Browsing by Author "Nanteza, Ann"
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Item Assessment of the Impact of Early Diagnosis and Early Treatment in the Integrated Control of East Coast Fever (ECF) Involving Acquired Immunity Induced by Natural Infection in Ankole Cattle(Pathogens, 2023) Nanteza, Ann; Nsubuga, Julius; Kazibwe, Anne; Terundaja, Clare; Matovu, Enock; Lubega, George WilliamThe integrated control of East Coast fever (ECF) by early diagnosis and treatment involving acquired immunity induced by natural infection in Ankole cattle was assessed. A longitudinal study was carried out in Kiruhura district, southwestern Uganda for six months on 244 Ankole breed of cattle from 18 herds under natural tick challenge and relaxed tick control measures. Calves aged three to six months old were recruited and monitored daily by farmers for detection of ECF clinical symptoms. The reported sick animals were treated using Buparvaquone and treatment outcome determined. Monthly follow-ups and blood collections were done to monitor ECF status. Blood was analyzed for Theileria parva parasites by microscopy, DNA by polymerase chain reaction (PCR) and antibodies by enzyme-linked immunosorbent assay (ELISA). The overall prevalence of ECF clinical disease within six months period was 30.3% (74). The major symptoms of early clinical ECF disease were fever and enlarged parotid or prescapular lymph nodes. Clinical cases were categorized as mild, 24% (18) or moderate, 76% (56). There was an overall recovery rate of 100% (74) of the ECF cases whereby 94.6% (70) recovered promptly and 5.4% (4) recovered slowly. Based on blood analysis, prevalence of ECF at baseline was 3.7% (9) by microscopy, 31.1% (76) by PCR and 38.1% (93) by ELISA. A significant increase (p < 0.05) was shown by the increased number of calves with T. parva specific antibodies in the sera from 38.1% at baseline to 68.8% after six months. High antibody levels (positive percentage ≥ 50%) were detected in all ECF-treated and recovered calves at the end of six months. The acquired immunity to ECF was high in treated and recovered cattle, indicating that natural exposure to infection, accurate early diagnosis and effective treatment enhance development of immune-protection in indigenous cattle in an endemic area. The prominent early clinical symptoms for ECF could be exploited in the development of decision support tools for chemotherapy and other integrated control measures.Item Association between serotonin transporter gene polymorphisms and increased suicidal risk among HIV positive patients in Uganda(BMC genetics, 2017) Kalungi, Allan; Seedat, Soraya; Hemmings, Sian M. J.; Merwe, Lize van der; Joloba, Moses L.; Nanteza, Ann; Nakassujja, Noeline; Birabwa, Harriet; Serwanga, Jennifer; Kaleebu, Pontiano; Kinyanda, EugenePersons living with HIV/AIDS (PLWHA) are at an increased risk of suicide. Increased suicidal risk is a predictor of future attempted and completed suicides and has been associated with poor quality of life and poor adherence with antiretroviral therapy. Clinical risk factors have low predictive value for suicide, hence the interest in potential neurobiological correlates and specific heritable markers of suicide vulnerability. The serotonin transporter gene has previously been implicated in the aetiology of increased suicidal risk in non-HIV infected study populations and its variations may provide a platform for identifying genetic risk for suicidality among PLWHA. The present cross-sectional study aimed at identifying two common genetic variants of the serotonin transporter gene and their association with increased suicidal risk among human immunodeficiency virus (HIV)-positive adults in Uganda. Results: The prevalence of increased suicidal risk (defined as moderate to high risk suicidality on the suicidality module of the Mini Neuropsychiatric Interview (M.I.N.I) was 3.3% (95% CI, 2.0–5.3). The 5-HTTLPR was found to be associated with increased suicidal risk before Bonferroni correction (p-value = 0.0174). A protective effect on increased suicidal risk was found for the 5-HTTLPR/rs25531 SA allele (p-value = 0.0046)- which directs reduced expression of the serotonin transporter gene (5-HTT). Conclusion: The SA allele at the 5-HTTLPR/rs25531 locus is associated with increased suicidal risk among Ugandan PLWHA. Further studies are needed to validate this finding in Ugandan