Molecular epidemiology of Streptococcus pneumoniae isolates causing invasive and noninvasive infection in Ethiopia.

Streptococcus pneumoniae, a medically important opportunistic bacterial pathogen of the upper respiratory tract, is a major public health concern, causing a wide range of pneumococcal illnesses, both invasive and noninvasive. It is associated with significant global morbidity and mortality, including pneumonia, meningitis, sepsis, and acute otitis media. The major purpose of this study was to determine the molecular epidemiology of Streptococcus pneumoniae strains that cause invasive and noninvasive infections in Ethiopia. A prospective study was undertaken in two regional hospitals between January 2018 and December 2019. Whole-genome sequencing was used to analyze all isolates. Serotypes and multilocus sequence types (MLST) were derived from genomic data. The E-test was used for antimicrobial susceptibility testing. Patient samples obtained 54 Streptococcus pneumoniae isolates, 33 from invasive and 21 from noninvasive specimens. Our findings identified 32 serotypes expressed by 25 Global Pneumococcal Sequence Clusters (GPSCs) and 42 sequence types (STs), including 21 new STs. The most common sequence types among the invasive isolates were ST3500, ST5368, ST11162, ST15425, ST15555, ST15559, and ST15561 (2/33, 6% each). These sequence types were linked to serotypes 8, 7 C, 15B/C, 16 F, 10 A, 15B, and 6 A, respectively. Among the noninvasive isolates, only ST15432, associated with serotype 23 A, had numerous isolates (4/21, 19%). Serotype 14 was revealed as the most resistant strain to penicillin G, whereas isolates from serotypes 3, 8, 7 C, and 10 A were resistant to erythromycin. Notably, all serotype 6 A isolates were resistant to both erythromycin and penicillin G. Our findings revealed an abnormally significant number of novel STs, as well as extremely diversified serotypes and sequence types, implying that Ethiopia may serve as a breeding ground for novel STs. Recombination can produce novel STs that cause capsular switching. This has the potential to influence how immunization campaigns affect the burden of invasive pneumococcal illness. The findings highlight the importance of continuous genetic surveillance of the pneumococcal population as a vital step toward enhancing future vaccine design.

Nasopharyngeal carriage, antimicrobial susceptibility patterns, and associated factors of Gram-positive bacteria among children attending the outpatient department at the University of Gondar Comprehensive Specialized Hospital, Northwest Ethiopia.

BACKGROUND: Gram-positive bacteria residing in the nasopharynx can lead to severe illnesses in children, such as otitis media, pneumonia, and meningitis. Despite the potential threat, there is a lack of comprehensive data regarding the carriage rates of these bacteria among children in outpatient departments in the study area. OBJECTIVE: This study aimed to assess the nasopharyngeal carriage, antimicrobial resistance patterns, and associated factors of Gram-positive bacteria among children attending the outpatient department at the University of Gondar Comprehensive Specialized Hospital, Northwest Ethiopia. METHODS: A hospital-based cross-sectional study was conducted from May 1, 2023, to August 30, 2023. A total of 424 nasopharyngeal swab samples were collected using sterile nasopharyngeal swabs, inoculated on Blood Agar and Mannitol Salt Agar plates, and identified through colony morphology, Gram stain, and biochemical tests. Antimicrobial susceptibility of the identified bacterial isolates was determined employing both the Kirby-Bauer and modified Kirby-Bauer methods. D-tests were conducted using clindamycin and erythromycin discs to detect inducible clindamycin resistance, while cefoxitin disc tests were utilized to ascertain methicillin resistance. Data entry was executed using Epi-Data version 4.6, and subsequent analysis was performed utilizing SPSS version 25. Bivariable and multivariable logistic regression analyses were employed to identify associated factors. An adjusted odds ratio at a 95% confidence interval with a P-value of < 0.05 was considered statistically significant. RESULTS: The overall nasopharyngeal carriage rate of Gram-positive bacteria was 296/424 (69.8%, 95% CI: 65.3-74.0). Staphylococcus aureus was the most prevalent 122/424 (28.8%), followed by Streptococcus pneumoniae 92/424 (21.7%). Methicillin resistance was observed in 19/122 (15.6%) of S. aureus and 3/60 (5%) of coagulase-negative staphylococcus (CoNS) species. Inducible clindamycin resistance was 10/122 (8.2%) in S. aureus and 4/53 (7.5%) in coagulase-negative staphylococcus species. Multidrug resistance was found in 146/296 (49.3%, 95% CI: 43.6-55.0) of the isolates. Associated factors with a bacterial carriage were large family size (AOR = 3.061, 95% CI: 1.595-5.874, P = 0.001), having siblings under five years old (AOR = 1.991, 95% CI: 1.196-3.313, P = 0.008), indoor cooking (AOR = 2.195, 95% CI: 1.275-3.778, P = 0.005), an illiterate mother (AOR = 3.639, 95% CI: 1.691-7.829, P = 0.001), and hospital visits (AOR = 2.690, 95% CI: 1.405-5.151, P = 0.003). CONCLUSION: The study found a high nasopharyngeal carriage of Gram-positive bacteria in outpatient children, including notable levels of methicillin-resistant S. aureus and multi-drug-resistant isolates. Clindamycin, rifampin, and erythromycin were the most effective antimicrobials for the tested isolates. Factors contributing to bacterial carriage include visits to healthcare facilities, larger family sizes, having younger siblings, maternal illiteracy, and indoor cooking. This emphasizes the need for methicillin-resistant S. aureus surveillance in pediatric outpatient settings and community health education, especially for children's guardians. Additionally, improving household ventilation by separating kitchens from sleeping areas and regular screening of younger siblings in healthcare environments were recommended to reduce bacterial transmission within family members. The study also called for studies with advanced procedures like minimum inhibitory concentration testing and molecular characterization to better comprehend the resistance patterns and genes in circulating bacteria.

