Reservoirs of Nosocomial Pathogens in Intensive Care Units: A Systematic Review.

Background: Nosocomial pathogens are known to exacerbate morbidity and mortality in contemporary critical healthcare. Hospital fomites, which include inanimate surfaces, have been identified as "breeding grounds" for pathogens that cause nosocomial infections. This systematic review aimed to deliver incisive insights on nosocomial pathogens in intensive care units (ICUs) and the role of fomites as potential reservoirs for their transmission. Method: An extensive exploration of electronic databases, including PubMed and Scopus, from 1990 to 2023, was carried out between 25th and 29th May 2023, per standard PRISMA guidelines. Information were extracted from articles that reported on fomites in the ICU. Studies that did not quantitatively report the fomite contamination, and those that exclusively took samples from patients in the ICU were excluded from the analysis. Results: About 40% of the total samples collected on fomites from all the studies yielded microbial growth, with species of Staphylococcus being the most predominant. Other prevalent microbes were Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, Candida spp., Enterococcus sp., and Enterobacter sp. The neonatal intensive care unit (NICU) had the highest proportion of contaminated fomites. Among known fomites, the sphygmomanometer exhibited a 100% detection rate of nosocomial pathogens. This included E. aerogenes, Staphylococcus aureus, coagulase-negative Staphylococci (CoNS), E. coli, and K. pneumoniae. Multidrug-resistant (MDR) bacteria, such as methicillin-resistant S. aureus (MRSA), vancomycin-resistant Enterococci (VRE), extended-spectrum beta-lactamase (ESBL)-producing E. coli, and MDR Pseudomonas aeruginosa were commonly isolated on fomites in the ICUs. Conclusion: Many fomites that are readily used in patient care in the ICU harbour nosocomial pathogens. The most common fomite appeared to be mobile phones, sphygmomanometers, and stethoscopes, with Staphylococcus being the most common contaminant. Consequently, the need for rigorous disinfection and sterilization protocols on fomites in the ICU cannot be overemphasized. Additionally, heightened awareness on the subject among health professionals is crucial to mitigating the risk and burden of nosocomial infections caused by drug-resistant bacteria.

Sepsis-Related Lung Injury and the Complication of Extrapulmonary Pneumococcal Pneumonia.

Globally, sepsis and pneumonia account for significant mortality and morbidity. A complex interplay of immune-molecular pathways underlies both sepsis and pneumonia, resulting in similar and overlapping disease characteristics. Sepsis could result from unmanaged pneumonia. Similarly, sepsis patients have pneumonia as a common complication in the intensive care unit. A significant percentage of pneumonia is misdiagnosed as septic shock. Therefore, our knowledge of the clinical relationship between pneumonia and sepsis is imperative to the proper management of these syndromes. Regarding pathogenesis and etiology, pneumococcus is one of the leading pathogens implicated in both pneumonia and sepsis syndromes. Growing evidence suggests that pneumococcal pneumonia can potentially disseminate and consequently induce systemic inflammation and severe sepsis. Streptococcus pneumoniae could potentially exploit the function of dendritic cells (DCs) to facilitate bacterial dissemination. This highlights the importance of pathogen-immune cell crosstalk in the pathophysiology of sepsis and pneumonia. The role of DCs in pneumococcal infections and sepsis is not well understood. Therefore, studying the immunologic crosstalk between pneumococcus and host immune mediators is crucial to elucidating the pathophysiology of pneumonia-induced lung injury and sepsis. This knowledge would help mitigate clinical diagnosis and management challenges.

The Potential of Bacteriophage-Antibiotic Combination Therapy in Treating Infections with Multidrug-Resistant Bacteria.

The growing threat of antibiotic resistance is a significant global health challenge that has intensified in recent years. The burden of antibiotic resistance on public health is augmented due to its multifaceted nature, as well as the slow-paced and limited development of new antibiotics. The threat posed by resistance is now existential in phage therapy, which had long been touted as a promising replacement for antibiotics. Consequently, it is imperative to explore the potential of combination therapies involving antibiotics and phages as a feasible alternative for treating infections with multidrug-resistant bacteria. Although either bacteriophage or antibiotics can potentially treat bacterial infections, they are each fraught with resistance. Combination therapies, however, yielded positive outcomes in most cases; nonetheless, a few combinations did not show any benefit. Combination therapies comprising the synergistic activity of phages and antibiotics and combinations of phages with other treatments such as probiotics hold promise in the treatment of drug-resistant bacterial infections.

In vitro antimicrobial activity of crude propolis extracts and fractions.

