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Background: Malaria remains the leading cause of hospitalization and death in the healthcare system. This study explored the malaria burden and community response to government malaria control programs in Omoro district. Method: This retrospective study involved 576 patient results from purposely selected health facilities data from health center III (HCIII) of Odek, Bobi, and Lapainat and health center IV (HCIV) of Lalogi. And prospective random section study involved 288 participants from Lutori and Lagude cells and Atyang A and Lagwaya villages who consented to answer the pretested questionnaire. Results: The prevalence of malaria in Omoro district in 2018 and 2019 was 81.6% and 97.2% for hospital record positivity and community surveys, respectively. The participants had 100% knowledge of malaria signs, symptoms, and cause. The average number of malaria attacks an individual received in the district from the health facility and community data was three. Nonadherence to government control programs was associated with an increased incidence of malaria infections. From questionnaire, the proportion of people that used local herbal remedies for treatment and prevention of malaria were 21.2% of the sampled 288 participants. Conclusion: The high rate of malaria attacks indicates that the area has a high prevalence of malaria-carrying mosquitoes. The increase in the proportion of malaria attacks in 2019 suggests that the burden of malaria increased compared with that in the previous year, with approximately 21% of local herbal remedies for malaria treatment and management. The findings of this study suggest that malaria attacks are associated with household size, age, sex, occupation, and the household head. Participants who did not respond positively to government programs experienced more malaria attacks. These findings can be used to develop interventions to reduce the incidence of malaria in this population., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2024 Divas Soyekwo et al.)

Background: Malaria remains one of the most critical disease causing morbidity and mortality in Uganda. Indoor residual spraying (IRS) and the use of insecticide-treated bed nets are currently the predominant malaria vector control interventions. However, the emergence and spread of insecticide resistance among malaria vectors threaten the continued effectiveness of these interventions to control the disease, particularly in high transmission areas. To inform decisions on vector control, the current study evaluated the Anopheles malaria vector species and their susceptibility levels to 0.1% bendiocarb and 0.25% pirimiphos-methyl insecticides used in IRS intervention program in Namutumba district, Eastern Uganda., Methods: Anopheles larvae were collected between March and May 2017 from different breeding sites in the parishes of Nsinze and Nawaikona in Nsinze sub-county and reared to adults to assess the susceptibility status of populations in the study area. Mosquitoes were identified using morphological keys and species-specific polymerase chain reaction (PCR) assays. Susceptibility tests were conducted on 2- to 5-day-old non-blood-fed adult female Anopheles that emerged using insecticide-impregnated papers with 0.1% bendiocarb and 0.25% pirimiphos-methyl following standard World Health Organization (WHO) insecticide susceptibility bioassays. A Log-probit regression model was used to derive the knock-down rates for 50% and 95% of exposed mosquitoes., Results: A total of 700 mosquito larvae were collected from different breeding sites. Morphological identification showed that 500 individuals that emerged belonged to Anopheles gambiae sensu lato (s.l.), the main malaria vector. The PCR results showed that the dominant sibling species under the A. gambiae complex was Anopheles arabiensis 99.5% (395/397). WHO bioassay tests revealed that the population of mosquitoes exhibited high levels of susceptibility (24-hour post-exposure mortality 98-100%) to both insecticides tested. The median knock-down time, KDT 50 , ranged from 6.6 to 81.4 minutes, while the KDT 95 ranged from 21.6 to 118.9 minutes for 0.25% pirimiphos-methyl. The KDT 50 for 0.1% bendiocarb ranged from 2.8 to 62.9 minutes, whereas the KDT 95 ranged from 36.0 to 88.5 minutes., Conclusions: These findings indicate that bendiocarb and pirimiphos-methyl are still effective against the major malaria vector, A. arabiensis in Nsinze sub-county, Namutumba district, Uganda and can be effectively used for IRS. The study has provided baseline information on the insecticide susceptibility status on malaria vectors in the study area. However, routine continuous monitoring program of insecticide susceptibility and malaria vector composition is required so as to guide future decisions on insecticide use for IRS intervention toward malaria elimination and to track future changes in vector population., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2023 Julius Iga et al.)