and other sub-Saharan samples.Item Associations between Aminoquinoline Resistance Genotypes and Clinical Presentations of Plasmodium falciparum Infection in Uganda(Antimicrobial Agents and Chemotherapy, 2020) Cuu, Gloria; Asua, Victor; Tukwasibwe, Stephen; Nsobya, Sam L.; Nanteza, Ann; Kimuda, Magambo Phillip; Mpimbaza, Arthur; Rosenthal, Philip J.Mutations that mediate resistance of Plasmodium falciparum to aminoquinoline antimalarials are selected by prior drug use and may alter parasite fitness, but associations with clinical presentations are uncertain. We evaluated genotypes in samples from a case-control study of determinants of severe malaria in Ugandan children 4 months to 10 years of age. We studied 274 cases with severe malaria matched by age and geography to 275 uncomplicated malaria controls and 179 asymptomatic parasitemic controls. The overall prevalence of mutations of interest (considering mixed results as mutant) was 67.0% for PfCRT K76T, 8.5% for PfMDR1 N86Y, 71.5% for PfMDR1 Y184F, and 14.7% for PfMDR1 D1246Y. Compared to asymptomatic controls, the odds of mutant PfCRT 76T were lower for uncomplicated (odds ratio, 0.42 [95% confidence interval, 0.24 to 0.72]; P < 0.001) or severe (0.56 [0.32 to 0.97]; P = 0.031) malaria; the odds of mutant PfMDR1 86Y were lower for uncomplicated (0.33 [0.16 to 0.65]; P < 0.001) or severe (0.21 [0.09 to 0.45]; P < 0.001) malaria; and the odds of mutant PfMDR1 1246Y were higher for uncomplicated (1.83 [0.90 to 3.98]; P = 0.076) or severe (2.06 [1.01 to 4.55]; P = 0.033) malaria. The odds of mutant PfMDR1 184F were lower in severe than asymptomatic (0.59 [0.37 to 0.92]; P = 0.016) or uncomplicated (0.61 [0.41 to 0.90]; P = 0.009) malaria. Overall, the PfCRT 76T and PfMDR1 86Y mutations were associated with decreased risk of symptomatic malaria, PfMDR1 1246Y was associated with increased risk of symptomatic malaria, and PfMDR1 184F was associated with decreased risk of severe malaria. These results offer insights into parasite genotypes in children with different presentations, although the basis for the identified associations is likely complex.Item Bacterial Aetiology and Antibiotic Susceptibility Profile of Post-Operative Sepsis among Surgical Patients in a Tertiary Hospital in Rural Eastern Uganda(Microbiology research journal international, 2018) Masifa, George; Jacob, Stanley Iramiot; Muhindo, Rita; Olupot-Olupot, Peter; Nanteza, AnnPost-operative wound sepsis remains a surgical challenge of public health concern constituting approximately 20% of the health care-associated nosocomial infections. This study aimed at determining the prevalence and antimicrobial resistance patterns of bacterial pathogens isolated from post-operative wound infections at Mbale Regional Referral Hospital. Materials and Methods: This was a descriptive cross-sectional study conducted from June to October 2015. Study participant samples were sub-cultured upon reception in the Microbiology laboratory and the isolated bacterial pathogens were analysed. Phenotypic antimicrobial susceptibility profiles were determined using the Kirby-Bauer method. Interpretation of the zone diameters was done following the Clinical and Laboratory Standards Institute guidelines. Phenotypic screening for Methicillin-resistant Staphylococcus aureus (MRSA) was performed using oxacillin (1 μg). D-test was also performed for phenotypic screening of inducible clindamycin resistant Staphylococcus aureus, Data were entered into Microsoft Excel and analysed using IBM SPSS statistics (version 16). Results: Overall post-operative sepsis was 69/80 (86.2%) with Staphylococcus aureus as the most predominant organism 41/104 (39.4%) followed by Escherichia coli 22/104 (21.2%) and Klebsiella species 15/104 (14.4%). Of the 41/104 isolated Staphylococcus aureus, 27/41(65.9%) were MRSA strains and 5/41 (12.2%) were inducible clindamycin resistant Staphylococcus aureus strains. The isolated Staphylococcus aureus was resistant to multiple drugs though susceptible to vancomycin and clindamycin. In addition, none of the isolated Enterococci species was vancomycin resistant. Although most of the isolated Gram-negative organisms were sensitive to imipenem, resistance was observed for tetracycline, trimethoprim/sulphamethoxazole, and ceftriaxone. Conclusion: Staphylococcus aureus was the most common causative agent associated with postoperative sepsis with most of the strains being MRSA. Multi-drug resistance was observed in 63/104 (60.6%) of the isolated organisms in our study. Hence the need to better develop and strengthen antimicrobial stewardship programs as well as to understand the carriage of antimicrobial resistance genes among these organisms.Item Bacteriophage activity against and characterisation of avian pathogenic Escherichia coli isolated from colibacillosis cases in Uganda(PLoS ONE, 2020) Kazibwe, George; Katami, Phionah; Alinaitwe, Ruth; Alafi, Stephen; Nanteza, Ann; Nakavuma, Jesca L.Avian 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.