The search for antimicrobials in propolis presents a new dimension for addressing the problem of antimicrobial drug resistance. The aim of this study was to determine the antimicrobial activity of extracts of crude propolis collected from different regions in Ghana and their active fractions. The antimicrobial activity of the extracts, as well as that of the chloroform, ethyl acetate, and petroleum ether fractions of the active samples were determined using the agar well diffusion method. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the most active fractions were determined. The various crude propolis extracts frequently produced zones of inhibition against Staphylococcus aureus (17/20) than Pseudomonas aeruginosa (16/20), and Escherichia coli (1/20) test isolates. Chloroform and ethyl acetate solvents produced fractions possessing greater antimicrobial activity than the petroleum ether fraction. The mean MIC range of the most active fractions was greatest for S. aureus (76.0 ± 34.8-48.0 ± 33.0 mg/ml) than for P. aeruginosa (40.8 ± 33.3-30.4 ± 6.7 mg/ml) and E. coli, as was the mean MBC. Propolis has antimicrobial potential, and hence should be exploited as an alternative for the treatment of bacterial infections.

In Vitro effects of petroleum ether, dichloromethane, methanolic and aqueous leaf extracts of Eucalyptus grandis on selected multidrug-resistant bacteria.

BACKGROUND: The emergence and spread of antimicrobial resistance is of grave concern, requiring the search for newer and more effective antimicrobials to combat infections caused by resistant microbes. This study assessed the antimicrobial effects of Eucalyptus grandis crude extracts against selected multidrug resistant bacteria. METHODOLOGY: Four different crude leaf extracts of E. grandis were prepared using petroleum ether, dichloromethane, methanol, and water, with the aid of the Soxhlet extraction method. These were screened against methicillin-resistant Staphylococcus aureus (MRSA), multidrug resistant Pseudomonas aeruginosa, and multidrug resistant Escherichia coli, using the agar well diffusion method. Phytochemical screening was carried out to evaluate the bioactive phytochemical constituents responsible for the antimicrobial effect. RESULTS: Each of the extracts, except for the one prepared from water, had antimicrobial activity against the screened bacteria. The non-polar petroleum ether extract had the highest antimicrobial activity (19.33-24.33 mm), including bactericidal effects, compared to the medium polar dichloromethane and polar methanol extracts, which recorded zone diameter ranges of 14.33-16.67 mm and 16.33-17.67 mm, respectively. The Gram-negative bacteria (E. coli and P. aeruginosa) were the least susceptible in comparison with the Gram-positive bacterium (MRSA), probably owing to differences in their cell wall structures. Furthermore, phytochemical screening indicated the presence of alkaloids, tannins, saponins, terpenoids, and flavonoids. CONCLUSION: The findings suggest that E. grandis could be potentially useful in the treatment of infections caused by multidrug resistant bacteria.

Multicenter Surveillance of Antimicrobial Resistance among Gram-Negative Bacteria Isolated from Bloodstream Infections in Ghana.

BACKGROUND: Antimicrobial resistance (AMR) in Gram-negative bacteria-causing bloodstream infections (BSIs), such as Klebsiella pneumoniae and non-typhoidal Salmonella (NTS), is a major public health concern. Nonetheless, AMR surveillance remains scarce in sub-Saharan Africa, where BSI treatment is largely empirical. The aim of the study was to determine the distribution and AMR patterns of BSI-causing NTS, K. pneumoniae, and other Gram-negative bacteria in Ghana. METHODS: A cross-sectional study was conducted between April and December 2021 at eleven sentinel health facilities across Ghana as part of a pilot study on the feasibility and implementation of the human sector AMR surveillance harmonized protocol in sub-Saharan Africa. Gram-negative bacteria recovered from blood specimens of febrile patients were identified using MALDI-TOF and evaluated for antimicrobial resistance using the BD Phoenix M50 analyzer and Kirby-Bauer disc diffusion. The Department of Medical Microbiology at the University of Ghana served as the reference laboratory. RESULTS: Out of 334 Gram-negative blood isolates, there were 18 (5.4%) NTS, 85 (25.5%) K. pneumoniae, 88 (26.4%) Escherichia coli, 40 (12.0%) Acinetobacter baumannii, 25 (7.5%) Pseudomonas aeruginosa, and 77 (23.1%) other Gram-negative bacteria. As a composite, the isolates displayed high resistance to the antibiotics tested-amoxicillin (89.3%), tetracycline (76.1%), trimethoprim-sulfamethoxazole (71.5%), and chloramphenicol (59.7%). Resistance to third-generation cephalosporins [ceftriaxone (73.7%), cefotaxime (77.8%), and ceftazidime (56.3%)] and fluoroquinolones [ciprofloxacin (55.3%)] was also high; 88% of the isolates were multidrug resistant, and the rate of extended-spectrum beta-lactamase (ESBL) production was 44.6%. Antibiotic resistance in K. pneumoniae followed the pattern of all Gram-negative isolates. Antibiotic resistance was lower in NTS blood isolates, ranging between 16.7-38.9% resistance to the tested antibiotics. Resistance rates of 38.9%, 22.2%, and 27.8% were found for cefotaxime, ceftriaxone, and ceftazidime, respectively, and 27.8% and 23.8% for ciprofloxacin and azithromycin, respectively, which are used in the treatment of invasive NTS. The prevalence of multidrug resistance in NTS isolates was 38.9%. CONCLUSIONS: Multicenter AMR surveillance of Gram-negative blood isolates from febrile patients was well-received in Ghana, and the implementation of a harmonized protocol was feasible. High resistance and multidrug resistance to first- or second-choice antibiotics, including penicillins, third-generation cephalosporins, and fluoroquinolones, were found, implying that these antibiotics might have limited effectiveness in BSI treatment in the country. Continuation of AMR surveillance in Gram-negative blood isolates is essential for a better understanding of the extent of AMR in these pathogens and to guide clinical practice and policymaking.

