Correction: Multidrug resistance pattern of Acinetobacter species isolated from clinical specimens referred to the Ethiopian Public Health Institute: 2014 to 2018 trend anaylsis.

[This corrects the article DOI: 10.1371/journal.pone.0250896.].

Antimicrobial resistance trends in clinical Escherichia coli and Klebsiella pneumoniae in Ethiopia.

Background: Clinicians rely on local antimicrobial resistance pattern data to guide empiric treatment for seriously ill patients when culture and antimicrobial susceptibility testing results are not immediately available. Objective: This study aimed to analyse 5-year trends in antimicrobial resistance profiles of Escherichia coli and Klebsiella pneumoniae isolates. Methods: Bacteriology reports from 2017 to 2021 at the Ethiopian Public Health Institute were analysed retrospectively. Isolates were identified using either the VITEK 2 Compact system, the BD Phoenix M50 instrument, or conventional biochemical tests. Antimicrobial susceptibility testing was conducted using either the Kirby-Bauer disk diffusion method or the VITEK 2 Compact system and BD Phoenix M50 systems available at the time of testing. The Cochran Armitage trend test was employed to test the significance of antimicrobial resistance trends over time. P-values less than 0.05 were considered statistically significant. Results: Of the 5382 bacteriology reports examined, 458 (9%) were on E. coli and 266 (5%) were on K. pneumoniae. Both K. pneumoniae (88%) and E. coli (65%) demonstrated high resistance to extended-spectrum cephalosporins. However, both K. pneumoniae (14%) and E. coli (5%) showed lower rates of resistance to carbapenems compared to other antimicrobials. In K. pneumoniae, resistance to carbapenems (from 0% to 38%; p < 0.001) and ciprofloxacin (from 41% to 90%; p < 0.001) increased significantly between 2017 and 2021. Conclusion: Both organisms showed very high resistance to broad-spectrum antibiotics. Additionally, K. pneumoniae demonstrated a statistically significant rise in ciprofloxacin and carbapenem resistance. What this study adds: This study emphasises the significance of regular reporting of local antimicrobial resistance patterns as this information can guide appropriate empiric therapy and efforts to address antimicrobial resistance issues.

Antimicrobial Susceptibility Patterns of Shigella Species among Children under Five Years of Age with Diarrhea in Selected Health Centers, Addis Ababa, Ethiopia.

Background: Shigella and parasitic infections are common public health problems throughout the world. Shigellosis is an acute gastroenteritis infection and one of Ethiopia's most common causes of morbidity and mortality, especially in children under five. High resistance rates to commonly used antibiotic agents have been documented in different locations in Ethiopia. Objective: This study aimed to characterize the antimicrobial features of the Shigella species isolated from children under five years of age with acute diarrhea in Addis Ababa, Ethiopia. Methods: Using a cross-sectional study, freshly passed fecal specimens were collected for intestinal parasite and bacterial isolation. Fecal samples for bacterial identification were placed immediately in Cary-Blair media and transported to the Ethiopian Public Health Institution (EPHI) laboratory. Antimicrobial susceptibility testing (AMST) was conducted using the disk diffusion method. Data were described using descriptive statistical tools. The association of independent and dependent variables was evaluated with logistic regression. A P value ≤0.05 was considered statistically significant. Results: The prevalence of intestinal parasites was 8.2% with seven different species. Among the 534 stool-cultured specimens, 47 (8.8%) were positive for Shigella species. Antimicrobial susceptibility testing (AMST) showed that 100%, 93.6%, 80.9%, 72.3%, and 57.5% were susceptible to norfloxacin, nalidixic acid, ciprofloxacin, gentamicin, and cefoxitin, respectively. However, 100% of the isolates were resistant to amoxicillin and erythromycin. More than 50% of the isolates were resistant to three and above antibiotics, while none of them were susceptible to all the antibiotics tested. All risk factors assessed did not show a statistically significant association with Shigella infection. Conclusion: The high levels of antibiotic resistance observed among the commonly prescribed antibiotics are alarming. The emerging resistance to ciprofloxacin and nalidixic acid signals a severe public health threat in the management of shigellosis. Raising awareness about resistance and educating health professionals, policymakers, and the public can help improve the quality of patient care and rational antibiotic use.

Antimicrobial Resistance and Virulence Gene Profile of Clinical Staphylococcus aureus: A Multi-Center Study from Ethiopia.

