Department of Medical Microbiology
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Item Covid-19 In Patients Presenting With Malaria-Like Symptoms At Korle Bu Polyclinic, Accra(University Of Ghana, 2022-08) Asamoah, I.Malaria is one of Ghana's most frequent illnesses and the most common cause of febrile sickness. The prevalence of malaria in patients who visit the Korle Bu Polyclinic in Accra with malaria-like symptoms is around 8%. It is therefore essential to look into COVID-19's contribution to malaria-like symptoms. Most infectious diseases including COVID-19 and arboviral infections mimic malaria due to the overlapping of non-specific symptoms they both share. This study investigated COVID-19 in patients presenting with malaria-like symptoms at the Korle Bu Polyclinic, Accra. A cross-sectional study was conducted among patients presenting with malaria-like symptoms at the Korle Bu polyclinic from June to August, 2021. A total of 300 patients who qualify for Malaria Rapid Diagnostic Testing, and met the criteria for the study, were selected using simple random sampling. Two to three millilitres of whole blood, nasopharyngeal and oropharyngeal swab samples was collected for screening of Plasmodium falciparum and SARS-CoV-2 respectively. The whole blood sample was also used for COVID-19 antibody test and full blood count. Sixty out of three hundred (20%) study patients were positive for SARS-CoV-2 using Real-time PCR and twenty six out of three hundred (8.7%) tested positive using COVID-19 Rapid Antigen test. Majority were females (42/60; 70%), and participants had mean age of 28 years. Seven out of three hundred (2.3%) tested positive for Plasmodium falciparum using Malaria RDT and microscopy and eight out of three hundred (2.7%) using Nested Conventional PCR. Two coinfection of Plasmodium falciparum and SARS-CoV-2 were reported. Most of the patients came from the Ablekuma South district. Most patients with CT’s below 30 (High viral load) presented with 5 or more symptoms (p value <0.01). The most common symptom experienced by the study patients at the polyclinic was headache (95%; 57/60) followed by general body weakness and pain (78.3%;47/60), tiredness (60%; 36/60) and fever ( 51.7%; 31/60) Comorbidities reported were hypertension, diabetes, Asthma, hypertension and diabetes and none had sickle cell diseases. Most of the study patients had been previously exposure to SARS CoV-2 (113/300) and 66.7% (34/51) Astrazeneca vaccinated patients had no antibody. Due to the synergy of symptoms, screening for COVID-19 in patients presenting with malaria-like symptoms is vital for immediate diagnosis and treatment. Keywords: COVID-19, Korle Bu, AccraItem In-Vitro Evaluation Of Antibacterial Properties Of Euphorbia Hirta Against Selected Multidrug-Resistant Bacteria In Ghana.(University of Ghana, 2022-06) Gyabeng, J.Background: Treatment of infections is an important area of public health concern as the prevalence of multidrug-resistant (MDR) bacteria is on the rise. MDR bacteria are associated with high morbidity and mortality worldwide. Medicinal plants including Euphorbia hirta have shown effectiveness in the treatment of infections and have been one area of interest worldwide for the treatment of diseases due to their high antimicrobial properties against MDR bacteria. In Ghana, the continuous spread of MDR bacteria has resulted in prolonged illness, increased healthcare costs and heightened fatalities which can suddenly cripple the country’s economy. One way to reduce the burden of MDR bacteria is to screen for new classes of antimicrobials from natural products and medicinal plants. Thus, this research aimed to evaluate the antimicrobial properties of E. hirta against selected MDR bacteria in Ghana. Methodology: Five solvents systems (methanol, distilled water, ethyl acetate petroleum ether and dichloromethane) with varying polarities were used to extract E. hirta via cold and Soxhlet extraction methods. The agar-well diffusion method was used to determine the antimicrobial activity of the various extracts against some selected MDR bacteria. Column chromatographic technique was used to the separate most potent crude extract into fractions and their antimicrobial activity were determined. Fractions that showed antimicrobial activity were further purified using column chromatography. Purified fractions were analyzed for the functional groups of compounds present using gas chromatography mass spectrometer (GCMS). