Resistance to pyrazinamide in Mycobacterium tuberculosis complex isolates from previously treated tuberculosis cases in Southwestern Oromia, Ethiopia.

Objective: Pyrazinamide (PZA) susceptibility testing is important to develop evidence-based algorithms for case management. We aimed to assess the prevalence of PZA-resistance and its impact on treatment outcomes in previously treated tuberculosis (TB) cases in southwestern Oromia, Ethiopia. Methods: A Phenotypic Drug Susceptibility Testing (DST) of PZA with BACTEC MGIT 960 was conducted at the Mycobacteriology Research Center of Jimma University (MRC-JU) from June to November 2021 on sixty-six Mycobacterium tuberculosis complex (MTBC) isolates from previously treated TB cases. SPSS software package version 21 was used. The differences in the proportion of PZA resistance between the groups were compared using the chi squared test. Logistic regression was used to identify the association between PZA resistance and treatment outcomes. Results: Among 66 MTBC isolates (49 rifampicin-resistant and 17 rifampicin-sensitive) included in this study, 31.8 % were resistant to PZA. The proportion of PZA resistance was almost three times higher in previously treated TB cases with rifampicin resistance than in rifampicin-sensitive patients (38.8 % vs. 11.8 %, p = 0.039). An unfavorable treatment outcome was documented for 23 % (15/65) of the participants. Patients with PZA resistance were almost four times more likely to have an unfavorable treatment outcome than patients with PZA sensitive (aOR 4.2, 95 % CI: 1.13-15.3). Conclusions: The prevalence of PZA resistance was high compared to the pooled PZA resistance estimated worldwide. The majority of TB cases with PZA resistance had an unfavorable treatment outcome. PZA susceptibility testing should be included in the multidrug-resistant TB diagnostic algorithm to improve management of these patients.

Assessment of Phage-Mediated Inhibition and Removal of Multidrug-Resistant Pseudomonas aeruginosa Biofilm on Medical Implants.

Purpose: Despite the growing interest in bacteriophage (phage) usage for the prevention, control, and removal of bacterial biofilms, few scientific data exist on phage applications on medical implant surfaces, while none exists on multiple implants. In this study, we aimed to isolate, biophysically characterize and assess phages as potential antibiofilm agents to inhibit and remove multidrug-resistant (MDR) Pseudomonas aeruginosa biofilm on catheter and endotracheal tube surfaces. Methods: The well-identified stored clinical isolates (n = 7) of MDR P. aeruginosa were obtained from Jimma Medical Center. Specific phages were isolated and characterized based on standard protocols. The phages were tested for their antibiofilm effects in preventing colonization and removing preformed biofilms of MDR P. aeruginosa, following phage coating and treatment of catheter and endotracheal tube segments. Results: Two P. aeruginosa-specific phages (ΦJHS-PA1139 and ΦSMK-PA1139) were isolated from JMC compound sewage sources. The phages were biophysically characterized as being thermally stable up to 40°C and viable between pH 4.0 and 11.0. The two phages tested against clinical MDR strains of P. aeruginosa showed broad host ranges but not on other tested bacterial species. Both phages reduced MDR bacterial biofilms during the screening step. The phage-coated segments showed 1.2 log10 up to 3.2 log10 inhibition relative to non-coated segments following 6 h coating of segments prior to microbial load exposure. In both phages, 6 h treatment of the segments with 106 PFU/mL yielded 1.0 log10 up to 1.6 log10 reductions for ΦJHS and 1.6 log10 up to 2.4 log10 reductions for ΦSMK. Conclusion: Our results suggest that phages have great potential to serve the dual purpose as surface coating agents for preventing MDR bacterial colonization in medical implants and as biofilm removal agents in implant-associated infections.

Community Engagement in the Fight Against COVID-19: Knowledge, Attitude, and Prevention Practices Among Dire Dawa Residents, Eastern Ethiopia.

