was at 7.5% (n = 200). The prevalence was high among participants aged 18–35 years (13.3%), muslims 12 (14.0%), those with no formal education (33.3%) and divorced 2 (14.3%). Consuming of raw milk (OR 2.162, 95% CI 0.021–1.379) and being a Muslim (OR 6.101, 95% CI 1.601–23.248) were associated with increased risk of Brucella abortus. It was concluded that human Brucella infection due to Brucella abortus is commonly associated with consumers of raw milk products and muslims in Wakiso district

Assessing short evolution brucellosis in a highly brucella endemic cattle keeping population of Western Uganda: a complementary use of Rose Bengal test and IgM rapid diagnostic test

Abstract Background Brucellosis is a worldwide and zoonotic disease often sadly misdiagnosed in endemic areas. Challenges of availability and accessibility of diagnostic tools are common in resource constrained populations where the most vulnerable are found, surveillance and diagnosis are limited too. Methods A cross-sectional study using a simple two stage cluster sampling method was conducted to measure short evolution brucellosis burden among cattle keeping households that are one of the highest risk populations to be exposed to Brucella infection. A total of 216 households were randomly selected from 18 rural villages from the Western Region of Uganda. Household blood samples were tested for Brucella antibodies using the highly sensitive Rose Bengal test (RBT) and IgM ELISA Lateral Flow Assay (LFA). Results Among the total tested population, 58.8% did not react with any of the tests, 13.4% reacted with both tests. Among those that reacted with both (N = 29), 62.1% had weak (+ 1) LFA staining, 34.5% had moderate (2+) LFA staining. Altogether, both weak and moderate staining (96.5%) are consistent with sub-acute disease, while only one (3.4%) had strong (3+) LFA staining consistent with acute infection. 19.4% of the samples tested positive only with RBT, consistent with chronic infection, eighteen samples (8.3%) reacted exclusively with IgM LFA. Conclusion We identified a high prevalence of short evolution brucellosis in the cattle keeping household members. Prevalence of chronic infection diagnosed with RBT only was higher than the prevalence of short evolution brucellosis. IgM LFA results depict possible cases of cross reaction with Salmonella spp., Plasmodium etc. Ultimately, we identified a consistent prevalence of short evolution brucellosis in the cattle keeping household members. Indeed, the use of a combined diagnostic with LFA and RBT is easy and amenable for an active disease surveillance and accurate diagnosis in rural settings

Circulation of bluetongue virus in goats in the Karamoja region of Uganda

The presence of bluetongue virus (BTV) in indigenous goats from the Karamoja region of northern Uganda was investigated. A total of 300 goats were sampled (serum and whole blood) from five districts within the Karamoja region. The samples were analysed for the presence of bluetongue (BT) antibodies using a commercial Enzyme-linked immunosorbent assay (ELISA) and for the presence of BTV viral RNA by real-time Reverse transcription polymerase chain reaction (RT-PCR), because BTV is an RNA virus. Of the 300 goats tested, 269 (90%) were positive for BTV antibodies, indicating high levels of BTV circulation within the region. Out of the 150 whole blood samples tested for the presence of the virus by real-time RT-PCR, 84 (56%) were positive for BTV RNA. This study, which is the first of its kind in Uganda, showed a high seroprevalence of BT antibodies and active circulation of BTV in a high proportion of goats in the Karamoja region

Epidemiology and preclinical management of dog bites among humans in Wakiso and Kampala districts, Uganda: Implications for prevention of dog bites and rabies.

