Role of CTX-M-15 gene in spread of extended-spectrum beta-lactamases among immunocompetent patients in Ghana.

Background: Patients with faecal carriage of extended-spectrum beta-lactamases (ESBL)-producing Enterobacterales serve as reservoirs and sources of dissemination and infection. Objective: This report examined immunocompetent patients for faecal carriage of ESBL-producing Enterobacterales in a district care hospital setting in Ghana. Methods: Between March 2019 and May 2020, cross-sectional sampling was performed to enrol patients and conduct questionnaire-structured interviews for factors that predispose patients to ESBL faecal carriage. Faecal samples from study patients were quantified for ESBL-producing Enterobacterales. The ESBL genes were characterised by polymerase chain reaction and sequencing. Results: The overall proportion of ESBL faecal carriage was 35.5% (n = 38/107). The blaCTX-M gene, mostly CTX-M-15, was detected in 89.5% (n = 34/38) of the ESBL-producing isolates. The other ESBL types included blaSHV (n = 3) and blaOXA (n = 1). The CTX-M-15-positive isolates, when present in a faecal sample compared to the non-ESBL-CTX-M-15 isolates, constituted the predominant faecal Enterobacterales, with significantly higher colony counts than all other enterobacteria in that sample. In multivariate regression, independent risk factors for faecal carriage of ESBL-producing Enterobacterales were hospitalisation in the past year, infections since admission, use of antibiotics in the past 6 weeks, and admission from another hospital. Conclusion: The study found that CTX-M-15-producing isolates were the predominant faecal Enterobacterales, and that further investigations are needed to determine the reasons behind this dominance. What this study adds: The CTX-M-15-producing isolates dominance in this study shows the misuse and abuse of antibiotics in an African medical facility and indicates the potential role of immunity in controlling ESBL spread, which is to be investigated further.

Similarities in Bacterial Uropathogens and Their Antimicrobial Susceptibility Profile in Diabetics and Their Non-Diabetic Caregivers at a National Diabetes Management and Research Centre, Accra-Ghana.

BACKGROUND: Diabetes mellitus has some damaging effects on the genitourinary system and has been found to have adverse effects on the host immune system, resulting in higher risk of infections including urinary tract infections (UTI). Incidences of UTI caused by bacteria have been increasingly reported globally and the abuse of antibiotics leading to evolving resistant strains of antibiotics is a public health challenge for the management of this condition. Information on the difference in types of bacterial agents causing UTI in diabetic patients and non-diabetic individuals, and their susceptibility profiles, will facilitate effective management among these groups of patients. Therefore, this study aimed at determining the difference in prevalence of UTI, the causative bacteria, and their antimicrobial susceptibility profiles in diabetic patients and non-diabetic individuals at a diabetes management center. METHODS: This was a prospective cross-sectional study conducted amongst 100 diagnosed diabetic patients and 100 non-diabetic individuals. Urine sample was collected aseptically and analyzed microbiologically for the presence of urinary tract bacterial pathogens. Drug susceptibility testing was conducted on the isolates by the Kirby Bauer method to ascertain the antibiotic susceptibility patterns. RESULTS: Among the diabetic and non-diabetic individuals, urinary tract bacterial pathogens were observed in 28.0% and 26.0% of samples, respectively. The organisms were in the following proportions for diabetic and non-diabetic individuals, respectively: E. coli (14/28, 50% and 8/26, 30.7%), S. aureus (2/28, 7.1% and 4/26, 15.4%), K. pneumoniae (4/28, 14.3% and 8/26, 30.7%), K. ozoenae (2/28, 7.14% and 0/26, 0%), K. oxytoca (0/28, 0% and 4/26, 15.4%) and C. urealyticum (6/28, 21.4% and 2/26, 7.69%). The difference between the proportions of bacteria isolated was, however, not statistically significant (p-value = 0.894). Bacteria isolated from both diabetic and non-diabetic individuals were highly susceptible to most of the antibiotics tested, especially nitrofurantoin, cefuroxime, ceftriaxone, and cefotaxime. CONCLUSION: This study has shown that similarities exist in prevalence of UTI, the causative bacteria, and their antimicrobial susceptibility patterns amongst diabetic patients and non-diabetic individuals at a diabetes management center. These data will help in the management of UTI among these individuals.