Phenotypic and genotypic detection of carbapenemase-producing Escherichia coli and Klebsiella pneumoniae in Accra, Ghana.

AIM: To describe the occurrence of carbapenem resistance among multidrug-resistant (MDR) Escherichia coli and Klebsiella pneumoniae isolated from clinical specimens in Accra using phenotypic and genotypic methods. METHODOLOGY: The study was cross-sectional, involving 144 clinical MDR E. coli and K. pneumoniae isolates recovered from the Central Laboratory of the Korle Bu Teaching Hospital (KBTH). The isolates were re-cultured bacteriologically, identified using standard biochemical tests, and subjected to antibiotic susceptibility testing using the Kirby-Bauer method. Carbapenem resistance was determined based on imipenem, meropenem, and ertapenem zones of inhibition, as well as minimum inhibitory concentrations (MICs). Carbapenemase production was determined phenotypically by modified Hodge test (MHT) and modified carbapenem inactivation method (mCIM), and genotypically with multiplex PCR targeting the blaKPC, blaIMP, blaNDM, blaVIM, and blaOXA-48 genes. RESULTS: Of the 144 MDR isolates, 69.4% were E. coli, and 30.6% were K. pneumoniae. The distribution of antimicrobial resistance rates among them was ampicillin (97.2%), cefuroxime (93.1%), sulfamethoxazole-trimethoprim (86.8%), tetracycline (85.4%), cefotaxime and cefpodoxime (77.1% each), amoxicillin-clavulanate (75%), ceftriaxone (73.6%), ciprofloxacin (70.8%), levofloxacin (66.0%), cefepime (65.3%), ceftazidime (64.6%), gentamicin (48.6), piperacillin-tazobactam (40.3%), cefoxitin (14.6%), amikacin (13.9%), ertapenem and meropenem (5.6% each), and imipenem (2.8%). In total, 5.6% (8/144) of them were carbapenem-resistant (carbapenem MIC range = 0.094-32.0 µg/ml), with 75% (6/8) of these testing positive by the phenotypic tests and 62.5% (5/8) by the genotypic test (of which 80% [4/5] carried blaOXA-48 and 20% (1/5) blaNDM). The blaVIM, blaIMP, and blaKPC genes were not detected. CONCLUSION: Although the rates of antibiotic resistance among the isolates were high, the prevalence of carbapenemase producers was low. The finding of blaOXA-48 and blaNDM warrants upscaling of antimicrobial resistance surveillance programmes and fortification of infection prevention and control programmes in the country.

Microbial Contamination of Herbal Preparations on the Ghanaian Market, Accra.

In developing countries, an estimated 80% of the population use traditional herbal medicines as part of their primary health care. As the market for herbal medicine expands in many African countries, partly due to their use in the treatment of COVID-19, there is the need to address all the associated safety issues. The aim of the study was to evaluate the microbial contamination of locally prepared, as well as imported foreign herbal products sold in Accra. Standard microbiological methods were employed in the enumeration of coliforms and the identification of pathogenic microbes in 60 herbal preparations that were sampled. A larger proportion (76.7%) of local herbal preparations was contaminated with bacteria as compared with imported ones (63.3%). Bacillus species and Pseudomonas aeruginosa were the predominant bacteria obtained from foreign and locally manufactured herbal preparations, respectively. A proportion of 36.7% (11) of the local samples were positive for coliform and the coliform counts ranged from 3.0 × 101 cfu/ml to 2.0 × 104 cfu/ml. Two foreign herbal samples (6.7%) were positive for coliforms; one had a count of 1.7 × 105 cfu/g while the other had 2 × 104 cfu/g. Herbal preparations sold in markets of Accra harbour several microbial pathogens; the risk is relatively higher for locally produced herbal preparations compared to imported herbal preparations. As a result, it is recommended that quality assurance in the production of local herbal preparations should be thoroughly monitored from the beginning of production to the final selling of the preparations. There is also the need to strengthen microbiological safety monitoring of imported herbal preparations.

Multidrug-Resistant Gram-Negative Bacteria Contaminating Raw Meat Sold in Accra, Ghana.

