Adebayo Lamikanra

Heterogeneous Virulence of Enteroaggregative Escherichia coli Strains Isolated from Children in Southwest Nigeria

Enteroaggregative Escherichia coli (EAEC) has been implicated in acute and persistent diarrhea, and most strains harbor a member of a partially-conserved plasmid family (called pAA). We studied EAEC isolated from Nigerian children aged <5 years to elucidate the roles of plasmid and chromosomal EAEC loci. We tested a total of 131 EAEC strains isolated from acute diarrhea case patients and control subjects for hybridization with 8 pAA plasmid-derived and 2 chromosomal gene probes, for several in vitro phenotypes and for resistance to antimicrobial agents. Using by multiple logistic regression, we found genes encoding the AAF/II fimbriae to be strongly associated with diarrhea in this population. EAEC strains appear to be of heterogeneous virulence, and data suggest that AAF/II may be a marker for pathogenic strains.

Etiology of Acute Diarrhea in Adults in Southwestern Nigeria

ABSTRACT Stool specimens from 113 adult outpatients with diarrhea in southwestern Nigeria and 63 controls were examined for bacterial and parasitic enteric pathogens. Enterohemorrhagic Escherichia coli (EHEC) (P < 0.02), enteroaggregative E. coli (EAEC) (P < 0.02), and Entamoeba histolytica (P < 0.0002) were significantly associated with diarrhea. Salmonella, Shigella, nontoxigenic Vibrio cholerae, other categories of diarrheagenic E. coli, as well as a variety of helminths were recovered more frequently from the stools of patients than from the stools of controls but did not show a significant association with disease. Multiple pathogens were recovered from 36.3% of specimens, and bloody diarrhea was commonly associated with E. histolytica and diarrheagenic E. coli infections. The majority of EHEC isolates were non-O157 strains that carried the stx2 gene. Of the 23 EHEC-infected patients, 12 (52.2%) presented during the 10th week of the study. EHEC strains isolated within this cluster were more likely to hybridize with the enterohemolysin gene probe, to be nonmotile and sorbitol positive, and to fail to agglutinate O157 antisera. Pulsed-field gel electrophoresis demonstrated that the only strains with XbaI profiles that occurred more than once were isolated during the 10th and 11th weeks of the study, suggesting an outbreak. The study has demonstrated that E. histolytica, EHEC, and EAEC are important diarrheal pathogens within the study area and that sporadic and epidemic EHEC infections occur in developing as well as developed countries. Routine surveillance for diarrheagenic E. coli, even only at the tertiary-care level, would be useful in identifying outbreaks and assist in identifying environmental reservoirs and transmission routes.

Antimicrobial Resistance in Escherichia coli Strains From Urinary Tract Infections

BACKGROUND An increase in resistance against many different drugs among urinary tract infection (UTI) E coli isolates has been observed in the last 2 decades. This study determined the trends of antimicrobial resistance in E coli to commonly used antibiotics. METHODS The study was conducted in Ile-Ife, southwest Nigeria. Patients with features suggestive of UTI were investigated for presence of significant bacteriuria. Urine isolates were identified. Antimicrobial susceptibility was evaluated in accordance with standard bacteriological methods. RESULTS Of 442 urine specimens, 158 (35.8%) yielded significant growth, including 41 (25.6%) with E coli. Among the E coil isolates, antimicrobial susceptibility varied in prevalence by agent in descending order as follows: nitrofurantoin (80%), ofloxacin (24%), ciprofloxacin (15%), nalidixic acid (10%), cotrimoxazole (5%), and amoxicillin/clavulanic acid (2%). No isolate was susceptible to amoxicillin, gentamicin, or tetracycline. All were also found to be resistant to at least 3 commonly used drugs. All 25 isolates tested for extended-spectrum beta-lactamase (ESBC) production were found to be presumptive ESBCs producers. CONCLUSION The results demonstrate the continued susceptibility of E coil to nitrofurantoin and their widespread and increasing resistance to amoxicillin, gentamicin, cotrimoxazole, ciprofloxacin, ofloxacin, and tetracycline. Nitrofurantoin is a--and, in this locale, perhaps the only--rational drug for empiric treatment of uncomplicated UTI. There is a need for a comprehensive study of the involvement of ESBC-producing E coli in UTI in this environment.

Antimicrobial Spectrum of Alchornea cordifolia Leaf Extract

The 50% aqueous ethanol extract of Alchornea cordifolia (Schum and Thonn) Muell. Arg. leaf was screened for activity against 74 microbial strains representing aerobic, facultative and anaerobic bacteria as well as fungi. The panel of test strains included organisms from culture collections as well as clinical and environmental isolates. A concentration of 5 mg/mL of extract inhibited 36.5% of the isolates and 95.9% were inhibited by a concentration of 20 mg/mL. Only three strains, all filamentous fungi, were not susceptible to 40 mg/mL of the extract, the highest concentration tested. The extract showed the best activity against gram‐positive bacteria and yeasts with inhibitory concentrations against these organisms being under 5 mg/mL. The results demonstrate that the A. cordifolia extract has a very broad spectrum of activity and suggests that it may be useful in the treatment of various microbial infections. Copyright

Regional dissemination of a trimethoprim-resistance gene cassette via a successful transposable element.