Magnitude and antimicrobial susceptibility profile of bacteria isolated from pediatric sepsis cases at University of Gondar Hospital, Northwest Ethiopia.

BACKGROUND: Sepsis is one of the major causes of morbidity and mortality among pediatric patients throughout the world. The varying microbiological pattern of sepsis warrants the need for researches on the causative organisms and their antimicrobial susceptibility pattern. The epidemiology of neonatal and pediatric sepsis in Ethiopia is under-research. The objective of this study was to evaluate the burden of bacterial pathogens and their antimicrobial susceptibility patterns among children suspected of sepsis. METHODS: An institutional-based prospective cross-sectional study was conducted on 370 pediatric(age birth-15 years) patients suspected of sepsis at the University of Gondar Comprehensive Specialized hospital from December 2020 to November 2021. Blood samples were collected aseptically and inoculated into Tryptone Soya Broth for culture. The organisms grown were identified by standard microbiological methods and subjected to antibiotic susceptibility testing by modified Kirby-Bauer disk diffusion method recommended by Clinical laboratory and standard institute. Methicillin resistance was confirmed using Cefoxitin disk diffusion method. Data entry and analysis were done using Statistical Package for Social Sciences (SPSS) version 26 software. A p-value less than 0.05 at 95% confidence interval was considered statically significant. RESULTS: Out of the total 370 study subjects, 21.6% (80/370) of them were culture positive. Of these, 43 (53.8%) and 37 (46.3%) were Gram-positive and Gram-negative bacterial pathogens, respectively. The most prevalent Gram-positive bacterial isolate was Staphylococcus aureus (n = 24; 30%) and coagulase negative staphylococci (n = 7; 8.8%). Among the Gram-negative bacterial isolates, the leading bacteria was Klebsiella pneumoniae (n = 20; 25%) followed by Escherichia coli (n = 7; 8.8%). Clindamycin, Chloramphenicol, Gentamicin and Ciprofloxacin were the most effective antibiotics against Gram-positive bacterial isolates while Amikacin, Meropenem and Chloramphenicol were effective against Gram-negative pathogens. Methicillin resistance was detected in 45.8% of Staphylococcus aureus. Multi-drug resistance (MDR) antimicrobial susceptibility pattern was observed in 76% of the bacterial isolates. CONCLUSION: Gram positive bacteria were the predominant isolates among pediatric sepsis cases and most of the bacterial isolates showed MDR. Staphylococcus aureus and Klebsiella pneumoniae were frequently isolated bacteria. The high prevalence of drug resistance warrants rational use of antibiotics and the need for regular antibiotic susceptibility surveillance studies.

Prevalence of Salmonella spp., Shigella spp., and intestinal parasites among food handlers working in University of Gondar student's cafeteria, Northwest Ethiopia.

Background: Food-borne infections continue to be a major public health problem at the international level. The issue becomes more serious in developing countries like Ethiopia. Objective: This study aimed to examine the prevalence of Salmonella and Shigella species and intestinal parasites, as well as antimicrobial resistance patterns and associated factors among food handlers at the University of Gondar cafeteria in northwest Ethiopia. Methods: An institutional-based cross-sectional study was conducted from February to June 2021 in the University of Gondar cafeterias. Data related to the socio-demographic characteristics and hygienic practices of study participants were collected using structured questionnaires. A total of 290 stool samples were collected from food handlers. Culture and conventional biochemical tests were used to isolate the Salmonella and the Shigella species. Wet mount, Formol-ether concentration, and Kato Katz techniques were applied to identify intestinal parasites. Additionally, drug susceptibility tests were performed using the disk diffusion method. Statistical analysis was done using SPSS version 26. Results: Of 290 food handlers' stool samples analyzed, Twenty-seven 27 (9.3%) were positive for both Salmonella and Shigella species. The prevalence of Salmonella and Shigella species was 16 (5.5%) and 11 (3.8%), respectively. Most of the isolated pathogens were resistant to tetracycline 19 (70.4%), and trimethoprim/sulphamethoxazole 19 (70.4%). The overall rate of multi-drug resistant Shigella and Salmonella isolate was 59.3%. Besides, Fifty-seven 57 (19.7%) of the participants were positive for one or more intestinal parasites. The most prevalent intestinal Parasitosis was E. histolytica/dispar 22 (7.6%), followed by G. lamblia 13 (4.5%), and Ascaris lumbricoides 11 (3.8) not washing hands after using the toilet (AOR: 4.42, 95% CI: 1.57, 10.56), and consuming unpasteurized milk (AOR: 3.14, 95% CI: 1.65, 3.96), were factors significantly associated with the prevalence of Salmonella, and Shigella infection. Similarly, not washing hands after using the toilet (AOR: 2.19, 95% CI: 1.0, 1.4), and consuming unpasteurized milk (AOR: 10.4, 95% CI: 3.8, 28.8), were factors significantly associated with the prevalence of intestinal parasites infection. Conclusion: The prevalence of intestinal parasites, Salmonella, and Shigella species was high. Therefore, it is imperative to implement a public health policy that includes ongoing microbiological surveillance.

Shiga toxin-producing Escherichia coli O157:H7 among diarrheic patients and their cattle in Amhara National Regional State, Ethiopia.

