Carbapenemase Production and Detection of Colistin-Resistant Genes in Clinical Isolates of Escherichia Coli from the Ho Teaching Hospital, Ghana.

Background: Effective and successful treatment of infectious diseases is a significant gain in clinical settings. However, resistance to antibiotics, especially the last-resort medicines, including carbapenems and colistin is on the rise. Aim: The aim of this study was to detect carbapenemase production and colistin-resistant genes in clinical isolates of Escherichia coli. Method. The study was a cross-sectional study carried out from July 2018 to June 2019. One hundred and thirty-five nonrepetitive E. coli isolates obtained from various clinical samples were screened for carbapenemase production using meropenem (10 µg) and imipenem (10 µg) disks. Screened-positive isolates were further subjected to a confirmatory test using modified carbapenem inhibition method (mCIM). Deoxyribonucleic acid (DNA) was extracted from all the isolates to detect colistin-resistant genes by polymerase chain reaction. Data were analyzed using GraphPad Prism version 8.00 for Windows and IBM SPSS version 26 (IMB Corp. New York, USA). Results: Of the 135 isolates, 2 were screened positive for carbapenemase production but tested negative to mCIM. With the colistin-resistant genes, only mcr-1 and mcr-2_700bp were detected in 3 of the E. coli isolates, representing 2.2%. The mcr-1 was detected in a high vaginal swab sample of a female aged between 65 and 84 years. Mcr-2_700bp was also detected in urine and blood samples of the patients. Conclusion: The study investigated the presence of carbapenemase and colistin-resistant genes in E. coli organisms. The absence of carbapenemase in the isolates and the detection of colistin-genes call for strict infection prevention and control practices to prevent their introduction and spread to other bacterial species, respectively.

Burden of Fluoroquinolone Resistance in Clinical Isolates of Escherichia coli at the Ho Teaching Hospital, Ghana.

BACKGROUND: The growing burden of antibiotic resistance is a threat to the management of infections. Infections by Escherichia coli are routinely treated with fluoroquinolone antimicrobial agents. Due to their frequent use, there has been increasing resistance to these drugs. We set out to determine the burden of fluoroquinolone resistance among clinical E. coli isolates at the Ho Teaching Hospital, Ghana. METHODS: This was a cross-sectional study conducted from July 2018 to June 2019. One hundred and thirty-five E. coli isolates were cultured from various clinical samples. Antimicrobial susceptibility testing was performed using the Kirby-Bauer disk diffusion method with discs of nalidixic acid (NAL), ciprofloxacin (CIP), norfloxacin (NOR) and levofloxacin (LEV). Deoxyribonucleic acid (DNA) was extracted from the resistant isolates for the detection of fluoroquinolone resistant genes by polymerase chain reaction. RESULTS: Ninety of the 135 isolates (66.7%) were resistant to at least one of the four fluoroquinolone drugs investigated. Resistance to NAL, CIP, NOR, and LEV was 51.0%, 51.1%, 38.8% and 35.7% respectively. Out of the fluoroquinolone resistant isolates, 69 carried one or more fluoroquinolone resistant genes. The predominant resistant genes were aac(6')-Ib-cr (48.9%) and qnrD (25.6%). Seven of the isolates carried both qnrS and aac(6')-Ib-cr genes. Two isolates carried 5 different fluoroquinolone resistant genes. CONCLUSION: High prevalence of resistance to 4 fluoroquinolone drugs was recorded with associated resistant genes. This is a threat to current efforts to control the spread of antibiotic resistance and calls for concerted efforts to curb the spread of these resistant organisms.

Plasmid-mediated quinolone resistance determinants in clinical bacterial pathogens isolated from the Western Region of Ghana: a cross-sectional study.