Background Antimicrobial resistance is a growing international problem. We observed a 50% increase in the prevalence of trimethoprim resistance among fecal Escherichia coli from healthy Nigerian students between 1998 and 2005, a trend to increase that continued in 2009. Methods and Findings A PCR-based screen revealed that 131 (43.1%) of isolates obtained in Nigeria in 2005 and 2009 carried integron-borne dfrA cassettes. In the case of 67 (51.1%) of these isolates, the cassette was a class 1-integron-borne dfrA7 gene, which has been reported at high prevalence from E. coli isolates from other parts of Africa. Complete sequencing of a 27 Kb dfrA7-bearing plasmid from one isolate located the dfrA7 gene within a Tn21-type transposon. The transposon also contained an IS26-derived bla/sul/str element, encoding resistance to β-lactams, sulphonamides and streptomycin, and mercury resistance genes. Although the plasmid backbone was only found in 12 (5.8%) of trimethoprim-resistant isolates, dfrA7 and other transposon-borne genes were detected in 14 (16.3%) and 32 (26.3%) of trimethoprim resistant isolates collected in Nigeria in 2005 and 2009, respectively. Additionally, 37 (19.3%) of trimethoprim-resistant E. coli isolates collected between 2006 and 2008 from Ghana were positive for the dfrA7 and a transposon marker, but only 4 (2.1%) harbored the plasmid backbone. Conclusions Our data point to transposition as a principal mechanism for disseminating dfrA7 among E. coli from Nigeria and Ghana. On-going intensive use of the affordable broad-spectrum antibacterials is likely to promote selective success of a highly prevalent transposable element in West Africa.

Antibiotic-resistant cell-detaching Escherichia coli strains from Nigerian children.

ABSTRACT The properties of 23 cell-detaching Escherichia coli strains that were isolated from stool specimens in Nigeria are described. Common properties of the strains included the presence of genes encoding α-hemolysin (100%), pyelonephritis-associated pili (100%), and cytotoxic necrotizing factor 1 (70%) as well as lactose negativity (70%) and multiple antibiotic resistance (74%). Antibiotic resistance was shown in most cases to be transferable and associated with the presence of class 1 integrons. Phenotypic properties and pulsed-field gel electrophoresis analysis demonstrated that the majority of the strains, particularly multiply resistant, lactose-negative O4:H40 strains, were closely related. Multiply-resistant cell-detaching E. coli strains may represent an important reservoir for antibiotic resistance genes.

Rapid evolution of fluoroquinolone-resistant Escherichia coli in Nigeria is temporally associated with fluoroquinolone use

BackgroundAntibiotic resistance has necessitated fluoroquinolone use but little is known about the selective forces and resistance trajectory in malaria-endemic settings, where selection from the antimalarial chloroquine for fluoroquinolone-resistant bacteria has been proposed.MethodsAntimicrobial resistance was studied in fecal Escherichia coli isolates in a Nigerian community. Quinolone-resistance determining regions of gyrA and parC were sequenced in nalidixic acid resistant strains and horizontally-transmitted quinolone-resistance genes were sought by PCR. Antimicrobial prescription practices were compared with antimicrobial resistance rates over a period spanning three decades.ResultsBefore 2005, quinolone resistance was limited to low-level nalixidic acid resistance in fewer than 4% of E. coli isolates. In 2005, the proportion of isolates demonstrating low-level quinolone resistance due to elevated efflux increased and high-level quinolone resistance and resistance to the fluoroquinolones appeared. Fluoroquinolone resistance was attributable to single nucleotide polymorphisms in quinolone target genes gyrA and/or parC. By 2009, 35 (34.5%) of isolates were quinolone non-susceptible with nine carrying gyrA and parC SNPs and six bearing identical qnrS1 alleles. The antimalarial chloroquine was heavily used throughout the entire period but E. coli with quinolone-specific resistance mechanisms were only detected in the final half decade, immediately following the introduction of the fluoroquinolone antibacterial ciprofloxacin.ConclusionsFluoroquinolones, and not chloroquine, appear to be the selective force for fluoroquinolone-resistant fecal E. coli in this setting. Rapid evolution to resistance following fluoroquinolone introduction points the need to implement resistant containment strategies when new antibacterials are introduced into resource-poor settings with high infectious disease burdens.