BACKGROUND: Shiga toxin-producing Escherichia coli O157:H7 (STEC O157:H7) is a zoonotic pathogen that causes diarrhea, hemorrhagic colitis, and hemolytic uremic syndrome worldwide. This study aimed to determine the prevalence, antibiotic susceptibility, and associated risk factors of STEC O157:H7 among diarrheic patients and their cattle. METHODS: A cross-sectional study was conducted among diarrheic patients and their cattle in Amhara National Regional State, Ethiopia from December- 2020 to June- 2022. A total of 1,149 diarrheic patients and 229 cattle were included in the study. STEC O157:H7 detection was done using culture, latex agglutination test, and polymerase chain reaction on diarrheic stool samples and recto-anal mucosal swabs of cattle. Antibiotic susceptibility tests were performed using disk diffusion techniques. Risk factors association were identified using binary and multivariable logistic regression analysis. RESULTS: The overall prevalence of STEC O157:H7 in diarrheic patients and their cattle was 11.1% (128/1149) and 14.4% (33/229) respectively. High percentage of the study subjects were found in under-five children (34.5%). Age less than 5 (AOR: 4.02, 95%CI:1.608-10.058,P = 0.003), and greater than 64 years old (AOR:3.36, 95% CI:1.254-8.986, P = 0.016), presence of diarrheic patient in the house (AOR:2.11, 95%CI:1.309-3.390, P = 0.002), availability of cattle in the house (AOR:2.52, 95%CI:1.261-5.049, P = 0.009), and habit of consuming raw foods (AOR:4.35, 95%CI:2.645-7.148, P = 0.000) were risk factors. Antibiotic resistance was shown in 109(85.2%), and 31(93.9%) isolates from diarrheic patients and their cattle respectively. The highest levels of antibiotic resistance were found to tetracycline (54.7%, 69.7%) in diarrheic patients and their cattle respectively. Multiple drug resistance was also observed among 56(43.8%) and 11(33.3%) isolates in diarrheic patients and their cattle respectively. CONCLUSION: Our study showed high prevalence of STEC O157:H7 in diarrheic patients and their cattle. Therefore, health education should be given to the community on how to care for animals, proper sanitation, and the impact of raw food consumption.

Multidrug-resistant and carbapenemase-producing critical gram-negative bacteria isolated from the intensive care unit environment in Amhara region, Ethiopia.

BACKGROUND: Intensive care units are units where healthcare-associated infections (HAIs) are common and antimicrobial resistance rates are increasing. Microbial contamination in hospital environment plays an important role in the development of HAIs. Intervention-based improvements in infection prevention and control at national and facility level are critical for the containment of antimicrobial resistance and prevention of HAIs. OBJECTIVES: This study aimed to determine the distribution of multidrug-resistant and carbapenemase-producing critical gram negative bacteria (Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa and Acinetobacter species) and their antibiotic resistance in intensive care unit environmental surfaces at the University of Gondar and Felege Hiwot Comprehensive Specialized Hospitals. METHODS: This was multicenter hospital-based cross sectional study. Environmental samples were swabbed from all intensive care units using a normal saline moistened-sterile cotton tip stick. Bacteria culturing and antibiotic susceptibility testing were performed following standard microbiological techniques. Selected meropenem-resistant isolates were phenotypically assessed for carbapenemase production using modified and simplified carbapenem inactivation methods. RESULTS: From a total of 384 environmental samples analyzed, 126 (32.8%) showed growth and 162 isolates were identified. K. pneumoniae (79/162, 48.8%) was the commonest isolate followed by Acinetobacter species (51/162, 31.5%), E. coli (19/162, 11.7%) and P. aeruginosa (13/162, 8.0%). Multidrug-resistant and carbapenemase-producing isolates were detected on most hospital environment surface types, especially from the baby bed sets and incubators. The most common multidrug-resistant and principal carbapenemase producer was K. pneumoniae, with rates of 71(89.9%) and 24(85.7%), respectively. CONCLUSION: This study revealed the distribution of multidrug-resistant and carbapenemase-producing critical gram negative bacteria in the environment of intensive care unit. Higher detection rate of multidrug-resistant and carbapenemase-producing K. pneumoniae on most environmental surfaces calls for urgent control action and further attention.

Prevalence of Methicillin-ResistantStaphylococcus aureus and Multidrug-Resistant Strains from Patients Attending the Referral Hospitals of Amhara Regional State, Ethiopia.

Background: Staphylococcus aureus (S. aureus) causes different types of human infections and can develop resistance to many antibiotics. There is a scarcity of data on the mecA gene and multidrug-resistant (MDR) strain distribution of this organism in developing countries, such as Ethiopia. This study investigated the presence of mecA gene and MDR profile of S. aureus among patients attending referral hospitals of Amhara regional state. Methods: Of the total of 110 isolates collected from Amhara regional referral hospitals, 70 MDR isolates were further processed for isolation of S. aureus mecA gene. Genomic DNA was isolated using a Sigma-Aldrich genomic DNA isolation kit for Gram-positive bacteria. Amplification of S. aureus mecA gene was performed with the amplicon size of 533 bp. Antimicrobial susceptibility test including methicillin resistance was determined by the Kirby-Bauer disc diffusion method. Results: The majority of the isolates were recovered from patients aged less than 5 years (51; 36.7%) and the least number of isolates was recorded in age group greater than 60 years (6; 4.3%). Most of the isolates were from blood (61; 43.9%), followed by wounds (45; 32.4%). A high resistance rate was observed in penicillin (81; 73.6%), followed by cotrimoxazole (78; 70.9%), ceftriaxone (76; 69%), erythromycin (66; 60%), and tetracycline (65; 59.1%). Phenotypically, considering cefoxitin as a surrogate marker, 38 (34.5%) of the isolates were methicillin-resistant. The overall MDR isolates were 80 (72.7%). The PCR amplification result of the mecA gene was 14 (20%). Conclusions and Recommendations. High rates of MDR and methicillin-resistantS. aureus were reported. PCR amplification indicated that 20% of MRSA isolates were the mecA gene carriers. Large-scale studies for the detection of MDR strains of S. aureus including MRSA using molecular techniques should be encouraged in the Amhara region.

Multidrug-resistant and extended-spectrum beta-lactamase-producing Enterobacteriaceae isolated from chicken droppings in poultry farms at Gondar City, Northwest Ethiopia.