Background: There is a global concern that coronavirus disease 2019 (COVID-19) cannot be addressed without the integration and active engagement of communities. We aimed to investigate the knowledge, attitude, and practices (KAP) of the residents in Eastern Ethiopia toward COVID-19. Method: A community-based cross-sectional study was conducted on May 1-30, 2020 among the residents of Dire Dawa, Eastern Ethiopia. Data were collected using a structured questionnaire on the awareness, knowledge, attitudes, and preventive practices toward COVID-19. We used random sampling to select the participants. Data was entered into the statistics and data (STATA) version for data cleaning and analysis. Binary logistic regression models with 95% CI were used to conduct bivariable [crude odds ratio (COR)] and multivariable [adjusted odds ratio (AOR)] analyses. Result: A total of 415 community residents responded to the questionnaire. Of those participants, 52.5% (95% CI 47.7-57) had adequate knowledge, 31.6% (95% CI 27-36) had favorable attitudes, and 49% (95% CI 45-50) had good practice toward the precautionary measures of COVID-19. Adequate knowledge had a significant association with urban residence (AOR = 5, 95% CI 3.1-8.4) and literacy (AOR = 3.1, 95% CI 1.5-6.7). Good preventions practices had a significant association with place of residence (AOR = 4.1, 95% CI 2.3-7.2), literacy (AOR = 2.9, 95% CI: 1,2-7.4), adequate knowledge (AOR = 3.5, 95% CI 2.3-5.8), and favorable attitude (AOR = 2.3, 95% CI 1.4-3.8) about the disease. Conclusion: The overall COVID-19-related KAP was inadequate in the majority of the residents of Dire Dawa that occupy irregular migration flows. These call for robust community-centered behavioral communication strategies that could bridge the gaps and help prevent and control COVID-19 and other future pandemics in their community.

Bacterial Biofilm Destruction: A Focused Review On The Recent Use of Phage-Based Strategies With Other Antibiofilm Agents.

Biofilms are bacterial communities that live in association with biotic or abiotic surfaces and enclosed in an extracellular polymeric substance. Their formation on both biotic and abiotic surfaces, including human tissue and medical device surfaces, pose a major threat causing chronic infections. In addition, current antibiotics and antiseptic agents have shown limited ability to completely remove biofilms. In this review, the authors provide an overview on the formation of bacterial biofilms and its characteristics, burden and evolution with phages. Moreover, the most recent possible use of phages and phage-derived enzymes to combat bacteria in biofilm structures is elucidated. From the emerging results, it can be concluded that despite successful use of phages and phage-derived products in destroying biofilms, they are mostly not adequate to eradicate all bacterial cells. Nevertheless, a combined therapy with the use of phages and/or phage-derived products with other antimicrobial agents including antibiotics, nanoparticles, and antimicrobial peptides may be effective approaches to remove biofilms from medical device surfaces and to treat their associated infections in humans.

Spoligotype-based population structure of Mycobacterium tuberculosis in the Jimma Zone, southwest Ethiopia.

BACKGROUND: To understand the population dynamics and propose more effective preventive strategies, defining the population structure of the circulating Mycobacterium tuberculosis strains is important. METHODS: A total of 177 M. tuberculosis complex isolates from pulmonary tuberculosis (TB) cases in southwest Ethiopia were genotyped by spoligotyping. Of the strains included in this study, 126 were pan-susceptible strains while the remaining 51 isolates were resistant to one or more first-line anti-TB drugs. The genotyping results were compared to the international spoligotyping (SITVIT) database of the Pasteur Institute of Guadeloupe and the newly revised publicly available international multi-marker database (SITVITWEB/SPOLDB4). An online tool Run TB-Lineage was also used to predict the major lineages using a conformal Bayesian network analysis. RESULTS: The spoligotyping of the 177 isolates resulted in 69 different spoligotype patterns of which 127 (71.8%) were clustered into 19 spoligoclusters (with clustering rate of 61.02%). Each cluster contains 2-29 isolates. Of the isolates with corresponding SIT in SITVIT/SDB4, the predominant strains identified were SIT 37 of the T3 subfamily with 29 isolates followed by SIT 53 of the T1 subfamily with 20 isolates. SIT 777 of the H4 subfamily and SIT 25 of the CAS1_DELHI subfamily each consisting of six isolates were identified. Eighty spoligotype patterns were orphan as they were not recorded in the SITVIT2/SPDB4 database. Further classification of the isolates on the basis of major lineages showed that 82.5% and 14.1% of the isolates belonged to Euro-American and East African Indian lineages, respectively, while 2.8% of the isolates belonged to Mycobacterium africanum and 0.6% to Indo-Oceanic. CONCLUSION: The ill-defined T and H clades were predominant around Jimma. The substantial number of orphans recorded in the study area warrants for additional studies with genotyping methods with better resolution and covering whole areas of southwest Ethiopia.

Tuberculosis among Jimma University Undergraduate Students: First Insight about the Burden of Tuberculosis in Ethiopia Universities-Cross-Sectional Study.