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 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 Comparative detection of foot-and-mouth disease virus by reverse transcription loop-mediated isothermal amplification assay and real time polymerase chain reaction in Uganda(Int J Biotechnol Food Sci, 2016) Mukasa, Hussein Kafeero; Mwiine, Frank Norbert; Atuhaire, David Kalenzi; Ochwo, Sylvester; Nanteza, AnnFoot-and-mouth disease (FMD) is a viral disease. FMD diagnosis in the field is based on clinical signs that are shared by other vesicular diseases, hence to confirm FMD a laboratory is needed. Laboratory diagnostic techniques including serology may fail to distinguish between vaccinated and new infection, virus isolation may take up to 4 days to yield results, while molecular techniques including PCR, which are accurate, sensitive, specific and rapid, are costly and require special training of the laboratory staff. These challenges limit laboratory diagnosis yet in Uganda FMD outbreaks are common since the disease is endemic. This work reports the comparative detection of Foot-and-mouth disease virus (FMDV) by reverse transcription-loop mediated isothermal amplification (RT-LAMP) and Real-Time polymerase chain reaction (rRT-PCR) in Uganda based on the 3D polymerase (3Dpol) gene. The rRT-PCR assay is considered as the gold standard. A total of 89 cattle samples that included epithelial tissues (16.9%) and oral swabs (83.1%) were collected from outbreak cases in Eastern Districts of Mbale and Budaka. These were applied to molecular assays of rRT-PCR and RT- LAMP using primers and probes targeting the 3Dpol gene. The diagnostic sensitivity and specificity of RT-LAMP as a screening test and rRT-PCR as the reference test was 94.44% (95% CI = 94.11 to 94.78%) and 98.59% (95% CI = 98.50 to 98.68%), respectively. The kappa value for diagnostic agreement between rRT-PCR and RT-LAMP was 93.0% (95% CI = 83.50 to 100%), showing a perfect agreement. In conclusion, the RT-LAMP assay had a very high sensitivity and specificity when compared to the reference test of rRT-PCR. It was also very rapid since it gave results in 45 to 60 min. Due to its simplicity, sensitivity and specificity, LAMP assay has the potential for use in routine surveillance of FMD in Uganda.Item East Coast Fever Carrier Status and Theileria parva Breakthrough Strains in Recently ITM Vaccinated and Non-Vaccinated Cattle in Iganga District, Eastern Uganda(Pathogens, 2023) Oligo, Stephen; Nanteza, Ann; Nsubuga, Julius; Musoba, Abubakar; Kazibwe, Anne; Lubega, George WillyEast Coast fever (ECF) is a tick-borne disease of cattle that hinders the development of the livestock industry in eastern, central and southern Africa. The ‘Muguga cocktail’ live vaccine, delivered by an infection and treatment method (ITM), remains the only immunisation strategy of controlling ECF. However, there are challenges of the live vaccine inducing ECF carrier status in immunised animals and the possibility of lack of protection from parasite strains that are antigenically different from the vaccine strains. In Uganda, there are insufficient data regarding the ECF carrier status and T. parva genetic diversity in vaccinated and associated non-vaccinated cattle to assess the effectiveness of ITM vaccination. Blood was collected from recently ECF vaccinated (98) and non-vaccinated (73) cattle from Iganga district in Eastern Uganda at 120 days post-vaccination. The p104 gene nested PCR was used to screen for T. parva DNA, 11 minisatellite and 3 microsatellite markers (SSR) were used for genotyping. Two minisatellite markers (MS7 and MS19) were used to determine whether ECF carrier status was due to the T. parva vaccine or local strains. The prevalence of T. parva based on p104 nPCR was 61.2% (60/98) (RR 