In rabies endemic areas, appropriate management of dog bites is critical in human rabies prevention. Victims must immediately wash bite wound for 15 minutes with water, soap, and a disinfectant before seeking medical care. This study investigated the epidemiology of dog bites and the determinants of compliance to these pre-clinical guidelines requirements among dog bite victims from high rabies-burden areas of Wakiso and Kampala, Uganda. An explanatory sequential mixed-methods study design was used. Quantitative data were collected from 376 dog-bite patients at two healthcare facilities. Qualitative data were also collected through 13 in-depth interviews with patients, healthcare workers, herbalists, and veterinarians. Qualitative data were analyzed using a deductive thematic approach. Generalized linear models were used to determine factors associated with compliance. Nearly half (190, 51%) of the patients were from Wakiso District and 293 (77.9%) had grade II wounds. Most of the wounds (171, 45.5%) were on the legs. Two-thirds of the bites occurred in public places. Only 70 (19%) of the bite patients had complied with pre-clinical guidelines. Nearly half of the patients had applied substances that were not recommended e.g. herbs (47/193), antiseptics (46/193), "black stone" (25/193), and unknown creams (10/193). Factors negatively associated with compliance included: being aged 15 years or older, adjPR = 0.70 (0.47-0.92) and knowing the dog owner, adjPR = 0.65 (0.36-0.93). However, attainment of secondary or higher education, adjPR = 1.76 (1.24-3.79), being in employment, adjPR = 1.48 (1.09-2.31), perception that the dog was sick, adjPR = 1.47 (1.02-2.72) and knowledge about the dog's subsequent victim(s) adjPR = 0.35 (0.17-0.70) were positively associated with compliance. High occurrence of dog bites in public places by free-roaming dogs suggests the need for deliberate promotion of responsible dog ownership. Additionally, targeted health education may be required to improve the low compliance to pre-clinical guidelines

"As long as the patient tells you it was a dog that bit him, why do you need to know more?" A qualitative study of how healthcare workers apply clinical guidelines to treat dog bite injuries in selected hospitals in Uganda.

Dog-mediated rabies is on the increase in Uganda despite the availability of post-exposure prophylaxis (PEP). PEP procedures are expounded in the Uganda Clinical Guidelines (UCG) of 2016. We assessed adherence by health workers to UCG while managing dog bites in two PEP centers and obtained insights into motivations of their practices. Using qualitative methods, we observed the health worker-patient encounters, reviewed medical records, and interviewed 14 health workers that were involved in managing dog bite injuries. We used deductive thematic analysis to identify codes in themes developed from UCG. We found that much of the history of the bites was taken, but it was neither verified nor written down on the patient's file. Classification of wounds was inaccurate and ancillary laboratory assessments like culture and sensitivity tests were not conducted in all cases. Although antibiotics were given for both treatment and prophylactic purposes, the prescription was based on availability and affordability, not UCG recommendations. Rabies immunoglobulin (RIG) was not administered to deserving patients due to unavailability and high costs to the patient. Anti-rabies vaccine (ARV) was prescribed indiscriminately and some health workers attributed this to pressure from patients. Health education regarding prevention of dog bites was not given to patients due to time constraints on the side of the providers as a result of high caseloads at the emergency departments. Challenges to adherence to guidelines were identified as frequent ARV stock outs; inadequate cooperation among health facilities; and insufficient knowledge and skills on how injuries and rabies should be managed. We conclude that clinical management of dog bites is not fully in line with UCG. We argue that adoption of an integrated bite case management and cost-saving strategies as well as continuing medical education programs on rabies control and management could improve the clinical management of dog bites

Africa-United States joint curriculum development of a Master of Science degree in international infectious disease management at Makerere University

Trans-boundary infectious diseases and zoonoses once thought to be limited by geographical demarcations are now a constant threat to global animal and human health, trade, food security, and socio-economic wellbeing. Many colleges and schools worldwide are training students to serve as professionals who will improve animal health, public health and food security globally. With this realization, Makerere University and North Dakota State University (NDSU) developed the first US-Africa trans-Atlantic degree addressing integrated disease management and international biosecurity. Both institutions received a grant from United States Department of Agriculture, Higher Education Challenge program to develop a joint Master of Science degree in International Infectious Disease Management (MS-IDM). This grant also funded four graduate students� stipend and research. Additionally, the two institutions received funds through �Capacity building in Integrated Management of Transboundary Animal Diseases and Zoonoses (CIMTRADZ)" project that supported students from Uganda. Faculty from CIMTRADZ participating institutions offered short term training to MS-IDM students, including workshops at The International Scientific Boma and conference in Uganda. A joint degree (MS-IDM) was developed in 2011. Ten MS-IDM students graduated and over 20 fellowships were awarded. Student exchange and joint mentorship of students by faculty at both institutions occurred. There were challenges experienced and valuable lessons learnt. The MS-IDM degree provided opportunities for human capacity development to manage transboundary animal diseases and zoonoses in East and Central Africa. This paper describes the curriculum development, challenges experienced and lessons learned, and informs future similar endeavors in internationalizing curricula in higher education