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.

Fecal Carriage of Extended-Spectrum ß-Lactamase and Carbapenemase-Producing Enterobacteriaceae Among Oncology and Non-Oncology Patients at Jimma Medical Center in Ethiopia: A Comparative Cross-Sectional Study.

Purpose: Extended-spectrum ß-lactamase (ESBL) and carbapenemase-producing Enterobacteriaceae (CPE) are among the major threats to global health because of their encoded protection against key antibiotics. Methods: A comparative cross-sectional study was conducted among oncology and non-oncology patient groups (1:1; n = 214) on a consecutive sampling approach. Stool or rectal swab was collected from June 2021 to November 2021 and screened for ESBL-PE and CPE using ChromID-ESBL media. Confirmation for the enzymes was made by using combination disc and modified carbapenem inactivation methods, respectively. Disk diffusion method was used to determine antimicrobial susceptibility testing following the recommendations of CLSI 2022. SPSS software version 23 was used for data analysis. Results: Fecal carriage prevalence of ESBL-PE was found in 90 (84.1%) of oncology participants and in 77 (71.9%) of non-oncology patients (p = 0.032). Escherichia coli was the most common ESBL-PE isolate in 82 (62.5%) and 68 (88.3%) of oncology and non-oncology patients, followed by Klebsiella oxytoca [15 (11.5%) versus 6 (7.8%)], respectively. Out of the total ESBL-PE isolates from both oncology and non-oncology patient groups, the maximum level of resistance was observed against ciprofloxacin 177 (86.3%), trimethoprim-sulfamethoxazole 103 (80.3%), tetracycline 97 (75.8%), whereas enhanced susceptibility was appreciated to tigecycline 200 (97.6%), meropenem 162 (79.0%), and ertapenem 145 (70.7%) with no significant difference between oncology and non-oncology group. Carbapenemase-producing isolates from oncology patients were 12 (11.2%), whereas it was 4 (3.7%) (p = 0.611) from non-oncology group. Bacterial isolates from oncology in this study showed a trend of multiple drug resistance of 113 (88.3%). Conclusion: The results revealed alarmingly high carriage rates of ESBL and CPE among all study participants. Moreover, the isolates showed increased resistance rates to alternative drugs and had multiple antibiotic-resistant patterns. Hence, it is important to emphasize strict adherence to antimicrobial stewardship program as well as infection prevention and control practices.

Uropathogenic bacterial profile and antibiotic susceptibility pattern of isolates among gynecological cases admitted to Jimma Medical Center, South West Ethiopia.

Urinary tract infection (UTI) is one of the most common bacterial infections in women; about 50% of women get during their life time. Moreover, it is a common health problem in patients with gynecological pathologies, which increases the chance of acquiring infection. The aim of this study was to determine the bacterial profile that causes UTI and their antibiotic susceptibility pattern among admitted gynecological cases. A cross-sectional study was conducted in south west Ethiopia region. A total of 386 patients admitted with gynecological cases were recruited by sequential sampling technique and structured questionnaire was used to collect socio-demographic and risk factor-related data. About 10 ml freshly voided midstream and catheterized urine specimens were collected using sterile containers. Identification of isolate was done using culture characteristics, gram staining, and a series of biochemical tests. The antibiotic susceptibility test was performed as per the Kirby-Bauer disc diffusion technique. The data obtained were entered into EpiData Version 3.1 and analyzed using SPSS Version 25. A P value of less than 0.05 was used as a level of significance. In this study, the overall prevalence of UTI was 25.4%. Escherichia coli was the most frequently isolated bacteria, which accounted for 38 (37.6%), followed by Klebsiella species 22 (21.8%), CONS 14 (13.9%), Staphylococcus aureus 10 (9.9%), Enterobacter species 6 (5.9%), Citrobacter species 5 (4.9%), Proteus mirabilis 4 (4%), and Pseudomonas aeroginosa 2(2%). Histories of UTI (AOR = 1.977, 95% CI 1.06, 3.68, P = 0.032) and catheterization (AOR = 2.38, 95% CI 1.28, 4.45, P = 0.006) were found to be statistically associated with significant bacteriuria. Gram-negative isolates showed a high level of resistance, 88.3% for ampicillin and 66.2% for tetracycline, and a relatively low level of resistance against ceftazidime, 22.1%, and meropenem, 3.9%. Gram-positive uropathogens showed a high level of resistance to penicillin, 91.6%, whereas all isolates were sensitive 100.0% to nitrofurantoin. Furthermore, 80 (79.2%) of the isolates had multidrug resistance, and 16 (26.7%) of both E. coli and Klebsiella spp. produced Extended spectrum ß-lactamase (ESBL). In this study, a high prevalence of uropathogenic bacteria and multidrug resistance for commonly prescribed drugs were observed with a significant number of ESBL producers. Therefore, screening admitted gynecological patients, especially for those who have history of catheterization and UTI, by urine culture and antimicrobial susceptibility testing is important.