2.234, 95% CI 1.49–3.35, p-value < 0.001) among recently vaccinated cattle and 27.4% (20/73) (RR 1.00) among associated non-vaccinated cattle. The Muguga cocktail vaccine strains were responsible for carrier status in 10 (58.8%) by MS7 and 11 (64.7%) by MS19 in vaccinated cattle. Genotypes of T. parva with different-sized alleles to the vaccine strains that could be potential ‘breakthroughs’ were detected in 2 (11.8%)) and 4 (23.5%) isolates from vaccinated cattle based on MS7 and MS19 minisatellite markers, respectively. Using 14 SSR markers, T. parva diversity was higher in vaccinated (Na = 2.214, Ne = 1.978, He = 0.465) than associated non-vaccinated (Na = 1.071, Ne = 1.048, He = 0.259) cattle. The principal component analysis (PCA) showed isolates from vaccinated cattle were closely related to those from non-vaccinated cattle. The analysis of molecular variance (AMOVA) revealed high genetic variation (96%) within T. parva isolates from vaccinated and non-vaccinated cattle but low variation (4%) between vaccinated and non-vaccinated cattle. This study reveals the role of ITM in inducing the carrier status and higher T. parva genetic diversity in vaccinated cattle. The low genetic variation between T. parva isolates in both vaccinated and non-vaccinated cattle may be suggestive of the protective role of vaccine strains against genetically related local strains in the study area.Item Epidemiological Overview of African Swine Fever in Uganda (2001–2012)(Journal of Veterinary Medicine, 2013) Atuhaire, David Kalenzi; Ochwo, Sylvester; Afayoa, Mathias; Mwiine, Frank Norbert; Arinaitwe, Eugene; Ademun-Okurut, Rose Anna; Okuni, Julius Boniface; Nanteza, Ann; Ayebazibwe, Christosom; Okedi, Loyce; Olaho-Mukani, William; Ojok, LonzyAfrican swine fever (ASF) is a contagious viral disease, which can cause up to 100% mortality among domestic pigs. In Uganda there is paucity of information on the epidemiology of the disease, hence a study was carried out to elucidate the patterns of ASF outbreaks. Spatial and temporal analyses were performed with data collected monthly by the district veterinary officers (DVOs) and sent to the central administration at MAAIF from 2001 to 2012. Additionally, risk factors and the associated characteristics related to the disease were assessed based on semistructured questionnaires sent to the DVOs. A total of 388 ASF outbreaks were reported in 59 districts. Of these outbreaks, 201 (51.8%) were reported in districts adjacent to the national parks while 80 (20.6%) were adjacent to international borders. The number of reported ASF outbreaks changed over time and by geographical regions; however, no outbreak was reported in the North-Eastern region. ASF was ranked as second most important disease of pigs, and it occurred mostly during the dry season (𝑃 = 0.01). Pig movements due to trade (OR 15.5, CI 4.9–49.1) and restocking (OR 6.6, CI 2.5–17.3) were the major risk factors. ASF control strategies should focus on limiting pig movements in UgandaItem Field Evaluation of LED Fluorescence Microscopy for Demonstration of Trypanosoma brucei rhodesiense in Patient Blood(Microscopy Research, 2019) Matovu, Enock; Edielu, Andrew; Ojom, James; Nanteza, Ann; Kato, Charles Drago; Ndung’u, Joseph MathuDiagnosis of Trypanosoma brucei rhodesiense human African trypanosomiasis requires demonstration of parasites in body fluids by microscopy. The microscopy methods that are routinely used are difficult to deploy in resource-limited settings due to practical challenges, including lengthy and tedious procedures, and the need for specific equipment to centrifuge samples in glass capillary tubes. We report here on a study that was conducted in a rural region of eastern Uganda to evaluate new methods that take advantage of a field-deployable LED fluorescence microscope. Examination of acridine orange-stained blood smears by LED fluorescence microscopy resulted in a diagnostic accuracy that was similar to that of routine methods, while the time needed to identify parasites was shortened significantly. These findings make these new microscopy methods attractive alternatives to procedures that are currently used for diagnosis of T. b. rhodesiense human African trypanosomiasis.Item Genome Sequences of Bacteriophages UPEC01, UPEC03, UPEC06, and UPEC07 Infecting Avian Pathogenic Escherichia coli(Microbiology Resource Announcements, 