Occurrence and Diversity of Biofilm Producing Multidrug Resistant Escherichia coli in Dug Wells Used for Domestic Purposes in Ile-Ife, Southwest Nigeria

Antibiotic resistance in Escherichia coli has emerged as a serious and growing threat to public and environmental health and domestic wells have been fingered as a potential reservoir of resistant E. coli due to faecal contamination, and are ideal for microbial biofilm formation. Therefore, this study was carried out to investigate biofilm production among 60 multidrug-resistant (MDR) E. coli recovered from domestic wells in Ile-Ife, Nigeria. All biofilm positive isolates were screened for seven associated (papC, iroN, biofilm, fimH, cna, fbna and bcsa) genes by multiplex polymerase chain reaction (PCR). The diversity of the isolates was investigated using (GTG)5-PCR, and a dendrogram was drawn with interactive-tree-of-life software. Overall, 45 of 60 (75%) isolates produced biofilms, comprising strong (n=15; 25%), moderate (n=16; 26.7%), and weak (n=14; 23.3%) producers. Moreover, 35 (77.8%) isolates had at least one biofilm gene, 30 (66.7%) had multiple genes, and 5 (11.1%) had one. The biofilm gene (biofilm, fimH, bcsa, iroN) combination was identified in 10 (28.7%) of the 60 isolates. The isolates (n=14; 40%) with moderate biofilm production had the highest number of genes dominated by the biofilm gene (biofilm, fimH, bcsa) combination, while isolates (n=8; 22.9%) with weak production had the least number of genes and comprised mainly the fimH and cna gene combination. The biofilm formers showed 32 distinct fingerprints and were grouped into nine clades, with clade 6 having the most strains (31.6%). The occurrence of MDR biofilm-producing organisms in domestic wells constitutes a public health concern. There is a need to develop strategies to curb faecal contamination of wells to prevent outbreaks of untreatable diseases.

Water quality and antimicrobial resistant profile of Escherichia coli isolated from dug wells used for domestic purposes

The rapid spread of antimicrobial resistance (AMR) poses a significant threat to public safety. The use of water containing resistant bacteria could increase the risk of spreading AMR. This study assessed the quality of 143 dug wells used for domestic purposes in some communities in Nigeria and determined the resistance profile of isolated Escherichia coli (E. coli). The MicrobactTM identification kit was used to identify the isolates, and the susceptibility profile was evaluated using the Kirby-Bauer disc diffusion method. The combination disc technique was used to test all isolates for extended spectrum beta lactamase (ESBL) production. Polymerase chain reaction was used to identify ESBL genes, Integrons, and plasmid-mediated quinolone resistance genes. A total of 110 (76.9%) wells were contaminated by coliform bacteria. Of these, 94 (84.45%) wells yielded 202 E. coli strains. The isolates were commonly resistant to ampicillin (60.9%) but were all susceptible to meropenem. Seventy-seven (38.1%) isolates were multi-drug resistant. Two isolates harbored blaCTX-M and blaTEM separately while four (19%) ciprofloxacin-resistant isolates carried the oqxAB/aac-lb-cr gene. All isolates with resistance genes harbored class 1 and/2 Integrons. Most wells had coliform counts far above the World Health Organization’s recommended limit, indicating that they are unsafe to drink. The presence of multidrug-resistant isolates in well water poses a serious risk to consumers since it might lead to outbreaks of untreatable water-borne diseases

Nasal carriage of multi-drug resistant panton valentine leukocidin positive Staphylococcus aureus in healthy individuals of Tudun-Wada, Gombe State, Nigeria