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of crude ethyl acetate extracts and active fraction was determined. Results: Results from this study showed that, soxhlet maceration process had higher yield than cold maceration but the antimicrobial activity of extracts from both methods were the same. The University of Ghana http://ugspace.ug.edu.gh xvi Antimicrobial susceptibility test (AST) results revealed that K. pneumoniae isolates. recruited in the study were resistant to all extracts used. Furthermore, all test organisms were resistant to dichloromethane and petroleum ether extracts. Out of the 15 test organisms used, methanol and aqueous extracts were potent against 5 test organisms. Phytochemical analysis revealed the presence of phytoconstituents such as reducing sugars, phenolic compounds, saponins, flavonoids, anthracenosides and phytosterols. GC-MS analysis shows that 1,2,3-Benzenetriol is the probable sugars present in the active fraction. MIC and MBC results indicated that ethyl acetate extracts and the active fraction had the same MBC values with 3.13 mg/ml as their lowest MBC concentration. The MIC value recorded for crude ethyl acetate was between 50- 6.25 mg/ml whiles that of the active fraction (CF5) was between 50 -12.5 mg/ml. Conclusion; Polar extracts of whole E. hirta plant have antimicrobial activity with ethyl acetate extracts being the highest. The plant has the potential to be used as an antimicrobial agent. Therefore, in-vivo studies should be conducted using different polar solvents to extract the whole plant of E. hirta to exploit its antimicrobial activity in living thing.Item Bacterial Aetiology And Risk Factors Associated With Childhood Otitis Media In Accra, Ghana(University of Ghana, 2021-09) Bannah, V.Background: Otitis media (OM), also known as middle ear infection, has a high prevalence among young children. Young children have horizontally aligned Eustachian tubes (ET), smaller and shorter middle ear anatomical features and more frequent upper respiratory tract infections than adults. These make them more vulnerable to OM. The complications of OM include hearing loss, delayed speech development, impaired intellectual development, and societal challenges in later life. Nonetheless, there is a paucity of data regarding the bacterial aetiologies responsible for the condition and their associated antimicrobial susceptibility patterns, and little is known about the risk factors associated with OM. Aim: This study aimed to investigate the bacterial aetiologies and risk factors associated with otitis media among children in Accra, Ghana Materials and method: The research was a cross-sectional study in which children below 13 years old attending the ear, nose and throat clinics (ENT) of the Princess Marie Louise Children’s Hospital (PMLH), 37 Military Hospital and Mamprobi Hospital with suppurative otitis media were recruited during the study period. Following standard bacteriological methods, sterile ear swabs were used to take middle ear discharge from study participants for culture and antimicrobial susceptibility testing. A standard questionnaire was also used to collect data on socio-demographic characteristics, potential risk factors and clinical information. Results: Out of the 100 swabs from the 100 study subjects, 97 swabs gave positive yields, out of which 87.3% were pathogens. The most commonly-isolated pathogen was Pseudomonas aeruginosa (38.5%), followed by Klebsiella pneumoniae (19.8%) and Proteus mirabilis (11.5%). A high level of resistance was recorded for ampicillin, piperacillin-tazobactam, cefuroxime, amoxicillin-clavulanate, trimethoprim-sulfamethoxazole, tetracycline and ceftriaxone. Of the potential risk factors of ASOM and CSOM evaluated, only child history of ASOM showed a statistically significant association with the development of CSOM. Conclusion: Pseudomonas aeruginosa, Klebsiella pneumonia, and Proteus mirabilis were the common bacteria aetiologic agents of childhood OM. Poor responses to first-line antibiotics such as ampicillin and amoxicillin-clavulanate were reported, hence ciprofloxacin and gentamicin are recommended to be used in the management of OM in children. A history of ASOM was the risk factor identified to be associated with developing CSOM.Item Carriage Of Multi-Drug Resistant Enterobacterales And Acinetobacter Baumannii Among Hospitalised Paediatric Patients At The Child Health Department, Korle-Bu Teaching Hospital(University Of Ghana, 2022-08) Nelson, H.BACKGROUND: Infection and