Isolation and Characterization of Lactic Acid Bacteria from Fermented Milk Produced in Jimma Town, Southwest Ethiopia, and Evaluation of their Antimicrobial Activity against Selected Pathogenic Bacteria.

Background: Raw milk is usually contaminated with pathogenic bacteria. Fermentation of milk is important to inhibit the growth of contaminants, spoilage, and pathogenic bacteria. The objective of this study was to isolate lactic acid bacteria from fermented milk and evaluate their antimicrobial activity against selected pathogenic bacteria. Methods: Laboratory-based experimental study design was conducted from May-July, 2021.Three samples of Ergo (each of 250 ml) were collected from Jimma town. Lactic acid bacteria (LAB) isolates were identified through integrated phenotypic techniques. Further identification was conducted through using API 50 CHL strips. Antimicrobial activities (AMAs) of LAB isolates were tested against clinical isolates of E. coli, S. aureus, and Salmonella spp. using agar well diffusion method. The data were analyzed by using SPSS software version 21 and Microsoft Excel spreadsheet. Tables and figures were applied to describe characteristics of data. Results: Twelve LAB isolates were identified. Those LAB isolates include six Lactococcus lactis subsp. lactis, Lactobacillus acidophilus (2), Lactiplantibacillus plantarum (1), Limosilactobacillus fermentum (2), and Leuconostoc lactis (1). Based on primary screening of LAB, isolates/strains ESCIa, ESBIa, and ESCIc show strong AMA against S. aureus, E. coli, and Salmonella spp. The CFS of ESCIc showed the highest AMA against S. aureus and Salmonella spp. with a zone of inhibition of 14.12 ± 1.6 mm and 12.9 ± 3.6 mm, respectively, while ESBIa showed the highest AMA against E. coli with a zone of inhibition of 13.5 ± 2.1 mm. The CFSs of selected LAB strains were heat tolerant at varying temperatures up to 100°C. The CFSs of selected LAB strains were inactivated by proteinase enzymes, but they are not inactivated with amylase enzymes. Conclusions and Recommendation. All 12 LAB isolates exhibited antimicrobial activity against tested bacterial strains. Lactobacillus isolates showed the highest antagonistic activity on tested indicator strains. Thus, they are possible alternatives to antibiotics in the era of antimicrobial resistance. S. aureus was the most sensitive to antimicrobial effects/agents of selected LAB isolates. Consumption of fermented foods is advisable since they support the growth of healthy GIT microbiota. Fermentation serves as biopreservation of food. However, analysis of probiotic features and in vivo probiotic effects of those LAB isolates will be subject of future research/study.

Stool-based Xpert MTB/RIF assay for the diagnosis of pulmonary tuberculosis in children at a teaching and referral hospital in Southwest Ethiopia.

