Vibrio cholerae O1 and Escherichia coli O157:H7 from drinking water and wastewater in Addis Ababa, Ethiopia.

BACKGROUND: In Addis Ababa, Ethiopia, open ditches along innner roads in residential areas serve to convey domestic wastewater and rainwater away from residences. Contamination of drinking water by wastewater through faulty distribution lines could expose households to waterborne illnesses. This prompted the study to assess the microbiological safety of wastewater and drinking water in Addis Ababa, identify the pathogens therein, and determine their antibiotic resistance patterns. RESULTS VIBRIO CHOLERAE: O1, mainly Hikojima serotype, was isolated from 23 wastewater and 16 drinking water samples. Similarly, 19 wastewater and 10 drinking water samples yielded Escherichia coli O157:H7. V. cholerae O1 were 100% resistant to the penicillins (Amoxacillin and Ampicillin), and 51-82% were resistant to the cephalosporins. About 44% of the V. cholerae O1 isolates in this study were Extended Spectrum Beta-Lactamase (ESBL) producers. Moreover, 26% were resistant to Meropenem. Peperacillin/Tazobactam was the only effective ß-lactam antibiotic against V. cholerae O1. V. cholerae O1 isolates showed 37 different patterns of multiple resistance ranging from a minimum of three to a maximum of ten antimicrobials. Of the E. coli O157:H7 isolates, 71% were ESBL producers. About 96% were resistant to Ampicillin. Amikacin and Gentamicin were very effective against E. coli O157:H7 isolates. The isolates from wastewater and drinking water showed multiple antibiotic resistance against three to eight antibiotic drugs. CONCLUSIONS: Open ditches for wastewater conveyance along innner roads in residence areas and underground faulty municipal water distribution lines could be possible sources for V. cholerae O1 and E. coli O157:H7 infections to surrounding households and for dissemination of multiple drug resistance in humans and, potentially, the environment.

Prevalence and Antimicrobial Susceptibility Profile of Salmonella and Shigella among Diarrheic Patients Attending Selected Health Facilities in Addis Ababa, Ethiopia.

Diarrhea is one of the important public health problems in developing countries. Salmonella and Shigella species are the major bacterial causal agents of diarrhea. The increasing burden of antimicrobial resistance is posing difficulty in the treatment of these pathogens. This study aimed to assess the occurrence of Salmonella and Shigella in the feces of diarrheic patients receiving health services in Addis Ababa, Ethiopia, and to determine their antimicrobial susceptibility profile. A cross-sectional study involving 13 health centers was conducted where 428 diarrheic patients were recruited. Standard microbiology techniques were used to isolate Salmonella and Shigella from stool samples. In addition, Salmonella isolates were confirmed by polymerase chain reaction (PCR). The Kirby-Bauer disc diffusion method was employed to assess susceptibility to 11 antimicrobials for each of the Salmonella and Shigella isolates. The prevalence of Salmonella and Shigella spp. among diarrheic patients was 8.4%; n = 36 and 5.6%; n = 24, respectively. Thirty (83.3%) of Salmonella isolates were susceptible to all antimicrobials tested, whereas 4 (10.8%) of isolates were resistant to 2 or more antimicrobials and 2 (5.6%) were multidrug resistant. Resistance to ampicillin was recorded in only one (2.7%) of Salmonella isolates; however, resistance to ampicillin was recorded in 12 (50%) of the Shigella isolates. Half of the Shigella isolates (n = 12) were resistant to 2 or more antimicrobials while 5 (20.8%) of them were resistant to 3 or more antimicrobials. The overall rate of resistance to antimicrobials was more common in Shigella compared to Salmonella isolates. In conclusion, Salmonella and Shigella were isolated from the feces of diarrheic patients, with a higher rate of antimicrobial resistance in Shigella isolates, which could make the treatment of shigellosis challenging. Therefore, increasing hygienic practices during food preparation to reduce the burden of Salmonella and Shigella infection and prudent use of antimicrobials are recommended to limit the spread of antimicrobial resistant strains.

Salmonella serotypes and their antimicrobial susceptibility in apparently healthy dogs in Addis Ababa, Ethiopia.