carriage of multidrug resistant (MDR) Enterobacterales and Acinetobacter baumannii are increasing globally and complicate the management of infections in children. Outbreaks of infections due to these MDR pathogens, particularly carbapenem-resistant Enterobacterales (CRE) and A. baumannii in hospitals are widespread and are a growing problem. Carriage of MDR pathogens is a precursor for invasive infections which are associated with high morbidity and mortality. This study determined the prevalence and epidemiology of MDR pathogens, with a focus on carbapenem-resistant Enterobacterales and A. baumannii among paediatric inpatients of the Korle-Bu Teaching Hospital. AIM: This study aimed at identifying the risk factors for carriage of carbapenem-resistant Enterobacterales and A. baumannii, and the molecular genotypes of carbapenemase-producing isolates, among paediatric inpatients at the Korle-Bu Teaching Hospital. METHOD: A cross-sectional study was conducted over 8 months period, from March to October 2021 at the Child Health Department, Korle-Bu Teaching Hospital. A systematic sampling method was used to recruit the participants. Relevant clinical data was extracted from participants’ medical records per a structured data collection instrument. Rectal swabs were collected from participants and inoculated onto MacConkey agar and incubated at 35-37°C for 18-24hrs. Different colonial morphotypes were identified by standard bacteriological techniques and confirmed with MALDI-TOF spectometry. Antimicrobial susceptibility testing was performed on all isolates. Carbapenem resistant isolates were screened for carbapenemase production using modified Hodge test. Multiplex polymerase chain reaction (PCR) and gel imaging techniques were used to evaluate the presence, and to characterise carbapenemase genes present. Frequency tables were used to University of Ghana http://ugspace.ug.edu.gh v summarize the prevalence and distribution of MDR organisms. Associations between risk factors and carriage of carbapenem resistant organisms were analysed using multinomial logistic regression. RESULTS: A total of 344 bacteria isolates; 331 Enterobacterales and 13 A. baumannii were isolated from rectal swabs of 299 paediatric inpatients ≤ 13years. The most common isolates were E. coli (60.5%, n = 208), K. pneumoniae (29.9%, n = 103) and A. baumannii (3.8%, n = 13). Prevalence of MDR among the isolated organisms were 75.6% (n = 260); E. coli (74.0%, n = 154), K. pneumoniae (76.7%, n = 79), and A. baumannii (100%, n = 13). Carriage of ESBL producing Enterobacterales was 72.6% (n = 217); with E. coli (46.8%, n = 140) and K. pneumoniae (25.1%, n = 75) being the most predominant ESBL phenotypes. Faecal carriage of carbapenem resistant bacteria was 23.1% (n = 69). E. coli (11%, n = 33), K. pneumoniae (7.4%, n = 22), A. baumannii (3.3%, n = 10) were the most common carbapenem resistant isolates. 52.2% (n = 36) of these carbapenem resistant isolates expressed phenotypic carbapenemase activity by the modified Hodge test (MHT). Thirty two (46.4%) were found to harbour at least one carbapenemase gene; blaOxa-48 (20.3%, n = 14), blaVIM (15.9%, n = 11), blaNDM (4.4%, n = 3), and blaIMP (5.8%, n = 4). Five (15.6%) harboured 2 carbapenemase genes, but none harboured 3 or more genes. Prior exposure to carbapenems and fluoroquinolones increased the odds of carriage of carbapenem-resistant Enterobacterales and A. baumannii by approximately two folds. CONCLUSION: This study reports high faecal carriage of MDR bacteria among paediatric inpatients of the Korle-Bu Teaching Hospital. This includes carbapenem-resistant Enterobacterales and A. baumannii with blaOxa-48 and blaVIM carbapenemase genes being the commonest. Prior antibiotic exposure to carbapenems and fluoroquinolones within the past year were significant risk factors for carriage of carbapenem-resistant isolates.Item Characterization Of Uropathogenic Escherichia Coli (Upec) In Hiv Seropositive Women With Asymptomatic Bacteriuria(University Of Ghana, 2022-08) Amegbletor, H.K.BACKGROUND: Asymptomatic bacteriuria (ASB), a precursor for Urinary tract infection (UTI), is high among People living with HIV (PLHIV). E. coli is the most implicated organism. An understanding of the risk factors for ASB, virulence genes and resistance profile of uropathogens among this population is important to the management and control. AIM: To characterize uropathogenic E. coli (UPEC) in HIV seropositive women with asymptomatic bacteriuria METHODOLOGY: This cross-sectional study was carried out at St. Martin de Pores Hospital, Eikwe in the Western Region of Ghana. A structured questionnaire was used to extract clinical information from the folders of 400 HIV seropositive women. The information included patient demographics, history of hospitalization, HAART treatment initiation date and WHO disease stage. Urine samples were obtained, cultured and identified with MALDI TOF biotyper. Antibiotic resistance pattern was determined, and genes coding for virulence and integrons were screened for using Multiplex PCR and gel imaging techniques. RESULTS: From the 400 samples cultured, 21.15% (85/400) were positive. The most prevalent organisms were; Escherichia coli 69.4% (59/85), Enterococcus faecalis 8.2% (7/85), Klebsiella pneumoniae 7.0% (6/85)), Proteus mirabilis 7.0% (6/85), Staphylococcus hemolyticus 3.5% (3/85). All uropathogenic E. coli isolates were resistant to ampicillin and 98.3% (58/59), resistant to trimethoprim sulfamethoxazole, followed by tetracycline 94.9% (56/59), cefuroxime 74.6% (45/59), amoxiclav 49.2% (30/59) and ciprofloxacin 32.2% (19/59). The lowest resistance was recorded to meropenem and fosfomycin at 1.7% and nitrofurantoin 6.8% (4/59). The commonest virulence genes observed were ChuA 66.1% (n=39/59), PapC 57.6% (n=34/59), cnf1 50.8% (n=30/59), kpsMTII 45.8% (n=27/59), iuAt 35.6% (n=21/59) and usp gene 8.5% (n=5/59). Two isolates (3.4%) harbored all 5 genes (iuAt, cnf1, papC, chuA, kpsMTII). Five isolates haboured iuAt, cnf1, papC and chuA (8.5%, n=5/59). Three genes (iuAt, cnf1, papC) were all observed in 6 isolates (10.2%, n=6/59) whilst 7 isolates (12%, n=7/59) were found to harbor genes iuAt and cnf1. The commonest integron was intI 42% (n=25/59) followed by intII 20% (n=12/59). Viral load [(OR=1.000, 95% CI, 1.000-1.000) (p = 0.295)], HAART duration [(OR=1.036, 95% CI, 0.287-1.042) (p=0.287)], age [(OR=1.020, 95% CI, 0.999-1.042) (p = 0.06)] and WHO disease stage [(OR=1.286, 95% CI, 0.535-1.6) (p = 0.885)] showed no significant association with the occurrence of asymptomatic bacteriuria (p≥0.05). HAART duration was observed to be a predictor of resistance to amoxiclav [(OR=1.329, 95% CI, 1.113-1.588) (p= 0.002)]. CONCLUSION: This study showed that asymptomatic bacteriuria is common among women living with HIV (WLHIV) visiting Eikwe district hospital, with no association with age, viral load, WHO disease stage and HAART duration. High prevalence of multidrug resistant UPEC coupled with the high carriage of virulence genes indicates that WLHIV are at a high risk of developing urinary tract infections with the potential for complications. There is a need for regular monitoring of bacteriuria and antibiotic susceptibility testing among this population.Item Phenotypic And Genotypic Detection Of Carbapenemase-Producing Escherichia Coli And Klebsiella Pneumoniae In Accra, Ghana(University Of Ghana, 2021-07) Dwomoh, F.P.Background: Escherichia coli and Klebsiella pneumoniae belong to the Enterobacteriaceae family, and are pathogens of high clinical significance. They are prone to acquiring antibiotic resistance genes, including those encoding carbapenemases, which confer resistance to carbapenems (the current drugs of last resort against infections with multidrug-resistant [MDR] Enterobacteriaceae). However, little is known about carbapenemase-producing E. coli and K. pneumoniae in Ghana. Aim: To investigate the occurrence of carbapenem resistance among MDR E. coli and K. 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 originating from 15 different sample types from the Central Microbiology Laboratory of the Korle Bu Teaching Hospital between the periods of December 2020 to March 2021. The isolates were re-cultured, subsequently 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 confirmed phenotypically with the Modified Hodge test (MHT) and Modified Carbapenem Inactivation Method (mCIM) and genotypically with multiplex PCR targeting blaKPC, blaIMP, blaNDM, blaVIM, and blaOXA-48. 