Background: Tsetse flies are vectors of the genus Trypanosoma that cause African trypanosomiasis, a serious parasitic disease of people and animals. Reliable data on the vector distribution and the trypanosome species they carry is pertinent for planning sustainable control strategies. This study was carried out to estimate the spatial distribution, apparent density, and trypanosome infection rates of tsetse flies in two districts that fall within a vector genetic transition zone in northern Uganda., Materials and Methods: Capturing of tsetse flies was done using biconical traps deployed in eight villages in Oyam and Otuke, two districts that fall within the vector genetic transition zone in northern Uganda. Trapped tsetse flies were sexed and morphologically identified to species level and subsequently analyzed for detection of trypanosome DNA. Trypanosome DNA was detected using a nested PCR protocol based on primers amplifying the internal transcribed spacer (ITS) region of ribosomal DNA., Results: A total of 717 flies (406 females; 311 males) were caught, all belonging to the Glossina fuscipes fuscipes species. The overall average flies/trap/day (FTD) was 2.20 ± 0.3527 (mean ± SE). Out of the 477 (201 male; 276 females) flies analyzed, 7.13% (34/477) were positive for one or more trypanosome species. Three species of bovine trypanosomes were detected, namely, Trypanosoma vivax , 61.76% (21/34), T. congolense , 26.47% (9/34), and T. brucei brucei , 5.88% (2/34), and two cases of mixed infection of T. congolense and T. brucei brucei , 5.88% (2/34). The infection rate was not significantly associated with the sex of the fly (generalized linear model (GLM), χ 2 = 0.051, p = 0.821, df = 1, n = 477) and district of origin ( χ 2 = 0.611, p = 0.434, df = 1, n = 477). However, trypanosome infection was highly significantly associated with the fly's age based on wing fray category ( χ 2 = 7.56, p = 0.006, df = 1, n = 477), being higher among the very old than the young., Conclusion: The relatively high tsetse density and trypanosome infection rate indicate that the transition zone is a high-risk area for perpetuating animal trypanosomiasis. Therefore, appropriate mitigation measures should be instituted targeting tsetse and other biting flies that may play a role as disease vectors, given the predominance of T. vivax in the tsetse samples., Competing Interests: The authors declare that they have no competing interests., (Copyright © 2022 Robert Opiro et al.)

Background: African trypanosomiasis, caused by protozoa of the genus Trypanosoma and transmitted by the tsetse fly, is a serious parasitic disease of humans and animals. Reliable data on the vector distribution, feeding preference and the trypanosome species they carry is pertinent to planning sustainable control strategies., Methodology: We deployed 109 biconical traps in 10 villages in two districts of northwestern Uganda to obtain information on the apparent density, trypanosome infection status and blood meal sources of tsetse flies. A subset (272) of the collected samples was analyzed for detection of trypanosomes species and sub-species using a nested PCR protocol based on primers amplifying the Internal Transcribed Spacer (ITS) region of ribosomal DNA. 34 blood-engorged adult tsetse midguts were analyzed for blood meal sources by sequencing of the mitochondrial cytochrome c oxidase 1 (COI) and cytochrome b (cytb) genes., Results: We captured a total of 622 Glossina fuscipes fuscipes tsetse flies (269 males and 353 females) in the two districts with apparent density (AD) ranging from 0.6 to 3.7 flies/trap/day (FTD). 10.7% (29/272) of the flies were infected with one or more trypanosome species. Infection rate was not significantly associated with district of origin (Generalized linear model (GLM), χ 2  = 0.018, P = 0.895, df = 1, n = 272) and sex of the fly (χ2 = 1.723, P = 0.189, df = 1, n = 272). However, trypanosome infection was highly significantly associated with the fly's age based on wing fray category (χ 2  = 22.374, P < 0.001, df = 1, n = 272), being higher among the very old than the young tsetse. Nested PCR revealed several species of trypanosomes: T. vivax (6.62%), T. congolense (2.57%), T. brucei and T. simiae each at 0.73%. Blood meal analyses revealed five principal vertebrate hosts, namely, cattle (Bos taurus), humans (Homo sapiens), Nile monitor lizard (Varanus niloticus), African mud turtle (Pelusios chapini) and the African Savanna elephant (Loxodonta africana)., Conclusion: We found an infection rate of 10.8% in the tsetse sampled, with all infections attributed to trypanosome species that are causative agents for AAT. However, more verification of this finding using large-scale passive and active screening of human and tsetse samples should be done. Cattle and humans appear to be the most important tsetse hosts in the region and should be considered in the design of control interventions., (© 2021. The Author(s).)