BACKGROUND: Ethiopian universities are facilities where students live in relative overcrowding condition. This might favor the chance of tuberculosis transmission among students. This study was done to determine the magnitude and associated factors of tuberculosis among Jimma University students. METHODS: A cross-sectional study was done from February 2015 to July 2015. Hundred twenty-nine consented participants were interviewed using structured questionnaire. Biological specimens were collected and cultured on Mycobacterium Growth Indicator Tube. Mycobacterium tuberculosis complex verification was done by SD BIOLINE TB Ag MPT64 Rapid test. Frequency distribution, logistic regression, and independent sample t-test were used to analyze the data using SPSS Version 20. RESULT: Magnitude of all forms of tuberculosis among Jimma University undergraduate students was 209.1 per 100000-student population. Contact history [AOR: 4.76, 95% CI (1.31-17.31)], smoking [AOR: 6.67, 95% CI (1.51-29.44)], khat chewing [AOR: 5.56, 95% CI (1.66-18.69)], and low body mass index [AOR: 5.37, 95% CI (1.46-19.78)] were determinants of tuberculosis. CONCLUSION: The magnitude of tuberculosis among Jimma University undergraduate students is high. TB is associated with previous tuberculosis patient contact and behavioral factors. Hence, students with these risk factors should be given enough attention for the control of TB in Jimma University.

Drug resistance-conferring mutations in Mycobacterium tuberculosis from pulmonary tuberculosis patients in Southwest Ethiopia.

OBJECTIVE/BACKGROUND: The nature and frequency of mutations in rifampicin (RIF) and isoniazid (INH) resistant Mycobacterium tuberculosis isolates vary considerably according to geographic locations. However, information regarding specific mutational patterns in Ethiopia remains limited. METHODS: A cross-sectional prospective study was carried out among confirmed pulmonary tuberculosis cases in Southwest Ethiopia. Mutations associated with RIF and INH resistances were studied using GenoType MTBDRplus line probe assay in 112 M. tuberculosis isolates. Culture (MGIT960) and identification tests were performed at the Mycobacteriology Research Center of Jimma University, Jimma, Ethiopia. RESULTS: Mutations conferring resistance to INH, RIF, and multidrug resistance were detected in 36.6% (41/112), 30.4% (34/112), and 27.7% (31/112) of M. tuberculosis isolates respectively. Among 34 RIF-resistant isolates, 82.4% (28/34) had rpoB gene mutations at S531L, 2.9% (1/34) at H526D, and 14.7% (5/34) had mutations only at wild type probes. Of 41 INH-resistant strains, 87.8% (36/41) had mutations in the katG gene at Ser315Thr1 and 9.8% (4/41) had mutations in the inhA gene at C15T. Mutations in inhA promoter region were strongly associated with INH monoresistance. CONCLUSION: A high rate of drug resistance was commonly observed among failure cases. The most frequent gene mutations associated with the resistance to INH and RIF were observed in the codon 315 of the katG gene and codon 531 of the rpoB gene, respectively. Further studies on mutations in different geographic regions using DNA sequencing techniques are warranted to improve the kit by including more specific mutation probes in the kit.

GeneXpert MTB/RIF Assay for the Diagnosis of Tuberculous Lymphadenitis on Concentrated Fine Needle Aspirates in High Tuberculosis Burden Settings.

INTRODUCTION: The diagnosis of tuberculous lymphadenitis (TBL) remains challenging. The routinely used methods (cytology and smear microscopy) have sub-optimal sensitivity. Recently, WHO recommends GeneXpert to be used as the initial diagnostic test in patients suspected of having extra-pulmonary tuberculosis (EPTB). However, this was a conditional recommendation due to very low-quality evidence available and more studies are needed. In this study we evaluated the performance of Xpert for the diagnosis of TBL on concentrated fine needle aspirates (FNA) in Southwest Ethiopia. METHODS: FNA was collected from presumptive TBL cases. Two smears were prepared from each aspirate and processed for cytology and conventional microscopy. The remaining aspirate was treated with N-acetyl-L-cysteine-NaOH and centrifuged for 15minutes at 3000g. The concentrated sediment was used for culture and Xpert test. Capilia TB-Neo test was used to differentiate M. tuberculosis complex (MTBC) from non-tuberculous mycobacteria (NTM). Composite bacteriological methods (culture and/or smear microscopy) were considered as a reference standard. RESULT: Out of 143 enrolled suspects, 64.3% (92/143) were confirmed TBL cases by the composite reference standard (CRS). Xpert detected M. tuberculosis complex (MTBC) in 60.1% (86/143) of the presumptive TBL cases. The sensitivity of Xpert compared to CRS was 87.8% [95% CI: 81.0-94.5] and specificity 91.1% [95% CI: 82.8-99.4]. The sensitivity was 27.8% for smear microscopy and 80% for cytology compared to CRS. Cytology showed the lowest specificity (57.8%). Xpert was positive in 4 out of 45 culture- and smear-negative cases. Among 47 cytomorphologically non-TBL cases, 15 were positive on Xpert. More than half of Xpert-positive cases were in the range of very low cut-off threshold values (28

Diagnostic performance of fluorescent light-emitting diode microscopy for tuberculous lymphadenitis in a high-burden setting.