BACKGROUND: The close bond between pet animals and family members poses risk of infection with zoonotic bacterial pathogens such as Salmonella. No data is available on occurrence of Salmonella in dogs in Ethiopia. The aim of this study was therefore to determine the prevalence, serotype distribution and antimicrobial resistance of Salmonella from feces of apparently healthy dogs in Addis Ababa, Ethiopia. RESULTS: Of the total 360 dogs examined, 42 (11.7%; 95% Confidence limit of 8.5%-15.4%) were positive for Salmonella. Fourteen serotypes were detected and the predominant ones were S. Bronx (n = 7; 16.7%), S. Newport (n = 6; 14.3%), followed by S. Typhimurium, S. Indiana, S. Kentucky, S. Saintpaul and S. Virchow (n = 4; 9.5%) each. Salmonella infection status was significantly associated with history of symptom of diarrhea during the past 60 days (OR = 3.78; CI = 1.76-8.13; p = 0). Highest resistance rates were found for oxytetracycline (59.5%), neomycin (50%), streptomycin (38.1%), cephalothin (33.3%), doxycycline (30.9%), ampicillin (30.9%) and amoxicillin + clavulanic acid (26.2%). Thirty eight (90.5%) of the isolates were resistant or intermediately resistant to at least one of the 16 antimicrobials tested. Resistance to two or more antimicrobials was detected in 30 (71.4%) of the isolates. Resistance to three or more antimicrobials was detected in 19 (45.2%) of the isolates. CONCLUSION: This study demonstrated high carriage rate of Salmonella serotypes known for causing human salmonellosis and large proportion of them were resistant to antimicrobials used in public and veterinary medicine for management of various bacterial infections, suggesting the possible risk of infection of human population in close contact with these dogs by drug resistant pathogens. Therefore, it is vital to work on raising public awareness on zoonotic canine diseases prevention measures and good hygienic practices.

Fecal prevalence, serotype distribution and antimicrobial resistance of Salmonellae in dairy cattle in central Ethiopia.

BACKGROUND: Salmonellae are major worldwide zoonotic pathogens infecting a wide range of vertebrate species including humans. Consumption of contaminated dairy products and contact with dairy cattle represent a common source of non-typhoidal Salmonella infection in humans. Despite a large number of small-scale dairy farms in Addis Ababa and its surrounding districts, little is known about the status of Salmonella in these farms. RESULTS: Salmonella was recovered from the feces of at least one animal in 7.6% (10/132) of the dairy farms. Out of 1203 fecal samples examined, 30 were positive for Salmonella resulting in a weighted animal level prevalence of 2.3%. Detection of diarrhea in an animal and in a farm was significantly associated with animal level (p = 0.012) and herd level (p < 0.001) prevalence of Salmonella. Animal level prevalence of Salmonella was significantly associated with age (p = 0.023) and study location; it was highest among those under 6 months of age and in farms from Adaa district and Addis Ababa (p < 0.001). Nine different serotypes were identified using standard serological agglutination tests. The most frequently recovered serotypes were Salmonella Typhimurium (23.3%), S. Saintpaul (20%), S. Kentucky (16.7%) and S. Virchow (16.7%). All isolates were resistant or intermediately resistant to at least one of the 18 drugs tested. Twenty-six (86.7%), 19 (63.3 %), 18 (60%), 16 (53.3%) of the isolates were resistant to streptomycin, nitrofurantoin, sulfisoxazole and tetracycline , respectively. Resistance to 2 drugs was detected in 27 (90%) of the isolates. Resistance to 3 or more drugs was detected in 21 (70%) of the isolates, while resistance to 7 or more drugs was detected in 11 (36.7%) of the isolates. The rate of occurrence of multi-drug resistance (MDR) in Salmonella strains isolated from dairy farms in Addis Ababa was significantly higher than those isolated from farms outside of Addis Ababa (p = 0.009). MDR was more common in S. Kentucky, S. Virchow and S. Saintpaul. CONCLUSION: Isolation of Salmonella serotypes commonly known for causing human salmonellosis that are associated with an MDR phenotype in dairy farms in close proximity with human population is a major public health concern. These findings imply the need for a strict pathogen reduction strategy.

Evaluation of the in vitro antibacterial activity of the solvent fractions of the leaves of Rhamnus prinoides L'Herit (Rhamnaceae) against pathogenic bacteria.

BACKGROUND: Medicinal plants play great roles in the treatment of various infectious diseases. Rhamnus prinoides is one of the medicinal plants used traditionally for treatment of bacterial diseases. The antibacterial activity of the crude extract of the plant had been shown by a previous study, but this study was undertaken to further the claimed medicinal use of the plant by screening its solvent fractions for the said activity so that it could serve as a basis for subsequent studies. METHODS: The solvent fractions of the plant were obtained by successive soxhlet extraction with solvents of increasing polarity, with chloroform and methanol, followed by maceration of the marc of methanol fraction with water. The antibacterial activity of the solvent fractions was evaluated on seven bacterial species using agar well diffusion method at different concentrations (78 mg/well, 39 mg/well and 19.5 mg/well) in the presence of positive and negative controls. The minimum inhibitory concentration of the solvent fractions was determined by micro-broth dilution method using resazurin as indicator. RESULT: Methanol and chloroform fractions revealed antibacterial activities against the growth of test bacterial strains with varying antibacterial spectrum and the susceptible bacterial species were Staphylococcus aureus, Streptococcus pyogen, Streptococcus pneumoniae and Salmonella typhi. The average minimum inhibitory concentration value of the methanol and chloroform fractions ranged from 8.13 mg/ml to 32.5 mg/ml and from 8.13 mg/ml to 16.25 mg/ml, respectively. CONCLUSION: The methanol and chloroform fractions demonstrated significant antibacterial activities against the growth of pathogenic bacteria but the aqueous fraction did not reveal antibacterial activity against any of the test bacteria.

