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Item Wide distribution of Mediterranean and African spotted fever agents and the first identification of Israeli spotted fever agent in ticks in Uganda(PLOS NEGLECTED TROPICAL DISEASES, 2023) Eneku, Wilfred; Erima, Bernard; Maranda Byaruhanga, Anatoli; Atim, Gladys; Tugume, Titus; Ukuli, Qouilazoni A.; Kibuuka, Hannah; Mworozi, Edison; Christina, Douglas; Jeffrey, W. Koehler; Nora, G. Cleary; Michael, E. von Fricken; Tweyongyere, Robert; Wabwire-Mangen, Fred; Karuhize Byarugaba, DenisRickettsia microorganisms are causative agents of several neglected emerging infectious diseases in humans transmitted by arthropods including ticks. In this study, ticks were collected from four geographical regions of Uganda and pooled in sizes of 1–179 ticks based on location, tick species, life stage, host, and time of collection. Then, they were tested by real-time PCR for Rickettsia species with primers targeting gltA, 17kDa and ompA genes, followed by Sanger sequencing of the 17kDa and ompA genes. Of the 471 tick pools tested, 116 (24.6%) were positive for Rickettsia spp. by the gltA primers. The prevalence of Rickettsia varied by district with Gulu recording the highest (30.1%) followed by Luwero (28.1%) and Kasese had the lowest (14%). Tick pools from livestock (cattle, goats, sheep, and pigs) had the highest positivity rate, 26.9%, followed by vegetation, 23.1%, and pets (dogs and cats), 19.7%. Of 116 gltA-positive tick pools, 86 pools were positive using 17kDa primers of which 48 purified PCR products were successfully sequenced. The predominant Rickettsia spp. identified was R. africae (n = 15) in four tick species, followed by R. conorii (n = 5) in three tick species (Haemaphysalis elliptica, Rhipicephalus appendiculatus, and Rh. decoloratus). Rickettsia conorii subsp. israelensis was detected in one tick pool. These findings indicate that multiple Rickettsia spp. capable of causing human illness are circulating in the four diverse geographical regions of Uganda including new strains previously known to occur in the Mediterranean region. Physicians should be informed about Rickettsia spp. as potential causes of acute febrile illnesses in these regions. Continued and expanded surveillance is essential to further identify and locate potential hotspots with Rickettsia spp. of concern.Item The danger of free-roaming dogs in peri urban settings: Two rabid dogs on rampage that bit 29 humans and 18 domestic animals in Wakiso and Luweero districts, Central Uganda(Research Square, 2024) Tayebwa, Dickson Stuart; Njalira, Kassim Rashid; Eneku, Wilfred; Ssekandi, Colin; Kyabarongo, Alex; Mutesasira, Jesse; Komugisha, Mariam; Atim, Stella Acaye; Kateregga, John; Muhangi, Denis; Tweyongyere, Robert; Biryomumaisho, Savino; Okwee Acai, JamesBackground Uganda averages 14,000 dog bites annually, with 86% caused by free-roaming dogs. Unfortunately, there is limited documentation to inform interventions. Here, we documented circumstances of attacks from free-roaming dogs on rampage in Central Uganda to identify critical gaps in the detection, preparedness, and response of the community. Methods We conducted an exploratory case study to actively survey attacks from rampaging dogs in Kampala Metropolitan Area, Central Uganda between November 2022 - November 2023. We traced the dogs responsible to immobilise and rule out rabies. We interviewed the victims and community to determine the circumstances of the bites, the healthcare sought and sentiments towards free-roaming dogs. We followed-up after 40 days, to assess adherence to rabies post exposure prophylaxis and to document any challenges encountered. Results Of the 8 cases reported, only two cases that occurred in Matugga and Mawale parishes fulfilled our criteria for rampage while the rest were predation, an incidental bite and straying into someone’s residence. The two rampaging dogs’ bit 29 humans, mostly children (75.8%) and 18 domestic animals. Immobilisation in Matugga failed while the dog in Mawale was killed by the community in a mob. Due to limited knowledge, 93.1% of the victims did not wash the wound and go the medical facility as recommended by WHO. Instead, they sought healthcare from the traditional healer (58.6%), stayed home or applied herbs (20.7%). Parallel with our advice, those victims sought medical care however they encountered challenges not limited to lack of funds and poor services at public health facilities. After inflicting bites, the dogs responsible and other free-roaming dogs were killed by hitting them with sticks in a mob. Conclusions Overall, our findings demonstrate the impact that a lone free-roaming dog with rabies can have on a community. They also reveal how poverty, limited awareness, reliance on traditional healers, and barriers to medical care access impede health care received by dog bite victims in this setting. Implementing educational campaigns and responsible ownership of free-roaming dogs should help reduce dog bites and rabies in Central Uganda, as well as in other socioeconomically similar settings.Item The danger of free roaming dogs in urban settings: A report of two suspected rabid dogs on rampage that inicted 47 bites to people and domestic animals in peri-urban Uganda(Reseaech Sqaure, 2023) Tayebwa, Dickson; Njalira, Kassim Rashid; Eneku, Wilfred; Ssekandi, Colin; Kyabarongo, Alex; Mutesasira, Jesse; Komugisha, Mariam; Atim, Stella Acaye; Kateregga, John; Muhangi, Denis; Tweyongyere, Robert; Tony, Lawrence Goldberg; Biryomumaisho, Savino; Okwee