Background: Panton-Valentine Leucocidin (PVL)-producing Staphylococcus aureus strains have been implicated in serious community-associated invasive infections and their increasing multidrug resistance is a major global health concern. Thus, we investigated the prevalence of the PVL gene and the antimicrobial resistance profile of nasal S.aureus isolates from healthy adults in Tundu-Wada, Gombe State of Nigeria.Methods and Materials: A total of 262 nasal samples from healthy adults were obtained and cultured. The isolates were identified as S. aureus by standard morphological and biochemical methods alongside with the Polymerase Chain Reaction (PCR) amplification of their 16S rRNA gene. Antimicrobial susceptibility testing was performed by the disc diffusion technique and the presence of mecA and PVL genes was determined by PCR analysis.Results: The overall nasal colonization of S. aureus was 17.6%. The prevalence of haemolysin and biofilm production among the isolates was 25(54.3%) and 42(91.3%), respectively. Only 2(4.3%) and 5(10.9%) possessed mecA and PVL genes respectively but none of the isolates harboured these two genes. All the isolates were resistant to amoxicillin but were highly susceptible (93.7%) to gentamicin. The prevalence of multi-drug resistance (MDR) among the isolates was M 45.7% and all PVL-producing isolates were MDR while one of the isolates with mecA gene exhibited extensive-drug resistance (XDR).Conclusion: This is the first report of nasal colonization of MDR PVL-producing S. aureus in healthy adults in Gombe, Northeastern Nigeria. This study highlights the importance of routine surveillance of healthy populations to provide useful strategies for controlling the spread of virulent multidrug-resistant organisms within the community

Prevalence and risk factors for extended-spectrum β-lactamaseproducing Gram-negative bacterial infections in hospitalized patients at a tertiary care hospital, southwest Nigeria

Background: Clinical infections caused by extended-spectrum β-lactamase (ESBL)-producing bacteria constitute great burden to healthcare delivery with these resistant pathogens contributing largely to the magnitude and spread of antimicrobial resistance globally. Hence, knowledge of the risk factors for acquisition of infection caused by ESBL-producing bacteria is crucial to instituting prompt and appropriate treatment as well as prevention and control measures. This study investigated the risk factors associated with the prevalence of ESBL-producing Gram-negative bacteria (GNB) infections among hospitalized patients in Uniosun Teaching Hospital (UTH), Osogbo, Nigeria. Methodology: A total of 359 hospitalized patients with clinical infections from whose clinical samples we isolated non-duplicate GNB were consecutively recruited. GNB were isolated following aerobic cultures of appropriate clinical samples and MicrobactTMGNB 24E kit was used for species identification. All isolates were screened for ESBL production by the combination disc method. Relevant clinical and demographic information was obtained using a designed data collection form, and multivariate logistic regression analysis was used to identify associated risk factors. Results: Ninety-four (26.2%) of the 359 patients had ESBL-producing GNB isolated from their clinical samples, with a preponderance of Escherichia coli (26.6%, n=25/94), although the most frequent ESBL-producer was Stenotrophomonas maltophilia (100%, n=2/2) and least frequent was Pseudomonas aeruginosa (2.6%, n=1/39). The study indicated that male gender, age group >60 years and farming were socio-demographic factors associated with significantly higher prevalence of ESBL-producing GNB infection. Other independent risk factors significantly associated with high prevalence of ESBL GNB infections were; (i) admission into intensive care unit and male surgical ward, (ii) presence of invasive devices such as intravenous line, endotracheal tube and urinary catheter, (iii) underlying conditions such as diabetes mellitus and benign prostatic hyperplasia, and (iv) immunocompromised state. Conclusion: The information obtained from this study can serve as baseline data for designing strategy to prevent drug-resistant infections and transmission in our hospital.   French title: Facteurs de prévalence et de risque pour les infections de bactéries gram-négatives de la β-lactamase prolongées de la β-lactamase chez les patients hospitalisés dans un hôpital de soins tertiaires, au sud-ouest du Nigéria Contexte: Les infections cliniques causées par des bactéries de la β-lactamase de spectre prolongée (ESBL) constituent une grande charge à la livraison des soins de santé avec ces agents pathogènes résistants contribuant en grande partie à la magnitude et à la propagation de la résistance antimicrobienne mondiale. Par conséquent, la connaissance des facteurs de risque d'acquisition d'une infection causée par les bactéries produisant des ESBL est essentielle à l'institution de traitement rapide et approprié, ainsi que des mesures de prévention et de contrôle. Cette étude a enquêté sur les facteurs de risque associés à la prévalence des bactéries gram-négatives de l'ESBL (GNB) parmi les patients hospitalisés dans l'hôpital d'enseignement Uniosun (Uth), Osogbo, Nigéria. Méthodologie: Un total de 359 patients hospitalisés avec des infections cliniques de laquelle les échantillons cliniques de laquelle nous avons isolé le GNB non dupliqué ont été recrutés consécutivement. GNB ont été isolés à la suite de cultures aérobies d'échantillons cliniques appropriés et de kit MicroBactTM GNB 24E a été utilisé pour l'identification des espèces. Tous les isolats ont été criblés pour la production ESBL par la méthode des disques combinées. Des informations cliniques et démographiques pertinentes ont été obtenues à l'aide d'un formulaire de collecte de données conçu et une analyse de régression logistique multivariée a été utilisée pour identifier les facteurs de risque associés. Résultats: Quatre-vingt-quatorze (26,2%) des 359 patients avaient des GNB producteurs de BLSE isolés de leurs échantillons cliniques, avec une prépondérance d'Escherichia coli (26,6%, n=25/94), bien que le producteur de BLSE le plus fréquent soit Stenotrophomonas maltophilia (100.0%, n=2/2) et la moins fréquente était Pseudomonas aeruginosa (2,6%, n=1/39). L'étude a indiqué que le sexe masculin, le groupe d'âge > 60 ans et l'agriculture étaient des facteurs sociodémographiques associés à une prévalence significativement plus élevée d'infections à GNB productrices de BLSE. D'autres facteurs de risque indépendants significativement associés à une prévalence élevée d'infections à BLSE GNB étaient; (i) admission en unité de soins intensifs et en salle de chirurgie pour hommes, (ii) présence de dispositifs invasifs tels qu'une ligne intraveineuse, un tube endotrachéal et un cathéter urinaire, (iii) conditions sous-jacentes telles que le diabète sucré et l'hyperplasie bénigne de la prostate, et (iv) immunodéprimé Etat. Conclusion: les informations obtenues à partir de cette étude peuvent servir de données de base pour la conception de la stratégie visant à prévenir les infections et la transmission résistantes à la drogue dans notre hôpital