Objective: The aim of the study was to determine the level of insecticide resistance and diversity in Anopheles mosquitoes in northern Uganda. Standard WHO insecticide susceptibility test assays were used to test for susceptibility to 0.5% malathion, 0.1% bendiocarb, 0.05% deltamethrin and 0.75% permethrin on 3-5 day old generation one progeny. We also screened for species diversity and knockdown resistance using PCR assay., Results: Anopheles gambiae s.s. is the predominant malaria vector in northern Uganda followed by An. arabiensis. An. gambiae s.s. was susceptible to malathion and bendiocarb with the observed mortality rate of 100% and 98-100% observed respectively while very high resistance was observed with deltamethrin and permethrin. Minimal KDR-eastern variant homozygous forms of 8.3% in An. gambiae s.s. were detected in Oyam district. In conclusion, this study confirms that An. gambiae s.s. females are susceptible to malathion and bendiocarb while high intensity of resistance was observed with deltamethrin and permethrin in the same area. Use of carbamate and organophosphate insecticides bendiocarb and malathion for indoor residual spraying activities in northern Uganda is highly recommended since high levels of pyrethroids resistance (deltamethrin and permethrin) was detected in the area.

Understanding the mechanisms that enforce, maintain or reverse the process of speciation is an important challenge in evolutionary biology. This study investigates the patterns of divergence and discusses the processes that form and maintain divergent lineages of the tsetse fly Glossina fuscipes fuscipes in Uganda. We sampled 251 flies from 18 sites spanning known genetic lineages and the four admixture zones between them. We apply population genomics, hybrid zone and approximate Bayesian computation to the analysis of three types of genetic markers: 55,267 double-digest restriction site-associated DNA (ddRAD) SNPs to assess genome-wide admixture, 16 microsatellites to provide continuity with published data and accurate biogeographic modelling, and a 491-bp fragment of mitochondrial cytochrome oxidase I and II to infer maternal inheritance patterns. Admixture zones correspond with regions impacted by the reorganization of Uganda's river networks that occurred during the formation of the West African Rift system over the last several hundred thousand years. Because tsetse fly population distributions are defined by rivers, admixture zones likely represent both old and new regions of secondary contact. Our results indicate that older hybrid zones contain mostly parental types, while younger zones contain variable hybrid types resulting from multiple generations of interbreeding. These findings suggest that reproductive barriers are nearly complete in the older admixture zones, while nearly absent in the younger admixture zones. Findings are consistent with predictions of hybrid zone theory: Populations in zones of secondary contact transition rapidly from early to late stages of speciation or collapse all together., (© 2018 John Wiley & Sons Ltd.)

Tsetse flies (genus Glossina ) are the only vector for the parasitic trypanosomes responsible for sleeping sickness and nagana across sub-Saharan Africa. In Uganda, the tsetse fly Glossina fuscipes fuscipes is responsible for transmission of the parasite in 90% of sleeping sickness cases, and co-occurrence of both forms of human-infective trypanosomes makes vector control a priority. We use population genetic data from 38 samples from northern Uganda in a novel methodological pipeline that integrates genetic data, remotely sensed environmental data, and hundreds of field-survey observations. This methodological pipeline identifies isolated habitat by first identifying environmental parameters correlated with genetic differentiation, second, predicting spatial connectivity using field-survey observations and the most predictive environmental parameter(s), and third, overlaying the connectivity surface onto a habitat suitability map. Results from this pipeline indicated that net photosynthesis was the strongest predictor of genetic differentiation in G. f. fuscipe s in northern Uganda. The resulting connectivity surface identified a large area of well-connected habitat in northwestern Uganda, and twenty-four isolated patches on the northeastern margin of the G. f. fuscipes distribution. We tested this novel methodological pipeline by completing an ad hoc sample and genetic screen of G. f. fuscipes samples from a model-predicted isolated patch, and evaluated whether the ad hoc sample was in fact as genetically isolated as predicted. Results indicated that genetic isolation of the ad hoc sample was as genetically isolated as predicted, with differentiation well above estimates made in samples from within well-connected habitat separated by similar geographic distances. This work has important practical implications for the control of tsetse and other disease vectors, because it provides a way to identify isolated populations where it will be safer and easier to implement vector control and that should be prioritized as study sites during the development and improvement of vector control methods.