Bacterial Contamination of Surgical Instruments Used at the Surgery Department of a Major Teaching Hospital in a Resource-Limited Country: An Observational Study.

Surgical instruments, be they disposable or reusable, are essential in any surgical procedure. Reusable surgical instruments should be properly sterilized or disinfected before use. However, the protocols are not always followed accordingly. This results in sterilization and disinfection failures, leading to a possible increase in the incidence of surgical site infections. This observational study report on bacterial contaminants identified instruments used for surgical procedures in a major teaching hospital in a resource-limited country. In total, 207 pre-sterilized surgical instruments and instrument parts used at three units-the general surgical theater, and the gastrointestinal (GI) endoscopy and urology endoscopy (uro-endoscopy) units-within the surgical department were randomly sampled and examined for bacterial contamination. Bacteria isolates were identified, and their antimicrobial susceptibility patterns were determined. Bacteria isolates that were identified included Citrobacter spp., Citrobacter freundii, Bacillus cereus, Staphylococcus hominis, and Staphylococcus aureus. Bacillus cereus was the most predominant bacteria isolated (30/61, 49.1%), and Staphylococcus hominis the least (1/61, 1.6%). In terms of the number of isolates from the three units examined, the uro-endoscopy unit recorded the highest followed by the general surgical theater and the GI endoscopy. However, there was no association between the various units and bacteria isolated, and no significant difference between the number of isolates among the various units (p = 0.9467, χ2 = 0.1095). In this study, even though CFU per device or device part counted was less than 20, bacteria isolated from the instruments used for a surgical procedure is of great concern considering that the setting of the study is a major teaching hospital. Multi-drug resistance was observed in almost all the isolated bacteria. Sterilization processes should be strictly adhered to, taking into consideration the length and temperature in order to reduce the risk of using contaminated instruments in these environments. It is therefore recommended that similar studies should be carried out in surgical departments at different levels of hospitals to ascertain the extent of this problem.

Aminoglycoside Therapy for Tuberculosis: Evidence for Ototoxicity among Tuberculosis Patients in Ghana.

Background: Hearing impairment (HI) is a major problem in Ghana; however, the few attempts at shedding light on its causes appear to overlook the adverse effects of some medications­a gap that this study sought to fill. Aminoglycoside therapy for tuberculosis (TB) treatment is one of these medications. Aim: The aim of this study was to establish the potential of aminoglycoside as a cause of hearing impairment among patients on anti-TB treatment. Method: This was a case−control study, involving patients receiving treatment for TB with aminoglycoside at the chest clinic of the Tema General Hospital and a control group of age- and gender-matched healthy volunteers. A structured questionnaire was administered to obtain the demographic data and case history of the participants. The hearing sensitivity of the participants was assessed using conventional pure tone audiometry and transient evoked otoacoustic emission tests. Results: A hearing loss prevalence of 20% (12/60) was found among patients receiving treatment for TB, with the intensity of impairment ranging from mild to severe. Hearing thresholds of patients receiving anti-TB medications were significantly elevated (p < 0.05) in comparison to the thresholds of the control group, especially at the high frequencies. Conclusion: This study shows that aminoglycoside therapy for tuberculosis may contribute to hearing impairment among tuberculosis patients in Ghana. Audiological management of these patients should therefore be an essential part of their therapeutic treatment plan.

Antimicrobial Activity of Selected Medicinal Plants from a Sub-Saharan African Country against Bacterial Pathogens from Post-Operative Wound Infections.