NASAL CARRIAGE OF MULTI-DRUG RESISTANT PANTON VALENTINE LEUKOCIDIN POSITIVE STAPHYLOCOCCUS AUREUS IN HEALTHY INDIVIDUALS OF TUDUN-WADA, GOMBE STATE, NIGERIA

Background: Panton-Valentine Leucocidin (PVL)-producing Staphylococcus aureus strains have been implicated in serious community-associated invasive infections and their increasing multidrug resistance is a major global health concern. Thus, we investigated the carriage of PVL gene and antimicrobial resistance profile of nasal S. aureus isolates from healthy adults in Tundu-Wada, Gombe State, Nigeria. Methods: A total of 262 nasal samples of healthy adults were obtained and cultured. The isolates were identified as S. aureus by standard conventional methods and Polymerase Chain Reaction (PCR) amplification of their 16S rRNA gene. Antimicrobial susceptibility testing was performed by the disc diffusion technique and the presence of mecA and PVL genes was determined by PCR analysis. Results: The overall nasal colonization of S. aureus was 17.6%. The prevalence of haemolysin and biofilm production among the isolates was 25 (54.3%) and 42 (91.3%) respectively. Only 2(4.3%) and 5(10.9%) possessed mecA and PVL genes respectively but none haboured these two genes. All the isolates were resistant to amoxicillin but were highly susceptible (93.7%) to gentamicin. The prevalence of multi-drug resistance (MDR) among the isolates was 45.7% and all PVL-producing isolates were MDR while one of the isolates with mecA gene exhibited extensively-drug resistance. The predominant antibiogram among MDR isolates was AML,SXT,E. Conclusion: This is the first report of nasal colonization of MDR PVL-producing S. aureus in healthy adults in Gombe, Northeastern Nigeria. It highlights the importance of routine surveillance of healthy populations to provide useful strategies for containing the spread of virulent multidrug-resistant organisms within the community