Acai, JamesBackground Human rabies cases associated with bites from rabid roaming dogs are prevalent in Uganda. Unfortunately, there is limited documentation of the circumstances surrounding those bites. Here, we documented dog attacks to humans and animals in two peri-urban areas in Central Uganda. Methods We responded to alerts of two suspected rabid dogs on rampage. We traced the dogs responsible for the attacks to diagnose the cause of the aggression and we interviewed the victims and community members to determine the circumstances of the bites and health care sought by the victims. We scheduled follow-up visits after 40 days, to assess compliance of the victims to rabies postexposure prophylaxis (PEP) and to document any challenges encountered in the process. Results The two rabid dogs inflicted 47 recorded attacks to 29 humans, mostly children (75.8%) and 18 domestic animals in Matugga and Mawale parishes in Wakiso and Luweero districts, respectively. Two individuals washed the wound with water and 7 individuals reported to the health facility for treatment in accordance with the WHO-guidelines for preclinical management of bites from suspected rabies cases. By contrast, 17 individuals sought treatment from a traditional healer while 5 individuals self-medicated with herbs at home. We advised those individuals to visit health facilities for proper medical care. All the victims started rabies PEP treatment; however, 2 individuals did not complete their prescription. Although rabies PEP at public health facilities was free, regularly the victims paid 3–5 USD to the clinician as an incentive while the cost at private health facilities was 13–15 USD for a single dose. Due to the unprovoked attacks, the victims and local leaders advocated for elimination of roaming dogs, but the roaming dog owners objected. Conclusions Overall, our results demonstrate the outsized impact that a single-rabid dog can have on a community. They also reveal how poverty, knowledge gaps, community trust in traditional healers, and barriers to healthcare access negatively affect medical care given to dog bite victims in this setting. Educational campaigns and control of roaming dogs should help improve the situation in Central Uganda, as well as in other socioeconomically similar settings.Item Spatio-temporal analysis of sheep and goat pox outbreaks in Uganda during 2011–2022(BMC Veterinary Research, 2023) Nizeyimana, Gerald; Vudriko, Patrick; Erume, Joseph; Mubiru, Frank; Eneku, Wilfred; Biryomumaisho, Savino; Mwebe, Robert; Arinaitwe, Eugene; Rose, Ademun; Atim, Stella; Ayebazibwe, Chrisostom; Muhanguzi, Dennis; Tweyongyere, RobertBackground 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.Item Seroprevalence of Q-fever, spotted fever, typhus group Rickettsia and Orientia among febrile patients visiting hospital-based sentinel sites in Uganda(PAMJ - One Health, 2023) Eneku, Wilfred; Erima, Bernard; Byaruhanga, Anatoli Maranda; Nora, Gillian Cleary; Atim, Gladys; Tugume, Titus; Ukuli, Qouilazoni Aquino; Kibuuka, Hannah; Mworozi, Edison; Christina, Douglas; Jeffrey, William Koehler; Michael, Emery von Fricken; Biryomumaisho, Savino; Tweyongyere, Robert; Wabwire-Mangen, Fred; Byarugaba, Denis KaruhizeIntroduction: rickettsioses are emerging zoonotic febrile illnesses transmitted to humans by ticks, fleas, lice, and mites. Q-fever, Spotted fever group (SFG), Typhus group (TG) rickettsia and Scrub typhus (STG) have been reported with varying prevalence across East Africa. However, little is known about the burden of exposure in Uganda. The aim of this study was to determine the seroprevalence and associated risk factors of rickettsial diseases in Uganda. Methods: a total of 460 archived serum samples collected from patients with fever of unknown origin after screening across five hospital-based sentinel sites were analysed. The samples were collected during 18-month period of active surveillance for acute febrile illnesses, from January 2018 through June 2019. We performed IgM ELISA tests on the 460 sera for SFG and TG rickettsia, IgM IFA for STG and Phase 2 IgG ELISA for Q-fever. We also assessed risk factors associated with the serostatus. Results: the population comprised predominantly children, had balanced gender proportions, with 66% coming from rural areas. The overall seroprevalence of SFG rickettsiosis was 6.3%; however, 11.5% and 10.8% prevalence rates were observed in Gulu and Bwera hospitals respectively. This was higher than the 3.7% observed in the capital city Kampala, although the differences were not statistically significant (Fisher's exact = 0.489). Overall seropositivity of Q-fever was 7.6%, although Bwera Hospital had the highest rate (12.5%) and Mulago had the lowest rate (2%). The differences were not considered statistically significant (Fishers exact= 0.075). Increasing age (OR-adjusted=1.4, 95%CI=1.0-1.9, p=0.026) and rural background (OR-adjusted=2.6, 95%CI=1.6- 6.4, p=0.037) were both significantly associated with seropositivity for Q-fever, while only increasing age had higher odds for seropositivity for SFG rickettsia (OR-adjusted= 1.9, 95% CI= 1.4- 2.6, p<0.001). One serum sample of a 10-monthold male from Bwera hospital was reactive to both SFG and Q-fever antibodies. We found four sera reactive cases to typhus group IgM and another four reactive to Orientia spp. IgM. However, we were not able to determine associating factors due to low seropositivity rates. Conclusion: here, we report for the first time the seroprevalence of Qfever, SFG and STG in febrile patients