Browsing by Author "Mugasa, Claire Mack"
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Item Comparative detection of African swine fever virus by loop-mediated isothermal amplification assay and polymerase chain reaction in domestic pigs in Uganda(African Journal of Microbiology Research, 2014) Atuhaire, David Kalenzi; Afayoa, Mathias; Katiti, Dianah; Mwiine, Frank Norbert; Nanteza, Ann; Mugasa, Claire Mack; Matovu, Enock; Olaho-Mukani, William; Ojok, LonzyAfrican swine fever (ASF) is a contagious viral disease, which can cause up to 100% mortality among domestic pigs. Pig production is growing rapidly in Uganda among East African countries and is not only a source of food but also an important income for many people living in the rural areas. Field diagnosis of ASF depends only on clinical signs and has to be confirmed in the laboratory since the clinical signs are not pathognomonic. Diagnostic techniques for ASF are focused on serological tests for detection of antigen and antibody, genomic DNA detection by polymerase chain reaction (PCR), and on virus isolation and localization in clinical samples. There have been many recent reports of ASF outbreaks in Uganda yet laboratory diagnosis is limited due to the high cost and expertise required. This work reports the evaluation and application of a loop-mediated isothermal amplification (LAMP) test for detecting African swine fever virus (ASFV) DNA based on the topoisomerase II gene. Thirty (30) tissue samples obtained from suspected ASF outbreaks were collected from different regions of Uganda. The tissue samples were found to have lesions consistent with ASF. One hundred and eighty eight (188) additional blood samples were obtained from the abattoir and field surveillance. Six primers targeting the topoisomerase II gene were used. The sensitivity and specificity of LAMP and OIE recommended diagnostic PCR were compared. The LAMP assay is rapid with results obtained within 1 h (45-60 min). The sensitivity of LAMP for the detection of ASFV was 100% (95% CI: 91.78-100) while the specificity was 44% (95% CI: 36.52-51.69). The Kappa statistic for level of agreement between PCR and LAMP test in the detection of ASFV was 23.7% (95% CI: 16.42-30.91). This Kappa value indicated a fair agreement between the two assays. No cross reaction was observed with Porcine circovirus type 2virus and E. coli isolated from pigs in Uganda. This is the first study evaluating and applying the LAMP assay in the detection of ASF in domestic pigs in Uganda. The LAMP assay was found to be more sensitive than PCR. Due to its simplicity, sensitivity and specificity, the LAMP assay has the potential for use in the diagnosis and routine surveillance of ASF in Uganda.Item Haemoparasitic Infections in Cattle from a Trypanosoma brucei Rhodesiense Sleeping Sickness Endemic District of Eastern Uganda(Tropical medicine and infectious disease, 2020) Matovu, Enock; Mugasa, Claire Mack; Waiswa, Peter; Kitibwa, Annah; Boobo, Alex; Mathu Ndung’u, JosephWe carried out a baseline survey of cattle in Kaberamaido district, in the context of controlling the domestic animal reservoir of Trypanosoma brucei rhodesiense human African trypanosomiasis (rHAT) towards elimination. Cattle blood was subjected to capillary tube centrifugation followed by measurement of the packed cell volume (PCV) and examination of the bu y coat area for motile trypanosomes. Trypanosomes were detected in 561 (21.4%) out of 2621 cattle screened by microscopy. These 561 in addition to 724 apparently trypanosome negative samples with low PCVs ( 25%) were transported to the laboratory and tested by PCR targeting the trypanosomal Internal Transcribed Spacer (ITS-1) as well as suspect Tick-Borne Diseases (TBDs) including Anaplasmamosis, Babesiosis, and Theileriosis. PCR for Anaplasma sp yielded the highest number of positive animals (45.2%), followed by Trypanosoma sp (44%), Theileria sp (42.4%) and Babesia (26.3%); multiple infections were a common occurrence. Interestingly, 373 (29%) of these cattle with low PCVs were negative by PCR, pointing to other possible causes of aneamia, such as helminthiasis. Among the trypanosome infections classified as T. brucei by ITS-PCR, 5.5% were positive by SRA PCR, and were, therefore, confirmed as T. b. rhodesiense. E orts against HAT should therefore consider packages that address a range of conditions. This may enhance acceptability and participation of livestock keepers in programs to eliminate this important but neglected tropical disease. In addition, we demonstrated that cattle remain an eminent reservoir for T. b. rhodesiense in eastern Uganda, which must be addressed to sustain HAT elimination.Item Impact of Intervention Practices on Malaria Treatment Outcomes Among Patients in Bushenyi District, Uganda(Research Square, 2021) Nyabayo Maniga, Josephat; Kalenzi Atuhaire, David; Mugasa, Claire MackMalaria remains a major vector borne disease causing both mortalities and morbidities in the