BACKGROUND: The poultry sector is one of the largest and fastest-growing agricultural sub-sector, especially in developing countries like Ethiopia. In poultry production, poultry farmers use sub-optimum doses of antibiotics for growth promotion and disease prevention purpose. This indiscriminate use of antibiotics in poultry farms contributes to the emergence of antibiotic-resistant bacteria, which has adverse implications for public health. Therefore, this study is aimed to assess multidrug resistance and extended-spectrum beta-lactamase-producing Enterobacteriaceae from chicken droppings in poultry farms. METHODS: A total of 87 pooled chicken-dropping samples were collected from poultry farms from March to June 2022. Samples were transported with buffered peptone water. Selenite F broth was used for the enrichment and isolation of Salmonella spp. Isolates were cultured and identified by using MacConkey agar, Xylose lysine deoxycholate agar, and routine biochemical tests. Kirby-Bauer disk diffusion technique and combination disk test were used for antibiotic susceptibility testing and confirmation of extended-spectrum beta-lactamase production, respectively. Data were entered using Epi-data version 4.6 and then exported to SPSS version 26 for analysis. RESULT: Out of 87 pooled chicken droppings, 143 Enterobacteriaceae isolates were identified. Of these, E. coli accounts for 87 (60.8%), followed by Salmonella spp. 23 (16.1%), P. mirabilis 18 (12.6%) and K. pneumoniae 11 (7.7%). A high resistance rate was observed for ampicillin 131 (91.6%), followed by tetracycline 130 (90.9), and trimethoprim-sulfamethoxazole 94 (65.7%). The overall multidrug resistance rate was 116/143 (81.1%; 95% CI: 74.7-87.5). A total of 12/143 (8.4%; CI: 3.9-12.9) isolates were extended-spectrum beta-lactamase producers, with 11/87 (12.6%) E. coli and 1/11 (9.1%) K. pneumoniae. CONCLUSION AND RECOMMENDATIONS: High prevalence of multi-drug resistant isolates was observed. This study alarms poultry as a potential reservoir of extended-spectrum beta-lactamase-producing Enterobacteriaceae, which might shed and contaminate the environment through faecal matter. Prudent use of antibiotics should be implemented to manage antibiotic resistance in poultry production.

Molecular diversity of hepatitis B virus among pregnant women in Amhara National Regional State, Ethiopia.

BACKGROUND: Despite the availability of effective vaccines and treatments for hepatitis B virus (HBV), it continues to be a major public health problem in sub-Saharan Africa including Ethiopia. Routine screening for HBV in pregnant women is widely recommended, but there is lack of screening for HBV during pregnancy in Ethiopia. Therefore, this study aimed to assess viral load, and genetic diversity among pregnant women in the Amhara National Regional State, Ethiopia. MATERIALS AND METHODS: Hepatitis B surface antigen (HBsAg) testing was performed on 1846 pregnant women, 85 of who tested positive were included in this study. HBV DNA was isolated from 85 positive sera, and the partial surface/polymerase gene was amplified and sequenced. HBV genotypes, sub-genotypes, serotypes and mutations in surface genes and polymerase were studied. RESULTS: Out of 85 pregnant women`s HBsAg positive sera, 59(69.4%) had detectable viral DNA. The median viral load was 3.4 log IU/ml ranging from 2.6 to7.6 and 46 samples were successfully sequenced and genotyped. Genotypes A and D were identified in 39 (84.8%) and 7 (15.2%); respectively. All genotype A isolates were further classified into sub-genotype A1 and serotype adw2 (84.8%) whereas genotype D isolates were further classified into three sub genotypes; 2 (4.3%) D2, 1(2.2%) D4, and 4 (8.7%) D10 with serotypes ayw2 (10.9%), and ayw3 (4.3%). There were 19 (41.3%) surface gene mutations in the major hydrophilic region (MHR). Six (13.1%) of them were discovered in MHR`s `a'-determinant region. Six polymerase gene mutations (13%) were identified. CONCLUSION: Genotype A was the predominant genotype in the Amhara National Regional State. The surface and polymerase gene mutations identified in this study may lead to immune therapy failure, diagnostics escape and drug resistance. Thus, the data generated in this study will contribute to the planning of HBV diagnosis, vaccination and treatment, and most importantly to the prevention of vertical transmission of HBV in Ethiopia. Therefore, further molecular studies on HBV are warranted and continuous surveillance is important for patient management and for the prevention and control of HBV infection in the country.

Extended-Spectrum Beta-Lactamase- and Carbapenemase-Producing Enterobacteriaceae Family of Bacteria from Diarrheal Stool Samples in Northwest Ethiopia.

Background: Resistance among the commensal flora is a serious threat because they are highly populated ecosystems like the gut, maybe a source of extraintestinal infections. Infections due to extended-spectrum beta-lactamase (ESBL)- and carbapenemase (CPM)-producing Enterobacteriaceae family of bacteria impose a major global issue because they are usually resistant to multiple antimicrobial agents. Data on the fecal ESBL- and CPM-producing group of bacteria in developing countries including Ethiopia are limited mainly due to resource constraints. Thus, this study aimed to determine the prevalence of multidrug-resistant (MDR)-, ESBL-, and CPM-producing Enterobacteriaceae family of bacteria from diarrheal stool samples at the University Hospital, Northwest Ethiopia. Materials and Methods: A hospital-based cross-sectional study was conducted involving a total of 384 study participants having gastrointestinal complaints from January to April 2019. A diarrheal stool sample was aseptically collected and inoculated on a MacConkey agar plate. After getting pure colonies, biochemical and antimicrobial susceptibility testing was done following standard microbiological techniques. ESBL production was screened using ceftazidime and cefotaxime and confirmed using a combined disk diffusion test. Carbapenemases were screened by meropenem disk and confirmed by the modified carbapenem inactivation method. Data were checked, cleaned, and entered using Epi Info version 7.1 and transferred to SPSS version 20 for analysis. Result: A total of 404 Enterobacteriaceae groups of bacteria were isolated from 384 diarrheal stool samples. The overall prevalence of fecal MDR-, ESBL-, and CPM-producing group of Enterobacteriaceae was 196 (48.5%), 66 (16.3%), and 4 (1%), respectively. Of the total ESBL-producing Enterobacteriaceae, E. coli (41/66 (62.1%)) and K. pneumoniae (18/66 (27.3%)) were the most predominant isolates. One half of CPE has been observed in Citrobacter species and the rest in E. coli (25%) and P. vulgaris (25%). Conclusion and Recommendation. Finding the high rate of ESBL-producing Enterobacteriaceae and CPE requires strict infection control measures and careful selection of empirical therapy in the study area. Therefore, active surveillance with large sample size and better infection prevention control is needed.