BACKGROUND: Globally, the application of medicinal plants in the management of acute and chronic wounds can be considered a common occurrence in most traditional medicine practices. In view of this, many plants in the tropical and subtropical regions have been screened for their wound-healing activities. Consequently, plants having antimicrobial activity against multidrug-resistant (MD-R) pathogens can be considered great assets. Therefore, this study evaluated ethanolic and aqueous extracts of five medicinal plants (Psidium guajava, Myrianthus arboreus, Alchornea cordifolia, Momordica charantia, and Justicia flava) for their antimicrobial activities against MD-R bacterial pathogens isolated from post-operative wounds; Methods: This involved the aqueous and ethanolic extraction of the selected medicinal plants. Preliminary phytochemical constituents of the plants were examined. The agar well diffusion method was then used to determine the antibacterial activity of the leaves against reference strains (Escherichia coli ATCC 25922, Salmonella typhi ATCC 19430, Pseudomonas aeruginosa ATCC 27853 and Staphylococcus aureus ATCC 25923, and a Methicillin-Resistant Staphylococcus aureus strain) as well as the MDR clinical isolates (E. coli, P. aeruginosa, S. aureus and CoNS) from the wounds; Results: The preliminary phytochemical analysis of the leaves showed the presence of saponins, phenolics, and reducing sugars in almost all the plants tested. All plant extracts were observed to have some antimicrobial activity against at least one reference strain. For the clinical MDR isolates, A. cordifolia from this study showed highest inhibition to growth of all bacteria used. Activity of J. flava against S. aureus was highest as compared to that of E.coli and P. aeruginosa. Similar observation was made for M. arboreus, P. guajava and M. charantia where the highest activity was observed against S. aureus; Conclusion: This study has mainly shown that P. guajava, M. arboreus, A. cordifolia, M. charantia, and J. flava exhibits antimicrobial activities against MD-R bacterial pathogens isolated from post-operative wounds. Also, these plants has bioactive phytochemical compounds with potential medicinal values for the treatment of numerous infections. Therefore, these plants may be helpful in the management of acute and chronic wounds, especially in traditional medicine practices.

High rates of multi-drug resistant gram-negative organisms associated with surgical site infections in a teaching hospital in Ghana.

BACKGROUND: There is limited data to guide the prevention and management of surgical site infections (SSI) in low- and middle-income countries. We prospectively studied aetiological agents associated with SSI and their corresponding antibiotic susceptibility patterns in a tertiary hospital in Ghana. METHODS: As part of a cohort study carried out at the surgical department of the Korle Bu Teaching Hospital (KBTH) from July 2017 to April 2019, wound swabs were collected from patients diagnosed with SSI. Isolates cultured from the wound swabs were identified by MALDI TOF and susceptibility testing was conducted according to EUCAST 2020 guidelines. Clinical data were monitored prospectively. RESULTS: Of 4577 patients, 438 developed an SSI and 352 microbial isolates were cultured. Isolates were predominantly Gram negative (286, 81%), a pattern seen for all kinds of surgery and all wound classes. The most common species included Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus and Acinetobacter baumannii. The majority of organisms were multi-drug resistant including 86% of E. coli, 52% of A. baumannii and 86% of K. pneumoniae; and 65% (17/26) of the cefotaxime-resistant K. pneumoniae were extended spectrum ß-lactamase producing. One of 139 E. coli, 15 of 49 P. aeruginosa, and 6 of 23 A. baumannii were meropenem resistant, but no clonal pattern was found. There was a 1% (5/428) prevalence of methicillin-resistant S. aureus. CONCLUSIONS: The predominance of Gram-negative organisms and the high level of multi-drug resistance indicate a need to re-evaluate antibiotic prophylaxis and treatment protocols in surgical practice in low- and middle-income countries.

Microscopic Observation Drug Susceptibility (MODS) Assay: A Convenient Method for Determining Antibiogram of Clinical Isolates of Mycobacterium tuberculosis in Ghana.

(1) Background: Present methods for drug susceptibility tests (DST) rely on culture methods that are sophisticated and relatively faster, or a slow and cheaper option. These methods frustrate disease control; therefore, there is a need for methods that incorporate key functions of microscopy and culture, with reduced cost burden and sophistry. Thus, the purpose of this study was to identify which, among the most commonly used (in Ghana) methods, can conveniently be used at health centers located in rural areas for effective DST determination of Mycobacterium tuberculosis (MTB). (2) Methods: Mycobacterium tuberculosis isolates were tested for their susceptibility to streptomycin, isoniazid, rifampicin, ethambutol (SIRE), and pyrazinamide by microscopic observation drug susceptibility (MODS) and BACTEC MGIT 960 methods. Evaluations were based on shorter turnaround periods, rapidity, ease of use, cost, etc. A comparative analysis was statistically expressed as kappa values. (3) Results: Endpoints for drug susceptibilities by MODS averaged 13 days (7-32), whilst that for BACTEC MGIT 960 was 10 days with a further 12 days to detect resistance. Therefore, a turnaround period of 22 days was needed for DST by BACTEC MGIT 960, compared to 13 days for MODS. There were differences in correlation levels between the two methods, as determined by their kappa values. (4) Conclusion: The MODS assay was found to be less costly, more user-friendly, and still able to be conveniently used at health centers located in rural areas known to be endemic for TB, particularly in Ghana.