Non-typhoidal Salmonella serotypes, antimicrobial resistance and co-infection with parasites among patients with diarrhea and other gastrointestinal complaints in Addis Ababa, Ethiopia.

BACKGROUND: Non-typhoidal Salmonella (NTS) is an important public health problem worldwide. Consumption of animal-derived food products and direct and/or indirect contact with animals are the major routes of acquiring infection with NTS. Published information, particularly on the serotype distribution of NTS among human patients with gastroenteritis and associated risk factors, is scarce in Ethiopia. This study investigated the prevalence, risk factors, serotype distribution and antimicrobial susceptibility of Salmonella species among diarrheic out-patients attending health centers in Addis Ababa and patients with various gastrointestinal complaints at Tikur Anbessa Specialized Hospital (TASH). METHODS: Stool samples were cultured for Salmonella species according to the WHO Global Foodborne Infections Network laboratory protocol. Salmonella serotyping was conducted using slide agglutination and microplate agglutination techniques. Antibiotic susceptibility testing was performed using the disk diffusion method according to Clinical and Laboratory Standards Institute guidelines. RESULTS: A total of 59 (6.2 %) stool samples, out of 957 were culture positive for Salmonella species. Fifty-five (7.2 %) of 765 diarrheic patients from health centers and 4 (2.1 %) of 192 patients from TASH were culture positive for Salmonella species. Multivariable logistic regression analysis after adjusting for all other variables revealed statistically significant association of Salmonella infection with consumption of raw vegetables (OR = 1.91, 95 % CI = 1.29-2.83, χ(2) = 4.74, p = 0.025) and symptom of watery diarrhea (OR = 3.3, 95 % CI = 1.23-8.88, χ(2) = 10.54, p = 0.005). Eleven serotypes were detected, and the most prominent were S. Typhimurium (37.3 %), S. Virchow (34 %), and S. Kottbus (10.2 %). Other serotypes were S. Miami, S. Kentucky, S. Newport, S. Enteritidis, S. Braenderup, S. Saintpaul, S. Concord and S. V:ROUGH-O. Resistance to three or more antimicrobials was detected in 27 (40.3 %) of the isolates. Resistance to five or more antimicrobials was detected in 17 (25.4 %). Resistance to individual antimicrobials was found at varying proportions: streptomycin (50; 74.6 %), nitrofurantoin (27; 40.3 %), sulfisoxazole (26; 38.8 %), kanamycin (23; 34.3 %), cephalothin (12; 17.9 %), and ampicillin (11; 16.4 %) respectively. Two S. Kentucky, one S. Typhimurium and one S. Concord isolates were multi-drug resistant to more than 10 antimicrobials. CONCLUSIONS: The study demonstrated significant association of Salmonella infection with consumption of raw vegetables. There was no significant association of Salmonella infection with co-occurring parasites. The study also showed the dominance of S. Typhimurium and S. Virchow in primary health care units. Overall, prevalence of MDR was low compared to previous studies. Although their proportion was low, S. Kentucky and S. Concord demonstrated wider spectrum of MDR. Continuous monitoring of circulating serotypes, antimicrobial resistance profile and characterization on molecular resistance determinants is essential for proper treatment of patients and for identifying potential environmental origins of antimicrobial resistance.

Antibiogram of Escherichia coli Isolated from Dairy Cattle and in-Contact Humans in Selected Areas of Central Ethiopia.

Background: Antimicrobial resistance (AMR) is a global threat to public and animal health. Escherichia coli is considered an indicator organism for monitoring AMR among gram-negative Enterobacteriaceae in humans and animals. The current study aims to assess the antibiogram profile of E. coli isolated from dairy cattle and in-contact humans in central Ethiopia and to identify risk factors associated with multidrug resistance (MDR). Methods: A cross-sectional study was conducted in which 58 farms were recruited from selected districts of central Ethiopia. E. coli was isolated using standard bacteriological techniques. A total of 200 representative isolates (140 from cattle and 60 from humans in contact) were randomly selected and tested for susceptibility to a panel of 13 antimicrobials using the Kirby-Bauer disc diffusion assay. Results: The highest rate of resistance was observed for sulfamethoxazole+trimethoprim (58.6%, 82/140) and amoxicillin+clavulanic acid (70.0%, 42/60) among E. coli isolates from cattle and hmans, respectively. In contrast, resistance rates in isolates from in contact humans with the cattle were 30%, 33.3%, and 66.7%, respectively. Resistance to tetracycline (p=0.02), streptomycin (p=0.03), and sulfamethoxazole+trimethoprim (p=0.007) was significantly high in E. coli isolated from cattle on commercial dairy farms than in those isolated from cattle on smallholder farms. There was no significant difference (p>0.05) in the rate of resistance between E. coli isolated from in contact humans with smallholder and commercial dairy farms. Antimicrobial use for treatment purpose (p=0.04) and non-compliance with the drug withdrawal period (p=0.03) were significantly associated with the farm-level occurrence of MDR. Conclusion: A high rate of resistance was detected in E. coli isolated from the feces of dairy cattle and in-contact humans. This necessitates an effective intervention through a one-health approach and further molecular studies are required to establish source attribution.