2022) Kazibwe, George; Ndekezi, Christian; Alinaitwe, Ruth; Alafi, Stephen; Nanteza, Ann; Magambo, Phillip K.; Nakavuma, Jesca L.Here, we present the genome sequences of four bacteriophages that infect avian pathogenic Escherichia coli. The phages were isolated from raw sewage in Kampala, Uganda. The genome sizes of the phages ranged between 143,140 bp and 178,307 bp, with an average G1C content of 41.25%. Phages infecting avian pathogenic Escherichia coli (APEC) have the potential to be applied as phage therapy in the management of avian colibacillosis, a devastating disease that is responsible for significant economic losses in the poultry industry (1). The emergence of multidrug-resistant pathogenic E. coli strains has sparked interest in the search for alternative control measures for bacterial pathogens, including, among others, the use of phages (2). In this study, whole-genome sequencing of bacteriophages was carried out to determine the genetic characteristics and the taxonomic identification or classification of these phages as part of a larger study aimed at identifying and establishing phage stocks that can be used to supplement the use of antibiotics in managing avian colibacillosis in Uganda. The bacteriophages in this study were isolated from sewage at the National Water and Sewerage Corporation treatment plant (Kampala, Uganda). Several E. coli field isolates (Table 1) obtained from chicken droppings were used as isolation hosts for the phages following previously described methods (3). Briefly, 10 mL of raw sewage was centrifuged (10,000 g for 10 min) to obtain a supernatant, which was added to 10 mL of 2 tryptic soy broth (TSB) containing 100 mL of overnight E. coli broth culture. The mixture was incubated (30°C for 48 h at 120 rpm) and centrifuged (7,000 rpm for 5 min at 4°C), and the supernatant was filtered (0.45mm). The phage lysate obtained was plaque purified three times to produce a uniform phage stock. The isolated phages that could infect the APEC isolates from chickens that had died from colibacillosis were selected (4). Genomic DNA was extracted from the phages using 2% SDS and purified using a Qiagen Genomic-tip 100/G kit according to the manufacturer’s instructions.Item Genome Sequences of Bacteriophages UPEC01, UPEC03, UPEC06, and UPEC07 Infecting Avian Pathogenic Escherichia coli(Microbiology Resource Announcements, 2022) Kazibwe, George; Ndekezi, Christian; Alinaitwe, Ruth; Alafi, Stephen; Nanteza, Ann; Kimuda, Magambo Phillip; Nakavuma, Jesca LukangaHere, we present the genome sequences of four bacteriophages that infect avian pathogenic Escherichia coli. The phages were isolated from raw sewage in Kampala, Uganda. The genome sizes of the phages ranged between 143,140 bp and 178,307 bp, with an average G+C content of 41.25%.Item High proportion of Ugandans with pre-pandemic SARS-CoV-2 cross-reactive CD4+ and CD8+ T-cell responses(medRxiv, 2023) Namuniina, Annemarie; Muyanja, Enoch S.; Biribawa, Victoria M.; Ssemaganda, Aloysious; Nanteza, Ann; Bagaya, Bernard Ssentalo; Galwango, Ronald M.; Redd, Andrew D.The estimated mortality rate of the SARS-CoV-2 pandemic varied greatly around the world with multiple countries in East, Central, and West Africa having significantly lower rates of COVID-19 related fatalities than many resource-rich nations with significantly earlier wide-spread access to life-saving vaccines. One possible reason for this lower mortality could be the presence of pre-existing cross-reactive immunological responses in these areas of the world. To explore this hypothesis, stored peripheral blood mononuclear cells (PBMC) from Ugandans collected from 2015-2017 prior to the COVID-19 pandemic (n=29) and from hospitalized Ugandan COVID-19 patients (n=3) were examined using flow-cytometry for the presence of pre-existing SARS-CoV-2 cross-reactive CD4+ and CD8+ T-cell populations using four T-cell epitope mega pools. Of pre-pandemic participants, 89.7% (26/29) had either CD4+ or CD8+, or both, SARS-CoV-2 specific T-cell responses. Specifically, CD4+ T-cell reactivity (72.4%) and CD8+ T-cell reactivity (65.5%) were relatively similar, and 13 participants (44.8%) had both types of cross-reactive types of T-cells present. There were no significant differences in response by sex in the population. The rates of cross-reactive T-cell populations in these Ugandans is higher than previous estimates from resource-rich countries like