Introduction: quinolones are critically important antibiotics that are reserved for treating very severe infections caused by multidrug-resistant bacterial pathogens. However, their indiscriminate uses have resulted in an increased number of resistant strains in many parts of the world including Ghana. We determined the quinolone resistance profile of Gram-negative bacterial pathogens and characterized the underlying molecular determinants of resistance. Methods: Gram-negative pathogens obtained from clinical specimens at three hospital laboratories were tested for resistance to quinolones and other commonly used antibiotics. ESBL production among the Enterobacterial isolates was confirmed using the combined disc diffusion method. We then used PCR to determine seven types of plasmid-mediated quinolone resistance genes present in the isolates resistant to nalidixic acid and ciprofloxacin. Results: in this study, 29.5% of the isolates were resistant to ciprofloxacin, with the highest of 50% among E. coli resistance to the other quinolones was levofloxacin (24.4%), norfloxacin (24.9%), and nalidixic acid (38.9%). Significant proportions of the quinolone-resistant isolates were ESBL producers (P-values < 0.001). The aac(6´)-Ib-cr, qnrS, oqxA, and qepA genes were present in 43 (89.6%), 27 (56.3%), 23 (47.9%), and one (2.1%) of the isolates, respectively. None of the isolates tested positive to qnrA, qnrB, and oqxB genes. The presence of the aac(6´)-Ib-cr gene positively correlated with resistance to ceftriaxone, cefotaxime, and gentamicin (P-values < 0.05). Conclusion: high proportions of Gram-negative bacterial isolates were resistant to quinolones and most of these isolates possessed multiple PMQR genes. There is a need to implement measures to limit the spread of these organisms.

Genotypic characterization of extended-spectrum ß-lactamase producing urinary isolates among pregnant women in Ho municipality, Ghana.

Objective: The case of antibiotic resistance has become a major global concern and Extended Spectrum ß-lactamase (ESBL) producing organisms have so far remained the biggest culprit. The consequences of urinary tract infection (UTI) and antibiotic resistance among pregnant women cannot be underestimated. We investigated UTI and ESBL production among urinary pathogens isolated from pregnant women. Method: We obtained non-repeat, clean catch midstream urine samples from 1345 pregnant women suspected of having UTI for bacterial identification at the Ho Teaching Hospital Laboratory between June 2013 and March 2015. The isolates were taken through relevant biochemical testing for identification and then subjected to antimicrobial agents for susceptibility testing using the disc diffusion method. We tested for ESBL production by the combined disc method and ESBL positive (+ESBL) phenotype isolates were genotyped for BlaTEM, BlaSHV, and BlaCTX-M using polymerase chain reaction (PCR). Data were analyzed using SPSS v24 and p-values < 0.05 were considered statistically significant. Results: Of the 1345 urine samples tested, 230 (17.1%, 95% CI: 15.1%-19.1%) yielded significant bacteriuria. The most common bacterium isolated was Staphylococcus aureus (29.6%) followed closely by Escherichia coli (28.7%) both of which were highest during the second trimester of gestation. We isolated 152 gram-negative isolates with 41.4% (63/230) being + ESBL. Of the 63 + ESBL, 45 (71.4%) possessed blaTEM, 42 (66.7%) had blaCTX-M and 2 (3.2%) possessed blaSHV genes; 38 possessed multiple ESBL genes comprising 2 with both SHV and TEM genes and 36 with both CTX-M and TEM genes. Conclusion: High prevalence of UTI and persistent transmission of ESBLs among pregnant women in the Ho Municipality is worrying and a course for public health concern. We recommend urine culture during pregnancy as a routine laboratory investigation to avoid birth-related complications.

Occurrence of Cryptosporidium Infection and Associated Risk Factors among HIV-Infected Patients Attending ART Clinics in the Central Region of Ghana.