Asymptomatic bacteriuria in an apparently healthy population and its relation to hypertension

Background: Hypertension is a major health problem in sub-Sahara Africa. Several studies have suggested a role of asymptomatic bacteriuria (ASB) in the aetiology of hypertension, but there is a dearth of information on this association in Africa where the burden of hypertension is high. We investigated the prevalence of asymptomatic bacteriuria, its association with hypertension and determined the antibiotic resistance patterns of implicated bacterial isolates in an urban community of Ile-Ife.Methods: One hundred and seventy-four apparently healthy individuals were investigated for ASB. Relevant information was obtained from them with standard proforma. Their Blood pressure was measured with a standard mercury sphygmomanometer. All samples were processed on cysteine lactose electrolyte deficient medium and chocolate agar. Antimicrobial susceptibility testing was done using Kirby-Bauer disk diffusion technique.Results:Fifty (28.7%) individuals were positive for ASB. ASB was commonly detected among the female subjects (X2=5.619; p-value = 0.01777), and among individuals in the age group of 50 59 years. Those that were hypertensive were two and a half times more likely to have ASB (Odd ratio=2.5; p-value=0.01369; CI=1.19-5.35). The highest percentage of hypertensive female participants with ASB was found in the age group of 30-39 years (33.3%) while among the male participants, the highest percentage was found in the age group of 60-69 years (9.5%). Escherichia coli (n=13; 26%) and Staphylococcus aureus (n=13; 26%) were the commonest organisms implicated in ASB. The majority of the isolates (>90%) were multidrug resistant. Isolates of Escherichia coli were commonly resistant to ampicillin (83.3%), ceftriaxone (72.7%) and cefepime (66.7%). Isolates of Staphylococcus aureus were all (100%) resistant to erythromycin, cloxacillin and streptomycin. All isolates were least resistant to cotrimoxazole (<8%).Conclusion: Women as well as men in the age group of 50-59 years were more likely to develop ASB. ASB could be contributing to the rising incidence of hypertension in this environment. Co-occurrence of hypertension and ASB portends a grave problem for apparently healthy individuals in this environment.Keywords: Asymptomatic bacteriuria, Hypertension, Escherichia coli, healthy individuals, urban communit

Isolation and evaluation of Candida species and their association with CD4+ T cells counts in HIV patients with diarrhoea

Background: Gastrointestinal infection is one of the most common infections among HIV patients. Candida spp have been implicated in the aetiology of chronic diarrhoea in HIV patients, but little is known about this in Nigeria.Objectives: We determined the prevalence of faecal candidiasis in HIV patients in relation to diarrhoea, CD4 counts, and other socio-demographic factors and the spectrum of Candida isolates involved.Methods: One hundred and fifty four HIV patients were investigated. Candida species were identified by standard techniques. Socio-demographic and clinical information was obtained from the patients using a structured questionnaire. The CD4 count was estimated using a single platform flow cytometer.Results: Candida overgrowth was detected in 61 (39.5%) HIV patients, and diarrhoea was associated with candidiasis in the subjects (P=0.001). Candidiasis was commonly detected among subjects in the 29-39 years’ age group. A CD4 count below 200 cells/mm2 (62.3%) was a risk factor for acquiring candidiasis among HIV patients (P=0.001). Candida albicans (65.6%) was the most frequently recovered species followed by Candida krusei (16.4%) and Candida tropicalis (14.8%).Conclusion: Candidiasis is an important opportunistic infection in HIV-patients in Ile-Ife. There is need for regular checks for opportunistic infections, including candidiasis in HIV patients to monitor disease progression and prevent subsequent complications.Keywords: Candida species, CD4+ T cells counts, HIV, diarrhoe

