Academic submissions (CoVAB)http://hdl.handle.net/10570/40892024-03-28T19:51:14Z2024-03-28T19:51:14ZAssaying for thermostable DNA polymerase activity in putative theromophilic bacteria isolates from selected garbage dumping sites and hot springs, in UgandaTusuubira, Sharifuhttp://hdl.handle.net/10570/70582019-01-04T08:47:17Z2018-01-01T00:00:00ZAssaying for thermostable DNA polymerase activity in putative theromophilic bacteria isolates from selected garbage dumping sites and hot springs, in Uganda
Tusuubira, Sharifu
Molecular and biotechnological research and training in Uganda is hampered by the high cost of acquisition of the required/appropriate chemicals and reagents, notably the DNA polymerase enzyme. Subsequently, there is insufficient practical training in molecular techniques, especially the Polymerase Chain Reaction (PCR) based, at tertiary institutions. This study set out to explore bacterial thermophiles from selected compost pits and hot springs in Uganda for production of thermostable polymerases as an alternative source for use in practical training and biotechnology. The study assessed the basic bacteria cell morphology and colony characteristics; and utilized genotypic methods of 16sRNA sequencing together with Naїve Bayesian Classification to offer a statistical test to flag taxa differentially represented between samples. Putative novel bacteria were isolated from selected garbage compost sites and geothermal hot springs in Uganda. The genotyping revealed that an isolate from compost UG-Wampewo belonged to Proteobacterium at 60% degree of confidence; while the hot spring isolates, Ug Kiga_Mud2,and Ug Kiga, are probably a novel species under Genus Aneurinibacillus (80 % degree of confidence) and Genus Pantoea (47%) respectively. Isolate Ug Kiga_Mud1 is most likely to be a strain of Aneurinibacillus thermoaurophillus as indicated by a degree of confidence at 100%. Characterization for polymerase enzyme activity revealed that the crude protein extracted from compost isolates showed negative results when used in PCR while the crude protein extract from the geothermal hot springs showed positive results. The phenotypic characterization of Ug-Wampewo indicated that this putative novel bacterium is a gram negative coccobacillus with very large cream colonies on nutrient agar. The study recommends comprehensive physiological, biochemical and chemotaxonomic characterization of the novel putative organisms. In addition, a detailed assessment of the organisms is recommended in order to explore the potential applications of the novel putative bacteria in industry and biotechnology.
2018-01-01T00:00:00ZAssessment of event mobile application for livestock disease reporting in KaramojaOkori, Ongom Edwardhttp://hdl.handle.net/10570/70042018-12-20T07:39:04Z2018-11-01T00:00:00ZAssessment of event mobile application for livestock disease reporting in Karamoja
Okori, Ongom Edward
The context of this study draws immensely from the global human and animal disease reporting perspectives, guided by the tripartite frameworks of WHO, FAO and OIE through their tools of IHR, EMPRES and PVS respectively down to the Pan African AU-IBAR, the East African Regional platform mandated by two Regional Economic Communities (RECs): IGAD and EAC. It further hinges on the premise that the success of disease reporting from the field depends on the competences to recognize clinical manifestations of the disease being reported, in addition to the use of diagnostic laboratories as key support to the diagnostic process.
The high rejection of disease reports collected using EMA-i tool by field data collectors from Karamoja region of Uganda was the main objective of this study. The study purpose was to assess the impact of level of education and profession on the knowledge of EMA-i protocols. Forty one (41) data collectors from Karamoja, comprising of CAHWs, veterinary paraprofessionals and veterinarians were purposively selected and administered structured questionnaires for quantitative data analysis. In addition, a total of 18 key informants’ interviews including all the 7 Karamoja DVOs, 3 Karamoja based Veterinarians; 3 Officials from NADDEC/MAAIF, 3 paraprofessionals and 2 CAHWs leaders from Karamoja. In addition, 2 FGDs were held in Kotido district.
The results from analysis of quantitative data, targeting education level, found out that out of the 41 reporters, only 1 primary reporter with education was knowledgeable while 5 were unknowledgeable; for Secondary education, 6 were knowledgeable and 4 unknowledgeable, whereas Diploma holders 6 were knowledgeable, and 1 not knowledgeable; while all the 18 University degree graduates were knowledgeable in most areas of the EMA-i protocol. Whereas for profession, 10 CAHWs were not knowledgeable and 9 knowledgeable while the 22 paraprofessionals and veterinarians were found to be knowledgeable in providing overall information on disease reporting using EMA-i.
The information from the quantitative and qualitative data was triangulated with the results of EMA-i data from Karamoja during July 2016 to September 2017 that showed 45% of disease reports were invalidated. Basing on saturated opinion by KIs, it was established that most data by CAHWs are invalidated due to wrong diagnosis, indicative of their inadequacy in knowledge of the EMA-i tool, thus leading to the concluding that the level of education, is the main factor to quality of disease data.