BACKGROUND: Diagnosis of tuberculosis (TB) in children is challenging mainly due to the difficulty of obtaining respiratory specimen and lack of sensitive diagnostic tests. The objective of this study was to evaluate the diagnostic performance of Xpert MTB/RIF (Xpert here after) for the diagnosis of pulmonary TB (PTB) from stool specimen in children. METHODS: A cross-sectional study was conducted among consecutively recruited children (less than 15 years old) with presumptive PTB at Jimma Medical Center, Ethiopia. One pulmonary specimen (expectorated sputum or gastric aspirate) was collected from each participant and tested for TB by Xpert and Lowenstein-Jensen (LJ) culture. In addition, one stool specimen per child was collected and tested by Xpert after a single step, centrifuge-free stool processing method adapted from KNCV TB Foundation. Diagnostic performance of Xpert was calculated with reference to LJ culture and to a composite reference standards (CRS) comprising of confirmed TB (positive by Xpert and/or culture) and unconfirmed TB (clinical diagnosis with improvement after anti-TB treatment). RESULTS: A total of 178 children were enrolled; 152 of whom had complete microbiological results. Overall, TB was diagnosed in 13.2% (20/152) of the children with presumptive TB. Of these, only ten had microbiologically confirmed TB (positive Xpert and/or culture) and the remaining ten were clinically diagnosed with positive response to anti-TB treatment and were classified as unconfirmed TB. Stool Xpert had sensitivity of 100% (95%CI: 66.4-100) and specificity of 99.3% (95%CI: 96.2-100) compared to culture; however, the sensitivity was decreased to 50% (95%CI: 27.2-72.8) when compared to CRS. The Xpert on gastric aspirate had sensitivity of 77.8% (95%CI: 40-97.2) compared to culture and 40% (95%CI: 19.1-64) compared to CRS. CONCLUSIONS: The sensitivity of Xpert for stool sample is comparable to that for gastric aspirate. Stool sample is a potential alternative to pulmonary specimen in the diagnosis of pulmonary TB in children using Xpert.

Antibody and cytokine levels in visceral leishmaniasis patients with varied parasitemia before, during, and after treatment in patients admitted to Arba Minch General Hospital, southern Ethiopia.

BACKGROUND: Visceral leishmaniasis is a disease caused by disseminated Leishmania donovani infection which affects almost half a million people annually. Most of the patients are reported from the Indian sub-continent, Eastern Africa and Brazil. In this study, we aimed to determine the levels of antibodies and cytokines in visceral leishmaniasis patients and to examine associations of parasitemia with the clinical states of patients. A prospective study was carried out, enrolling a total of 48 active VL patients who were evaluated before, during different time points and, three months after treatment. Serum cytokine concentrations, antibody levels, parasitemia, laboratory (hematologic and biochemical) measurements, and clinical parameters were assessed. RESULTS: Counts of WBC and platelets, and measurements of hemoglobin (Hb) increased during treatment (P ≤ 0.05). Elevated levels of circulating IL-10, IFN-γ, and TGF-ß1 were measured before treatment. The observed increase in serum IL-10 remarkably declined within 7 days after the start of treatment. Anti-leishmanial antibody index (AI) was high in all VL patients irrespective of spleen aspirate parasite grade before treatment and at different times during treatment. However, a significant (P ≤ 0.05) decrease of AI was observed 120 days post-treatment. IL-2 serum levels were below the detection limit at all sampling points. CONCLUSIONS: The present results suggest that IL-10, IFN-γ, and TGF-ß1 can be used as markers of active visceral leishmaniasis. In addition, measuring circulating cytokines concentrations, particularly IL-10, in combination with other clinical evaluations, could be used as criteria for the cure. The observation that a high serum concentration of IFN-gamma at baseline was associated with low parasitemia deserves further investigations.

Antimicrobial Resistance Profile of Different Clinical Isolates against Third-Generation Cephalosporins.

BACKGROUND: Drug resistant microorganisms lead to an increase in morbidity and mortality as they boost the risk of inappropriate therapy. Hence, data on antimicrobial resistance help define the best possible treatment for individual patients. Therefore, this study aimed to screen the antimicrobial resistant profile of 3rd generation cephalosporin drugs in Jimma University Specialized Teaching Hospital. METHODS: A hospital based prospective cross-sectional study was conducted in Jimma University Specialized Hospital (JUSH) from April to August 2016. The clinical samples such as wound swab, urine, sputum, and stool were collected from hospitalized patients. Then, bacterial species were isolated and identified as per the standard microbiological methods. Antimicrobial susceptibility tests were carried out using various antimicrobial discs by Kirby-Bauer disc diffusion method. RESULTS: Totally, 248 bacterial isolates were obtained from 154 (62.1%) male and 94 (37.9%) female patients. Escherichia coli (25.4%) and Staphylococcus aureus (19.0 %) were the predominant organisms isolated from specimens. About 140 (56.5%) and 149 (60.1%) of the total bacterial isolates were found to be resistant to ceftriaxone and ceftazidime, respectively. The majority of Escherichia coli isolates 46 (73%) were resistant to ceftriaxone and 41 (65%) of them were resistant to ceftazidime. Staphylococcus aureus, which accounted 19% of the total bacterial isolates, showed 23.4% and 34% resistance to ceftriaxone and ceftazidime, respectively. Among the bacterial strains revealing resistant to ceftriazone and ceftazidime, about 109 (44%) and 108 (43.5%) of them were resistant to two, three, or four other drugs, respectively. CONCLUSION: Bacterial resistance towards third-generation cephalosporin (ceftriaxone and ceftazidime) is escalating as more than half of the isolated strains demonstrated resistance to these drugs. Moreover, these strains also revealed multidrug resistance mainly against clinically used drugs which could render therapy unsuccessful. Therefore, in clinical use appropriate medications should be selected based on the data obtained from antimicrobial susceptibility tests.