Prevalence and Antimicrobial Susceptibility Profile of Salmonella from Poultry Farms and In-Contact Humans and Associated Risk Factors in Addis Ababa, Ethiopia.

Poultry and poultry products are the common sources of Salmonella,which is one of the serious food-borne bacterial diseases in humans. Little is known about the status of Salmonella and their antimicrobial susceptibility in poultry farms in Addis Ababa. This study was conducted to estimate the prevalence and antimicrobial susceptibility of Salmonella isolates and to investigate possible risk factors for the occurrence of Salmonella in poultry farms in Addis Ababa. We recruited 58 poultry farms, from which 471 poultry-related samples and 44 stool samples from in-contact humans were collected. The isolates were tested for their susceptibility to 11 antimicrobials using the Kirby-Bauer disk diffusion assay. The farm-level prevalence of Salmonella was 36.2% and the sample-level prevalence was 6.4% for samples taken from poultry farms and 4.5% in human stool samples who have contact with poultry. On-farm waste disposal practices and chicken being purchased from different multiplication farms were significantly associated with Salmonella positivity of the farms (p < 0.05). Eleven (34.4%) Salmonella isolates were resistant to streptomycin, and nine (28.1%) were resistant to tetracycline. Thirteen (40.6%) Salmonella isolates were resistant to two or more antimicrobials tested in this study, whereas resistance to 3 or more antimicrobials was detected in seven (21.9%) isolates. In conclusion, a high prevalence of Salmonella and a high rate of resistance to multiple antimicrobials were detected in poultry farms in Addis Ababa. Hence, implementation of strong biosecurity measures and rational use of antimicrobials are recommended.

Urinary tract infection among people living with human immunodeficiency virus attending selected hospitals in Addis Ababa and Adama, central Ethiopia.

Background: Urinary tract infections (UTIs) and antibacterial resistance (ABR) are important public health problems, but they are not well-studied among people living with human immunodeficiency virus (PLHIV) globally, especially in low-income countries. Therefore, it is important to regularly measure the extent of UTIs and ABR in the most susceptible populations. This study aimed to investigate the prevalence of UTIs, associated factors, bacterial causal agents, and their antibiotic susceptibility profile among PLHIV in central Ethiopia. Methods: A hospital-based cross-sectional study was conducted to recruit 688 PLHIV by a simple random sampling method. Background information was gathered through interviews, while clinical information was gathered from recent information sheets of patient charts using organized, pretested, and validated study tools. Midstream urine was collected aseptically and transported to the Microbiology Laboratory of Aklilu Lemma Institute of Pathobiology within 4 h of collection, maintaining its cold chain. Standard conventional microbial culture methods and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry were used to identify the bacterial isolates at the species level. Kirby Bauer's disk diffusion method was used to determine the antibiotic susceptibility profile of the bacterial isolates based on the interpretation guidelines of the Clinical Laboratory Standard Institute. Logistic regression models were used to examine factors associated with the occurrence of UTIs among PLHIV attending selected hospitals in Addis Ababa, and Adama. Results: Out of 688 PLHIVs involved in the current study, 144 (20.9%) were positive for UTIs, whereas the majority were asymptomatic for UTIs. In the multivariable logistic regression analysis, only HIV RNA ≥ 200 copies/ml [AOR = 12.24 (95% CI, 3.24, 46.20), p < 0.01] and being symptomatic for UTIs during the study period [AOR = 11.57 (95% CI, 5.83, 22.97), p < 0.01] were associated with the occurrence of UTIs. The dominant bacterial species isolated were Escherichia coli (E. coli; n = 65; 43%), followed by Enterococcus faecalis (E. faecalis; n = 16; 10.6%) and Klebsiella pneumoniae (K. pneumoniae; n = 11; 7.3%). Over half of the E. coli isolates were resistant to antibiotics such as gentamicin (GM; n = 44; 67.7%), amikacin (AN; n = 46; 70.8%), nalidixic acid (NA; n = 42; 64.6%), ciprofloxacin (CIP; n = 40; 61.5%), and azithromycin (AZM; n = 45; 69.2%). All of the K. pneumoniae isolates (n = 11; 100%), (n = 6; 54.5%), and (n = 7; 63.6%) were resistant to [amoxicillin as well as amoxicillin + clavulanic acid], ceftriaxone, and sulfamethoxazole + trimethoprim, respectively. All the Staphylococcus aureus (S. aureus) isolates were resistant to cefoxitin, which implies methicillin-resistant S. aureus (MRSA). Conclusion: The high prevalence of UTIs and antibiotic resistance revealed in the current study needs public health interventions such as educating the population about preventive measures and the importance of early treatment of UTIs. Our findings also highlight the need to provide UTI screening services for PLHIV, and healthcare providers should adopt antibiotic stewardship programs to promote and ensure their appropriate and judicious use.