2018-11-01T00:00:00ZAssociation between malaria preventive treatment regimens in pregnancy and plasmodium falciparum drug resistance mediating polymorphisms in Busia district, Eastern UgandaNayebare, Patiencehttp://hdl.handle.net/10570/82792021-04-01T07:18:00Z2020-06-01T00:00:00ZAssociation between malaria preventive treatment regimens in pregnancy and plasmodium falciparum drug resistance mediating polymorphisms in Busia district, Eastern Uganda
Nayebare, Patience
Intermittent preventive treatment in pregnancy (IPTp) with monthly sulfadoxine-pyrimethamine (SP) is recommended for malaria-endemic parts of Africa, but efficacy is compromised by resistance and, in recent trials, dihydroartemesinin-piperaquine (DP) has shown better antimalarial protective efficacy. We utilized blood samples from a recent trial to evaluate selection by IPTp with dihydroartemesinin-piperaquine or sulfadoxine-pyrimethamine of Plasmodium falciparum genetic polymorphisms that alter sensitivity to these drugs. The prevalence of known genetic polymorphisms associated with altered drug sensitivity was determined in parasitemic samples, including 375 collected before IPTp drugs were administered, 125 randomly selected from those receiving sulfadoxine-pyrimethamine, and 80 from those receiving dihydroartemesinin-piperaquine. For women receiving dihydroartemesinin-piperaquine, the prevalence of mixed/mutant sequences was greater in samples collected during IPTp compared to that in samples collected prior to the intervention for PfMDR1 N86Y (19.7% vs. 4.1%, p<0.001),PfMDR1 Y184F (73.7% vs. 53.6%, p=0.002), and PfCRT K76T (45.6% vs. 24.4%, p=0.002).Considering sulfadoxine-pyrimethamine, prior to IPTp the prevalence of all five common antifolate mutations was over 92%, and this prevalence increased following exposure to sulfadoxine-pyrimethamine, although none of these changes were statistically significant. The prevalence of two additional mutations associated with high-level sulfadoxine-pyrimethamine resistance was greater, but not significantly so, in samples collected during IPTp compared to those collected before the intervention (PfDHFR 164L: 9.3% vs.3.9%, p=0.07; PfDHPS 581G: 3.4% vs. 2.9%, p=0.73). Use of IPTp in Uganda selected for parasites with mutations associated with decreased sensitivity to IPTp regimens. Principal component analysis suggested that gravidity, parasite density, gestational age, participant age and treatment regimen contributed to the differences in prevalence of polymorphisms before and after IPTp.
A dissertation submitted to the Directorate of Research and Graduate Training in partial fulfillment of the requirements for the award of the Degree of Master of Science in Molecular Biology and Biotechnology of Makerere University.
2020-06-01T00:00:00ZCharacterisation of integrons which potentially mediate dissemination of antibiotic resistance in enterobacteriaceae isolated from healthy community membersAto, Adera Doreenhttp://hdl.handle.net/10570/90922021-11-23T12:00:01Z2020-10-01T00:00:00ZCharacterisation of integrons which potentially mediate dissemination of antibiotic resistance in enterobacteriaceae isolated from healthy community members
Ato, Adera Doreen
Antibiotic resistance has become a global public health concern, by limiting the number of drugs used to treat bacterial infections. Antimicrobial resistance and multidrug resistance (MDR) therefore increase morbidity and mortality of people having bacterial infections in the community. Integrons are known as the genetic agents responsible for the capture and spread of antibiotic resistance determinants among diverse Gram-negative bacteria isolates. The activities of integrons have assisted the introduction of antibiotic resistance into genes pathogens and commensals of humans. Several studies have shown that integrons play a significant role in the spread of antibiotic resistance. This study examined the role of integrons in the spread of antibiotic resistance in Enterobacteriaceae from faeces of healthy community members. The antibiotic resistance patterns of 32 Escherichia coli and 8 Klebsiella pneumoniae isolates were determined by Kirby disk diffusion and conjugation was used to determine the transfer rates of antibiotic resistance. The 40 bacteria isolates were then tested for presence of integrons by polymerase chain reaction and the integrons present characterised by amplifying the integron variable region. The PCR amplicons of the integron variable region were then sequenced to determine the antibiotic resistance gene cassettes present. The isolates were found to have high resistance to beta-lactams (100%), sulphonamides (100%) and fluoroquinolones (88%). A high frequency of transfer ranging from 9×10-2 to 1.1×103 transconjugants per recipient E. coli J53 strain was recorded. The prevalence of class 1, class 2 and class 3 integrons were 100%, 35% and 0%. The integron cassette profiles were aadA1 (70%), dfrA17 (50%) and dfrA7 (10%) and they confer resistance to aminoglycosides and trimethoprim. The study revealed that 23 (58%) Escherichia coli and 2 (5%) Klebsiella pneumoniae isolates were MDR. The study also revealed that integrons with antibiotic resistance genes aadA1, dfrA7 and dfrA17are present in the isolates from healthy community members, thus they have a potential to capture genes which confer resistance to various antibiotics. The study recommends proper use of antibiotics, improved personal hygiene and sanitation to minimize the oral faecal spread of antibiotic resistance in the community. The study also recommends characterisation of transconjugants to determine which resistance gene cassettes are most disseminated.
A dissertation submitted to the School of Research and Graduate Studies in partial fulfilment of the requirements for the award of the Degree of Master of Science in Molecular Biology and Biotechnology of Makerere University.
2020-10-01T00:00:00Z