Background: Staphylococcus aureus causes a wide range of infections from mild skin and soft tissue to severe life-threatening bacteremia. The pathogenicity of S. aureus infections is related to various bacterial surface components and extracellular proteins such as toxic-shock syndrome (TSS) toxin and Panton-Valentine leukocidin (PVL). In this study we determine the antimicrobial resistance of isolated strains and their virulence genes in Ethiopia. Methods: A total of 190 archived S. aureus isolates from four Ethiopia Antimicrobial Resistance (AMR) Surveillance sites were analyzed. The identification of S. aureus was done by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF Biotyper) and antimicrobial susceptibility test (AST) was done using VITEK® 2. Multiplex PCR was used to detect mecA, mecC, pvl and spa genes and super-antigens (sea, seb, sec, seh and sej staphylococcal enterotoxins). Results: A total of 172 isolates were confirmed as S. aureus, 9 (5.23%) were methicillin-resistant S. aureus (MRSA) and 163 (94.76%) were methicillin-susceptible S. aureus (MSSA). AST showed that 152 (88.4%) isolates were resistant to penicillin; 90 (52.32%) resistant to trimethoprim-sulfamethoxazole; and 45 (26.16%) resistant to tetracycline. A total of 66 (38.37%) isolates harbored at least one staphylococcal enterotoxin gene and 31 (46.96%) isolates had more than one. The most frequent enterotoxin gene encountered was seb 28 (16.28%). The TSST-1 gene was detected in 23 (13.37%). Presence of staphylococcal enterotoxin gene showed significant association with antibiotic resistance to cefoxitin, benzylpenicillin, oxacillin, erythromycin, clindamycin, tetracycline and SXT. The pvl gene was detected in 102 (59.3%) of isolates. Isolates from patients below 15 years of age showed significantly high numbers of pvl gene (P = 0.02). Presence of sej (P = 0.011) and TSST-1 (P <0.001) genes were associated with the presence of pvl gene. Conclusion: In this study, isolates were highly resistant to oral antibiotics and the pvl, seb, sea and TSST-1 genes were prevalent.

A High Level of Antimicrobial Resistance in Gram-Positive Cocci Isolates from Different Clinical Samples Among Patients Referred to Arsho Advanced Medical Laboratory, Addis Ababa, Ethiopia.

Background: Gram-positive cocci are clinically important pathogens that cause infections and their development of antibiotic resistance continues to pose a severe threat to public health. Therefore, this study aims to investigate the level of antimicrobial resistance among Gram-positive cocci isolated from different clinical samples among patients referred to Arsho Advanced Medical Laboratory, Addis Ababa, Ethiopia. Methods: From January to April 2018, a cross-sectional study was conducted at Arsho Advanced Medical Laboratory. Seven hundred ninety-two (792) different clinical samples were obtained from 792 individuals and inoculated into blood culture bottles and Blood Agar base. Bacterial identification was done using the number, type, and morphology of colonies, as well as Gram staining, catalase testing, and coagulase test after isolation of pure growth on culture media using the standard operating procedure. VITEK 2 compact system was used for bacterial identification and drug susceptibility testing. The information entry and analysis were performed by using SPSS version 20. Results: Out of 792 clinical samples cultured, the prevalence of Gram-positive cocci was 12.6% (n=100/792). The most frequent one is S. aureus 54% (n=54/100) followed by coagulase-negative Staphylococcus species 42% (n=42/100), S. agalactiae 1% (n=1/100) and E. faecalis 3% (n=3/100). Penicillin showed the highest resistance rate 85% (n=85/100), followed by sulfamethoxazole/trimethoprim (47%), and oxacillin (38%); however, highest sensitivity was seen towards linezolid 97% (n=97/100) and vancomycin 94% (n=94/100). The total multi-drug resistance (MDR) Gram-positive cocci were 44% (n=44/100). Conclusion: This study demonstrated high antimicrobial resistance and multi-drug resistance. This suggests that the importance of continuous monitoring of antimicrobial resistance patterns is crucial for selecting the suitable drug for treatment and infection prevention.

First PCR Confirmed anthrax outbreaks in Ethiopia-Amhara region, 2018-2019.