Antibiotic use in surgical units of selected hospitals in Ghana: a multi-centre point prevalence survey.

BACKGROUND: Improper use of antibiotics leads to the emergence of resistant microorganisms as well as drug toxicity, increased healthcare costs, morbidity and mortality. Globally, an estimated 25-68% of hospitalized patients receive suboptimal antibiotic regimes. Information on the extent of this problem in Ghana is currently limited, particularly in surgical units. To strategize for interventions, we estimated the antibiotic use prevalence in surgical departments in a country-wide point prevalence survey (PPS) in Ghana. METHODS: Between October 2016 and December 2016, we conducted a cross-sectional multi-center country-wide PPS. This involved an audit of in-patients' records from all units/departments of ten systematically selected hospitals in Ghana. Data were collected with a standardized questionnaire, adopted from the European Centre for Disease Prevention and Control. In this report, we present data on antibiotic use from the surgical units. RESULTS: Of 2107 eligible patients included in the PPS, 540 patients were identified in surgical units, of which 70.7% (382/540) received antibiotic therapy. A total of 636 antibiotic prescriptions were issued to these surgical patients; 224 (58.6%) for treatment, including 50 for treatment of hospital-acquired infections, and 144 (37.7%) for prophylaxis (medical and surgical). Median duration of antibiotic therapy prior to the survey was 5 days (interquartile range (IQR): 3-8 days). Surgical prophylaxis was administered for longer than the recommended one day in 107 of 144 (88.4%) patients. The choice of antibiotics was largely similar for community- and hospital-acquired infections as well as for prophylaxis. Only 3.7% of patients had microbiological analysis done on clinical samples. CONCLUSION: We found a high prevalence of antibiotic use, with the choice of antibiotics, in some cases, inconsistent with the country's treatment guidelines. Antibiotics were administered for long duration including antibiotics for prophylactic purposes and the majority was started without supporting microbiological analysis. Prescription practices that encourage rational use of antibiotics guided by microbiology and enforcement of antibiotic policy guidelines should be the target for future interventions.

Plasma Levels of Cytokines (IL-10, IFN-γ and TNF-α) in Multidrug Resistant Tuberculosis and Drug Responsive Tuberculosis Patients in Ghana.

The emergence of multidrug-resistant tuberculosis (MDR⁻TB) and more recently, extensively drug-resistant (XDR) TB has intensified the need for studies aimed at identifying factors associated with TB drug resistance. This study determined the differences in plasma concentrations of pro-inflammatory (IFN-γ and TNF-α) and anti-inflammatory (IL-10) cytokines in MDR-TB and drug-susceptible (DS) TB patients, in addition to some socio-economic factors. Plasma levels of IL-10, IFN-γ and TNF-α were measured in 83 participants (comprising 49 MDR-TB and 34 DS-TB patients) using sandwich ELISA. Levels of the three cytokines were elevated in MDR-TB patients compared to DS-TB patients. The mean level of IL-10 (7.8 ± 3.61 ρg/mL) measured in MDR-TB cases was relatively higher than those of TNF-α and IFN-γ, and statistically significant (p = 0.0022) when compared to the level of IL-10 (4.8 ± 4.94 ρg/mL) in the DS-TB cases. There were statistically significant associations between MDR-TB and factors such as education level (X² = 9.895, p = 0.043), employment status (X² = 19.404, p = 0.001) and alcoholism (X² = 3.971, p = 0.046). This study adds to the knowledge that IFN-γ, TNF-α and IL-10 play a role in the host response to Mycobacterium tuberculosis (MTB). Alcohol intake can be considered as an important MDR-TB risk factor.

Antibiotic prescribing in paediatric inpatients in Ghana: a multi-centre point prevalence survey.