Cryptosporidium species are intestinal protozoan parasites that infect and cause diarrhoea in animals and humans. The current study was conducted to determine the prevalence and risk factors of Cryptosporidium infection among HIV-infected patients in the Central region of Ghana. In this cross-sectional study, four hundred eighteen documented HIV-infected participants from four health facilities that provide antiretroviral therapy (ART) services across the Central region of Ghana were selected by systematic random sampling. An enzyme-linked immunosorbent assay (CoproELISATM, Cryptosporidium Savyon® Diagnostics Ltd., Ashdod, Israel) was used to detect Cryptosporidium antigens in stool samples obtained from participants. Information regarding participants' sociodemographic characteristics and clinical symptoms as well as potential environmental and behavioral risk factors were collected using a structured questionnaire. Chi-square or Fisher's exact tests were used to determine associations between Cryptosporidium infections and explanatory variables, while risk factors were assessed using multivariate logistic regression analysis. The overall prevalence of Cryptosporidium infection among HIV-infected participants in this study was 6.2% (95% CI: 3.90-8.54). Cryptosporidium was not significantly associated with any of the sociodemographic variables, patient clinical symptoms, and environmental factors. However, the prevalence of the parasite was significantly higher 25% (95% CI: 1.17-48.83; p = 0.013) among participants who did not always wash their hands before meals and those who did not always wash vegetables before eating them, 23.5% (95% CI: 1.05-46.01; p = 0.016). Multivariate logistic regression analysis showed that participants who used public water closet facilities were approximately 9 times more likely to become infected with the parasite than those who practised open defecation (OR: 8.83; 95% CI: 1.22-64.13; p = 0.031). In conclusion, Cryptosporidium is prevalent among HIV-infected patients in the Central region of Ghana. An important risk factor identified was the use of the public water closet toilet facility. More attention should be given to ensuring cleanliness at shared water closet facilities in addition to adequate disinfection of hands after using such facilities.

Cryptosporidium Infection and Associated Risk Factors among Cattle in the Central Region of Ghana.

Cryptosporidium species infects a wide number of animals including livestock all over the world. The current study was done to determine the prevalence and risk factors of Cryptosporidium infection among cattle in the Central Region of Ghana. Two hundred and eighty-seven (287) faecal samples were randomly collected from animals on eight cattle farms in four districts across two agroecological zones. A commercial enzyme-linked immunosorbent assay kit (CoproELISA, Savyon® Diagnostics Ltd., Israel) for Cryptosporidium was used in the detection of Cryptosporidium antigens in faecal samples. Characteristics of the animals such as age, sex, and location, as well as consistency of faecal samples, were collected. Pearson's chi-square or Fisher's exact test was used to determine the association between explanatory variables and Cryptosporidium infection while a logistic regression model was also used to determine the risk of infection. The overall prevalence of Cryptosporidium infection was 23.7% (95% CI, 18.7-28.6). Prevalence was significantly higher (p = 0.049) among cattle aged 12-month old and above compared to those under 12 months of age. Among the four districts in the study area, Cape Coast metropolis recorded a significantly higher prevalence (60.5%; CI, 49.3-71.8), (p < 0.001) compared to the other three. Furthermore, a significant association was observed between the consistency of faecal samples and Cryptosporidium infection (p = 0.042). The prevalence of Cryptosporidium infection was also significantly higher among cattle from the coastal savanna zone (26.9%; 95% CI, 21.0-32.8) compared to those from the semideciduous forest area (p = 0.017). Cattle in the forest zone had a lower risk of being infected with the parasite compared to those from the coastal savanna zone (OR 0.408; 95% CI, 0.182-0.915). In conclusion, Cryptosporidium was prevalent among cattle in the Central Region of Ghana. A higher prevalence of Cryptosporidium infection occurred in older animals and among animals in the coastal agroecological zone. The area of location and age of animals were identified as risk factors for Cryptosporidium infection in the Central Region of Ghana.

Occurrence and distribution of extended-spectrum ß-lactamase in clinical Escherichia coli isolates at Ho Teaching Hospital in Ghana.