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 were 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 were low. There is evidence that blaOXA-48 and blaNDM may be circulating in Ghana, warranting upscaling of antimicrobial resistance surveillance programmes and fortification of infection prevention and control programmes in the country.Item Antibiotic Residues And Multidrug-Resistant Gram-Negative Bacteria Contaminating Raw Meat Sold In Accra, Ghana(University Of Ghana, 2021-07) Baah, D.A.Background: Antimicrobial resistance (AMR) is considered by the World Health Organization as one of the greatest threats to health and economies in recent times. Multidrug resistance limits treatment options and has serious implications for human and animal health. Efforts to combat AMR should be based on the One Health approach and involve human health, animal health, and the environment. In Ghana, previous studies on AMR have given little attention to animal source food, which is a major route of transmission of antibiotic-resistant zoonotic pathogens. Aim: The aim of the study was to investigate the occurrence of antibiotic residues and multidrug-resistant bacteria in meat sold in Accra. Methods: This was a cross-sectional study in which 270 meat samples were collected. The presence of antibiotic residues in the meat samples was detected using microbiological inhibition assays. Standard microbiological methods were employed in cultural isolation and identification of bacterial pathogens present in the meat samples. Bacteria isolated from the samples were subjected to antimicrobial susceptibility testing (using the Kirby-Bauer method) against the following antimicrobials: amikacin (30 μg), ampicillin (10 μg), amoxicillinclavulanate (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: The prevalence of antibiotic residues among the meat samples was 7.7% [beef (0.0%), goat meat (0.0%), and chicken (23.3%, n = 21)]. Furthermore, thirty-two (32) different types of bacterial agents, totaling 588, were isolated from the samples. The predominant ones were Escherichia coli [262; Beef = 30.5%, n = 80; Goat meat = 30.5%, n = 80; Chicken = 38.9%, n = 102], Aeromonas hydrophila [117; Beef = 35.9%, n = 42; Goat meat = 53.0%, n = 62; Chicken = 11.1%, n = 13], Vibrio cholerae [20; Beef = 50.0%, n = 10; Goat meat = 50.0%, n = 10; Chicken = 0.0%, n = 0], Aeromonas veronii [19; Beef = 63.1%, n = 12; Goat meat = 36.8%, n = 7; Chicken = 0.0%, n = 0], and Klebsiella pneumoniae [18; Beef = 22.2%, n = 4; Goat meat = 16.7%, n = 3; Chicken = 61.1%, n = 11]. The prevalence of MDR among the contaminating bacteria was 14.9% (n = 83), and the distribution was beef (3.8%, n = 21), goat meat = (5.0%, n = 28), and chicken (6.1%, n = 34). Also, the MDR distribution among the predominant bacteria was E. coli (Overall = 18.7%, n = 49; Beef = 5.7%, n = 15; Goat meat = 5.7%, n = 15; Chicken = 7.3%, n = 19), A. hydrophila (Overall = 11.1%, n =13; Beef = 2.3%, n = 3; Goat meat = 7.7%, n = 9; Chicken = 0.9%, n = 1), V. cholerae and A. veronii (0.0% each), and K. pneumoniae (Overall = 5.6%, n = 1; Beef = 0.0%, n = 0; Goat meat = 0.0%, n = 0; Chicken = 5.6%, n = 1). Moreover, 2.0% (n = 11) of the contaminating bacteria were ESBL producers, all of which occurred in 11 of the chicken samples, and their distribution was: E. coli (1.3%, n = 7), K. pneumoniae, Pantoea spp., E. cloacae, and Serratia plymuthica (0.2% each, n = 1). Conclusion: The prevalence of antibiotic residues in the meat samples was low, and the occurrence was restricted to chicken. The major bacterial contaminants were E. coli, A. hydrophila, V. cholerae, A. veronii, and K. pneumoniae. The prevalence of multidrug resistance was moderate, while that of ESBL producers was low.Item Baseline Susceptibility Of Malaria Vectors To Clothianidin In The Northern Region Of Ghana(University Of Ghana, 2022-06) Zong, C.M.P.The emergence and spread of insecticide resistance coupled with the widespread distribution of malaria vectors in Ghana makes malaria control a cumbersome task. Currently in Ghana and most of the West African countries, malaria vectors have developed resistance to all four (4) classes of insecticides. Therefore, a new insecticide, clothianidin, which has a different mode of action, has been deployed for Indoor Residual spraying (IRS) in Ghana since March 2021. A detailed understanding of current phenotypic susceptibility to clothianidin used for IRS and the effect of this on mosquito mortality, as well as underlining resistance mechanisms is vital to inform management strategies. The aim of this study was to document the sub lethal effects of clothianidin on malaria vector susceptibility and longevity. This was a cross-sectional study. WHO susceptibility bioassays were conducted to determine phenotypic susceptibility of Anopheles gambiae complex to clothianidin whilst cone bioassays were conducted to determine the effect of exposure on mosquito mortality. Conventional PCR was done to