in Uganda. This report also provides the second study in over five decades since the earliest report of TG rickettsia. Testing for these pathogens in patients with acute febrile illness with unknown etiology may hold value, however more studies are required to provide information on disease ecology, risk factors, and transmission dynamics of these pathogens in Uganda.Item Retrospective study on cattle and poultry diseases in Uganda(International Journal of Veterinary Science and Medicine, 2017) Byaruhanga, Joseph; Tayebwa, Dickson S.; Eneku, Wilfred; Afayoa, Mathias; Mutebi, Francis; Ndyanabo, Susan; Kakooza, Steven; Okwee-Acai, James; Tweyongyere, Robert; Wampande, Eddie M.; Vudriko, PatrickCattle and poultry enterprises are among the major contributors to food security and socioeconomic empowerment of households in Uganda. However, various diseases constrain their productivity. A two-year retrospective study between April 2012 and March 2014 was conducted using records for cattle and poultry diseases diagnosed at the Central Diagnostic Laboratory (CDL) to determine prevalent diseases in Uganda. The laboratory received 836 samples from poultry (36.3%) and cattle (63.7%). Of the 836 samples, 47.5% had a definitive diagnosis of disease causation. Most of the cattle and poultry diseases diagnosed were protozoan diseases (39.3%) followed by bacterial (21.4%), viral (17.1%), helminthiasis (11.1%), nutritional diseases (4%) and others (7.1%). For poultry, viral diseases (29.5%) and protozoan diseases (27.1%) especially newcastle disease (44.3%) and coccidiosis (100%) respectively, were the most diagnosed. While for cattle, hemo-protozoan parasites (52.1%) were the most prevalent, of which 92.9% were east coast fever infection. Bacterial infection (20.5%) in cattle were the second most diagnosed diseases and mastitis was the most diagnosed (46.2%). In summary, coccidioisis, collibacillosis, newcastle disease, gumboro disease, and avian helminthiasis were the most prevalent poultry diseases while in cattle, east coast fever, helminthiasis, mastitis, brucellosis and rabies were the most frequently diagnosed diseases. This study has identified the major diseases that hinder poultry and cattle production in Uganda. The data generated by CDL could be used for surveillance, monitoring and designing strategic interventions for control of poultry and cattle diseases in Uganda.Item Reflections on Drivers for the Emergence and Spread of Antimicrobial Resistant Bacteria Detected from Chickens reared on Commercial Layer Farms in Mukono District, Uganda(Veterinary Medicine: Research and Reports, 2023) Kakooza, Steven; Tayebwa, Dickson Stuart; Njalira, Kassim Rashid; Kayaga, Edrine Beatrice; Asiimwe, Ismail; Komugisha, Mariam; Wanyana, Mariam; Kisekka, Raymond; Kyabarongo, Alex; Kiryabwire, David H.; Nabatta, Esther; Eneku, WilfredPurpose We investigated the fecal carriage of antimicrobial resistant Escherichia coli and potential practices influencing antimicrobial resistance (AMR) dynamics among poultry farm settings in Mukono District, Uganda. Methods Twenty-nine commercial layer farms were visited and samples collected from 10 birds. The samples were then subjected to culture and sensitivity testing. The investigative framework for antimicrobial stewardship practices (IFAP) was used as a participatory tool to generate data through interviews and observations on antimicrobial use, drivers for use, players, and actions following non-responsive treatment outcomes. Results The cultures done on 290 cloacal swabs yielded a total of 273 Escherichia coli isolates (94.1% recovery rate) which were tested in vitro for their sensitivity to different antibiotics. The prevalence of multi-drug resistant E. coli was 59.3% (162/273). A high prevalence of resistance to tetracycline (91.6%, n = 250) and trimethoprim sulphamethoxazole (70.3%, n = 192) was noted. In this collection of isolates, the prevalence of molecular determinants associated with the predominant phenotypes was; tetA (79.3%; 138/174), tetB (17.2%; 30/174), tetC (7.5%; 13/174), sul1 (11.5%; 20/174), and sul2 (60.3%; 105/174). Responses derived using the IFAP revealed several vices related to misuse and overuse of antibiotics, a threat to the poultry industry. The farmers also reported habits of selling off sick birds for slaughter when treatment outcomes were non-responsive. Such a practice could drive dissemination of antimicrobial resistant organisms and antibiotic residues to the consumers of those poultry products. Conclusion The IFAP tool was useful and can be modified, and adopted for use in engaging agricultural communities in participatory AMR surveillance. A high carriage of multi-drug resistant E. coli was detected in the birds. On these farms, the worrying antimicrobial stewardship practices discovered could be sponsoring the emergence and spread of antimicrobial resistant bacteria in the Ugandan context.Item Porcine Circovirus type 2 – Systemic disease on pig farms and associated knowledge of key players in the pig industry in Central Uganda(International Journal of Veterinary Science and Medicine, 2018) Eneku, Wilfred; Mutebi, Francis; Mwiine, Frank Norbert; Okwee-Acai, James; Ojok, LonzyPorcine Circovirus type 2 (PCV2) infections and associated diseases have been rarely studied in Africa. There is no report of PCV2 infection-associated morbidity and the level of awareness of stakeholders has never been investigated in Uganda. This cross sectional survey investigated the occurrence of Porcine Circovirus type 2 – systemic disease (PCV2-SD) among pigs and the associated level of awareness of stakeholders in Central Uganda. Data were collected using questionnaires, Focus Group Discussions (FGDs), key informant interviews and laboratory investigations. All respondents (n = 131) and farmers attending FGDs (n = 31) had never heard of PCV2-SD and only 16.7% (n = 2) of the interviewed animal health workers (n = 12) knew about the disease. Among the farms, 20 piglets presenting with a chronic wasting and a persistent diarrhea were detected and sampled for laboratory investigations. Severe lymphoid depletion with histiocytic and macrophage infiltration in lymphoid organs (n = 8), shortening of intestinal villi (n = 9), abscesses in various organs (n = 15) and granulomatous pneumonia (n = 2) were the major histopathological lesions described. Immunohistochemistry and PCR assays on organs with implicating lesions confirmed PCV2 infection in 25% (n = 5) of the 20 pigs. The study confirmed the occurrence of PCV2 infections among piglets with persistent diarrhea on pig farms in central Uganda and revealed a low level of associated knowledge among farmers and veterinary practitioners. The study arouses the need for systematic studies on prevalence of PCV2 infections and sensitization of stakeholders on occurrence of PCV2 infections in Uganda.Item Molecular detection of Coxiella burnetii in ticks collected from animals and the environment in Uganda(Zoonoses and Public Health, 2024) Eneku, Wilfred; Erima, Bernard; Maranda Byaruhanga, Anatoli; Nora, Cleary; Atim, Gladys; Tugume, Titus; Ukuli, Qouilazoni Aquino; Kibuuka, Hannah; Mworozi, Edison; Tweyongyere, Robert; Christina, E. Douglas; Jeffrey, W. Koehler; Michael, E. von Fricken; Wabwire-Mangen, Fred; Byarugaba, Denis K.Aims Coxiella burnetii is a highly infectious organism that is easily spread through aerosols causing Q fever in humans. Ticks can harbour and transmit C. burnetii to animals, contributing to disease maintenance. Our aim was to examine the presence of C. burnetii in ticks in Uganda. Methods and Results In this study, ticks were collected from five Ugandan districts and tested by real-time PCR for C. burnetii (Coxiella outer membrane protein 1 gene). A total of 859 tick pools (9602 individual ticks) were tested, and pool positivity for C. burnetii was 5.5% (n = 47). Pooled prevalence differed by district; the highest was Luwero (7.3%), then Gulu (6.6%), and Kasese had the lowest (1.3%). However, district variation was not statistically significant (Fisher's exact = 0.07). Ticks collected from dogs and cats had the highest positivity rates [23/47, (48.9%)] followed by livestock (cattle, goats, sheep, and pigs) [18/47, (38.3%)] and vegetation [6/47, (12.8%)]. Haemaphysalis elliptica had the highest infection rates, followed by Rhipicephalus appendiculatus, Amblyomma variegatum and Rhipicephalus decoloratus had similar prevalence. Conclusions Although ticks are not the primary transmitters of C. burnetii to humans, pathogen detection in ticks can be an indirect indicator of risk among animal hosts. Vulnerable populations, including occupations with close animal contact such as farming, butchery, and veterinary practice, have an increased risk of C. burnetii exposure. Veterinarians and clinicians should be aware that C. burnetii may cause human and animal illness in these regions.Item Molecular detection and characterization of emerging pathogens of Rickettsia- felis and felis-like organisms from peri-domestic eas in Uganda(Research Article, 2023) Eneku, Wilfred; Erima, Bernard; Maranda Byaruhanga, Anatoli; Nora, G. Cleary; Atim, Gladys; Tugume, Titus; Ukuli, Qouilazoni A.; Kibuuka, Hannah; Mworozi, Edison; Christina Douglas; Jeffrey W. Koehler; Michael E. Fricken; Biryomumaisho, Savino; Matovu, Enock; Tweyongyere, Robert; Wabwire-Mangen, Fred; Byarugaba, Denis K.Background: Flea-borne spotted fever is an emerging zoonosis caused by Rikecttsia felis, a Gram-negative obligate intracellular bacterium. The agent is believed to be cosmopolitan, following the distribution pattern of its host and reservoir, Ctenocephalides felis. However, the epidemiology and public health risk it poses remains poorly understood in sub-Saharan Africa, including Uganda. Yersinia pestis, is primarily transmitted by rodent fleas, Xenopsylla cheopis, but other fleas, particularly C. felis, have vectoral capacity. They are neglected in Ugandan entomological surveillance and public health practices, particularly outside endemic foci of bubonic plague. Methods: We collected 14,641 fleas from domestic animals, rodents and homestead environment; compared their diversity and abundance. Pooled into 714 flea pools by species, collection time, host, and host species, 172 pools were selected based on seasons and analyzed for Yersinia pestis Pla genes, while 62 pools were tested for Rickettsia species gltA, ompA, and 17kDA genes by qPCR and Sanger sequencing. Results: Five flea species were identified from the collections: Ctenocephalides canis, C. felis, Echidnophaga gallinacea, Pulex irritans, and Xenopsylla cheopis. Ctenocephalides was the predominant genus, accounting for 84.8% of fleas collected, mostly found on dogs and goats. Except for P. irritans (which was found in Gulu district) the other four flea species were found across all districts, year-round, with higher numbers collected in dry seasons compared to rainy seasons (c2=47.64, df=20, p<0.001). Rattus rattus constituted 74% of rodents captured from human dwellings and was the only rodent species with fleas, where X. cheopis