Objective: This study determined the occurrence and distribution of Extended Spectrum ß-Lactamase (ESBL) genotypes of E. coli isolates in Ho Teaching Hospital, Ghana. Design: A cross-sectional study. Setting: A single centre study was conducted at Ho Teaching Hospital of Ghana. Participants: Patients who visited Ho Teaching Hospital Laboratory with the request for culture and susceptibility testing. Main outcome measure: Escherichia coli were isolated, and Extended-Spectrum ß-Lactamase genes were detected. Results: Of the 135 isolates, 56(41.5%,95% CI: 33.1% - 50.3%) were ESBL producers. More males, 14(58.3%), produced ESBL than females, 42(37.8%). The ESBL prevalence was highest among the elderly who were 80 years and above 3(100.0%), with the least prevalence among patients within 50-59 years and 0-9 years age bracket, representing 4(25.0%) and 3(27.3%), respectively. The total prevalence of ESBL was marginally higher among out-patients (41.8% 95% CI: 31.9% - 52.2%) compared to in-patients [40.5% 95% CI: 24.8% - 57.9]. BlaTEM-1 was the predominant ESBL genotype obtained from 83.9% (47/56) of the confirmed ESBL producing isolates, with the least being TOHO-1 4(7.1%). The co-existence of 2 different ESBL genes occurred in 19(33.9%) of the isolates. The single and quadruple carriage were 16(28.6%) and 3(5.4%), respectively. The highest co-existence of the ESBL genotypes was recorded for blaTEM-1 and blaCTXM-1 15(26.8%), followed by blaTEM-1, blaCTXM-1 and blaSHV-73 [12(21.4%)]. Conclusion: The high prevalence of ESBL-producing E. coli isolates with multiple resistant gene carriage is a threat to healthcare in the study area. Funding: This research received no external funding.

Estimated Burden of Serious Fungal Infections in Ghana.

Fungal infections are increasingly becoming common and yet often neglected in developing countries. Information on the burden of these infections is important for improved patient outcomes. The burden of serious fungal infections in Ghana is unknown. We aimed to estimate this burden. Using local, regional, or global data and estimates of population and at-risk groups, deterministic modelling was employed to estimate national incidence or prevalence. Our study revealed that about 4% of Ghanaians suffer from serious fungal infections yearly, with over 35,000 affected by life-threatening invasive fungal infections. Incidence of cryptococcal meningitis, Pneumocystis jirovecii pneumonia, and disseminated histoplasmosis cases in AIDS was estimated at 6275, 12,610 and 724, respectively. Oral and esophageal candidiasis collectively affect 27,100 Ghanaians and 42,653 adult asthmatics are estimated to have fungal asthma. We estimate a prevalence of 12,620 cases of chronic pulmonary aspergillosis (CPA and an incidence of 1254 cases of invasive aspergillosis (IA). Estimated cases of candidemia and candida peritonitis cases were 1446 and 217, respectively. The estimated prevalence of recurrent vulvovaginal candidiasis (RVVC) and tinea capitis was 442,621 and 598,840, respectively. Mucormycosis and fungal keratitis each may affect 58 and 810 Ghanaians. These data highlight the urgent need for intensified awareness to improve diagnosis and management.

Characterization of highly virulent multidrug resistant Vibrio cholerae isolated from a large cholera outbreak in Ghana.

OBJECTIVE: The purpose of this study was to investigate the virulent factors of Vibrio cholerae which caused an unprecedented large cholera outbreak in Ghana in 2014 and progressed into 2015, affected 28,975 people with 243 deaths. RESULTS: The V. cholerae isolates were identified to be the classical V. cholerae 01 biotype El Tor, serotype Ogawa, responsible for the large cholera outbreak in Ghana. These El Tor strains bear CtxAB and Tcp virulent genes, making the strains highly virulent. The strains also bear SXT transmissible element coding their resistance to antibiotics, causing high proportions of the strains to be multidrug resistant, with resistant proportions of 95, 90 and 75% to trimethoprim/sulfamethoxazole, ampicillin and ceftriaxone respectively. PFGE patterns indicated that the isolates clustered together with the same pattern and showed clusters similar to strains circulating in DR Congo, Cameroun, Ivory Coast and Togo. The strains carried virulence genes which facilitated the disease causation and spread. This is the first time these virulent genes were determined on the Ghanaian Vibrio strains.