OBJECTIVE: Diagnosis of tuberculous lymphadenitis using fine-needle aspiration cytology is a simple and safe but low-specificity method, whereas conventional smear microscopy has variable sensitivity due to low bacterial load. We evaluated the diagnostic performance of fluorescent light-emitting diode (LED) microscopy on routinely collected fine-needle aspirates from tuberculous lymphadenitis presumptive cases. METHODS: Fine-needle aspirates were collected from patients clinically suspected of having tuberculous lymphadenitis as part of routine diagnosis. Smear preparation was performed from the aspirate and processed for cytology, conventional Ziehl-Neelsen and LED microscopy. The remaining aspirate was processed for culture on Lowenstein-Jensen media. Capilia TB-Neo test was used to differentiate M. tuberculosis complex from non-tuberculous mycobacteria. RESULT: A total of 144 tuberculous lymphadenitis presumptive cases were included. 66.7% (96/144) were positive for M. tuberculosis complex on culture. Only one isolate was identified as non-tuberculous mycobacteria. The detection rates of Ziehl-Neelsen and LED microscopy were 18.8% (27/144) and 34% (49/144), respectively. As compared to culture, sensitivity was 25.0% [95% CI: 16.3-33.7] for Ziehl-Neelsen microscopy and 45.8% [95% CI: 35.9-55.8] for LED microscopy. The specificity was 93.8% [95% CI: 86.9-100] for Ziehl-Neelsen microscopy and 89.6% [95% CI: 80.9-98.2] for LED microscopy. LED microscopy showed a statistically significant increase in sensitivity and similar specificity compared to Ziehl-Neelsen microscopy. Mean reading time of positive slides was 2.62 min/slide for Ziehl-Neelsen and 1.60 min/slide for LED microscopy. Cytology showed sensitivity of 82.3% and specificity of 54.2%. LED microscopy detected TB bacilli in 33.3% of cases cytologically classified as suppurative abscess. CONCLUSION: The LED microscopy for tuberculous lymphadenitis had significantly higher sensitivity and shorter screening time than Ziehl-Neelsen microscopy. Use of LED microscopy among cases classified as suppurative abscess on fine-needle aspirate cytology improves evidence-based diagnosis of presumptive tuberculous lymphadenitis cases. Moreover, LED microscopy could be considered as an alternative approach in settings where fine-needle aspirate cytology is impractical.

Drug resistance patterns of Mycobacterium tuberculosis complex and associated factors among retreatment cases around Jimma, Southwest Ethiopia.

BACKGROUND: Information on the pattern of drug resistant tuberculosis (TB) among re-treatment cases is crucial to develop appropriate control strategies. Therefore, we conducted this study to assess the drug resistance pattern of M. tuberculosis complex (MTBC) isolates and associated factors among re-treatment cases in Jimma area, Southwest Ethiopia. METHODS: Health facility-based cross-sectional study was conducted between March 2012 and April 2013 in Jimma area, Southwest Ethiopia. We included 79 re-treatment cases selected conveniently. Socio demographic and clinical data were collected using structured questionnaire. Sputum sample processing, mycobacterial culture, isolation and drug susceptibility testing (DST) were done at Mycobacteriology Research Centre (MRC) of Jimma University. All data were registered and entered in to SPSS version 20. Crude odds ratio (COR) and adjusted odds ratios (AOR) were calculated. P-values less than 0.05 were considered statistically significant. RESULTS: Seventy-nine re-treatment cases included in the study; 48 (60.8%) were males. Forty-seven (59.5%) study participants were from rural area with the mean age of 31.67 ± 10.02 SD. DST results were available for 70 MTBC isolates. Majority (58.6% (41/70)) isolates were resistant to at least one of the four first line drugs. The prevalence of multidrug-resistant TB (MDR-TB) was 31.4% (22/70). Place of residence (AOR = 3.44 (95 % CI: 1.12, 10.60), duration of illness (AOR = 3.00 (95 % CI: 1.17, 10.69) and frequency of prior TB therapy (AOR = 2.99, (95 % CI: 1.01, 8.86) were significant factors for any drug resistance. Moreover, history of treatment failure was found to be associated with MDR-TB (AOR = 3.43 (95 % CI: 1.14, 10.28). CONCLUSION: The overall prevalence of MDR-TB among re-treatment cases around Jimma was high. The rate of MDR-TB was higher in patients with the history of anti-TB treatment failure. Timely identification and referral of patients with the history of treatment failure for culture and DST need to be strengthened.