BACKGROUND: Anthrax is a disease that affects humans and animals. In Ethiopia, anthrax is a reportable disease and assumed to be endemic, although laboratory confirmation has not been routinely performed until recently. We describe the findings from the investigation of two outbreaks in Amhara region. METHODS: Following reports of suspected outbreaks in Wag Hamra zone (Outbreak 1) and South Gondar zone (Outbreak 2), multi-sectoral teams involving both animal and public health officials were deployed to investigate and establish control programs. A suspect case was defined as: sudden death with rapid bloating or bleeding from orifice(s) with unclotted blood (animals); and signs compatible with cutaneous, ingestion, or inhalation anthrax ≤7 days after exposure to a suspect animal (humans). Suspect human cases were interviewed using a standard questionnaire. Samples were collected from humans with suspected anthrax (Outbreak 1 and Outbreak 2) as well as dried meat of suspect animal cases (Outbreak 2). A case was confirmed if a positive test was returned using real-time polymerase chain reaction (qPCR). RESULTS: In Outbreak 1, a total of 49 cows died due to suspected anthrax and 22 humans developed symptoms consistent with cutaneous anthrax (40% attack rate), two of whom died due to suspected ingestion anthrax. Three people were confirmed to have anthrax by qPCR. In Outbreak 2, anthrax was suspected to have caused the deaths of two livestock animals and one human. Subsequent investigation revealed 18 suspected cases of cutaneous anthrax in humans (27% attack rate). None of the 12 human samples collected tested positive, however, a swab taken from the dried meat of one animal case (goat) was positive by qPCR. CONCLUSION: We report the first qPCR-confirmed outbreaks of anthrax in Ethiopia. Both outbreaks were controlled through active case finding, carcass management, ring vaccination of livestock, training of health professionals and outreach with livestock owners. Human and animal health authorities should work together using a One Health approach to improve case reporting and vaccine coverage.

Phenotypic characterization of carbapenem non-susceptible gram-negative bacilli isolated from clinical specimens.

BACKGROUND: Multidrug resistant, extremely drug-resistant, pan-drug resistant, carbapenem-resistant, and carbapenemase-producing gram-negative bacteria are becoming more common in health care settings and are posing a growing threat to public health. OBJECTIVE: The study was aimed to detect and phenotypically characterize carbapenem no- susceptible gram-negative bacilli at the Ethiopian Public Health Institute. MATERIALS AND METHODS: A prospective cross-sectional study was conducted from June 30, 2019, to May 30, 2020, at the national reference laboratory of the Ethiopian Public Health Institute. Clinical samples were collected, inoculated, and incubated for each sample in accordance with standard protocol. Antimicrobial susceptibility testing was conducted using Kirby-Bauer disk diffusion method. Identification was done using the traditional biochemical method. Multidrug-resistant and extensively drug-resistant isolates were classified using a standardized definition established by the European Centre for Disease Prevention and Control and the United States Centers for Disease Prevention and Control. Gram-negative organisms with reduced susceptibility to carbapenem antibiotics were considered candidate carbapenemase producers and subjected to modified carbapenem inactivation and simplified carbapenem inactivation methods. Meropenem with EDTA was used to differentiate metallo-ß-lactamase (MBL) from serine carbapenemase. Meropenem (MRP)/meropenem + phenylboronic acid (MBO) were used to differentiate Klebsiella pneumoniae carbapenemase (KPC) from other serine carbapenemase producing gram-negative organisms. RESULTS: A total of 1,337 clinical specimens were analyzed, of which 429 gram-negative bacterial isolates were recovered. Out of 429 isolates, 319, 74, and 36 were Enterobacterales, Acinetobacter species, and Pseudomonas aeruginosa respectively. In our study, the prevalence of multidrug-resistant, extensively drug-resistant, carbapenemase-producing, and carbapenem nonsusceptible gram-negative bacilli were 45.2%, 7.7%, 5.4%, and 15.4% respectively. Out of 429 isolates, 66 demonstrated reduced susceptibility to the antibiotics meropenem and imipenem. These isolates were tested for carbapenemase production of which 34.8% (23/66) were carbapenemase producers. Out of 23 carbapenemase positive gram-negative bacteria, ten (10) and thirteen (13) were metallo-beta-lactamase and serine carbapenemase respectively. Three of 13 serine carbapenemase positive organisms were Klebsiella pneumoniae carbapenemase. CONCLUSION: This study revealed an alarming level of antimicrobial resistance (AMR), with a high prevalence of multidrug-resistant (MDR) and extremely drug-resistant, carbapenemase-producing gram-negative bacteria, particularly among intensive care unit patients at the health facility level. These findings point to a scenario in which clinical management of infected patients becomes increasingly difficult and necessitates the use of "last-resort" antimicrobials likely exacerbating the magnitude of the global AMR crisis. This mandates robust AMR monitoring and an infection prevention and control program.