BACKGROUND: Efforts to combat antimicrobial resistance (AMR) should be based on the One Health approach, involving human health, animal health, and the environment. In Ghana, previous studies on AMR have given little attention to animal source food, a major route of transmission of antibiotic-resistant zoonotic pathogens. The aim of this study was to investigate the occurrence of multidrug-resistant (MDR) bacteria in meat sold in Accra. METHODS: This was a cross-sectional study in which 270 meat samples (90 each of beef, goat meat, and chicken) were collected, and investigated for contamination with multidrug-resistant bacteria. The bacteria were subjected to susceptibility testing against amikacin (30 µg), ampicillin (10 µg), amoxicillin-clavulanate (20/10 µg), cefuroxime (30 µg), ceftriaxone (30 µg), ceftazidime (30 µg), cefepime (30 µg), ciprofloxacin (5 µg), trimethoprim-sulfamethoxazole (1.25/23.75 µg), ertapenem (10 µg), meropenem (10 µg), imipenem (10 µg), tigecycline (15 µg), and gentamicin (10 µg). RESULTS: Thirty-two different types of bacteria, totalling 558, were isolated, the predominant being Escherichia coli (44.6%), Aeromonas hydrophila (19.9%), Vibrio cholerae (3.4%), Aeromonas veronii (3.2%), and Klebsiella pneumoniae (3.1%). The prevalence of MDR among the contaminating bacteria was 14.9%. The MDR distribution among the predominant bacteria was Escherichia coli (18.7%), Aeromonas hydrophila (11.1%), Vibrio cholerae and Aeromonas veronii (0.0% each), and K. pneumoniae (5.6%). Moreover, 2.0% of the contaminating bacteria were extended-spectrum beta-lactamase (ESBL) producers, all of which occurred in the chicken samples, and their distribution was: Escherichia coli (1.3%), Klebsiella pneumoniae, Pantoea spp., Enterobacter cloacae, and Serratia plymuthica (0.2% each). CONCLUSIONS: The meat samples were heavily contaminated with Escherichia coli and Aeromonas hydrophila, and less frequently, with Vibrio cholerae, Klebsiella pneumoniae, and other organisms. The prevalence of multidrug-resistant bacteria was moderate (14.9%), while that of ESBL producers was low (2%).

Staphylococcus aureus Nasopharyngeal Carriage and Antimicrobial Resistance among Adults with Sickle Cell Disease at the Korle Bu Teaching Hospital in Accra, Ghana.

Background: Sickle cell disease (SCD) patients are an important risk group for Staphylococcus aureus (S. aureus) carriage and infections. Little is, however, known about the nasopharyngeal carriage epidemiology of the pathogen in this vulnerable population. Aim: The aim of this study was to evaluate S. aureus and methicillin-resistant S. aureus (MRSA) nasopharyngeal carriage prevalence, carriage determinants, and antimicrobial resistance among SCD adults in Ghana. Methodology: Nasopharyngeal swabs, obtained from 200 SCD adults recruited at the Korle Bu Teaching Hospital, were cultured for S. aureus, and these isolates were subjected to antimicrobial susceptibility testing via the Kirby-Bauer method. Results: The prevalence of S. aureus carriage was 41.5% (n = 83), and that of MRSA carriage was 1.0% (n = 2). Moreover, carriage of coagulase-negative Staphylococcus (CoNS) was the only determinant of S. aureus carriage identified (OR = 0.012, P < .0001). However, neither this variable nor the other features of the participants emerged as a determinant of MRSA carriage. The antimicrobial resistance rates decreased across penicillin (98.8%, n = 82), tetracycline (54.2%, n = 45), gentamicin (32.5%, n = 27), ciprofloxacin (21.7%, n = 18), erythromycin (18.1%, n = 15), clindamycin (10.8%, n = 9), amoxicillin-clavulanic acid (10.8%, n = 9), teicoplanin (1.2%, n = 1), and linezolid (0.0%, n = 0), and the multidrug resistance rate was 45.8% (n = 38). Conclusion: The nasopharyngeal carriage prevalence of S. aureus in the current study was high, while that of MRSA was low. The isolates were highly resistant to several of the antibiotics tested, but not teicoplanin and linezolid, making these antibiotics suitable for treatment of S. aureus infections among the SCD population.

High prevalence of methicillin-resistant Staphylococcus aureus carriage among infants at the Children's Hospital, Accra, Ghana.

INTRODUCTION: Infants are at risk of Staphylococcus aureus (S. aureus) colonization and infection. The aim of this study was to investigate S. aureus and methicillin-resistant S. aureus (MRSA) colonization among infants, including the prevalence, predictors of colonization, and antibiogram. METHODOLOGY: The study was cross-sectional, and involved infants aged less than one year recruited at the Princess Marie Louise Children's Hospital in Accra, Ghana. Sociodemographic and clinical data of the participants were gathered with a structured questionnaire. Nasal swabs were also obtained from them and bacteriologically cultured. S. aureus was confirmed with the coagulase test, and MRSA was confirmed by polymerase chain reaction (PCR) of the mecA gene. Antimicrobial susceptibility testing of S. aureus was done using the Kirby-Bauer method. RESULTS: The carriage prevalence of S. aureus and MRSA were 34.9% (45/129) and 17.10% (22/129), respectively. Colonization with coagulase-negative Staphylococci (CoNS) was protective of both S. aureus (OR = 0.008; p < 0.001) and MRSA (OR = 0.052; p = 0.005) carriage. Maintenance of good hand hygiene prevented S. aureus carriage (OR = 0.16; p < 0.001). S. aureus resistance to antibiotics decreased across penicillin (96%), trimethoprim-sulfamethoxazole (61%), tetracycline (61%), erythromycin (39%), gentamicin (39%), fusidic acid (26%), rifampicin (17%), clindamycin (7%), and linezolid (0%); 68.8% S. aureus were multidrug resistant. CONCLUSIONS: S. aureus and MRSA prevalence were high among the infants. Colonization with CoNS and good hand hygiene maintenance were predictive of MRSA and methicillin-sensitive S. aureus (MSSA) colonization, respectively.