Insecticide decay rate on different wall surfaces is of importance to indoor residual spray (IRS) programs used as a malaria control intervention. Past IRS operations showed increasing populations of endophilic malaria vectors resting on indoor surfaces from various sites in Uganda following use of Ficam VC (bendiocarb) insecticide; variability of insecticide life was believed to be primarily due to wall surface type. Bendiocarb longevity was tested in the northern Uganda districts of Amuru, Apac, and Pader to assess its residual efficacy on three commonly encountered wall surfaces. Wall types included mud and wattle, plain brick, and painted plaster. A susceptible mosquito strain (Anopheles gambiae Kisumu) was used in all trials. Nine houses in each of the three districts were set with three test cones and one control cone per house, divided evenly among the three wall surfaces. Bioassays were run monthly through 6 mo. Painted plastered surfaces produced 100% mortality (at 24 h) through 6 mo. Plain brick surfaces killed 100% of test mosquitoes through 4 mo, while mud and wattle wall surfaces produced a 98% mortality rate at 3 mo post spray. The KD60 (knockdown at 60 min) for painted plastered surfaces was 100% for 6 mo, plain brick surface KD60 was 80% at 6 mo, and the mud and wattle surface KD60 was >80% at 3 mo. There was a significant effect on Ficam VC longevity by wall type and evidence of a relationship between test period and wall type on the KD60., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Uganda is the only country where the chronic and acute forms of human African Trypanosomiasis (HAT) or sleeping sickness both occur and are separated by < 100 km in areas north of Lake Kyoga. In Uganda, Glossina fuscipes fuscipes is the main vector of the Trypanosoma parasites responsible for these diseases as well for the animal African Trypanosomiasis (AAT), or Nagana. We used highly polymorphic microsatellite loci and a mitochondrial DNA (mtDNA) marker to provide fine scale spatial resolution of genetic structure of G. f. fuscipes from 42 sampling sites from the northern region of Uganda where a merger of the two disease belts is feared. Based on microsatellite analyses, we found that G. f. fuscipes in northern Uganda are structured into three distinct genetic clusters with varying degrees of interconnectivity among them. Based on genetic assignment and spatial location, we grouped the sampling sites into four genetic units corresponding to northwestern Uganda in the Albert Nile drainage, northeastern Uganda in the Lake Kyoga drainage, western Uganda in the Victoria Nile drainage, and a transition zone between the two northern genetic clusters characterized by high level of genetic admixture. An analysis using HYBRIDLAB supported a hybrid swarm model as most consistent with tsetse genotypes in these admixed samples. Results of mtDNA analyses revealed the presence of 30 haplotypes representing three main haplogroups, whose location broadly overlaps with the microsatellite defined clusters. Migration analyses based on microsatellites point to moderate migration among the northern units located in the Albert Nile, Achwa River, Okole River, and Lake Kyoga drainages, but not between the northern units and the Victoria Nile drainage in the west. Effective population size estimates were variable with low to moderate sizes in most populations and with evidence of recent population bottlenecks, especially in the northeast unit of the Lake Kyoga drainage. Our microsatellite and mtDNA based analyses indicate that G. f. fuscipes movement along the Achwa and Okole rivers may facilitate northwest expansion of the Rhodesiense disease belt in Uganda. We identified tsetse migration corridors and recommend a rolling carpet approach from south of Lake Kyoga northward to minimize disease dispersal and prevent vector re-colonization. Additionally, our findings highlight the need for continuing tsetse monitoring efforts during and after control.