world. Uganda as a country has currently scaled out major campaigns to reduce and eliminate the disease using different interventions. However, there is no clear data on the impact of such interventions on malaria treatment outcomes. Therefore, this study was aimed at assessing the impact of malaria intervention practices on Artemether- Lumefantrine (AL) treatment outcomes among the residents of Bushenyi district, Uganda, a high intensity malaria transmission area. Methods This was a descriptive cross-sectional study carried out among 184 study participants for a period of one year (August 2017 to August 2018) in four selected health centers in Bushenyi district, Uganda. The investigative methods used included a researcher administered questionnaire, laboratory and clinical evaluations of participants. Data analysis was done by using statistical package for social sciences (SPSS version 10 windows) for descriptive statistics. Results Statistically significant factors for treatment outcome at p ≤0.05 were; practicing indoor residual spraying (IRS) at home (𝑃 = 0.001; CI), source of prescription (𝑃 = 0.018; CI), finishing dosage (𝑃 = 0.006; CI), frequency of malaria infection (𝑃 = 0.028; CI), Frequency of antimalaria usage (𝑃 = 0.042; CI) and sleeping under insecticide treated nets (ITNs) (𝑃 = 0.039; CI) respectively. Conclusions IRS and ITNs were found to be the major intervention practice of malaria reduction after treatment with ACTs.Item Molecular Epidemiology of Anaplasmosis in Small Ruminants along a Human-Livestock-Wildlife Interface in Uganda(Heliyon, 2021) Kasozi, Keneth Iceland; Welburn, Susan Christina; Nalumenya, David Paul; Namayanja, Monica; Matama, Kevin; Zalwango, Kelly Katenta; Matovu, Wycliff; Zirintunda, Gerald; Ekou, Justine; Kembabazi, Stellamaris; Mugasa, Claire Mack; Kitibwa, Annah; Tayebwa, Dickson Stuart; Musinguzi, Simon Peter; Mahero, Michael; Ssengendo, Ibrahim; Nanteza, Anne; Matovu, Enock; MacLeod, Ewan ThomasInformation as regards the epidemiology of the Anaplasmataceae in small ruminants in several low- and middle-income countries is scarce. In this study a total of 712 DNA samples collected from small ruminants were analyzed for Anaplasmataceae and Anaplasma ovis using the 16S rRNA and MSP4 genes respectively. Infection risk was assessed by location, sex and age of the animals and qGIS® was used to construct spatial maps. The prevalence of Anaplasmataceae spp was 89.1% (95% CI: 77.5–95.9) and 79.1% (95% CI: 75.9–82.1) in ovines and caprines respectively (RR = 1.1, 95% CI: 1.0–1.3); higher than those previously reported in other eastern African countries. The prevalence of A. ovis was 26.1% and 25.4% for both ovines and caprines respectively with ovines showing significantly higher levels of infection than caprines (P < 0.05). The risk of Anaplasma ovis infections was not affected by age (OR = 1.2, 95% CI: 0.9–1.7) or sex (OR = 1.1, 95% CI: 0.6–2.0). Small ruminants located at the forest edge (<0.3 km) showed higher A. ovis prevalence than those found inland with infections present in the midland regions associated with increased agricultural activity. Anaplasma ovis remains a major challenge for small ruminant husbandry in Uganda and infections are under-reported. Policy efforts to prioritize management of Anaplasmataceae for small ruminant health would promote livestock productivity in vulnerable communities, improving livelihoods and ecosystem health.Item Preliminary evaluation of a Trypanosoma brucei FG-GAP repeat containing protein of mitochondrial localization [version 1; peer review: 1 approved, 1 approved with reservations](AAS Open Research, 2019) Namyanja, Monica; Xu, Zhi-Shen; Mugasa, Claire Mack; Lun, Zhao-Rong; Matovu, Enock; Chen, Zhengjun; Lubega, George W.Trypanosoma brucei, a causative agent of African Trypanosomiasis, is known to cross the blood brain barrier during the second stage of the disease. It was previously suggested that this parasite crosses the blood brain barrier in a manner similar to that of lymphocytes. This would imply that trypanosomes possess integrins that are required to interact with adhesion molecules located on the blood brain barrier microvascular endothelial cells, as a first step in traversal. To date, no T. brucei integrin has been described. However, one T. brucei putative FG-GAP repeat containing protein (typical of integrins) encoded by the Tb927.11.720 gene, was predicted to be involved in cell-cell/cell-matrix adhesion. Therefore, this study sought to characterize a putative FG-GAP repeat containing protein (FG-GAP RCP) and to determine its cellular localization as a basis for further exploration of its potential role in cell-cell or cell-matrix adhesion. Methods: In this study, we successfully cloned, characterized, expressed and localized this protein using antibodies we produced against its VCBS domain in T. brucei. Results: Contrary to what we initially suspected, our data showed that this protein is localized to the mitochondria