Multidrug-Resistant Bacteria in Hospital Wastewater of the Korle Bu Teaching Hospital in Accra, Ghana.

Background: Antimicrobial resistance (AMR) is one of the top 10 public health threats. One approach to tackling the AMR menace could involve expanding the range of AMR surveillance domains to include hospital wastewater (HWW), a domain that has largely been overlooked by researchers. Aim: To evaluate the occurrence of multidrug-resistant bacteria in hospital wastewater of the Korle Bu Teaching Hospital (KBTH). Methodology: This was a longitudinal study involving 288 HWW samples consecutively collected across 12 weeks from the pool of wastewater emanating from 2 critical care units of KBTH-The Child Health Unit and the Maternity Unit-on Mondays and Thursdays, each week. The samples were cultured for bacteria, which were identified using the Matrix-Assisted Laser Desorption/Ionization-Time of Flight (MALDI-TOF) technique and subjected to antimicrobial susceptibility testing via the Kirby-Bauer method. Results: In total, 294 bacteria of 23 different types, all being Gram-negative, were isolated from the 288 samples. The predominant ones were Escherichia coli (30.6%, n = 90), Klebsiella pneumoniae (11.2%, n = 33), Citrobacter freundii (10.9%, n = 32), Alcaligenes faecalis (5.8%, n = 17), and Pseudomonas mendocina (5.4%, n = 16). The prevalence of multidrug resistance among the isolates was 55.4% (n = 163). Moreover, the prevalence of extended-spectrum beta-lactamase (ESBL) producers was 15.6% (n = 46). E. coli accounted for the most ESBL-producing organisms (28.9%, n = 26). Conclusion: The wastewater generated by the Maternity and Child Health Units of KBTH harbored a wide range of multidrug resistant bacteria, with a good proportion of these being ESBL producers, and the predominant one being E. coli. The study thus identifies the wastewater of KBTH as an important source of multidrug resistant organisms, and underscores the significance of appropriate treatment of wastewater of the hospital and other clinical, and related settings prior to its discharge.

Distribution and Antimicrobial Resistance Profiles of Bacterial Aetiologies of Childhood Otitis Media in Accra, Ghana.

Background: Otitis media (OM), also known as middle ear infection, is a clinically significant childhood disease. In sub-Saharan Africa, there is a paucity of contemporary reports on it is bacterial aetiologies and antimicrobial resistance among them. Aim: To investigate the OM bacterial aetiologies and their antimicrobial resistance patterns among children visiting the Ear, Nose, and Throat clinics of 3 healthcare facilities in Accra, Ghana - Princess Marie Louise Children's Hospital, 37 Military Hospital, and Mamprobi Hospital. Methods: This cross-sectional study involved 100 children below 13 years old with suppurative otitis media. Following standard bacteriological methods, sterile ear swabs were used to take middle ear discharges from the study participants for culture and antimicrobial susceptibility testing. A standard questionnaire was also used to collect data on socio-demographic and clinical characteristics. Results: The major OM bacterial aetiologies were Pseudomonas aeruginosa (38.5%), Klebsiella pneumoniae (19.8%), Proteus mirabilis (11.5%), and Staphylococcus aureus (10.4%). The majority of the bacteria demonstrated low to moderate resistance (0%-33.3%) to most of the antibiotics. Eight of the bacteria (4 each of Klebsiella pneumoniae and Escherichia coli) were extended-spectrum beta-lactamase (ESBL) producers; 6 ampicillinase (Amp C)-producing organisms (4 Citrobacter spp. and one each of Morganella morganii and Serratia marcescens) were also identified, and they showed high antibiotic resistance. Conclusions: The predominant OM aetiologies were Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus mirabilis, and Staphylococcus aureus, and they were generally susceptible to most of the antibiotics tested. Amikacin, cefepime, ciprofloxacin, and meropenem could be valuable in the empirical management of childhood OM.

A Streptococcus pneumoniae lineage usually associated with pneumococcal conjugate vaccine (PCV) serotypes is the most common cause of serotype 35B invasive disease in South Africa, following routine use of PCV.