discriminate the sibling species of the Anopheles gambiae complex and also to detect target-site mutations. WHO susceptibility bioassay results showed significantly effective but prolonged mortality of mosquitoes. Mortality rates recorded with the insecticide demonstrated a delayed effect with optimum mortality requiring up to 144hrs, six (6) days. The mortality rate for wild Anopheles mosquitoes exposed in WHO cone bioassays for 5-min, 24hrs after was 63%, showing resistance. Mortality increased to 97% after 120hrs, indicating possible resistance but after 144hrs, mortality of 100% was reached indicative of vector susceptibility. Mosquitoes exposed for 10-min had a mortality of 72% after 24hrs.This increased to 97% after 96hrs indicating possible resistance, but 120hrs was taken to reach full susceptibility of 100%. After 24hrs, mosquitoes exposed for 20-min reached 82% mortality, which increased to 97% after 96hrs indicative of possible resistance, this increased slightly to 98% after 120hrs showing susceptibility. A 95% mortality was reached for mosquitoes exposed for 30-min after 24hrs, thus giving an indication of suspected resistance, however, after 48hrs mortality reached was 98% suggesting susceptibility The PCR results revealed that Anopheles gambiae s. was the most abundant malaria vector accounting for 63% followed by Anopheles coluzzii also accounting for 25%, with Anopheles arabiensis accounting for 9% of the samples. There was high allele frequency of Kdr west (L1014F) in all vector species identified (0.89-0.94) with Ace-1 occurring in moderation (0.32-0.44). In this study, it was found that malaria vectors were susceptible to clothianidin. There were also a lot of resistant mutations in wild Anopheles mosquitoes. The most abundant malaria vector species was Anopheles gambiae s.s. Continued surveillance for susceptibility of malaria vectors to clothianidin should be conducted yearly to monitor possible resistance.Item Multidrug-Resistant Bacteria In Hospital Wastewater Of The Korle Bu Teaching Hospital(University Of Ghana, 2022-07) Addae-Nuku, D.S.Background: Antimicrobial resistance (AMR) is one of the top ten (10) public health threats. It is, therefore, imperative to scale up efforts aimed at tackling the AMR menace. One approach could involve expanding the range of AMR surveillance targets to include hospital wastewater (HWW), a target that has largely been overlooked by researchers. General Aim: The aim of this study was 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 a total of 288 hospital wastewater samples consecutively collected across twelve weeks from the pool of wastewater emanating from two critical care units of the Korle Bu Teaching hospital, that is, the Child Health Unit and the Maternity Unit on Mondays and Thursdays, each week. The samples were immediately transported on ice to the laboratory and 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). Escherichia coli was the only organism whose proportion significantly differed between the two units studied [Maternity Unit = 23.8%, n = 37; Child Health Unit = 38.0%, n = 53, p = 0.02], and it persisted for weeks 1 to 6 in the wastewater from the Maternity Unit and Weeks 6 to11 in the wastewater from the Child Health Unit, while occurring intermittently during the other periods. The prevalence of multidrug resistance among the isolates was 55.4% (n = 163) [Maternity Unit = 53.4%, n = 87; Child Health Unit = 46.6%, n = 76, p = 0.22]. Moreover, the prevalence of extended-spectrum beta lactamase (ESBL) producers was 15.6% (n = 46) [Maternity Unit = 18.4%, n = 26; Child Health Unit = 13.1%, n = 20, p = 0.21]. 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 harboured a wide range of multidrug resistant bacteria, with a good proportion of these being ESBL producers, and the predominant and persistent one being Escherichia coli. The study thus identifies the wastewater of KBTH as an important source of infection transmission, and underscores the significance of appropriate treatment of wastewater of the hospital and other clinical and related settings prior to its discharge.Item Anaptimicrobial Resistance And Genetic Diversity Of Sthylococcus Aureus From Surgical Site Infections At Two Hospitals In Accra(University of Ghana, 2019-07) Bentum, J.N.Background Surgical site infections (SSIs) are the most common healthcare-associated infections affecting surgical patients. Such infections are often caused by methicillin-susceptible as well as methicillin-resistant