BACKGROUND: Excessive and inappropriate use of antibiotics in hospitalised patients contributes to the development and spread of antibiotic resistance. Implementing a stewardship programme to curb the problem requires information on antibiotic use. This study describes a multicentre point prevalence of antibiotic use among paediatric inpatients in Ghana. METHODS: Data were extracted from a multicentre point prevalence survey of hospital acquired infections in Ghana. Data were collected between September 2016 and December 2016 from ten hospitals through inpatient folder and chart reviews using European Centre for Disease Control (ECDC) adapted data collection instrument. From each site, data were collected within a 12-h period (8 am to 8 pm) by a primary team of research investigators and a select group of health professionals from each participating hospital. RESULTS: Among 716 paediatric inpatients, 506 (70.6%; 95% confidence interval (CI): 67.2 to 74.0%) were on antibiotics. A significant proportion of antibiotics (82.9%) was prescribed for infants compared to neonates (63.9%) and adolescents (60.0%). The majority of patients (n = 251, 49.6%) were prescribed two antibiotics at the time of the survey. The top five classes of antibiotics prescribed were third generation cephalosporins (n = 154, 18.5%) aminoglycosides (n = 149, 17.9%), second generation cephalosporins (n = 103,12.4%), beta lactam resistant penicillins (n = 83, 10.0%) and nitroimidazoles (n = 82, 9.9%). The majority of antibiotics (n = 508, 61.0%) were prescribed for community acquired infections. The top three agents for managing community acquired infections were ceftriaxone (n = 97, 19.1%), gentamicin (n = 85, 16.7%) and cefuroxime (n = 73, 14.4%). CONCLUSION: This study points to high use of antibiotics among paediatric inpatients in Ghana. Cephalosporin use may offer an important target for reduction through antibiotic stewardship programmes.

In Vitro Susceptibility of Mycobacterium ulcerans Isolates to Selected Antimicrobials.

Background. The current definitive treatment of Buruli ulcer with antibiotics makes the issue of antimicrobial drug resistance an unavoidable one. This is as a result of drug misuse by health personnel and patients' noncompliance to treatment regimen. Monitoring of these factors and screening for new effective antimicrobials are crucial to effective management of Buruli ulcer disease. This study therefore investigated the inhibitory activity of some antibiotics against isolates of Mycobacterium ulcerans. Methods. Activity of eight antibiotics was tested against twelve M. ulcerans isolates (2 reference strains and 10 clinical isolates). The anti-M. ulcerans activities were determined by the agar dilution method and the minimum inhibitory concentrations (MICs) were determined by the agar proportion method. Results. All antimicrobials investigated had activity against M. ulcerans isolates tested. The MICs ranged from 0.16 µg/mL to 2.5 µg/mL. Azithromycin recorded the highest inhibitory activity at a mean MIC of 0.39 µg/mL, whilst clofazimine a second-line antileprosy drug, recorded the lowest at a mean MIC of 2.19 µg/mL. Among the four antituberculosis drugs, rifampicin had the highest activity with a mean MIC of 0.81 µg/mL. Conclusion. Azithromycin could be considered as a lucrative alternative to existing treatment methods for inhibiting M. ulcerans in Ghana.

Evaluating decontamination protocols for the isolation of Mycobacterium ulcerans from swabs.

BACKGROUND: Mycobacterium ulcerans (M. ulcerans) is the causative agent of Buruli Ulcer (BU) disease. In order to inhibit the growth of the microbial contaminants during culture of M. ulcerans, it is necessary to decontaminate BU samples with effective chemical agents. This study aimed at investigating some selected chemicals as potential decontamination agents for the isolation of M. ulcerans from swabs. RESULTS: Povidone iodine at 0.5 and 1% exhibited the lowest contamination and recovery rate for microbial contaminants and M. ulcerans. The most effective decontamination method was the protocol using 2% cetylpyridinium chloride/4% sodium chloride (recovery rate = 53%, contamination rate = 14%). The observed difference between the recovery rate of 2% CPC/4% NaC and the other protocols was however not statistically significant (p = 0.76). CONCLUSIONS: Two percent (2%) cetylpyridinium chloride/4% sodium chloride can be conveniently used as an alternative decontamination method for the isolation of M. ulcerans from swabs.

Characterization of Staphylococcus aureus from Human Immunodeficiency Virus (HIV) patients in Accra, Ghana.