was the predominant species and E. gallinacea found on only three rodents. All 172 pools of fleas tested negative for Yersinia pestis. Of the 62 pools tested for Rickettsia spp., 29 (46.8%) were positive. Twenty-five PCR amplicons were successfully sequenced for 17kDa and two for ompA genes. Based on 17kDa, two were identified as R. felis from C. canis and 23 were R. asembonensis from multiple flea species, including C. canis collected goats and C. felis from cats. Conclusion: Our survey identified a high pooled detection rate (~50%) of Rickettsia spp. in fleas tested, suggesting a potential risk of human exposure and infection. Rickettsia felis and R. asembonensis were the predominant flea-borne Rickettsia spp. identified, with this study also representing the first report of Rickettsia spp. in E. gallinacea in Uganda.Item Integrating multi-wet laboratory diagnostics to study staphylococci in animals in Uganda(BMC Microbiology, 2024) Kakooza, Steven; Eneku, Wilfred; Nabatta, Esther; Wampande, Eddie M.; Ssajjakambwe, Paul; Wanyana, Mariam; Munyiirwa, Damien F. N.; Ndoboli, Dickson; Namuyinda, Dorcus; Athieno, Grace; Kayaga, Edrine; Okwasiimire, Rodney; Sayaka, Tsuchida; Kazunari, Ushida; Ken’ichi, Sakurai; Mutebi, FrancisBackground Several diagnostic environments in Uganda lack real-time, robust and high-throughput technologies for comprehensive typing of microbes, which is a setback to infectious disease surveillance. This study combined various wet laboratory diagnostics to understand the epidemiology of pathogenic staphylococci isolated from animals in Uganda and the implications for global health security priorities. Methods A retrospective study was conducted employing records and pathogenic staphylococci (from animals) archived at the Central Diagnostic Laboratory (CDL), Makerere University, Uganda, between January 2012 and December 2019. The bacteria were speciated by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and tested for virulence factors [beta lactamases, lecithinase, deoxyribonuclease (DNase), haemolysins] and resistance to ten antimicrobials of clinical and veterinary relevance. Tetracycline and methicillin resistance genes were also tested. Results The prevalent diseases were mastitis in cattle and skin infections in dogs. Of the 111 staphylococci tested by MALDI-TOF MS, 79 (71.2%) were Staphylococcus aureus, 27 (24.3%) were Staphylococcus pseudintermedius and 5 (4.5%) were Staphylococcus schleiferi. All these strains expressed haemolysins. The prevalence of strains with lecithinase, penicillinase, cephalosporinase and DNase was 35.9% (14/39), 89.7% (35/39), 0.0% (0/39) and 87.2% (34/39), respectively. Staphylococci were primarily resistant to early penicillins (over 80%), tetracycline (57.7%), and chloramphenicol (46.2%). Minimal resistance was noted with cloxacillin (0.0%), ciprofloxacin (9.6%), and cefoxitin (3.8%). The prevalence of multidrug resistance (MDR) was 78.8% for general staphylococci, 82.2% for S. aureus, 73.1% for S. pseudintermedius, and 60.0% for S. schleiferi. Multidrug resistant staphylococci were significantly more prevalent in the cattle isolates than in the dog isolates (P < 0.05). The prevalence of methicillin-resistant staphylococci (MRS) tested by resistance to cefoxitin and mecA carriage was 3.8%. These four strains were all isolated from dog skin infections. The tetK gene was the most predominant (35.4%), followed by tetM (25.0%). Conclusion In resource-constrained settings, the approach of integrated diagnostics promises sustainable disease surveillance and the addressing of current capacity gaps. The emergence of MRS (zoonotic bacteria) in companion animals creates a likelihood of reduced treatment options for related human infections, a threat to global health.Item In vivo aphrodisiac efficacy of aqueous and hydroalcoholic extracts of the leaf and root bark of Citropsis articulata in male Wistar rats(Research Article, 2024) Amutuhaire, Treasure Angie; Tusiimire, Jonans; Amito, Vivian Sharon; Ajayi, Clement Olusoji; Angupale, Jimmy Ronald; Abdelgadir, Alamin Abdelgadir; Anywar, Godwin; Ogwang, Patrick Engeu; Amaza, Anyase RonaldBackground The African cherry orange tree Citropsis articulata Swingle & Kellerman (family Rutaceae) is traditionally used to manage erectile dysfunction. This study aimed to determine the aphrodisiac activity of C. articulata leaves and root bark in male Wistar rats. Methods Aqueous and hydroalcoholic extracts of leaves and root bark were prepared via decoction and soxhlation, respectively. Extracts were administered to the animals at three dose levels, i.e., 100, 500 and 1000 mg/kg. Six male animals (aged 8–12 weeks, weighing 140–200 g) were used in each of the groups. Three animals per group (group B), unlike those immediately sacrificed at the end of 28 days (group A), were left for an extra 14 days without dosing to look out for delayed or sustained efficacy signs. Male sexual behavior, including attraction towards the female, penile erection, mount frequency, and mount latency, was monitored on days 0, 3, 7, 14, 21, 28, 35 and 42. At the end of both 28 days and the extra 14 days, the animals were humanely sacrificed via anesthesia with halothane, and their blood was collected for testosterone level analysis, while the sexual organs harvested were assessed grossly and histologically. Results Citropsis articulata root bark and leaves increased the attraction towards the female, penile erection, and mount frequency while lowering the mount latency. The plant extracts also increased testosterone