Pneumococcal serotype 35B is an important non-conjugate vaccine (non-PCV) serotype. Its continued emergence, post-PCV7 in the USA, was associated with expansion of a pre-existing 35B clone (clonal complex [CC] 558) along with post-PCV13 emergence of a non-35B clone previously associated with PCV serotypes (CC156). This study describes lineages circulating among 35B isolates in South Africa before and after PCV introduction. We also compared 35B isolates belonging to a predominant 35B lineage in South Africa (GPSC5), with isolates belonging to the same lineage in other parts of the world. Serotype 35B isolates that caused invasive pneumococcal disease in South Africa in 2005-2014 were characterized by whole-genome sequencing (WGS). Multi-locus sequence types and global pneumococcal sequence clusters (GPSCs) were derived from WGS data of 63 35B isolates obtained in 2005-2014. A total of 262 isolates that belong to GPSC5 (115 isolates from South Africa and 147 from other countries) that were sequenced as part of the global pneumococcal sequencing (GPS) project were included for comparison. Serotype 35B isolates from South Africa were differentiated into seven GPSCs and GPSC5 was most common (49 %, 31/63). While 35B was the most common serotype among GPSC5/CC172 isolates in South Africa during the PCV13 period (66 %, 29/44), 23F was the most common serotype during both the pre-PCV (80 %, 37/46) and PCV7 period (32 %, 8/25). Serotype 35B represented 15 % (40/262) of GPSC5 isolates within the global GPS database and 75 % (31/40) were from South Africa. The predominance of the GPSC5 lineage within non-vaccine serotype 35B, is possibly unique to South Africa and warrants further molecular surveillance of pneumococci.

Preliminary Investigation into Plasmodium-like Piroplasms (Babesia/Theileria) among Cattle, Dogs and Humans in A Malaria-Endemic, Resource-Limited Sub-Saharan African City.

Babesia and Theileria are protozoan parasites belonging to the order piroplasmida, transmitted by hard ticks, and can cause diseases known as piroplasmosis. Human infections are usually asymptomatic, except in immuno-compromised persons who present malaria-like symptoms. Moreover, microscopically, the morphologies of Babesia and Theileria can resemble that of the malaria parasite, Plasmodium. In malaria-endemic areas with limited resources, these similarities can increase the possibility of misdiagnosing a patient as having malaria instead of piroplasmosis, which may further lead to inappropriate choice of disease management. This preliminary investigation aimed at detecting Babesia/Theileria in cattle, dogs and humans in some parts of Accra. Whole blood samples were taken from febrile cattle (n = 30) and dogs (n = 33), as well as humans diagnosed with malaria (n = 150). Blood samples of all study subjects were microscopically screened for possible presence of haemoparasites. Samples whose smears had features suggestive of possible piroplasmic infection were all given the label "suspected Babesia/Theileria-infected" samples. Nested polymerase chain reaction (PCR) was performed on extracted deoxyribonucelic acid (DNA) from all the "suspected" samples of cattle, dogs and humans, with primer sets that can detect 18S rRNA genes of Babesia/Theileria spp. In addition to this, amplification was performed on the "suspected" dog samples using the BcW-A/BcW-B primer set which detects the 18S rRNA genes of B. canis, while the BoF/BoR primer set which targets the rap-1 region of B. bovis and another primer set which detects the 18S rRNA genes of most bovine Babesia spp. (including B. divergens) were used on the suspected cattle samples. For the human samples, however, additional amplification was done on the extracted DNA using primers for the three other Babesia targeted (B. divergens, B. bovis and B. canis). Microscopy showed possible Babesia/Theileria infection suspected in all three groups of subjects in the following proportions: cattle (10/30; 33%), dogs (3/33; 9%) and humans (6/150; 4%). DNA from one-third of the "suspected" dog samples yielded amplification with Babesia canis primers. Moreover, a broad-detecting set of primers (that can amplify some Babesia and Theileria species) amplified DNA from nine (9/30; 30%) of the "suspected" cattle samples, but none from those of the humans. Although for this study conducted in the city, the Babesia/Theileria primers used did not amplify DNA from the six "suspected" human samples; the possibility of Babesia/Theileria infection in humans in other parts of the country cannot be overruled. There is therefore a need for further studies on possible emergence of human babesiosis/theileriosis in other parts of Ghana and sequencing for specific identification of any circulating strain.

Dynamics in Morbidity Markers and Cytological Observations Made in Urine of Schistosoma haematobium-Infected Children: Pre- and Post-Praziquantel Treatment in an Endemic Setting.