Prevalence and antimicrobial susceptibility profile of Salmonella isolated from vegetable farms fertilized with animal manure in Addis Ababa Ethiopia.

The resistance of foodborne pathogens to antimicrobial agents is a potential danger to human health. Hence, establishing the status of good agricultural practices (GAPs) and the antimicrobial susceptibility of major foodborne pathogens has a significant programmatic implication in planning interventions. The objective of this study was to assess the gap in attaining GAP and estimate the prevalence and antimicrobial susceptibility profile of Salmonella in vegetable farms fertilized with animal manure in Addis Ababa, Ethiopia. A total of 81 vegetable farms from four sub-cities in Addis Ababa were visited, and 1119 samples were collected: soil (n = 271), manure (n = 375), vegetables (n = 398), and dairy cattle feces (n = 75). Additional data were collected using a structured questionnaire. Isolation of Salmonella was done using standard microbiology techniques and antimicrobial susceptibility testing was conducted using disk diffusion assays. Carriage for antimicrobial resistance genes was tested using polymerase chain reaction (PCR). Among the 81 vegetable farms visited, 24.7% used animal manure without any treatment, 27.2% used properly stored animal manure and 80.2% were easily accessible to animals. The prevalence of Salmonella was 2.3% at the sample level, 17.3% at the vegetable farm level, and 2.5% in vegetables. The highest rate of resistance was recorded for streptomycin, 80.7% (21 of 26), followed by kanamycin, 65.4% (17 of 26), and gentamicin, 61.5% (16 of 26). Multidrug resistance was detected in 61.5% of the Salmonella isolates. Vegetable farms have a gap in attaining GAPs, which could contribute to increased contamination and the transfer of antimicrobial resistance to the vegetables. The application of GAPs, including proper preparation of compost and the appropriate use of antimicrobials in veterinary practices, are recommended to reduce the emergence and spread of antimicrobial resistance.

Escherichia coli isolates from vegetable farms in Addis Ababa, Ethiopia: Antimicrobial susceptibility profile and associated resistance genetic markers.

The use of animal manure to fertilize soil is an emerging concern contributing to the transfer of antimicrobial-resistant pathogens to vegetables. Hence, assessing antimicrobial susceptibility profile of Escherichia coli in vegetable farms is essential to design appropriate interventions against antimicrobial resistance (AMR) in the food chain. This study assessed antimicrobial resistance profile and associated genetic markers among E. coli isolated from vegetable farms fertilized with animal manure in Addis Ababa, Ethiopia. A total of 1044 samples were collected using convenience sampling: soil (n = 271), manure (n = 375), and vegetables (n = 398) from 81 vegetable farms in Addis Ababa, Ethiopia. Antimicrobial susceptibility test was conducted for 100 E. coli isolates and antimicrobial resistance genes (ARGs) were tested by polymerase chain reaction (PCR). Of the 1044 collected samples, 25.3% were positive for E. coli, with significantly higher prevalence in the manure sample and samples collected from Akaki Kality sub-city (p < .05). The highest resistance rate was recorded for tetracycline (72%), followed by streptomycin (63%), and sulfamethoxazole +trimethoprim (56%). Multidrug resistance was detected in 61% of the E. coli isolates. The aac(3)-IV (76.9%), bla TEM (65.4%), aadA (60.3%), tet(A) (58.3%), and sulI (51.7%) were the commonly detected resistance genes. The current study showed a high burden of antimicrobial resistance among E. coli isolated from manure-amended vegetable farms, with potential of playing a significant role in the dissemination of antimicrobial resistance in the food chain. Efforts should be made to reduce the burden of resistant organisms and ARGs through prudent use of antimicrobials in livestock and application of appropriate composting techniques before using manure as fertilizer.

Genomic Characterization of Extended-Spectrum ß-Lactamase-Producing and Third-Generation Cephalosporin-Resistant Escherichia coli Isolated from Stools of Primary Healthcare Patients in Ethiopia.