levels after 28 days of daily extract administration but greatly lowered the testosterone levels in the animals that were left for an extra 14 days without dosing. Even histologically, there was the first evidence of very active spermatogenesis, which disappeared when the administration of the extract ceased. The administration of C. articulata leaf decoction at a dose of 500 mg/kg significantly increased the testosterone level (14.50 ± 2.53 ng/ml) compared with that of the positive control (4.00 ± 2.70 ng/ml), with a p-value of 0.021 (confidence interval: -19.45 to -1.533). Conclusion This study revealed that both the root bark and leaf of Citropsis articulata have aphrodisiac efficacy in male Wistar rats in vitro.Item High intensity of Tunga penetrans infection causing severe disease among pigs in Busoga, South Eastern Uganda(BMC Veterinary Research, 2017) Mutebi, Francis; Jürgen, Krücken; Hermann, Feldmeier; Waiswa, Charles; Norbert, Mencke; Eneku, Wilfred; Georg, von Samson-HimmelstjernaBackground: Towards the improvement of stakeholders’ awareness of animal tungiasis, we report 10 unusual severe clinical cases of pig tungiasis which were associated with very high infection intensities of T. penetrans in an endemic area. Results: Morbidity of ten pigs with high sand flea intensities detected during high transmission seasons in an endemic area in Busoga sub region, Uganda is described in detail. The cases of pigs presented with a very high number of embedded sand fleas (median = 276, range = 141–838). Acute manifestations due to severe tungiasis included ulcerations (n = 10), abscess formation (n = 6) and lameness (n = 9). Chronic morpho pathological presentations were overgrowth of claws (n = 5), lateral deviation of dew claws (n = 6), detachment (n = 5) or loss of dew claws (n = 1). Treatment of severe cases with a topical insecticidal aerosol containing chlorfenvinphos, dichlorvos and gentian violet resolved acute morbidity and facilitated healing by re-epithelialisation. Conclusions: The presentations of tungiasis highlighted in this report show that high intensities of embedded T. penetrans can cause a severe clinical disease in pigs. Effective tungiasis preventive measures and early diagnosis for treatment could be crucial to minimize its effects on animal health. T. penetrans can cause a severe clinical disease in pigs. Effective tungiasis preventive measures and early diagnosis for treatment could be crucial to minimize its effects on animal health.Item Bacteriophage activity against and characterisation of avian pathogenic Escherichia coli isolated from colibacillosis cases in Uganda(PLoS ONE, 2020-12) Kazibwe, George; Katami, Phionah; Alinaitwe, Ruth; Alafi, Stephen; Nanteza, Ann; Nakavuma, Jesca LukangaAvian Pathogenic Escherichia coli (APEC) cause colibacillosis leading to significant economic losses in the poultry industry. This laboratory-based study aimed at establishing stocks of avian pathogenic Escherichia coli lytic bacteriophages, for future development of cocktail products for colibacillosis management. The study determined the antibiotic susceptibility; phylogenetic categories, occurrence of selected serotypes and virulence genes among Escherichia coli stock isolates from chicken colibacillosis cases; and evaluated bacteriophage activity against the bacteria. Escherichia coli characterization was done through phenotypic and multiplex PCR methods. Bacteriophage isolation and preliminary characterization was achieved using the spot assay and overlay plating techniques. Fifty-six (56) isolates were phenotypically confirmed as E. coli and all exhibited resistance to at least one antimicrobial agent; while multi-drug resistance (at least three drugs) was encountered in 50 (89.3%) isolates. The APEC isolates mainly belonged to phylogroups A and D, representing 44.6% and 39.3%, respectively; whereas serotypes O1, O2 and O78 were not detected. Of the 56 isolates, 69.6% harbored at least one virulence gene, while 50% had at least four virulence genes; hence confirmed as APEC. Virulence genes, ompT and iutA were the most frequent in 33 (58.9%) and 32 (57.1%) isolates respectively; while iroN least occurred in 23 (41.1%) isolates. Seven lytic bacteriophages were isolated and their host range, at 1×108 PFU/ml, varied from 1.8% to 17.9% of the 56 APEC isolates, while the combined lytic spectrum was 25%. Phage stability was negatively affected by increasing temperatures with both UPEC04 and UPEC10 phages being undetectable at 70˚C; whereas activity was detected between pH 2 and 12. The high occurrence of APEC isolates resistant against the commonly used antibiotics supports the need for alternative strategies of bacterial infections control in poultry. The low host range exhibited by the phages necessitates search for more candidates before in-depth phage characterization and application. control in poultry. The low host range exhibited by the phages necessitates search for more candidates before in-depth phage characterization and application.Item Animal and human tungiasis-related knowledge and treatment practices among animal keeping households in Bugiri District, South-Eastern Uganda(Acta tropica, 2018-10) Mutebi, Francis; Jürgen, Krückenb; Georg, von Samson-Himmelstjernab; Waiswa, Charles; Norbert, Mencke; Eneku, Wilfred; Tamale, Andrew; Hermann, FeldmeierBackground: Zoonotic tungiasis caused by Tunga penetrans remains a serious public and animal health problem among endemic villages in Uganda and many sub Saharan African countries. Studies on human and animal