Nasopharyngeal Carriage and Antimicrobial Susceptibility Patterns of Streptococcus pneumoniae among Children under Five in Southwest Ethiopia.

Nasopharyngeal carriage of Streptococcus pneumoniae is found to play an important role in the development and transmission of pneumococcal diseases. In this study, we assessed the nasopharyngeal carriage, antimicrobial susceptibility patterns and associated risk factors of S. pneumoniae among children under five. A total of 361 children under five attending the outpatient department of Shanan Gibe Hospital in Jimma, Southwest Ethiopia were enrolled from June to September 2014. Nasopharyngeal specimens were collected using sterile plastic applicator rayon tipped swab and inoculated on tryptone soy agar supplemented with 5% sheep blood and 5 µg/mL gentamycin. Antimicrobial susceptibility testing was performed using the modified disk diffusion method. The overall prevalence of S. pneumoniae carriage was 43.8% (158/361) among children under five. Resistance to tetracycline, cotrimoxazole, penicillin, chloramphenicol and erythromycin was observed in 53.2% (84/158), 43.7% (69/158), 36.1% (57/158), 13.3% (21/158) and 8.9% (14/158) of isolates respectively. Multidrug resistance was seen in 17.7% (28/158) of isolates. In multivariate logistic regression analysis, children living with sibling(s) < 5 years old (adjusted odds ratio (AOR) = 1.798; 95% confidence interval (CI), 1.169-2.766) and malnutrition (AOR = 2.065; 95% CI, 1.239-3.443) were significantly associated with S. pneumoniae carriage. A high nasopharyngeal carriage of S. pneumoniae was observed among children under five in Southwest Ethiopia. There should be a strategy to prevent S. pneumoniae nasopharyngeal colonization and identify the appropriate antibiotic to the individual child.

Resistance to first-line anti-TB drugs is associated with reduced nitric oxide susceptibility in Mycobacterium tuberculosis.

BACKGROUND AND OBJECTIVE: The relative contribution of nitric oxide (NO) to the killing of Mycobacterium tuberculosis in human tuberculosis (TB) is controversial, although this has been firmly established in rodents. Studies have demonstrated that clinical strains of M. tuberculosis differ in susceptibility to NO, but how this correlates to drug susceptibility and clinical outcome is not known. METHODS: In this study, 50 sputum smear- and culture-positive patients with pulmonary TB in Gondar, Ethiopia were included. Clinical parameters were recorded and drug susceptibility profile and spoligotyping patterns were investigated. NO susceptibility was studied by exposing the strains to the NO donor DETA/NO. RESULTS: Clinical isolates of M. tuberculosis showed a dose- and time-dependent response when exposed to NO. The most frequent spoligotypes found were CAS1-Delhi and T3_ETH in a total of nine known spoligotypes and four orphan patterns. There was a significant association between reduced susceptibility to NO (>10% survival after exposure to 1 mM DETA/NO) and resistance against first-line anti-TB drugs, in particular isoniazid (INH). Patients infected with strains of M. tuberculosis with reduced susceptibility to NO showed no difference in cure rate or other clinical parameters but a tendency towards lower rate of weight gain after two months of treatment, independent of antibiotic resistance. CONCLUSION: There is a correlation between resistance to first-line anti-TB drugs and reduced NO susceptibility in clinical strains of M. tuberculosis. Further studies including the mechanisms of reduced NO susceptibility are warranted and could identify targets for new therapeutic interventions.