Diversity of antibiotic resistance genes and staphylococcal cassette chromosome mec elements in faecal isolates of coagulase-negative staphylococci from Nigeria

BackgroundCoagulase-negative staphylococci (CoNS) are opportunistic pathogens found as colonisers of the human gut. This study was carried out to examine the genetic resistance mechanisms in faecal isolates of CoNS. The study investigated 53 non-duplicate CoNS isolates obtained from the fresh stool samples of apparently healthy subjects in the community of Ile-Ife, South-Western Nigeria. Antibiotic susceptibility testing was assessed by the disc diffusion test while antibiotic resistance genes were analysed by PCR. mecA positive isolates were analysed by Staphylococcal Chromosome Cassette mec (SCCmec) and cassette chromosome recombinase (ccr) complex typing methods.ResultsResistance genes were detected only in isolates that showed resistance by phenotypic screening. The aac(6′)–aph(2″) gene was detected in all the three isolates resistant to gentamicin. Four of the five erythromycin resistant isolates were positive for the ermC gene, the remaining isolate carried the msrA gene. The tetK gene was detected in 6 of the 7 tetracycline resistant isolates while 4 possessed the tetM gene. Three of the isolates (S. haemolyticus, S. xylosus and S. capitis) had both genes. Several SCCmec types were found: SCCmec I- ccrAB β2-α2 (4 isolates: 3 S. epidermidis, 1 S. warneri), SCCmec IVb- ccrAB β2-α3 (1 isolate: S. epidermidis), SCCmec IVd- ccrAB β2-α3 (8 isolates: 3 S. epidermidis, 2 S. xylosus, 1 S. saprophyticus, 1 S. warneri, 1 S. capitis), and untypable (2 isolates: S. epidermidis).ConclusionThis genetic background could be a reservoir for interspecies gene transfer among CoNS and S. aureus in the intestinal tract.

A study of the prevalence of diarrhoeagenic Escherichia coli in children from Gwagwalada, Federal Capital Territory, Nigeria.

Introduction Diarrhoeagenic Escherichia coli (DEC) are major causes of diarrhoea in Nigeria. This study was conducted to determine the prevalence of diarrhoea caused by DEC within the Federal Capital Territory, Abuja, Nigeria. Methods A total of 730 rectal swabs obtained from 201 children with diarrhoea and 529 healthy controls aged 0-24 months were cultured for the isolation of Escherichia coli. All E. coli isolates were investigated by PCR to determine their pathotype. Results A total of 61 DEC strains were recovered at a rate of 18.4% and 2.6% from children with diarrhoea and healthy controls respectively. The DEC strains recovered were Enteroaggregative Escherichia coli (34.4%), Shiga-toxin producing Escherichia coli (31.1%), Enterotoxigenic Escherichia coli(18.0%), typical enteropathogenic Escherichia coli (15.0%) and Enteroinvasive Escherichia coli (1.6%). Shiga-toxin producing Escherichia coli andEnteroinvasive Escherichia coli were recovered only from children suffering from diarrhoea and the overall prevalence of DEC strains was significantly higher among the children with diarrhoea (P < 0.0001). The number of DEC strains obtained during the dry season was significantly higher (P = 0.012) than the number obtained in the rainy season. Conclusion Diarrhoea caused by E. coli in the Nigerian children studied is associated with several diarrhoeagenic pathotypes and a significant proportion of the healthy children were found to harbour EAEC and ETEC strains. These asymptomatic carriers may be regarded as potential transmitters of infection to vulnerable children in the study area.

Dissemination of the Transmissible Quinolone-Resistance Gene qnrS1 by IncX Plasmids in Nigeria

The plasmid-encoded quinolone resistance gene qnrS1 was recently found to be commonly associated with ciprofloxacin resistance in Nigeria. We mapped the qnrS1 gene from an Escherichia coli isolate obtained in Nigeria to a 43.5 Kb IncX2 plasmid. The plasmid, pEBG1, was sufficient to confer ciprofloxacin non-susceptibility, as well as tetracycline and trimethoprim resistance, on E. coli K-12. Deletion analysis confirmed that qnrS1 accounted for all the ciprofloxacin non-suceptibility conferred by pEBG1 and tetracycline and trimethoprim resistance could be attributed to tetAR and dfrA14 genes respectively. While it contained a complete IncX conjugation system, pEBG1 was not self-transmissible likely due to an IS3 element inserted between the pilX5 and pilX6 genes. The plasmid was however efficiently mobilizable. pEBG1 was most similar to another qnrS1-bearing IncX2 plasmid from Nigeria, but both plasmids acquired qnrS1 independently and differ in their content of other resistance genes. Screening qnrS1–positive isolates from other individuals in Nigeria revealed that they carried neither pEBG1 nor pNGX2-QnrS1 but that IncX plasmids were prevalent. This study demonstrates that the IncX backbone is a flexible platform that has contributed to qnrS1 dissemination in Nigeria.