The global spread of antimicrobial resistance genes (ARGs) in Escherichia coli is a major public health concern. The aim of this study was to investigate the genomic characteristics of extended-spectrum ß-lactamase (ESBL)-producing and third-generation cephalosporin-resistant E. coli from a previously obtained collection of 260 E. coli isolates from fecal samples of patients attending primary healthcare facilities in Addis Ababa and Hossana, Ethiopia. A total of 29 E. coli isolates (19 phenotypically confirmed ESBL-producing and 10 third-generation cephalosporin-resistant isolates) were used. Whole-genome sequencing (NextSeq 2000 system, Illumina) and bioinformatic analysis (using online available tools) were performed to identify ARGs, virulence-associated genes (VAGs), mobile genetic elements (MGEs), serotypes, sequence types (STs), phylogeny and conjugative elements harbored by these isolates. A total of 7 phylogenetic groups, 22 STs, including ST131, and 23 serotypes with different VAGs were identified. A total of 31 different acquired ARGs and 10 chromosomal mutations in quinolone resistance-determining regions (QRDRs) were detected. The isolates harbored diverse types of MGEs, with IncF plasmids being the most prevalent (66.7%). Genetic determinants associated with conjugative transfer were identified in 75.9% of the E. coli isolates studied. In conclusion, the isolates exhibited considerable genetic diversity and showed a high potential for transferability of ARGs and VAGs. Bioinformatic analyses also revealed that the isolates exhibited substantial genetic diversity in phylogenetic groups, sequence types (ST) and serogroups and were harboring a variety of virulence-associated genes (VAGs). Thus, the studied isolates have a high potential for transferability of ARGs and VAGs.

Antimicrobial Susceptibility and Characterization of Extended-Spectrum ß-Lactamase-Producing Escherichia coli Isolated from Stools of Primary Healthcare Patients in Ethiopia.

Antimicrobial resistance of Escherichia coli is a growing problem in both developed and developing countries. This study aimed to investigate the phenotypic antimicrobial resistance of E. coli isolates (n = 260) isolated from the stool specimen of patients attending public health facilities in Addis Ababa and Hossana. This study also aimed to characterize phenotypically confirmed extended-spectrum beta-lactamase (ESBL)-producing E. coli isolates (n = 22) using whole-genome sequencing. Resistance to 18 different antimicrobials was assessed using the disc diffusion method according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines. The highest resistance rate among the E. coli isolates was found for ampicillin (52.7%), followed by trimethoprim-sulfamethoxazole (29.6%). Of all isolates, 50 (19.2%) were multidrug-resistant and 22 (8.5%) were ESBL producers. ESBL genes were detected in 94.7% of the sequenced E. coli isolates, and multiple ß-lactamase genes were detected in 57.9% of the isolates. The predominant ESBL gene identified was blaCTX-M-15 (78.9%). The blaTEM-1B gene was detected in combination with other ESBL genes in 57.9% of the isolates, while only one of the sequenced isolates contained the blaTEM-1B gene alone. The blaCTX-M-3 gene was detected in three isolates. The genes blaCTX-M-15 and blaTEM-1B as well as blaCTX-M-15 and blaTEM-169 were confirmed to coexist in 52.6% and 10.5% of the sequenced E. coli isolates, respectively. In addition, blaOXA-1 was identified together with blaCTX-M-15 and blaTEM-1B in one isolate, and in one isolate, blaTEM-169 together with blaCTX-M-15 and blaTEM-1B was found. The results obtained show that measures need to be taken to reduce the spread of drug resistance and ensure the long-term use of available antimicrobials.

Antidiarrheal and Antibacterial Activities of Calpurnia aurea: Benth Seed Different Extracts.

Background: Calpurnia aurea is believed to have antidiarrheal potential but with limited scientific evidence. This study aimed investigating antidiarrheal and antibacterial activity of aqueous and 80% methanol seed extracts of the plant in mice and selected diarrhea-causing bacterial strains, respectively. Methods: Castor oil-induced diarrhea, prostaglandin-induced enteropooling, and castor oil-induced charcoal meal test models in mice of either sex using three dose levels (60, 120, and 240 mg/kg) were applied to evaluate antidiarrheal activity. Parameters, including onset, number, wet stool weight, weight and volume of secretion, and intestinal motility, were taken into consideration. The antibacterial activity was assessed on Shigella soni, Salmonella typhimurium, Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa using disk diffusion and microdilution techniques. Results: Compared to controls, pretreatment of mice at the graded dose (60, 120, and 240 mg/kg) resulted in a significant (p < 0.05) drop in frequency of wet stools and watery content of diarrhea as well as in delaying onset of diarrhea. Both extracts exhibited inhibition of diarrhea in a dose-dependent manner in all models used. The extracts also showed significant (p < 0.05) reduction in intestinal motility in castor oil-induced models. Both extracts showed a marginal activity against the selected bacterial strains; a better effect was seen with 80% methanol seed extract. Conclusion: Both extracts of the plant have beneficial effect in controlling diarrhea. This finding supports the use of the plant as a traditional antidiarrheal remedy.

Nontyphoidal Salmonella and Their Antimicrobial Susceptibility among Diarrheic Patients Attending Private Hospitals in Addis Ababa, Ethiopia.