tungiasis-related knowledge and treatment practices in endemic communities have never been undertaken, a limitation to development of sustainable control measures. Methods: A cross sectional study using semi-structured questionnaires (Supplementary file S1) was conducted among 236 animal rearing households in 10 endemic villages in Bugiri District, South-Eastern Uganda. Focus group discussions and observation checklists were used to validate and clarify the findings. Results: Most respondents knew the aetiology (89.4%), clinical signs (98%) and the ecology of T. penetrans as well as the major risk factors of human tungiasis (65.2%). In contrast, very few respondents were aware of animal tungiasis. Only 4.8% of those with infected animals on the compound knew that some of their animals were infected and 13.6% of the respondents had ever seen tungiasis-affected animals. Pigs (13.1%, n = 31) and dogs (0.85%, n = 2) were the only T. penetrans animal hosts known to animal owners. Affected humans were treated by extraction of embedded sand fleas using non-sterile sharp instruments in all households that reported occurrence of human tungiasis at least once (n = 227). Also, affected animals were mainly treated by mechanical removal of embedded sand fleas in households that have ever experienced animal tungiasis (four out of 12; 33.3%). In a few instances, plant and animal pesticides (n = 3) and other chemicals such as grease, paraffin and wood preservative (n = 3) were also used to treat animal tungiasis. Conclusion: The study revealed a high level of knowledge on human tungiasis but inadequate knowledge on the zoonotic nature of tungiasis. Commonly applied methods for treatment of human and animal tungiasis are a health hazard by themselves. Concerted i.e. One Health-based efforts aiming at promoting appropriate treatment of tungiasis, adequate living conditions and increased awareness on tungiasis in the communities are indicated in order to eliminate tungiasis-associated disease.Item A retrospective analysis of antimicrobial resistance in pathogenic Escherichia coli and Salmonella spp. isolates from poultry in Uganda(International Journal of Veterinary Science and Medicine, 2021-05) Kakooza, Steven; Muwonge, Adrian; Nabatta, Esther; Eneku, Wilfred; Dickson, Dickson; Wampande, Eddie; Munyiirwa, Damian; Kayaga, Edrine; Tumwebaze, Maria Agnes; Afayoa, Mathias; Ssajjakambwe, Paul; Tayebwa, Dickson Stuart; Sayaka, Tsuchida; Okubo, Torahiko; Kazunari, Ushida; Sakurai, Ken’ichi; Mutebi, FrancisThere are increasing reports of antimicrobial treatment failures for bacterial diseases of poultry in Uganda. The paucity of data on antimicrobial resistance (AMR) of pathogenic bacteria in Uganda is a major setback to AMR control. This study investigated the occurrence of fowl typhoid, colibacillosis, and AMR in associated pathogens from 2012 to 2018. Laboratory records from the Central Diagnostic Laboratory (CDL), a National Veterinary Diagnostic Facility located at Makerere University, were reviewed. Archived isolates of the causative bacteria for the two diseases were also evaluated for AMR. The frequencies of the two disease conditions, their clinical and necropsy presentations and the demographic data of the diagnostic samples were summarized from the records. Archived bacterial isolates were revived before antimicrobial susceptibility testing. This was done on Mueller Hinton agar using the disk diffusion method, against 16 antimicrobials of medical and veterinary importance according to the Clinical Laboratory Standards Institute guidelines. A total of 697 poultry cases were presented for bacteriological investigations in the review period. Colibacillosis and salmonellosis had prevalence rates of 39.7% (277/697) and 16.2% (113/697), respectively. A total of 63 and 92 isolates of Escherichia coli and Salmonella spp., respectively, were archived but 43 (68.3%) E. coli and 47 (51.1%) Salmonella spp. isolates were recovered and evaluated for AMR. Multidrug resistance was more frequent in E. coli (38; 88.4%) than salmonellae (25; 53.2%), (p < 0.001). The high prevalence of colibacillosis, salmonellosis and the AMR of associated pathogens warrants immediate institution of appropriate disease control measures.Item Structure and milk hygiene of dairy cooperative value chains in an intensive production area of Uganda—A bottleneck of intervention(Frontiers in Sustainable Food Systems, 2023) Yoshiharu, Sugino; Bugeza, James; Bahame, David; Byaruhanga, Joseph; Haruka, Shimazaki; Masahiko, Anzai; Taishi, Kayano; Mwebembezi, William; Akashaba, Andrew; Shimada, Taku; Yasukazu, Muramatsu; Kohei, MakitaIn Uganda, informal raw milk sales dominate for domestic dairy consumption. This study was implemented to identify the structure of the dairy value chain starting from farms that participated in the Japan International Cooperation Agency Safe Milk Promotion in Mbarara project conducted between and, to assess the hygiene conditions along the chain, and thereby identify the bottleneck of dairy hygiene intervention. A longitudinal study was conducted in dairy farms in Mbarara District to compare the practice, prevalence of subclinical mastitis, and level of milk hygiene in– hygiene intervention in and, before and after the milking. California Mastitis Test was used for diagnosis with subclinical mastitis. Bulk milk samples were collected and a checklist was used to examine hygiene practices by observation. Across sectional study was conducted in milk collecting centers using a structured questionnaire to quantify