COMPARATIVE EVALUATION OF GENOTYPING AND CULTURE-BASED TECHNIQUES FOR FUNGAL KERATITIS DETECTION

The study aims to compare the direct polymerase chain reaction with microbial culture for the detection and fungal pathogens in infectious keratitis.  A total of 81 corneal ulcers were culture and analyzed prospectively. PCR was performed with all corneal scrapping with fungal and bacteria specific primers. PCR products were analysed and compared with the culture results using standard methods. Of the 81 samples, 80 were positive by PCR, 51 for fungi and 29 for bacteria. Out of 51 PCR positive samples, 22 samples were culture positive and 29 were culture negative. The majority of PCR genotyped samples matched the positive culture results. The positive detection rate of 80/81 (98.8%) with high suspicion of fungal keratitis and positive detection rate of direct PCR 50/51(98.0%) were observed. The sensitivities for the diagnosis of fungal keratitis with direct PCR and culture were 98.0% (50/51) and 43.1% (22/51) (p< 0.001) whereas the specificities were 100.0% (2/2) and 100.0% (1/1) respectively. The time required to complete the direct PCR was only 3 hours. The direct PCR assay is a rapid diagnostic technique with high sensitivity and specificity for infectious keratitis and it is expected to have impact on the diagnosis and treatment of infectious keratitis. &nbsp

Isolation and evaluation of Candida species and their association with CD4+ T cells counts in HIV patients with diarrhoea.

Background: Gastrointestinal infection is one of the most common infections among HIV patients. Candida spp have been implicated in the aetiology of chronic diarrhoea in HIV patients, but little is known about this in Nigeria. Objectives: We determined the prevalence of faecal candidiasis in HIV patients in relation to diarrhoea, CD4 counts, and other socio-demographic factors and the spectrum of Candida isolates involved. Methods: One hundred and fifty four HIV patients were investigated. Candida species were identified by standard techniques. Socio-demographic and clinical information was obtained from the patients using a structured questionnaire. The CD4 count was estimated using a single platform flow cytometer. Results: Candida overgrowth was detected in 61 (39.5%) HIV patients, and diarrhoea was associated with candidiasis in the subjects (P=0.001). Candidiasis was commonly detected among subjects in the 29-39 years\u2019 age group. A CD4 count below 200 cells/mm2 (62.3%) was a risk factor for acquiring candidiasis among HIV patients (P=0.001). Candida albicans (65.6%) was the most frequently recovered species followed by Candida krusei (16.4%) and Candida tropicalis (14.8%). Conclusion: Candidiasis is an important opportunistic infection in HIV-patients in Ile-Ife. There is need for regular checks for opportunistic infections, including candidiasis in HIV patients to monitor disease progression and prevent subsequent complications

The profiles of packed cells volume, plasma electrolytes and glucose levels in malarial infected patients

Fifty patients (18 males, 32 females) with malaria infection and 50 apparently healthy control subjects (22 males, 28 females) were recruited for the study. Hematocrit level (PCV) was determined using Heco C haematology analyzer. Plasma electrolytes (Na+, K+, HCO3 -, Cl-), and glucose were respectively analyzed by SM23A Spectrophotometer, using TECO DIAGNOSTICS and RANDOX enzymatic glucose methods respectively. The results showed a significant reduction in the mean values of PCV (30.04±5.31%), Na+ (131.56±6.63 mmol/L), and glucose (85.92±13.85 mg/dL) in the malaria-infected subjects compared with the mean values of PCV (38.74±3.12%), Na+ (134.14±5.95 mmol/L), and glucose (92.40±13.99 mg/dL) (P<0.05) obtained from the control subjects. We observed higher significant mean values of K+ (3.93±0.79 mmol/L) and HCO3 - (23.56±2.55 mmol/L) in the malariainfected subjects compared with the control mean values of K+ (3.62±0.51mmol/L) and HCO3 - (23.48±2.02 mmol/L) (P<0.05). The mean values for chloride observed in the malaria-infected subjects, Cl- (99.52±7.44) was higher than the observed mean in the control subjects, Cl- (99.50±6.33), but was not statistically significant P>0.05. The mean (±standard deviation) of PCV, Na+, K+, HCO3 -, Cl- and glucose in malariainfected patients of different age groups were compared with the age-matched controls, and there were significant differences only in the age groups involving PCV and potassium in the 1-20 and >40 age brackets. This study has shown the importance of electrolyte management in patients with severe malaria to prevent attendant physiological failure during complications