Staphylococcus aureus. Methicillin-resistant Staphylococcus aureus (MRSA) is resistant to the entire class of beta-lactam antimicrobials; which are largely used in clinical medicine. Patients infected with MRSAs therefore have limited therapeutic options, and this may lead to prolonged periods of hospitalisation and high heath care cost. In Ghana, information on SSI as well as the occurrence and prevalence of MRSA and MSSA from such infections are scarce. Data on bacteria species recovered from SSI is key for effective surveillance and selection of appropriate antimicrobial therapy. This study therefore, investigated the proportions of MRSA and MSSA using phenotypic and molecular detection tools among patients diagnosed of surgical site infections in two hospitals. Aim The aim of this study was to determine the antimicrobial resistance patterns and molecular characteristics of Staphylococcus aureus detected in patients with surgical site infections at the Korle-Bu Teaching Hospital and 37-Military Hospital in Accra. Method This was a hospital-based cross-sectional study conducted from June to November 2018 at the Korle-Bu Teaching Hospital (KBTH) and 37-Military Hospital in Accra. Surgical patients diagnosed of SSI were recruited using the Centers for Disease Control (CDC) case definition for surgical site infection. Patient demographic data (age, sex, type of operation etc.) and wound swabs or aspirates were collected after receiving an informed consent. S. aureus was identified using colonial morphology, coagulase testing and the Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF-MS). Antimicrobial susceptibility testing was performed using the Kirby-Bauer disk diffusion method; measured zone sizes were interpreted according the CLSI guidelines. Multiplex PCR was performed to detect mecA (methicillin-resistant gene), spa (S. aureus specific gene) and pvl (Panton Valentine Leukocidin toxin gene) in the S. aureus isolates. Libraries for illumina sequencing were prepared using the Nextera DNA Flex Library preparation kit. Whole genome sequencing was done with the MiSeq Illumina sequencer at Noguchi Memorial Institute for Medical Research (NMIMR). Genomes were assembled using an in-house pipeline; assembled sequences were then uploaded to the centre for genomic epidemiology website (http://www.genomicepidemiology.org/) to determine the spa types, sequence types and virulence gene content of the S. aureus isolates. Results A total of 110 patients were recruited into the study, 34 (12.5%) were male and 76 (69.1%) were female. Patients between the ages of 25-44 years were highest in the number among the patients enrolled. Overall, 13 S. aureus isolates (11.8%; 13/110) were recovered and all were resistant to penicillin and susceptible to gentamicin and vancomycin. Cefoxitin resistance (4/13; 30.77%) was detected only in isolates from 37-Military Hospital. On the other hand, tetracycline (46.15%; 6/13) and norfloxacin (15.38%; 2/13) resistance was recorded at both hospitals. Sensitivity of isolates to linezolid (84.62%; 11/13), clindamycin (76.92%; 10/13), rifampicin (92.31%; 12/13), co-trimoxazole (92.31%; 12/13) and erythromycin (53.85%; 7/13) was very high. The four (30.76; 4/13) isolates resistant to cefoxitin (MRSA) and were also positive for mecA by PCR. The predominant S. aureus genotype found in the study was ST152-t355. The four MRSAs detected belonged to ST152-t355 and ST5-t586 clone. Eight (61.53%; 8/13) isolates were positive for the Panton Valentine Leukocidin toxin. Twelve other virulence genes were detected with haemolysin A and B (hlgA and hlgB) being the most prevalent. Conclusion S. aureus isolates recovered were genetically diverse. The detection of ST152 MRSA among surgical patients is particularly of interest; this global clone has also been reported in Central Europe, the Balkan, Switzerland and Denmark as a community acquired MRSA. Continuous surveillance may be required to monitor the spread of these pandemic clones in the hospital setting. expressions and future work will look at evaluating the performance of these algorithms on a created database of Africans as well as employing these algorithms to detect anger in a persuasive space and persuade the individual from angry to another emotion for example happy. Keywords: anger, persuasive spaces, anger recognition, facial expression, facial expression recognition, facial expression algorithms, deep learning, machine learning, ensemble learning algorithm, SVM, CNN