High prevalence of fecal carriage of Extended-spectrum beta-lactamase and carbapenemase-producing Enterobacteriaceae among food handlers at the University of Gondar, Northwest Ethiopia.

BACKGROUND: Fecal carriage of extended-spectrum beta-lactamase and Carbapenemase-producing Enterobacteriaceae is a potential risk for the transmission of infection with resistant strains. Understanding the burden of these resistant strains in asymptomatic people is essential to reduce the chain of infection transmission. However, data on the fecal carriage of Extended-spectrum Beta-lactamase and Carbapenemase-producing Enterobacteriaceae among food handlers were limited in developing countries especially in Ethiopia. The aim of the present study is, therefore, to assess fecal carriage rate, associated factors, and antimicrobial resistance patterns of Extended-spectrum Beta-lactamase and Carbapenemase-producing Enterobacteriaceae among food handlers at the University of Gondar Cafeterias, Northwest Ethiopia. MATERIALS AND METHODS: An institution-based cross-sectional study was conducted from February to June 2021 at the University of Gondar cafeterias. A total of 290 stool samples were collected, transported using Cary Blair transport medium, and processed. All isolates were cultured and identified by using MacConkey agar, and routine biochemical tests. Antimicrobial susceptibility testing was done to each isolate following the Kirby Bauer disk diffusion method. If the zone of inhibition was ≤ 22 mm for ceftazidime, ≤25 mm for ceftriaxone, and ≤27 for cefotaxime they were considered as potential ESBL strain and selected for a further phenotypic confirmatory. Moreover, the double-disc diffusion test and the modified carbapenem inactivation method were used for confirmations of Extended-spectrum ß-lactamase and Carbapenemase-producing Enterobacteriaceae respectively. If a ≥5mm difference in zone diameter for either antimicrobial agent in combination with clavulanic acid versus the zone diameter of the agent when tested alone (without B-lactamase inhibitor), was confirmed as ESBL-PE and if the zone of inhibition diameter between 6-15mm and 16- 18mm with a pinpoint colony, it was considered as carbapenem resistance Enterobacteriaceae. Data were entered using Epi-data version 4.6 and then exported to SPSS version 26 for analysis. Potential risk factors were assessed using multivariable logistic regression and a p-value less than 0.05 was considered statistically significant. RESULTS: Out of 290 stool samples, 63 (21.7%) and 7 (2.4%) were confirmed as Extended-spectrum ß-lactamase and Carbapenemase-producing Enterobacteriaceae. The most predominant ESBL-PE was E. coli 43 (14.8%) followed by K. pneumoniae 17 (5.9%). Most of the Extended-spectrum ß-lactamase and Carbapenemase-producing isolates were resistant to tetracycline, cefotaxime, ceftazidime, and ceftriaxone (100% each). In contrast, a low resistance level was recorded for Meropenem and cefoxitin. The overall Multi-drug resistant Enterobacteriaceae (MDR) was 147 (42.3%). Antibiotics usage in the last 3 months and drinking unpasteurized milk were associated with the carriage of the Extended-spectrum beta-lactamase-Producing Enterobacteriaceae. CONCLUSIONS AND RECOMMENDATIONS: The high fecal carriage rate of Multi-drug resistance isolate, Extended-spectrum ß-lactamase, and Carbapenemase-producing Enterobacteriaceae were recorded among food handlers. Therefore, this study gives signals in the spread of drug-resistant bacteria easily to the community. Hence, the need for adjusting and promotion of infection prevention measures to prevent the spread of drug-resistant bacteria should not be underestimated.

Prevalence and antimicrobial resistance pattern of Clostridium difficile among hospitalized diarrheal patients: A systematic review and meta-analysis.