the United States (20-50% reactivity). It is unclear what role, if any, this cross-reactivity played in decreasing COVID-19 related mortality in Uganda and other African countries, but does suggest that a better understanding of global pre-existing immunological cross-reactivity could be an informative data of epidemiological intelligence moving forward.Item Identification of coding sequences from a freshly prepared Trypanosoma brucei brucei expression library by polymerase chain reaction(International Journal of Biochemistry and Molecular Biology, 2013) Okalang, Uthman; Nanteza, Ann; Matovu, Enock; Lubega, George W.Animal African trypanosomiasis (AAT) also known as Nagana is a devastating disease among domestic animals in large parts of Sub-Saharan Africa causing loses in milk and meat production as well as traction power. However, there is currently no commercial vaccine against AAT. The parasites have also developed resistance to some of the drugs in use. Moreover, the use of affordable computer-aided wet bench methods in the search for vaccine and/or new drug targets against this disease have not yet been fully explored in developing countries. This study, therefore, explored the use of PCR to screen a freshly prepared bloodstream form Trypanosoma brucei brucei (T. b. brucei) expression library for coding sequences followed by bioinformatics analyses specifying the functions and importance of these proteins to parasite survival. Eleven protein coding sequences were identified from twenty nine purified clones. The putative retro transposon hot spot protein 4 (RHSP 4) was the only protein with a fully annotated DNA sequence. All the others were hypothetical or had partial or unqualified annotations. RHSP 4 and pyruvate dehydrogenase E1 component, alpha sub-unit (PDE1α) are involved in aerobic respiration whereas succinyl-Co A-3-ketoacid-coenzyme A transferase mitochondrial precursor (SKTMP) is predicted to be involved in ketone body catabolism. Cystathionine beta-synthase (CBS) and alpha-1,3-mannosyltransferase (αMT) have been predicted in cysteine biosynthesis and vesicular transport respectively. The functions of the hypothetical proteins encountered have neither been experimentally determined nor predicted. We hypothesize that both CBS and PDE1α are good drug targets. Overall, about 300 plates are required to PCR screen the entire Trypanosoma brucei genome in approximately eight months. This method is therefore, applicable and affordable in the search for new drug targets under conditions of limited resources among developing countries.Item Identification of Staphylococcus aureus: DNase and Mannitol salt agar improve the efficiency of the tube coagulase test(Annals of clinical microbiology and antimicrobials, 2010) Kateete, David P.; Kimani, Cyrus N.; Katabazi, Fred A.; Okeng, Alfred; Okee, Moses S.; Nanteza, Ann; Joloba, Moses L.; Najjuka, Florence C.The ideal identification of Staphylococcus aureus clinical isolates requires a battery of tests and this is costly in resource limited settings. In many developing countries, the tube coagulase test is usually confirmatory for S. aureus and is routinely done using either human or sheep plasma. This study evaluated Mannitol salt agar and the deoxyribonuclease (DNase) test for improving the efficiency of the tube coagulase test in resource limited settings. The efficiency of human and sheep plasma with tube coagulase tests was also evaluated.Item Inspiring Anti-Tick Vaccine Research, Development and Deployment in Tropical Africa for the Control of Cattle Ticks: Review and Insights(Vaccines, 2022) Kasaija, Paul D.; Kirunda, Halid; Nanteza, Ann; Kabi, Fredrick; Mugerwa, Swidiq; Fuente, José de laTicks are worldwide ectoparasites to humans and animals, and are associated with numerous health and economic effects. Threatening over 80% of the global cattle population, tick and tick-borne diseases (TTBDs) particularly constrain livestock production in the East, Central and Southern Africa. This, therefore, makes their control critical to the sustainability of the animal industry in the region. Since ticks are developing resistance against acaricides, anti-tick vaccines (ATVs) have been proposed as an environmentally friendly control alternative. Whereas they have been used in Latin America and Australia to reduce tick populations, pathogenic infections and number of acaricide treatments, commercially