Nontyphoidal Salmonella (NTS) is one of the major causes of bacterial foodborne infection. It is mainly manifested by self-limiting gastroenteritis in healthy individuals but can also cause severe complications including blood stream infection and mortality. The emergence of multidrug-resistant strains of Salmonella is becoming a global public health concern. This study is aimed at estimating the prevalence of Salmonella, identifying serotypes involved, and investigating antimicrobial susceptibility of the isolates among diarrheic patients attending private hospitals in Addis Ababa. We collected a total of 298 stool samples from diarrheic patients attending five private hospitals in Addis Ababa and isolated Salmonella according to standard microbiological techniques; the isolates were serotyped using slide agglutination and microplate agglutination techniques. Antimicrobial susceptibility test of the isolates was carried out using Kirby-Bauer disc diffusion assay according to Clinical Laboratory Standards Institute guidelines. Fourteen stool samples (4.7%) were positive for Salmonella, and Salmonella Kiambu was the most dominant serovar (n = 7, 50%) followed by S. Saintpaul (n = 4, 28.6%) and S. Haifa (n = 2, 14.3%). Three (21.4%) of the isolates were resistant to sulfisoxazole and tetracycline each and 2 (14.3%) to ampicillin. Resistance to two antimicrobials was detected only in 2 (14.3%) of the isolates, and none of the isolates were resistant to more than two antimicrobials. In conclusion, the current study showed low prevalence of NTS in diarrheic patients attending private hospitals in Addis Ababa. Although multidrug resistance to several antimicrobials was not detected in the isolates, prudent use of antimicrobials is recommended to guaranty the long-term use of the available antimicrobials.

Prevalence and Antimicrobial Susceptibility Profile of Staphylococcus aureus in Milk and Traditionally Processed Dairy Products in Addis Ababa, Ethiopia.

Staphylococcus aureus is a contagious pathogen that can cause various diseases in both humans and animals. Antimicrobial-resistant S. aureus is becoming an extremely important global health problem. A cross-sectional study was conducted from December 2019 to May 2020 to assess the occurrence of S. aureus and its antimicrobial susceptibility profiles in milk and traditionally processed dairy products in selected subcities of Addis Ababa. A total of 255 dairy product samples (175 raw milk and 80 traditionally processed dairy products) were collected from farms and retail markets. Samples were cultured for S. aureus according to standard microbiology techniques, and the Kirby-Bauer disk diffusion method was used to assess antimicrobial susceptibility of isolates to a panel of 12 antimicrobials. Susceptibility to methicillin was determined based on the sensitivity of isolates to cefoxitin, and resistant isolates were investigated for the presence of mecA and mecC genes using PCR. Staphylococcus aureus was isolated from 43 (24.6%) of milk, 7 (17.5%) of yogurt, and 2 (5%) of cottage cheese. A significantly higher rate of contamination with S. aureus was recorded among milk samples compared to yogurt and cottage cheese (p = 0.019). Out of 52 S. aureus isolates investigated for susceptibility to 12 antimicrobials, 49 (94.2%) of the isolates were resistant to ampicillin and 42 (80.8%) to amoxicillin+clavulanic acid. Twenty (38.5%) of the isolates were methicillin-resistant S. aureus (MRSA) based on susceptibility to cefoxitin. However, only one of these isolates (5%) was positive for mecA gene, and none of them were positive for the mecC gene. There was no significant difference (p > 0.05) in the rate of occurrence of MRSA among isolates from different sources. In conclusion, this study demonstrated a significant level of contamination of milk and dairy products with S. aureus and most isolates were multidrug resistant. The occurrence of MRSA in raw milk and dairy products signifies a serious public health threat as the practice of consuming raw dairy products in the study area is widespread. The lack of agreement between phenotypic and genotypic detection of MRSA suggests the need for further study to identify the genetic basis for the observed resistance phenotype.

Maternal Complications and Adverse Pregnancy Outcomes among Pregnant Women who Acquired Asymptomatic Bacteriuria in Addis Ababa, Ethiopia.

In this study, we aimed to document adverse pregnancy outcomes and maternal complications among pregnant women who acquired asymptomatic bacteriuria in Addis Ababa, Ethiopia. We used hospital-based prospective cohort study design in which we followed 44 pregnant women with asymptomatic bacteriuria confirmed by urine culture result of ≥105cfu/ml of urine. We documented adverse pregnancy outcomes and maternal complications in terms of frequency, percentage, mean, and standard deviation. Additionally, we used Pearson's correlation coefficient to investigate associations of selected variables with perinatal death as one of adverse pregnancy outcomes. Of the 44 pregnant women enrolled in the study, complete data was collected from 43 participants with one lost to follow-up. Six (14%) of women developed fever and were treated with antibiotic during pregnancy, 26 (60.5%) delivered with cesarean section, two (4.3%) perinatal deaths within seven days of delivery, one miscarriage, and 4 (9.3%) newborns were found underweight. The mean birth weight of the newborns was 3.1 kg ± 0.60. Almost half 21(48.8%) were born before 37 weeks of gestational age. Fourteen (32.6%) of newborns were born asphyxiated. Twenty-two (51.2%) of newborns developed early neonatal fever within 48 hours of delivery and treated with antibiotic. Correlation coefficient analysis revealed that weight and gestational age of newborns at birth, Apgar score at 1st and 5th minutes of birth and miscarriage were positively correlated and significantly associated with perinatal death. The occurrence of unsought pregnancy outcomes were frequent, and substantial number of pregnant women developed maternal complications. Therefore, screening pregnant women for asymptomatic bacteriuria and treating may reduce the possible maternal complications and adverse pregnancy outcomes.