Multidrug resistance pattern of Acinetobacter species isolated from clinical specimens referred to the Ethiopian Public Health Institute: 2014 to 2018 trend anaylsis.

BACKGROUND: Acinetobacter species have been a leading cause of nosocomial infections, causing significant morbidity and mortality over the entire world including Ethiopia. The most important features of A. baumannii are its ability to persist in the hospital environment and rapidly develop resistance to a wide variety of antibiotics. This study aimed to determine trend of antimicrobial resistance in Acinetobacter species over a five years period. METHOD: A retrospective data regarding occurrence and antimicrobial resistance of Acinetobacter species recovered from clinical specimens referred to the national reference laboratory was extracted from microbiology laboratory data source covering a time range from 2014 to 2018. Socio-demographic characteristics and laboratory record data was analyzed using SPSS 20. RESULTS: A total of 102 strains of Acinetobacter species were analyzed from various clinical specimens. Majority of them were from pus (33.3%) followed by blood (23.5%), urine (15.6%) and body fluid (11.7%). Significant ascending trends of antimicrobial resistance was shown for meropenem (12.5% to 60.7%), ceftazidime (82.1% to 100%), ciprofloxacin (59.4% to 74.4%), ceftriaxone (87.1% to 98.6%), cefepime (80.0% to 93.3%) and pipracillin- tazobactam (67.8% to 96.3%). However, there was descending trend of antimicrobial resistance for tobramycin (56.5% to 42.8%), amikacin (42.1% to 31.4%) and trimethoprim-sulfamethoxazole (79.0 to 68.2%). The overall rate of carbapenem non-susceptible and multidrug resistance rates in Acinetobacter species were 56.7% and 71.6%.respectively. CONCLUSION: A five year antimicrobial resistance trend analysis of Acinetobacter species showed increasing MDR and resistance to high potent antimicrobial agents posing therapeutic challenge in our Hospitals and health care settings. Continuous surveillance and appropriate infection prevention and control strategies need to be strengthened to circumvent the spread of multidrug resistant pathogens in health care facilities.

Establishment of COVID-19 testing laboratory in resource-limited settings: challenges and prospects reported from Ethiopia.

The Coronavirus pandemic is recording unprecedented deaths worldwide. The temporal distribution and burden of the disease varies from setting to setting based on economic status, demography and geographic location. A rapid increase in the number of COVID-19 cases is being reported in Africa as of June 2020. Ethiopia reported the first COVID-19 case on 13 March 2020. Limited molecular laboratory capacity in resource constrained settings is a challenge in the diagnosis of the ever-increasing cases and the overall management of the disease. In this article, the Ethiopian Public Health Institute (EPHI) shares the experience, challenges and prospects in the rapid establishment of one of its COVID-19 testing laboratories from available resources. The first steps in establishing the COVID-19 molecular testing laboratory were i) identifying a suitable space ii) renovating it and iii) mobilizing materials including consumables, mainly from the Malaria and Neglected Tropical Diseases (NTDs) research team at the EPHI. A chain of experimental design was set up with distinct laboratories to standardize the extraction of samples, preparation of the master mix and detection. At the commencement of sample reception and testing, laboratory contamination was among the primary challenges faced. The source of the contamination was identified in the master mix room and resolved. In summary, the established COVID-19 testing lab has tested more than 40,000 samples (August 2020) and is the preferred setting for research and training. The lessons learned may benefit the further establishment of emergency testing laboratories for COVID-19 and/or other epidemic/pandemic diseases in resource-limited settings.

High Prevalence of Multidrug-Resistant and Extended-Spectrum ß-Lactamase-Producing Enterobacteriaceae: A Cross-Sectional Study at Arsho Advanced Medical Laboratory, Addis Ababa, Ethiopia.