registered ATVs have not been adopted in tropical Africa for tick control. This is majorly due to their limited protection against economically important tick species of Africa and lack of research. Recent advances in various omics technologies and reverse vaccinology have enabled the identification of many candidate anti-tick antigens (ATAs), and are likely to usher in the next generation of vaccines, for which Africa should prepare to embrace. Herein, we highlight some scientific principles and approaches that have been used to identify ATAs, outline characteristics of a desirable ATA for vaccine design and propose the need for African governments to investment in ATV research to develop vaccines relevant to local tick species (personalized vaccines). We have also discussed the prospect of incorporating anti-tick vaccines into the integrated TTBDs control strategies in the sub-Saharan Africa, citing the case of Uganda.Item Interleukin (IL)-6 and IL-10 Are Up Regulated in Late Stage Trypanosoma brucei rhodesiense Sleeping Sickness(PLoS Neglected Tropical Diseases, 2015) Kato, Charles D.; Alibu, Vincent P.; Nanteza, Ann; Mugasa, Claire M.; Matovu, EnockSleeping sickness due to Trypanosoma brucei rhodesiense has a wide spectrum of clinical presentations coupled with differences in disease progression and severity across East and Southern Africa. The disease progresses from an early (hemo-lymphatic) stage to the late (meningoencephalitic) stage characterized by presence of parasites in the central nervous system. We hypothesized that disease progression and severity of the neurological response is modulated by cytokines.Item Phylogenetic Groups and Antimicrobial Susceptibility Patterns of Escherichia coli from Healthy Chicken in Eastern and Central Uganda(Journal of Veterinary Medicine, 2018) Kabiswa, Winston; Nanteza, Ann; Tumwine, Gabriel; Majalija, SamuelAntimicrobial resistance is an emerging problem in both humans and animals due to misuse and excessive use of drugs. Resistance in commensal E. coli isolates can be used to predict emergence of resistance in other gut microfora. Te aim of this study is to determine the phylogenetic groups and antimicrobial resistance patterns of E. coli from healthy chickens in Uganda. Te phylogenetic grouping of 120 fecal E. coli isolates from eastern and central Uganda was derived using the triplex PCR assay and their susceptibility patterns determined by agar disc difusion method to 5 antimicrobial drugs. Most E. coli is segregated into phylogenetic group A comprising 84%, while 12% and 4% were in groups D and B1, respectively. Similarly most E. coli from central (87%) and eastern Uganda (82%) belonged to group A. Overall, 85 (70%) of E. coli were resistant to antimicrobial drugs, of which 72/101 (70%) are in PG A, 10 of 14 (71.4%) in PG D, and 3 of 5 (60%) in PG B1. Signifcantly, most of the isolates in PG A from both central (66.7%) and (60.6%) eastern Uganda were resistant to one antimicrobial. Resistance to tetracycline alone or in combinationwith other drugs for central and eastern Uganda in PG A is 51% and 55%, respectively. Multidrug resistance to tetracycline and ciprofoxacin or nalidixic acid was 10% and 18% in isolates from central and 10% and 12% in isolates from eastern region, respectively. Phylogenetic group A accounts for most of the E. coli in chicken from Uganda. No diference in the resistance rates between the phylogenetic groups of E. coli has been observed. Te high prevalence of resistant E. coli strains from diferent phylogenetic groups in healthy chickens suggests antimicrobial drug selection pressure due to excessive drug in the rearing layer chickens.Item Plasma cytokine profiles associated with rhodesiense sleeping sickness and falciparum malaria co-infection in North Eastern Uganda(Allergy, Asthma & Clinical Immunology, 2019) Nsubuga, Julius; Kato, Charles Drago; Nanteza, Ann; Matovu, Enock; Alibu, Vincent PiusImmunological Human African Trypanosomiasis (HAT) studies often exclude malaria, although both infections overlap in specific endemic areas. During this co-infection, it is not known whether this parasitic interaction induces synergistic or antagonistic cytokine response among humans. This study determined prevalence of Plasmodium falciparum malaria among Trypanosoma brucei rhodesiense HAT and plasma cytokine profile levels associated with HAT and/or malaria infections.