Browsing by Author "Kassaza, Kennedy"

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    Investigating metabolic and molecular ecological evolution of opportunistic pulmonary fungal coinfections: protocol for a laboratory-based cross-sectional study
    (JMIR Publications, 2023-04-09) Njovu, Israel Kiiza; Nalumaga, Pauline Petra; Ampaire, Lucas; Nuwagira, Edwin; Mwesigye, James; Musinguzi, Benson; Kassaza, Kennedy; Taseera, Kabanda; Mukasa, James Kiguli; Bazira, Joel; Iramiot, Jacob Stanley; Baguma, Andrew; Bongomin, Felix; Kwizera, Richard; Achan, Beatrice; Cox, Michael J; King, Jason S; May, Robin; Ballou, Elizabeth R; Itabangi, Herbert
    Background: Fungal-bacterial cocolonization and coinfections pose an emerging challenge among patients suspected of having pulmonary tuberculosis (PTB); however, the underlying pathogenic mechanisms and microbiome interactions are poorly understood. Understanding how environmental microbes, such as fungi and bacteria, coevolve and develop traits to evade host immune responses and resist treatment is critical to controlling opportunistic pulmonary fungal coinfections. In this project, we propose to study the coexistence of fungal and bacterial microbial communities during chronic pulmonary diseases, with a keen interest in underpinning fungal etiological evolution and the predominating interactions that may exist between fungi and bacteria. Objective: This is a protocol for a study aimed at investigating the metabolic and molecular ecological evolution of opportunistic pulmonary fungal coinfections through determining and characterizing the burden, etiological profiles, microbial communities, and interactions established between fungi and bacteria as implicated among patients with presumptive PTB. Methods: This will be a laboratory-based cross-sectional study, with a sample size of 406 participants. From each participant, 2 sputa samples (one on-spot and one early morning) will be collected. These samples will then be analyzed for both fungal and bacterial etiology using conventional metabolic and molecular (intergenic transcribed spacer and 16S ribosomal DNA–based polymerase chain reaction) approaches. We will also attempt to design a genome-scale metabolic model for pulmonary microbial communities to analyze the composition of the entire microbiome (ie, fungi and bacteria) and investigate host-microbial interactions under different patient conditions. This analysis will be based on the interplays of genes (identified by metagenomics) and inferred from amplicon data and metabolites (identified by metabolomics) by analyzing the full data set and using specific computational tools. We will also collect baseline data, including demographic and clinical history, using a patient-reported questionnaire. Altogether, this approach will contribute to a diagnostic-based observational study. The primary outcome will be the overall fungal and bacterial diagnostic profile of the study participants. Other diagnostic factors associated with the etiological profile, such as incidence and prevalence, will also be analyzed using univariate and multivariate schemes. Odds ratios with 95% CIs will be presented with a statistical significance set at P<.05. Results: The study has been approved by the Mbarara University Research Ethic Committee (MUREC1/7-07/09/20) and the Uganda National Council of Science and Technology (HS1233ES). Following careful scrutiny, the protocol was designed to enable patient enrollment, which began in March 2022 at Mbarara University Teaching Hospital. Data collection is ongoing and is expected to be completed by August 2023, and manuscripts will be submitted for publication thereafter. Conclusions: Through this protocol, we will explore the metabolic and molecular ecological evolution of opportunistic pulmonary fungal coinfections among patients with presumptive PTB. Establishing key fungal-bacterial cross-kingdom synergistic relationships is crucial for instituting fungal bacterial coinfecting etiology.