INTRODUCTION: The aims of this study were to: a) determine the nasal carriage prevalence of Stahpylococcus aureus among HIV patients, b) to characterize S. aureus strains isolated. METHODOLOGY: Characterization of S. aureus isolates was done by antibiotyping, spa typing, and detection of Panton-Valentine leukocidin (PVL) genes. RESULTS: S. aureus isolated (10/124; 8%)  belonged to spa types t084 (n = 3), t10828 (n = 2), t311, t304, t774, t645, and t091. The isolates were resistant to penicillin (100%), tetracycline (40%), rifampicin (10%), fucidic acid (10%), norfloxacin (10%), erythromycin (10%), and sulfamethoxazole trimethoprim (10%). Multidrug resistance (MDR) was detected in 30% of the isolates. CONCLUSION: The finding of MDR S. aureus among HIV-positive patients suggests that surveillance of antimicrobial resistant S. aureus among this patient group could be considered as an infection control measure in the hospital.

Susceptibility Profiles of Mycobacterium ulcerans Isolates to Streptomycin and Rifampicin in Two Districts of the Eastern Region of Ghana.

Background. Drug resistance is a major challenge in antibiotic chemotherapy. Assessing resistance profiles of pathogens constitutes an essential surveillance tool in the epidemiology and control of infectious diseases, including Buruli ulcer (BU) disease. With the successful definitive management of BU using rifampicin and streptomycin, little attention had been paid to monitoring emergence of resistant Mycobacterium ulcerans (M. ulcerans) isolates in endemic communities. This study investigated the susceptibility profiles of M. ulcerans isolates from two BU endemic areas in Ghana to streptomycin and rifampicin. Methods. The antibiotic susceptibility of seventy (70) M. ulcerans isolates to rifampicin and streptomycin was determined simultaneously at critical concentrations of 40 µg/mL and 4 µg/mL, respectively, by the Canetti proportion method. Results. Resistance to rifampicin was observed for 12 (17.1%) M. ulcerans isolates tested, whilst 2 (2.9%) showed resistance to streptomycin. None of the isolates tested showed dual resistance to both rifampicin and streptomycin. Conclusion. Outcomes from this study may not be reflective of all BU endemic communities; it, however, provides information on the resistance status of the isolates, which is useful for monitoring of M. ulcerans, as well as BU disease surveillance and control.

Maximizing microscopy as a diagnostic tool in peripheral health centres of BU endemic areas in Ghana.

BACKGROUND: Buruli ulcer (BU) disease, a skin condition caused by Mycobacterium ulcerans (M. ulcerans) is endemic in remote rural areas. Disease diagnosis on clinical basis alone can be misleading, requiring definitive diagnosis based on laboratory tests. Resource constraints in BU endemic areas make microscopy for the detection of acid fast bacilli (AFB) an important and useful method. It is rapid, user-friendly, convenient and cheap. Despite its usefulness, its performance is relatively low. This study investigated modifications of the current method aimed at improving its performance. Forty (IS) 2404 polymerase chain reactions (PCR) positive BU samples were processed by eight physical (centrifugation and overnight sedimentation) and chemical (phenol ammonium sulphate and sodium hypochlorite) modifications of the current direct method. Assessments were based on standard AFB evaluation coupled with in house criteria; positivity (P), clarity and contrast (C) release of bacilli from specimen (R). Overall AFB positivity rate was 64% (409/640). Each protocol had 80 smears. The percentage positivity (P) for the conventional method was 58% (46/80) smears. The highest positivity rate of 57/80 (%) was by protocol 7 (5% phenol in 4% ammonium sulphate (PhAS) and concentrated by overnight gravitational sedimentation). The least positivity rate at 35% (28/80) was by protocol 1 (smears from direct application of swab tips). The differences in performance between the two chemical tested; 5% phenol in 4% ammonium sulphate (PhAS) and 3.5% NaHOCl was significant (p<0.05). The differences between the two physical methods were however not significant (p>0.05). This study concluded that BU samples treated with a solution of 5% phenol in 4% ammonium sulphate and concentrated by either centrifugation or overnight sedimentation is useful for maximizing AFB detection by bright field microscopy. This can be useful in rural health facilities with resource constraints.