BACKGROUND: Clostridium difficile is the leading cause of infectious diarrhea that develops in patients after hospitalization during antibiotic administration. It has also become a big issue in community-acquired diarrhea. The emergence of hypervirulent strains of C. difficile poses a major problem in hospital-associated diarrhea outbreaks and it is difficult to treat. The antimicrobial resistance in C. difficile has worsened due to the inappropriate use of broad-spectrum antibiotics including cephalosporins, clindamycin, tetracycline, and fluoroquinolones together with the emergence of hypervirulent strains. OBJECTIVE: To estimate the pooled prevalence and antimicrobial resistance pattern of C. difficile derived from hospitalized diarrheal patients, a systematic review and meta-analysis was performed. METHODS: Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guideline was followed to review published studies conducted. We searched bibliographic databases from PubMed, Scopus, Google Scholar, and Cochrane Library for studies on the prevalence and antimicrobial susceptibility testing on C. difficile. The weighted pooled prevalence and resistance for each antimicrobial agent was calculated using a random-effects model. A funnel plot and Egger's regression test were used to see publication bias. RESULTS: A total of 15 studies were included. Ten articles for prevalence study and 5 additional studies for antimicrobial susceptibility testing of C. difficile were included. A total of 1967/7852 (25%) C. difficile were isolated from 10 included studies for prevalence study. The overall weighted pooled proportion (WPP) of C. difficile was 30% (95% CI: 10.0-49.0; p<0.001). The analysis showed substantial heterogeneity among studies (Cochran's test = 7038.73, I2 = 99.87%; p<0.001). The weighed pooled antimicrobial resistance (WPR) were: vancomycin 3%(95% CI: 1.0-4.0, p<0.001); metronidazole 5%(95% CI: 3.0-7.0, p<0.001); clindamycin 61%(95% CI: 52.0-69.0, p<0.001); moxifloxacin 42%(95% CI: 29-54, p<0.001); tetracycline 35%(95% CI: 22-49, p<0.001); erythromycin 61%(95% CI: 48-75, p<0.001) and ciprofloxacin 64%(95% CI: 48-80; p< 0.001) using the random effect model. CONCLUSIONS: A higher weighted pooled prevalence of C. difficile was observed. It needs a great deal of attention to decrease the prevailing prevalence. The resistance of C. difficile to metronidazole and vancomycin was low compared to other drugs used to treat C. difficile infection. Periodic antimicrobial resistance monitoring is vital for appropriate therapy of C. difficile infection.

Hepatitis B and Hepatitis C Viral Infections and Associated Factors Among Prisoners in Northeast Ethiopia.

BACKGROUND: Hepatitis is an inflammation of the liver and often caused by viruses. Hepatitis viruses are the leading causes of liver-related morbidity and mortality worldwide, with Hepatitis B and C viruses share the great majority. Studies have shown that prison settings are one of the high-risk environments for the transmission of these viruses. However, there is limited information on the seroprevalence and associated factors of hepatitis B and C viral infection among Ethiopian prisoners. METHODS: A facility-based cross-sectional study was conducted among 339 prisoners in Dessie town, Ethiopia from February to April 2020. Hepatitis B surface antigen and antibody against hepatitis C virus in serum were determined using Enzyme-Linked Immunosorbent Assay. We imputed the data using "EpiData 3.1" software and exported it to Statistical Package for Social Sciences version 20.0 for analysis, and a p-value of <0.05 was considered statistically significant. RESULTS: The overall seroprevalence of hepatitis B surface antigen and anti-hepatitis C virus among prisoners was 22/339 (6.5%) (95% CI = 3.8-9.4), and 4/339 (1.2%) (95% CI = 0.0-2.4), respectively. Multiple sexual partners, previous imprisonment, body tattooing, and contact with the jaundiced patient were independently associated with hepatitis B virus infection. Prisoners who had a history of blood transfusion, and dental extraction were independently associated with hepatitis C virus infection. CONCLUSION: The seroprevalence of hepatitis B and hepatitis C viral infection among Dessie town prisoners was intermediate and low, respectively. The finding of a significant association between the presence of Hepatitis B surface antigen and hepatitis C virus antibodies among prisoners and factors calls for the need of serological testing for both Hepatitis B and C viruses to high-risk individuals. Strengthening screening strategies and prevention programs in prison settings is advisable to prevent disease transmission.

Alteration of Gut Microbiota and Its Impact on Immune Response in Patients with Chronic HBV Infection: A Review.

Chronic hepatitis B virus infection is a source of substantial global health problems, particularly in economically underdeveloped and/or developing countries. It is the primary cause of severe liver disorders such as liver fibrosis, cirrhosis, and hepatocellular carcinoma. The liver is connected by the bile duct to the small intestine that carries bile produced in the liver to the intestine. The liver is the initial organ exposed to materials originating from the gut including dietary compounds, bacteria, and their products. Human intestines harbor a wide diversity of the community of microbes which are collectively termed as gut microbiota. In chronic infection with the hepatitis B virus, microbial alteration of the gut is a source of systemic immune activation. Besides, gut permeability is altered in hepatitis B virus-infected patients with an increased bacterial translocation and endotoxin load in the portal vein that caused toll-like receptor activation in the liver, which facilitates immune-mediated liver injury. Toll-like receptors further triggered the host-wide inflammatory response by inducing signaling cascades such as nuclear factor-kappa B-linked pathways and by accelerating cytokine secretion like tumor necrosis factor-alpha, which evokes chronic inflammation and leads to liver lesion formation, fibrosis progression, and cirrhosis and hepatocellular carcinoma development. In conclusion, changes in intestinal flora play an important role in encouraging the production of chronic infection with the hepatitis B virus. Therefore, careful attention should be given to the maintenance of intestinal microecology of patients which can provide a sound foundation for the treatment of chronic infection with the hepatitis B virus.

Occurrence and Antibiogram of Escherichia coli O157 : H7 in Raw Beef and Hygienic Practices in Abattoir and Retailer Shops in Ambo Town, Ethiopia.