Asymptomatic Bacteriuria among Pregnant Women in Addis Ababa, Ethiopia: Prevalence, Causal Agents, and Their Antimicrobial Susceptibility.

Asymptomatic bacteriuria (ASBU) is an important health problem among pregnant women, particularly in low-income countries. This study aimed to estimate the prevalence of ASBU and potential risk factors among pregnant women attending antenatal care centers in Addis Ababa. It also aimed to identify causal bacterial pathogens and to assess their antimicrobial susceptibility. A health facility-based cross-sectional study was conducted from March to June 2019. Urine samples from a total of 281 pregnant women with no symptoms of urinary tract infection were tested for ASBU. Women whose urine samples carried greater than or equal to 105 colony-forming units (CFU) of bacteria per milliliter of urine when grown on plate count agar were considered positive for ASBU. Bacterial pathogens were isolated from urine samples of women with ASBU using standard microbiological techniques. Antimicrobial susceptibility of isolates was investigated using the Kirby-Bauer disk diffusion method on Muller-Hinton agar plates. Of 281 pregnant women examined, 44 (15.7%) were positive for ASBU. Logistic regression analysis of the putative risk factors tested in the current study showed that none of them were significantly associated with the occurrence of ASBU (p > 0.05). The most frequently isolated bacterial species were Escherichia coli 17 (30.2%), Proteus 13 (23.2%), and Enterococcus 11 (19.6%). All of the E. coli, Citrobacter, and Klebsiella isolates and 84.6% of Proteus were resistant to ampicillin. All bacterial isolates were resistant to at least one of the antimicrobials tested. Resistance to three or more antimicrobials was detected in 15 (88.2%) of E. coli, 13 (100%) of Proteus, and 8 (72.7%) of Enterococcus isolates. Resistance to as many as 7 antimicrobials among E. coli, 8 antimicrobials among Proteus, and 7 antimicrobials among Enterococcus isolates was recorded. Detection of ASBU in a substantial number of pregnant women in this study warrants the need for a detailed study on possible risks of developing symptomatic urinary tract infection (UTI) and associated complications. Multidrug resistance to several antimicrobials was observed in the majority of bacterial isolates. Regular assessment of antimicrobial susceptibility of uropathogens to commonly prescribed antimicrobials and implementation of prudent use of antimicrobials are recommended.

Prevalence and antimicrobial susceptibility of Salmonella in poultry farms and in-contact humans in Adama and Modjo towns, Ethiopia.

Consumption of contaminated poultry and poultry products represents a common source of nontyphoidal Salmonella infection. Little is known on the status of Salmonella and their antimicrobial susceptibility in poultry farms in Ethiopia. This study investigated the prevalence, serotype distribution, and antimicrobial susceptibility of nontyphoidal Salmonella among poultry farms in Adama and Modjo towns. Three hundred thirty-four cloacal swabs, 384 fecal droppings of birds, 59 feed, 59 floor swabs, and 36 stools from in-contact humans were collected and processed for Salmonella isolation. Isolates were tested for their susceptibility to 15 antimicrobials using Kirby-Bauer disk diffusion assay. Seventeen (28.8%) of the farms and 24 (2.9%) of the samples from poultry farms and 2.8% (1/36) of stool samples of humans in-contact with poultry were positive for Salmonella. Most of the isolates (n = 21) were recovered from fecal droppings of birds while the remaining isolates were recovered from floor swab samples (n = 2) and cloacal swab sample (n = 1). Only three Salmonella serovars: S. Haifa (n = 14, 56%), S. Anatum (n = 7; 28%), and S. Give (n = 4; 16%) were detected. Poultry farms in Adama town, large flock sized farms, and farms that used antimicrobials were significantly associated with the occurrence of Salmonella (p < .05). Twenty (80%) and 19 (76%) of Salmonella isolates were resistant to streptomycin and tetracycline, respectively. Nineteen (76%) of the isolates were resistant to two or more antimicrobials. Detection of multidrug-resistant strains of Salmonella in poultry farms suggests the need for detailed epidemiological and molecular studies to establish sources of acquisition of resistant Salmonella strains.