BACKGROUND: Multidrug-resistant Enterobacteriaceae particularly extended-spectrum beta-lactamase producers have become a major public health threat. Despite efforts to limit their spread, rates of multidrug-resistance members of the Enterobacteriaceae continue to increase throughout the world causing increased morbidity and mortality and raised costs for medical care. OBJECTIVE: The aim of this study was to determine the prevalence of multidrug resistance and extended-spectrum ß-lactamase-producing Enterobacteriaceae. METHODS: Four hundred forty Enterobacteriaceae isolates from outpatients referred to Arsho Advanced Medical Laboratory were identified and assessed for their antimicrobial resistance pattern by using the automated VITEK 2 compact system. Extended-spectrum ß-lactamase production was determined by the VITEK 2 automated compact system using the extended-spectrum ß-lactamase test panel as per the instruction of the manufacturer. RESULTS: The overall resistance rates of Enterobacteriaceae against cephalosporins, aminoglycosides, and fluoroquinolones were high. Nitrofurantoin with a resistance rate of 14.3% and piperacillin/tazobactam combination with a resistance rate of 17.3% were better active against this group of Gram-negative bacteria. Out of 440 isolates of Enterobacteriaceae, 42.1% were multidrug-resistant of which 34.3% and 8.95% were extensively drug-resistant and pan-drug resistant, respectively. Among 185 multidrug-resistant Enterobacteriaceae, 63.9% of the isolates produced extended-spectrum ß-lactamase of which 75.4%, 19.5%, 1.7%, 2.5%, and 0.8% were E. coli, K. pneumoniae, C. freundii, E. cloacae, and P. mirabilis, respectively. CONCLUSIONS: The present study demonstrated high prevalence rates of multidrug-resistant and extended-spectrum-beta-lactamase-producing Enterobacteriaceae. In order to combat these problems, infection control strategy and proper antibiotic policies should be formulated.

Bacterial urinary tract infection among adult renal transplant recipients at St. Paul's hospital millennium medical college, Addis Ababa, Ethiopia.

BACKGROUND: Despite significant advances in surgical techniques, immunosuppression protocols, follow up periods and antimicrobial stewardship in modern medicine; post-renal transplantation urinary tract infection remained a major public health problem globally. This multiple serious squeals includes asymptomatic bacteriuria, cystitis and pyelonephritis. Among these, the bacterial origin of infection complications accounts for the most significant clinical, socio-economic impacts in many countries of the world. Therefore, the aim of the study was to investigate the prevalence of bacterial isolates that cause urinary tract infections, assess antibiotic susceptibility pattern among symptomatic and asymptomatic renal transplant recipients attending at St. Paul's Hospital Millennium Medical College, Addis Ababa, Ethiopia. METHODS: A hospital-based cross-sectional study was conducted from December 2017 to August 2018 among 74 renal transplant recipients St. Paul's Hospital Millennium Medical College, Addis Ababa, Ethiopia. A first morning voided clean-catch mid-stream urine specimens were collected and 0.001 ml inoculated onto blood and MacConkey agar plates following the standard bacteriological protocols. It was incubated aerobically at 35-37 °C for 24-48 h. Cultural characteristics and series of biochemical tests were used for the identification of isolates to species level based on the standard bacteriological protocols. RESULTS: A hospital-based cross-sectional study has shown that significant bacteriuria was found in 11/74 (14.9, 95% CI =8.2-24.7) patients. The prevalence among females 6/32 (18.75%) was higher among males 5/42 (11.9%) without significant association (COR = 2.09, 95% CI = 1.04-8.45, P = 0.253). Urinary tract infection was higher in the age group of 35-49 years old (19.3%). Age was statistically significant and stronger independent associated risk factor with crude odds ratio = 3.67, 95% CI = 2.89-20.07 and P = 0.003, respectively. The most prevalent bacteria isolates were Escherichia coli 2(18.2%), Staphylococcus aureus 2(18.2%), Acinetobacter spp. 2(18.2%), Enterococcus spp. 2(18.2%), Coagulase-negative Staphylococci 2(18.2%) followed by Porteus mirabilis 1(9.1%). The majority (80%) of Gram-negative bacteria were resistant to ciprofloxacin, chloramphenicol, and trimethoprim/sulfamethoxazole. Simultaneously, the multidrug-resistant bacterial isolates accounts for 82% among tested kidney allograft recipients. CONCLUSIONS: In conclusion, the overall prevalence of urinary tract infection in the study participants was relatively low with a prevalence of 14.9%. Majority of the study participants were asymptomatic and a higher percentage of females were involved. The multidrug-resistant bacterial isolates in the present study account for 82%.