Foodborne infections are widespread and growing public health problems in the world. Shiga toxin-producing Escherichia coli O157 : H7 is one of the most significant foodborne pathogens. This study was conducted to assess the occurrence and antibiogram of E. coli O157 : H7 from raw beef as well as hygienic and sanitary practices of meat handling in abattoir and retailer shops. Systematic random sampling technique and census methods were used to collect samples from abattoir and retailer shops, respectively. All tryptone soya broth preenriched carcass samples were subcultured onto MacConkey agar. Then, the bacterium confirmed as Escherichia coli using biochemical tests was streaked onto Sorbitol-MacConkey agar and incubated at 37°C for 24 hrs. Escherichia coli O157 : H7 was confirmed by latex agglutination kit. In vitro antimicrobial susceptibility test of Escherichia coli O157 : H7 isolates was done against 13 antimicrobials. Hygiene and sanitation data were collected using a pretested structured questionnaire and observational checklist. Pearson Chi-square and Fisher's exact two-tailed tests were performed and differences were considered significant at P ≤ 0.05. Out of 197 meat samples, 23.4% (95% confidence interval (CI): 17.6-29.9%) and 9.1% (95% CI: 5.5-14.1%) were contaminated with Escherichia coli and Escherichia coli O157 : H7, respectively. There was a significant variation in the occurrence of Escherichia coli O157 : H7 between retailer shops (19.1%) and abattoir (7.2%) (P = 0.03). The study revealed that the municipal abattoir and retailer shops in Ambo town did not adhere to the required sanitation and hygienic standards. All Escherichia coli O157 : H7 isolates were susceptible to norfloxacin, sulfamethoxazole-trimethoprim, chloramphenicol, and ceftazidime. However, all isolates were resistant to amoxicillin. Multidrug resistance was widespread and was found in 66.3% of Escherichia coli O157 : H7 isolates. The occurrence of Escherichia coli O157 : H7 was high. Therefore, fulfilling national and international meat safety requirements, training and monitoring of meat handlers, and rational use of antimicrobials are recommended.

Antimicrobial resistance profile and multidrug resistance patterns of Streptococcus pneumoniae isolates from patients suspected of pneumococcal infections in Ethiopia.

BACKGROUND: Antimicrobial-resistant strains of Streptococcus pneumoniae have become one of the greatest challenges to global public health today and inappropriate use of antibiotics and high level of antibiotic use is probably the main factor driving the emergence of resistance worldwide. The aim of this study is, therefore, to assess the antimicrobial resistance profiles and multidrug resistance patterns of S. pneumoniae isolates from patients suspected of pneumococcal infections in Ethiopia. METHODS: A hospital-based prospective study was conducted from January 2018 to December 2019 at Addis Ababa city and Amhara National Region State Referral Hospitals. Antimicrobial resistance tests were performed from isolates of S. pneumoniae that were collected from pediatric and adult patients. Samples (cerebrospinal fluid, blood, sputum, eye discharge, ear discharge, and pleural and peritoneal fluids) from all collection sites were initially cultured on 5% sheep blood agar plates and incubated overnight at 37 °C in a 5% CO2 atmosphere. Streptococcus pneumoniae was identified and confirmed by typical colony morphology, alpha-hemolysis, Gram staining, optochin susceptibility, and bile solubility test. Drug resistance testing was performed using the E-test method according to recommendations of the Clinical and Laboratory Standards Institute. RESULTS: Of the 57 isolates, 17.5% were fully resistant to penicillin. The corresponding value for both cefotaxime and ceftriaxone was 1.8%. Resistance rates to erythromycin, clindamycin, tetracycline, chloramphenicol and trimethoprim-sulfamethoxazole were 59.6%, 17.5%, 38.6%, 17.5 and 24.6%, respectively. Multidrug resistance (MDR) was seen in 33.3% isolates. The most common pattern was co-resistance to penicillin, erythromycin, clindamycin, and tetracycline. CONCLUSIONS: Most S. pneumoniae isolates were susceptible to ceftriaxone and cefotaxime. Penicillin has been used as a drug of choice for treating S. pneumoniae infection. However, antimicrobial resistance including multidrug resistance was observed to several commonly used antibiotics including penicillin. Hence, it is important to periodically monitor the antimicrobial resistance patterns to select empirical treatments for better management of pneumococcal infection.

Pertussis among patients with clinically compatible illness in the Amhara Regional State, Ethiopia.

BACKGROUND: Pertussis is an acute respiratory tract disease caused by Bordetella pertussis. In 2014, 24.1 million pertussis cases, resulting in 160,700 deaths, were estimated to have occurred worldwide. This study aimed to determine the epidemiology of pertussis among patients with clinically compatible illness who visited selected hospitals in the Amhara Regional State of Ethiopia. METHODS: A cross-sectional study design was used to review pertussis patients with clinically compatible illness. Nasopharyngeal swabs were collected from 515 patients from July 2018 through February 2019. DNA was extracted from all nasopharyngeal swabs and samples were analyzed using real-time (RT-) PCR. Crude and adjusted odds ratios with corresponding 95% confidence intervals were estimated using bivariable and multivariable logistic regression analysis, respectively. RESULTS: The overall prevalence of Bordetella species among the study participants was 156 of 515 (30.3%) [95% CI = 26.4-34.6] as determined by Bordetella RT-PCR, including: 65 (41.7%) B. pertussis, 89 (57.1%) indeterminate B. pertussis, one (0.6%) Bordetella holmesii and one (0.6%) Bordetella parapertussis. CONCLUSIONS: This study found that pertussis is potentially endemic and a common health problem among patients visiting health institutions in the Amhara Regional State of Ethiopia. More data regarding pertussis in Ethiopia could inform development of effective prevention strategies.

Serotype Distribution of Streptococcus pneumoniae Isolates Causing Invasive and Non-Invasive Infections Using Whole-Genome Sequencing in Ethiopia.