BACKGROUND: Schistosomiasis is a neglected tropical disease caused by helminths of the genus Schistosoma. Morbidity markers and cytological observations such as squamous metaplastic cells, inflammatory cells, and hyperkeratotic cells in the urine of S. haematobium-infected children may suggest disease severity. They may also help predict severe forms of clinical presentation, such as bladder cancer in later years, among infected ones who miss out on early detection and treatment. Insights into possible changes in the morbidity markers and cytological observations in the urine of these S. haematobium-infected children before and after treatment would be of high clinical importance. AIM: The aim of this study was to identify changes/dynamics in morbidity markers and cytological abnormalities in the urine deposits of S. haematobium-infected children, pre- and post-praziquantel treatment. METHODOLOGY: This was a longitudinal study involving baseline and follow-up sampling among basic school children living in schistosomiasis-endemic communities. Urine samples were collected from 520 children at baseline and examined for S. haematobium ova by microscopy, while urine chemistry analyses were used for the examination of morbidity markers. The cytological analyses involved cytopathological examination of the urine deposits. Children whose urine showed positivity for S. haematobium eggs were treated with a single oral dose of praziquantel (40 mg/kg), after which urine chemistry and cytological analyses were repeated weekly for comparison with baseline, until the eighth week. RESULTS: Morbidity markers such as hematuria, proteinuria, and leukocyturia were detected both at baseline and post-treatment among the infected children (30/520). Hematuria was the predominant parameter (90%, 27/30) detected at baseline, followed by proteinuria (53.3%, 16/30). Leukocyturia was the rarest parameter detected at baseline (13.3%, 4/30). However, almost all these parameters declined gradually post-treatment. Regarding cytological analyses, inflammatory cells were observed most (70.0%, 21/30) at baseline. For hyperkeratotic cells and squamous metaplastic cells, 46.7% and 26.7% were respectively observed at baseline, all of which gradually declined during the weekly follow-ups. Notably, squamous metaplastic cells persisted in all the participants from Week 1 through Week 3 post-treatment, but declined gradually thereafter. CONCLUSIONS: Morbidity markers and cytological observations in the children gradually decreased after treatment. Therefore, we continue to recommend routine cytological screening for urogenital schistosomiasis patients at hospitals in S. haematobium-endemic locations using both baseline and follow-up samples to detect these abnormalities early and monitor changes that may be occurring after treatment. Such changes may be useful in assessing treatment progress in infected persons.

Contrasting Hygiene-Related Gastrointestinal Infections and Stress-Related Diseases at a Primary Health Care Facility within a Sub-Saharan African City: Before and during the COVID-19 Pandemic.

BACKGROUND: With the advent of the COVID-19 pandemic caused by SARS-CoV-2, protocols such as social distancing and upscaling of hygiene practices were implemented to limit the spread of the disease. Meanwhile, along with COVID-19 came stress due to restrictions on movement, trade and transport, and closure of schools, among others. AIM: This study compared the prevalence of hygiene-related gastrointestinal infections and stress-related diseases before (March 2019-February 2020) and during (March 2020-February 2021) the COVID-19 pandemic. METHODOLOGY: This was a retrospective single-center review of deidentified patient data from the Korle Bu Polyclinic, Accra, Ghana. RESULTS: Comparing the pre-COVID-19 era to the COVID-19 era, there was a statistically nonsignificant change in the number of cases and prevalence of gastroenteritis and enteric fever (p = 0.084 and 0.081, respectively), although for gastroenteritis, the prevalence was higher for the pre-COVID-19 era compared to during COVID-19 by 1.8 per 1000 cases, while that of enteric fever was higher during the COVID-19 era compared to the pre-COVID-19 era by 1.0 per 1000 cases. Of the stress-related diseases, statistically significant increases in the prevalence of anxiety disorders (p = 0.028), insomnia (p = 0.001), and headache (p = 0.010), were noted, with 2.3, 5.5, and 2.4 per 1000 cases, respectively. There were more female cases than male cases recorded for depression (p = 0.001), headache (p = 0.010), and hypertension (p = 0.001) during the pandemic, and these were statistically significant. CONCLUSION: During the pandemic, a significant increase in the prevalence of stress-related diseases was observed. However, a statistically nonsignificant change was recorded for gastrointestinal infections, with females reporting more of these disorders. Consequently, it is important to strengthen the capacity for managing stress-related conditions alongside diseases that cause pandemics when they arise.

Methicillin-resistant Staphylococcus aureus (MRSA) nasal carriage among patients with diabetes at the Korle Bu Teaching Hospital.

AIM: To investigate the epidemiology of S. aureus and MRSA nasal carriage among people with diabetes at the Korle Bu Teaching Hospital in Accra, including the prevalence, predictors of carriage, and antibiotic resistance. METHODOLOGY: This study was cross-sectional, involving 300 diabetes patients and 106 non-diabetic individuals. Swab specimens of the nares were obtained from the participants and bacteriologically-cultured. Identification and characterization of S. aureus and MRSA were based on standard bacteriological methods; antimicrobial susceptibility testing was by the Kirby-Bauer method. RESULTS: The prevalence of staphylococcal carriage, the diabetes group relative to the non-diabetes group, were 31.0% and 10.4% (S. aureus), and 3.3% and 0.0% (MRSA). Presence of diabetes predisposed to S. aureus carriage, but not MRSA nor coagulase-negative staphylococci (CoNS) carriage (OR = 3.88; p < 0.0001). Colonization with CoNS was protective of S. aureus (OR = 0.039, p < 0.001) and MRSA (OR = 0.115, p = 0.043) colonization among the diabetics. The antimicrobial resistance patterns recorded among the S. aureus isolated from the diabetic individuals relative to the non-diabetics were as follows: penicillin (95% vs. 91%), tetracycline (37% vs. 27%), cotrimoxazole (30% vs. 36%), erythromycin (17% vs. 0%), norfloxacin (13% vs. 0%), clindamycin (12% vs. 0%), gentamicin (9% vs. 0%), fusidic acid (10% vs. 9%), linezolid (4% vs. 0%), and rifampicin (5% vs. 0%). The proportion of multidrug resistant S. aureus was 41% (n = 38) in the diabetes group and 0% in the non-diabetes group; this difference was statistically significant (p = 0.01). CONCLUSIONS: The presence of diabetes predisposed the participants to S. aureus carriage by almost four folds, but not MRSA carriage. Colonization with CoNS was protective of S. aureus and MRSA carriage in the diabetes group. Finally, linezolid remains a good therapeutic agent for anti-MRSA therapy.