but not the plasma membrane. Our data showed that it contains putative calcium binding motifs within the FG-GAP repeats suggesting it could be involved in calcium signaling/binding in the mitochondrion of T. brucei. Conclusion: Based on its localization we conclude that this protein is unlikely to be a trypanosomal integrin and thus that it may not be involved in traversal of the blood brain barrier. However, it could be involved in calcium signaling in the mitochondrion.Item The prevalence and genetic characterisation of Cryptosporidium isolates from cattle in Kiruhura district, South Western Uganda(Journal of Parasitic Diseases, 2021) Witto, Sarah Gift; Kankya, Clovice; Akurut, Gloria; Mugasa, Claire Mack; Kazibwe, Anne; Ochwo, SylvesterCryptosporidium is an emerging opportunistic zoonotic pathogen that causes diarrheal illness in a wide range of hosts including livestock and humans. This study set out to establish the prevalence of Cryptosporidium as well as the circulating genotypes in order to elucidate the potential role of cattle in the spread of human cryptosporidiosis. Rectal coprological samples from 363 cattle in 11 households in Kiruhura district, Southwestern Uganda were collected and screened for the presence of Cryptosporidium oocysts using the phenol auramine staining method followed by fluorescent microscopy. DNA was extracted from the microscopy positive samples and the COWP gene amplified using PCR. PCR products were sequenced and subjected to phylogenetic analysis. Additionally a multiplex realtime PCR was used to identify the Cryptosporidium spp. Multivariable mixed effect logistic regression models were used to identify potential risk factors for Cryptosporidium infection. The overallItem Towards Point-of-Care Diagnostic and Staging Tools for Human African Trypanosomiaisis(Journal of tropical medicine, 2012) Matovu, Enock; Kazibwe, Anne Juliet; Mugasa, Claire Mack; Ndungu, Joseph Mathu; Kithingi Njiru, ZablonHuman African trypanosomiasis is a debilitating disease prevalent in rural sub-Saharan Africa. Control of this disease almost exclusively relies on chemotherapy that should be driven by accurate diagnosis, given the unacceptable toxicity of the few available drugs. Unfortunately, the available diagnostics are characterised by low sensitivities due to the inherent low parasitaemia in natural infections. Demonstration of the trypanosomes in body fluids, which is a prerequisite before treatment, often follows complex algorithms. In this paper, we review the available diagnostics and explore recent advances towards development of novel point-ofcare diagnostic tests.Item Transcriptome analysis of peripheral blood of Schistosoma Mansoni Infected Children from the Albert Nile Region in Uganda Reveals Genes Implicated in Fibrosis Pathology.(bioRxiv, 2023) Namulondo, Joyce; Nyangiri, Oscar Asanya; Kimuda, Magambo Phillip; Nambala, Peter; Nassuuna, Jacent; Egesa, Moses; Nerima, Barbara; Biryomumaisho, Savino; Nabukenya, Immaculate; Drago, Kato; Tweyongyere, Robert; Matovu, Enock; Mulindwa, Julius; Mugasa, Claire MackOver 290 million people are infected by schistosomes worldwide. Schistosomiasis control efforts focus on mass drug treatment with praziquantel (PZQ), a drug that kills the adult worm of all Schistosoma species. Nonetheless, re-infections have continued to be detected in endemic areas with individuals living in the same area presenting with varying infection intensities. Our objective was to characterize the transcriptome profiles in peripheral blood of children between 10 - 15 years with varying intensities of Schistosoma mansoni infection living along the Albert Nile in Uganda. RNA extracted from peripheral blood collected from 44 S. mansoni infected (34 high and 10 low by circulating anodic antigen [CAA] level) and 20 uninfected children was sequenced using Illumina NovaSeq S4 and the reads aligned to the GRCh38 human genome. Differential gene expression analysis was done using DESeq2 and enriched pathways in differentially expressed genes (DEGs) were identified using REACTOME. Principal component analysis revealed clustering of gene expression by gender when S. mansoni infected children were compared with uninfected children. In addition, we identified 14 DEGs between S. mansoni infected and uninfected individuals, 56 DEGs between children with high infection intensity and uninfected individuals, 33 DEGs between those with high infection intensity and low infection intensity and no DEGs between those with low infection and uninfected individuals. We also observed upregulation and downregulation of some DEGs that are associated with fibrosis and its regulation. These data suggest expression of fibrosis associated genes as well as genes that regulate fibrosis in S. mansoni infection. The relatively few significant DEGS observed in children with schistosomiasis suggests that chronic S. mansoni infection is a stealth infection that does not stimulate a strong immune response.