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    Molecular characterization of extended-spectrum beta-lactamase- producing bacteria isolated from pregnant women’s urine at Itojo Hospital, South Western Uganda
    (Microbiology Society, 2026-03-11) Twinomujuni, Muzafaru; Musinguzi, Benson; Asiimwe, Moses; Mpiima, Stephen Samuel; Zamarano, Henry; Orikushaba, Isaac; Muhanguzi, Deus; Twinamatsiko, Crinad; Mallya, Sarapia Paul; Samiri, Jamiru; Kamugisha, Joseph; Nalumaga, Pauline Petra; Kabanda, Taseera; Kassaza, Kennedy; Bagenda, Charles Nkubi; Tuhamize, Barbra; Bazira, Joel; Ricciardelli, Rosemary; Mpeirwe, Moses
    Background: Extended-spectrum β-lactamase (ESBL)-producing bacteria pose a global challenge because of resistance developing against a wide range of antimicrobial agents, complicating available treatment options. Thus, identifying the prevalent bacterial species producing ESBL enzymes and understanding how they are susceptible to antibiotics is necessary to inform effective treatment guidelines. Objective: We sought to characterize ESBL-producing bacteria isolated from pregnant women’s urine at Itojo Hospital, Ntungamo district, Southwestern Uganda. Methods: We conducted a cross-sectional study where we collected and analysed 340 urine samples from 340 pregnant women. We did antimicrobial susceptibility testing using the Kirby–Bauer disc diffusion method. Isolates were screened for ESBL production and confirmed using the combination disc test. Genotypic characterization was confirmed using multiplex PCR to detect blaTEM, blaCTX-M and blaSHV genes. Results: The prevalence of ESBL-producing bacteria was 29.7% (101/340). Escherichia coli 36/101 (35.6%) and Klebsiella species 33/101 (32.7%) were predominant ESBL producers. Genotypic analysis revealed blaTEM 50/101 (49.5%) and blaCTX-M 31/101 (30.7%) as the most prevalent genes, while blaSHV was less common, 8/101 (7.9%) Conclusion. The high prevalence of ESBL-producing bacteria and their resistance to commonly used antibiotics highlighted the need for targeted antibiotic therapy, antimicrobial stewardship and regular molecular surveillance.
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    Status of pulmonary fungal pathogens among individuals with clinical features of pulmonary tuberculosis at Mbarara University Teaching Hospital in Southwestern Uganda
    (SAGE Publications, 2021-08-31) Njovu, Israel Kiiza; Musinguzi, Benson; Mwesigye, James; Kassaza, Kennedy; Turigurwa, Joseph; Nuwagira, Edwin; Bazira, Joel; Kabanda, Taseera; Mpeirwe, Moses; Ampaire, Lucas; Mutekanga, Andrew; Kiguli, James; Achan, Beatrice; Itabangi, Herbert
    Pulmonary mycoses are important diseases of the respiratory tract caused by pulmonary fungal pathogens. These pathogens are responsible for significant morbidity and mortality rates worldwide; however, less attention has been paid to them. In this study we determined the prevalence of pulmonary fungal pathogens among individuals with clinical features of pulmonary tuberculosis at Mbarara Regional Referral Hospital. This was a hospital based cross sectional survey. Sputum samples were collected from each study participant. For each sample, the following tests were performed: Sabouraud dextrose agar for fungal culture, GeneXpert for Mycobacteria tuberculosis (MTB) and potassium hydroxide for fungal screening. Filamentous fungal growth and yeasts were further examined with lactophenol cotton blue staining and germ tube respectively. Out of 113 study participants, 80 (70.7%) had pulmonary fungal pathogens whilst those with pulmonary tuberculosis numbered five (4.4%). Candida albicans [21 (22.58%)] and Aspergillus species [16 (17.20%)] were the pathogens most identified among others. Two (1.7%) TB GeneXpert positive participants had fungal pathogens isolated from their sputum samples. We established a prevalence of 57 (71.3%) for pulmonary fungal pathogen (PFP) isolates, three (60.0%) for MTB in HIV positive patients and 18 (22.5%) for PFP, and zero (0.0%) for MTB in HIV negative patients. On the other hand, two (100%) HIV positive patients had both PFP isolates and MTB.Our findings highlight the diversity of neglected pulmonary fungal pathogens whose known medical importance in causing pulmonary mycoses cannot be overemphasised. Therefore this presents a need for routine diagnosis for pulmonary mycoses among TB suspects and set-up of antimicrobial profile for pulmonary fungal isolates to support clinical management of these cases.