Methicillin-Resistant Staphylococcus aureus (MRSA) Infection of Diabetic Foot Ulcers at a Tertiary Care Hospital in Accra, Ghana.

AIM: This study investigated the spectrum of bacteria infecting the ulcers of individuals with diabetes at the Korle Bu Teaching Hospital in Accra, Ghana, focusing on Staphylococcus aureus (S. aureus) and methicillin-resistant S. aureus (MRSA), with respect to their prevalence, factors predisposing to their infection of the ulcers, and antimicrobial resistance patterns. METHODOLOGY: This cross-sectional study was conducted at The Ulcer Clinic, Department of Surgery, Korle Bu Teaching Hospital, involving 100 diabetic foot ulcer patients. The ulcer of each study participant was swabbed and cultured bacteriologically, following standard procedures. Antimicrobial susceptibility testing was done for all S. aureus isolated, using the Kirby-Bauer method. RESULTS: In total, 96% of the participants had their ulcers infected-32.3% (n = 31) of these had their ulcers infected with one bacterium, 47.9% (n = 46) with two bacteria, 18.8% (n = 18) with three bacteria, and 1.0% (n = 1) with four bacteria. The prevalence of S. aureus and MRSA were 19% and 6%, respectively. The distribution of the other bacteria was as follows: coagulase-negative Staphylococci (CoNS) (54%), Escherichia coli (24%), Pseudomonas spp. (19%), Citrobacter koseri and Morganella morgana (12% each), Klebsiella oxytoca (11%), Proteus vulgaris (8%), Enterococcus spp. (6%), Klebsiella pneumoniae (5%), Proteus mirabilis and Enterobacter spp. (4%), Klebsiella spp. (2%), and Streptococcus spp. (1%). The resistance rates of S. aureus decreased across penicillin (100%, n = 19), tetracycline (47.4%, n = 9), cotrimoxazole (42.1%, n = 8), cefoxitin (31.6%, n = 6), erythromycin and clindamycin (26.3% each, n = 5), norfloxacin and gentamicin (15.8% each, n = 3), rifampicin (10.5%, n = 2), linezolid (5.3%, n = 1), and fusidic acid (0.0%, n = 0). The proportion of multidrug resistance was 47.4% (n = 9). Except for foot ulcer infection with coagulase-negative Staphylococci, which was protective of S. aureus infection of the ulcers (OR = 0.029, p = 0.001, 95% CI = 0.004-0.231), no predictor of S. aureus, MRSA, or polymicrobial ulcer infection was identified. CONCLUSIONS: The prevalence of S. aureus and MRSA infection of the diabetic foot ulcers were high, but lower than those of the predominant infector, coagulase-negative Staphylococci and the next highest infecting agent, E. coli. Diabetic foot ulcers' infection with coagulase-negative Staphylococci protected against their infection with S. aureus. The prevalence of multidrug resistance was high, highlighting the need to further intensify antimicrobial stewardship programmes.

Nucleoside Analogues Are Potent Inducers of Pol V-mediated Mutagenesis.

Drugs targeting DNA and RNA in mammalian cells or viruses can also affect bacteria present in the host and thereby induce the bacterial SOS system. This has the potential to increase mutagenesis and the development of antimicrobial resistance (AMR). Here, we have examined nucleoside analogues (NAs) commonly used in anti-viral and anti-cancer therapies for potential effects on mutagenesis in Escherichia coli, using the rifampicin mutagenicity assay. To further explore the mode of action of the NAs, we applied E. coli deletion mutants, a peptide inhibiting Pol V (APIM-peptide) and metabolome and proteome analyses. Five out of the thirteen NAs examined, including three nucleoside reverse transcriptase inhibitors (NRTIs) and two anti-cancer drugs, increased the mutation frequency in E. coli by more than 25-fold at doses that were within reported plasma concentration range (Pl.CR), but that did not affect bacterial growth. We show that the SOS response is induced and that the increase in mutation frequency is mediated by the TLS polymerase Pol V. Quantitative mass spectrometry-based metabolite profiling did not reveal large changes in nucleoside phosphate or other central carbon metabolite pools, which suggests that the SOS induction is an effect of increased replicative stress. Our results suggest that NAs/NRTIs can contribute to the development of AMR and that drugs inhibiting Pol V can reverse this mutagenesis.