the dairy value chain, and to sample milk from cooler tanks in Microbiological examinations of bulk milk from farms and collection centers were conducted using six point blood agar scoring and M Petri film, respectively. Participatory online appraisals with farmers and dairy cooperatives union were conducted to better understand the overall dairy value chains. The cooperatives sold milk to both formal and informal chains, but the sale of raw milk to Kampala was conducted by independent private traders. Within herd prevalence of subclinical mastitis significantly decreased from . % before the intervention to . % after (p < . ).However, the farm bulk milk score did not change ( . vs. . , p = . ). A significant increase in the total bacterial count was observed in the milk from collection centers (mean: . log CFU/ml) when compared to farm bulk milk (mean: . log CFU/ml; p < . ). Only . % of the samples from the centers met the microbiological criteria for processing for human consumption. Our findings suggest that intervention targeted only at mastitis does not lead to better public health due to the low level of hygiene in transportation and milk handling in milk collection centers. Systematic interventions are needed to improve post harvest dairy hygiene in Uganda.Item Sero-prevalence and risk factors associated with occurrence of anti-Brucella antibodies among slaughterhouse workers in Uganda(PLOS Neglected Tropical Diseases, 2024-03) Bugeza, James Katamba; Alinaitwe, Lordrick; Kristina, Roesel; Mugizi, Denis Rwabiita; Kivali, Velma; Kankya, Clovice; Ignacio, Moriyon; Elizabeth, AnneJessieCookBrucellosis is a febrile zoonosis occurring among high-risk groups such as abattoir workers and is a public health priority in Uganda. Whereas bacteriological isolation is conclusive, and some molecular methods have been found useful for diagnosis of brucellosis, they are technically complex and may delay commencement of treatment. Therefore, clinicians from resource poor settings rely on clinical examination and serology for diagnosis of human brucellosis. However, brucellosis lacks pathognomonic signs, and clinically resembles other endemic febrile illnesses which complicates diagnosis. Poor quality Brucella antigens, serological tests not validated for human use, and lack of consensus on the tests of choice complicate diagnosis of brucellosis. Previously, many studies employed a variety of tests to estimate sero-prevalence of brucellosis in high-risk groups leading in some cases to over estimation of the disease in Uganda. Here we applied the RBT and BrucellaCapt tests in a serial testing scheme to detect contacts, short and long evolution cases of brucellosis and report a sero-prevalence ranging from 7.3% to 9.0% among slaughter house workers in Uganda. Brucella seropositivity was associated with the region where participants worked, the slaughter of cattle, small ruminants, and grazing animals as an activity outside the work of the slaughterhouse.Item Sero-prevalence and Risk Factors Associated with Bovine Brucellosis in Central Equatoria State, South Sudan(Scholars Journals of Agriculture and Veterinary Science,, 2016) Lita, Emmanuel P.; Nasinyama, George W.; Ochi, Erneo B.; Bugeza, James; Erume, JosephBrucellosis is one of the most important zoonotic diseases of livestock and human in South Sudan. A cross sectional study was conducted in peri-urban Juba town and rural Terekeka County to estimate sero-prevalence and determine risk factors of bovine brucellosis. Ninety (90) respondents were randomly interviewed on demographic information and potential risk factors to brucellosis. Out of the 90 respondents interviewed, 44 were from 44 cattle herds in peri-urban Juba and 46 from 46 cattle herds in rural Terekeka County. Sera of 502 cattle were screened for Brucella antibodies using Rose Bengal Plate test. The positive sera on RBPT were then confirmed by competitive Enzyme-linked Immunosorbent Assay. SPSS version-18 and frequencies were used for sero-prevalence. Chi square and binomial logistic regressions were used for analysis of risk factors. RBPT and c-ELISA tests showed positive reactors and an overall individual animal sero-prevalence of 23.2%, 95% confidence interval (CI): (18.4 - 28.8) and 19.2%, 95% CI: (2.5 – 14.0), respectively in peri-urban Juba town. However, in rural Terekeka County the respective results showed seroprevalence of 40.5%, 95% CI: (34.5 – 46.4) and 39.3%, 95% CI: (33.3– 45.2). The overall herd level sero-prevalence on c-ELISA revealed 61.4% and 90.0% for peri-urban Juba town and rural Terekeka County cattle herds, respectively. The individual animal level risk factors in the study area revealed abortion history (OR= 4.941 and (CI): 2.077-11.753) and (OR= 6.251 and (CI): 2.920-13.379) significantly associated with brucella sero-positivity, respectively. No risk factor determined at herd level in peri-urban Juba town. However, in rural Terekeka County herds, number of cattle above 300 in a herd (p= 0.005, OR= 44.934) was significantly associated with brucellosis sero-positivity. Further epidemiological studies are needed for developing appropriate control strategies against bovine brucellosis in South Sudan.Item Sero-prevalence and factors associated with anti-Brucella antibodies in slaughter livestock in Uganda(Frontiers in Epidemiology, 2023-06) Bugeza, James; Kristina, Roesel; Ignacio, Moriyon; Mugizi, Denis; Alinaitwe, Lordrick; Kivali, Velma; Kankya, Clovice; Elizabeth, Anne Jessie CookBrucellosis 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.