Background: Glossina fuscipes fuscipes is a tsetse species of high economic importance in Uganda where it is responsible for transmitting animal African trypanosomiasis (AAT) and both the chronic and acute forms of human African trypanosomiasis (HAT). We used genotype data from 17 microsatellites and a mitochondrial DNA marker to assess temporal changes in gene frequency for samples collected between the periods ranging from 2008 to 2014 in nine localities spanning regions known to harbor the two forms of HAT in northern Uganda., Results: Our findings suggest that the majority of the studied populations in both HAT foci are genetically stable across the time span sampled. Pairwise estimates of differentiation using standardized FST and Jost's DEST between time points sampled for each site were generally low and ranged between 0.0019 and 0.1312 for both sets of indices. We observed the highest values of FST and DEST between time points sampled from Kitgum (KT), Karuma (KR), Moyo (MY) and Pader (PD), and the possible reasons for this are discussed. Effective population size (Ne) estimates using Waple's temporal method ranged from 103 (95% CI: 73-138) in Kitgum to 962 (95% CI: 669-1309) in Oculoi (OC). Additionally, evidence of a bottleneck event was detected in only one population at one time point sampled; Aminakwach (AM-27) from December 2014 (P < 0.03889)., Conclusion: Findings suggest general temporal stability of tsetse vectors in foci of both forms of HAT in northern Uganda. Genetic stability and the moderate effective population sizes imply that a re-emergence of vectors from local residual populations missed by control efforts is an important risk. This underscores the need for more sensitive sampling and monitoring to detect residual populations following control activities.

Background: Livestock trypanosomiasis, transmitted mainly by tsetse flies of the genus Glossina is a major constraint to livestock health and productivity in the sub-Saharan Africa. Knowledge of the prevalence and intensity of trypanosomiasis is important in understanding the epidemiology of the disease. The objectives of this study were to (a) assess the prevalence and intensity of trypanosome infections in cattle, and (b) to investigate the reasons for the heterogeneity of the disease in the tsetse infested districts of Amuru and Nwoya, northern Uganda., Methods: A cross-sectional study was conducted from September, 2011 to January, 2012. Blood samples were collected from 816 cattle following jugular vein puncture, and screened for trypanosomes by HCT and ITS-PCR. A Pearson chi-squared test and logistic regression analyses were performed to determine the association between location, age, sex, and prevalence of trypanosome infections., Results: Out of the 816 blood samples examined, 178 (22 %) and 338 (41 %) tested positive for trypanosomiasis by HCT and ITS-PCR, respectively. Trypanosoma vivax infection accounted for 77 % of infections detected by ITS-PCR, T. congolense (16 %), T. brucei s.l (4 %) and mixed (T. vivax/ T. congolense/T.brucei) infections (3 %). The risk of trypanosome infection was significantly associated with cattle age (χ (2)   = 220.4, df = 3, P < 0.001). The highest proportions of infected animals were adult males (26.7 %) and the least infected were the less than one year old calves (2.0 %). In addition, the risk of trypanosome infection was significantly associated with sex (χ (2)  = 16.64, df = 1, P < 0.001), and males had a significantly higher prevalence of infections (26.8 %) than females (14.6 %)., Conclusion: Our results indicate that the prevalence and intensity of trypanosome infections are highly heterogeneous being associated with cattle age, location and sex.