BACKGROUND: In Ethiopia, pneumococcal conjugate vaccine 10 (PCV10) was introduced in 2011 in the national vaccination program. This study was aimed to assess serotype distribution of invasive and non-invasive Streptococcus pneumoniae isolates using whole-genome sequencing. METHODS: A hospital-based prospective study was conducted from 2018 to 2019 at Addis Ababa and Amhara region referral hospitals, from all patients. Clinical Samples were collected and initially cultured onto 5% sheep blood agar at 37°C in a 5% CO2 atmosphere. Sequencing was done using the Illumina NextSeq 500 and SeroBA was used to predict serotypes from whole-genome sequencing raw data. RESULTS: Of the 57 S. pneumoniae isolates, there were 32 circulating serotypes. The most common serotypes were 15A/B/C (n=5, 8.8%), 6A (n=4, 7.0%), 10A/F (n=4, 7.0%), 23A (n=4, 7.0%) and 7C (n=3, 5.3%). The serotype coverage of PCV10 and PCV13 were 12.3% and 26.3% respectively. The most common invasive serotypes were 15A/B/C (n=5, 8.8%) and 6A (n=4, 7.0%), and non-invasive serotypes were 23A (n=4, 7.0%) and 10A/F (n=3, 5.3%). The most prevalent serotype obtained from PCV10 eligible children was 3 (n=2, 3.5%). The prevalent serotype obtained from PCV10 non-eligible patients were type 23A (n=4, 7%) and type 6A (n=3, 5.2%). The most common serotypes among children ≤18 years old were 10A/F, 7C, 35A/B, 16F, 19A, 3 and 38. However, the proportions of some non-vaccine serotypes (11A/B and 15A/B/C) were higher in adult patients. CONCLUSION: In this study a shift in the distribution of non-vaccinated S. pneumoniae serotypes increases in the population, and PCV10 serotype coverage was reduced as compared to PCV13. Therefore, it is important to continue monitoring serotype changes among all patients in addition to assessing the impact and effectiveness brought by vaccines and provides a foundation for prevention strategies and vaccine policies.

Multidrug resistance and extended-spectrum beta-lactamase producing Gram-negative bacteria from three Referral Hospitals of Amhara region, Ethiopia.

BACKGROUND: Multidrug resistance (MDR), extended-spectrum beta-lactamase (ESBL) and carbapenemase-producing Gram-negative bacteria (GNB) has become a public health threat worldwide. This threat is worse in developing countries where there is high infectious disease burden and spread of antimicrobial resistance co-exist. The aim of the present study was, therefore, to assess MDR, ESBL and carbapenemase producing GNB from patients attending three selected referral hospitals in Amhara region. METHODS: A cross-sectional study was conducted from December 2017- April 2018 at the University of Gondar Comprehensive Specialized Hospital, Dessie Referral Hospital and Debre Markos Referral Hospital of Amhara national regional state. A total of 833 study subjects were recruited using a convenient sampling technique. Clinical samples such as blood, urine, stool, wound, abscess, ear discharge, nasal discharge, cervical discharge and body fluid specimens were aseptically collected. Culturing for identification of bacteria and determination of drug susceptibility testing were done following standard microbiological techniques. Selected MDR isolates were phenotypically assessed for ESBL and carbapenemase production. RESULTS: Of the 833 clinical samples cultured for bacterial growth, 141 (16.9%) were positive for GNB. The most common GNB identified were E. coli 46 (32.6%), Klebsiella spp. 38 (26.5%) and Proteus spp. 13 (9.2%). The overall MDR prevalence was 121 (85.8%). Among the total isolates, 137 (97.2%) were resistant to ampicillin followed by cotrimoxazole 115 (81.6%), amoxicillin-clavulanic acid 109 (77.3%), cefixime 99 (70.2%), cefepime 93 (66.0%) and tetracycline 91 (64.5%). The extended-spectrum beta-lactamase producing GNB were 69/124 (55.6%). Of which Klebsiella spp. 19 (15.3%) and E. coli 17 (13.7%) were common ESBL producers. Carbapenemase-producing isolates were 8/51(15.7%). Of which Enterobacter, Klebsiella and E. coli were common carbapenemase producers. CONCLUSION AND RECOMMENDATION: Multi-drug resistance and ESBL producing isolates in the present study were high. E. coli and Klebsiella spp. were the most common ESBL producing GNB. Klebsiella spp., Enterobacter spp., E. coli and Citrobacter spp. were typical carbapenemase-producing isolates. Continuous monitoring, antibiotic stewardship and molecular detection of the gene responsible for drug resistance are important means to reduce the spread of drug-resistant pathogens.

Multiple drug resistance bacterial isolates and associated factors among urinary stone patients at the University of Gondar Comprehensive Specialized Hospital, Northwest Ethiopia.

BACKGROUND: The urinary stone and urinary tract infection (UTI) are invariably associated and are frequent causes of morbidity. Date on burden of UTI among urinary stone patients is lacking in Ethiopia. This study was aimed to assess bacterial profile, antimicrobial susceptibility and associated factors among urinary stone patients at the University of Gondar Comprehensive Specialized Hospital. METHODS: An institution based cross sectional study was conducted. Basic sociodemographic data were collected using a structured questionnaire. Bacterial identification of uropathogens and drug susceptibility testing were done following standard microbiological techniques. The data were entered and analyzed using SPSS version-23. Bivariate and multivariate logistic regressions were used to identify possible associated risk factors. Results with P value < 0.05 was considered statistically significant. RESULT: A total of 300 urinary stone patients were enrolled. Of these, 153 (51%) were male and 261(87%) were urban residents. The overall prevalence of urinary tract infection was 49 (16.3%) (95% CI 12-21%). A high level of resistance was observed to ampicillin, penicillin and trimethoprim-sulfamethoxazole while majority of isolates were most sensitive to nitrofurantoin and ciprofloxacin. Multi-drug resistant isolates were 16/49 (32.7%), 75% of them being Enterobacteriaceae isolates. More than one-third 9/26 (34.6%) of Gram-negative isolates were Extended Spectrum Beta-Lactamase (ESBL) producing E. coli and K. pneumoniae. Being female, history of urinary tract infection and history of drug use were the independent risk factors. CONCLUSION: Most of the bacterial isolates from urinary stone patients were resistant to ampicillin, penicillin and trimethoprim-sulfamethoxazole. E. coli and K. pneumoniae were the most common extended spectrum beta-lactamase producing isolates. Sex, history of urinary tract infection and previous drug use were found to be risk factors. Routine diagnosis of urinary stone patients for urinary tract infection should be promoted and further researches are encouraged.