A rapid, visually read, dot-ELISA developed for the detection and differentiation of trypanosome species in tsetse flies (Glossina spp.), was field tested alongside the standard fly dissection method on a range in south eastern Kenya. Of 104 G. pallidipes dissected, 2 were found to be infected with trypanosomes in their midguts. By the dissection method the infecting trypanosome species could not be identified, as both flies had no salivary gland infections. However, using the dot-ELISA, the 2 flies were shown to be infected with Trypanosoma congolense in their midguts. The midguts of an additional 6 (5.8%) of the 104 G. pallidipes tested positive for T. congolense int he dot-ELISA, even though no trypanosomes were seen on dissection. The infection rate for this fly species as determined using the dot-ELISA, therefore, was 7.7% for T. congolense in midgut infections compared to 1.9% identified by fly dissection. The salivary glands and mouthparts of the 6 additional tsetse flies identified by dot-ELISA were all negative as determined by the 2 techniques. None of 390 G. longipennis flies dissected and examined for trypanosomes in the midgut, salivary glands and mouthparts was shown, by this method, to be infected. Using the dot-ELISA, however, 17 (4.4%) of the flies tested positve for T. congolense in the midgut, whilst the salivary glands and mouthparts of the same flies were negative. Thus, the dot-ELISA appears to be more sensitive than the fly dissection method under field conditions. Moreover, the dot-ELISA can be performed in the field without electricity. It is simple to perform, and was not affected by high ambient temperatures (22-32 degrees C), or by contamination of reactants with dust.

During an outbreak of Rhodesian sleeping sickness in the Lambwe Valley in 1980 initial tsetse control measures consisted of applications of dieldrin to the periphery of the Ruma National Park. This activity had a marked effect on the prevalence of sleeping sickness. Concern about the use of dieldrin caused the cessation of this programme and justified an aerial spray programme using endosulfan. Although the Lambwe Valley did not appear to be a good candidate for aerial spray, the endosulfan had a marked effect on tsetse fly levels and on the prevalence of sleeping sickness. Sleeping sickness cases were detected in decreasing numbers for eight months following the endosulfan programme, but the subsequent five months yielded no cases of sleeping sickness in the area. Some flies persisted, however, and they had regained high levels in about a year. As the prevalence of sleeping sickness increased another aerial spray programme was initiated in 1983, using pyrethrum as insecticide. The pyrethrum aerial spray programme did not make significant reductions in the Glossina pallidipes population or in the prevalence of sleeping sickness. A subsequent ground control programme using insecticides (dieldrin and cypermethrin) and bush clearing, conducted primarily within the National Park, has subsequently limited the prevalence of sleeping sickness to low levels.

Trypanosoma vivax isolated from dairy cattle undergoing a haemorrhagic disease was inoculated into Ayrshire steers. Five of six infected animals experienced brief periods of diarrhoea and sublingual and gastro-intestinal haemorrhage. Gastro-intestinal bleeding coincided with markedly reduced numbers of thrombocytes and a high level of parasitaemia in the peripheral blood. Prothrombin times were extended and fibrinogen levels were elevated in infected animals. Plasma paracoagulation tests were positive for the presence of fibrin monomers and/or clottable early fibrin degradation products during the course of infection.

Trypanosoma simiae, although similar to T. congolense in morphology and mode of development in the tsetse vector, is regarded as a separate species mainly because it is highly pathogenic to the domestic pig and fails to infect rodents. To establish whether the two species are distinct biochemically, we compared by isoenzyme electrophoresis 2 isolates of T. simiae with 7 stocks of T. congolense, together with one of T. brucei. All isoenzyme patterns of the 6 enzymes examined differed in T. simiae from T. congolense and T. brucei stocks. This supports the designation of T. simiae as a separate species. However, a comparison involving a much larger collection of T. simiae is necessary in order to get conclusive evidence.

Five crossbred cattle infected with Trypanosoma vivax (Likoni) by Glossina morsitans developed capillary haemorrhages at the onset of parasitaemia, followed by the presence of occult blood in faecal samples and eventually melena. Two animals required treatment to survive, on days 13 and 38 respectively. The other three animals cleared their parasitaemias without treatment. Packed cell volume (PCV) levels decreased in all animals to levels ranging from 7.5 to 17%. Relapse in a treated animal initiated marked haemorrhage and a loss of 14 PCV units during a six-day period. Thrombocytopenia was common to all animals, and thrombocytes decreased to levels of 4000/microliters of blood. All animals developed increased levels of fibrinogen and fibrin monomer. Prolonged prothrombin times were found in all animals, and activated partial thromboplastin times were also extended in the two animals with high parasitaemias.