Occurrence of virulence gene signatures associated with diarrhoeagenic and non-diarrhoeagenic pathovars of Escherichia coli isolates from some selected rivers in South-Western Nigeria.

BACKGROUND: Diarrhoeal diseases are attributable to unsafe water stemming from improper sanitation and hygiene and are reportedly responsible for extensive morbidity and mortality particularly among children in developed and developing countries. METHODS: Water samples from selected rivers in Osun State, South-Western Nigeria were collected and analyzed using standard procedures. Escherichia coli isolates (n=300) were screened for 10 virulence genes using polymerase chain reaction for pathotyping. RESULTS: While the virulence gene (VG) lt for enterotoxigenic E. coli had the highest prevalence of 45%, the enteropathogenic E. coli genes eae and bfp were detected in 6 and 4% of the isolates respectively. The VGs stx1 and stx2 specific for the enterohemorrhagic E. coli pathotypes were detected in 7 and 1% of the isolates respectively. Also, the VG eagg harboured by enteroaggregative pathotype and diffusely-adherent E. coli VG daaE were detected in 2 and 4% of the isolates respectively and enteroinvasive E. coli VG ipaH was not detected. In addition, the VGs papC for uropathogenic and ibeA for neonatal meningitis were frequently detected in 19 and 3% of isolates respectively. CONCLUSIONS: These findings reveal the presence of diarrhoeagenic and non-diarrhoeagenic E. coli in the selected rivers and a potential public health risk as the rivers are important resources for domestic, recreational and livelihood usage by their host communities.

Isolation of bacteria with plant growth-promoting properties from microalgae-bacterial flocs produced in high-rate oxidation ponds.

Exploring plant growth-promoting (PGP) bacterial activity of microbial components aggregated by wastewater treatment can reduce dependence on fossil fuel-derived fertilisers. This study describes the isolation and identification of bacteria from microalgae-bacteria flocs (MaB-flocs) generated in high-rate algal oxidation ponds (HRAOP) of an integrated algal pond system (IAPS) remediating municipal wastewater. Amplified 16S rRNA gene sequence analysis determined the molecular identity of the individual strains. Genetic relatedness to known PGP rhizobacteria in the NCBI GenBank database was by metagenomics. Isolated strains were screened for the production of indoles (measured as indole-3-acetic acid; IAA) and an ability to mineralise NH4+, PO43-, and K + . Of the twelve bacterial strains isolated from HRAOP MaB-flocs, four produced indoles, nine mineralised NH4+, seven solubilised P, and one K. Potential of isolated strains for PGP activity according to one-way ANOVA on ranks was: ECCN 7b > ECCN 4b > ECCN 6b > ECCN 3b = ECCN 10b > ECCN 1b = ECCN 5b > ECCN 8b > ECCN 2b > ECCN 12b > ECCN 9b = ECCN 11b. Further study revealed that cell-free filtrate from indole-producing cultures of Aeromonas strain ECCN 4b, Enterobacter strain ECCN 7b, and Arthrobacter strain ECCN 6b promoted mung bean adventitious root formation suggestive of the presence of auxin-like biological activity.

Prevalence of integrons in multidrug-resistant Escherichia coli isolates from waters and vegetables in Nsukka and Enugu, Southeast Nigeria.

Irrigation of fresh produce with poorly treated wastewater or contaminated freshwater sources can lead to produce contamination and foodborne illnesses, as well as the dissemination of antimicrobial resistance determinants. In this study, we assessed the presence of integrons in multidrug-resistant Escherichia coli isolated from the University of Nigeria, Nsukka Wastewater Treatment Plant effluent, tap water, vegetables from irrigated gardens and vegetables sold in selected markets from Nsukka and Enugu cities. E. coli was isolated following standard laboratory procedure and confirmed through beta-glucuronidase (uidA)-targeted polymerase chain reaction (PCR). The antibiotic resistance of the isolates was determined using Bauer-Kirby disk diffusion assay, and multiplex PCR was used to determine the presence of class 1 and 2 integrons. Our result revealed a total of 188 E. coli isolates from WWTP effluent (n = 41), tap water (n = 10) and vegetables from greenhouse (n = 46), farms (n = 55) and market (n = 36). Multidrug resistance was detected in all the isolates, ranging from three-drug resistance in a single isolate to 7-drug resistance patterns in two different isolates. Of the total isolates, class 1 integrons were abundantly detected in 175 (93.1%) and class 2 in 5 (2.7%). All the class 2 integrons were found in isolates that were positive for class 1. The abundance of multidrug-resistant E. coli harbouring class 1 integrons in the effluent and vegetable samples is a potential public health risk. Therefore, the appropriate measures for the safe use of poorly treated wastewater for vegetable farm irrigation are required to be put in place to reduce the microbial load of the discharged effluent. Also, education of farmers and the community on the dangers of wastewater effluent-grown plants and proper methods for cleaning harvested vegetable is recommended.

Treated wastewater used in fresh produce irrigation in Nsukka, Southeast Nigeria is a reservoir of enterotoxigenic and multidrug-resistant Escherichia coli.

BACKGROUND: Occurrences of pathogens in environmental and irrigation waters, as well as the use of inadequately treated sewage for fresh produce constitute potential public health threats worldwide. OBJECTIVE: To investigate the treated wastewater used in fresh produce irrigation in Nsuskka, Southeastern Nigeria, as a reservoir enterotoxigenic and multidrug-resistant Escherichia coli. METHODS: Treated wastewater (from the sewage treatment facility at Nsukka, Southeast Nigeria), soil and irrigated vegetable samples were collected and analyzed using standard procedures. Escherichia coli isolated from the samples were screened for the presence of enterotoxigenic E. coli strain encoding lt gene and profiled for antibiotic resistance using the conventional PCR and standardized agar disk diffusion assays respectively. RESULTS: Of the total presumptive 103 isolates, PCR detected uidA gene in 87 (84 %), of which 23 (26 %) harboured the lt encoding ETEC gene. Generally, imipenem, cefuroxime and norfloxacin proved to be most effective of all the antibiotics employed. Wastewater isolates were variously susceptible to ciprofloxacin (95 %), norfloxacin (95 %), cefuroxime (93 %), chloramphenicol (93 %), trimethoprim and tetracycline (88 %), soil isolates to streptomycin (75 %) and vegetable isolates to cefuroxime (90 %), norfloxacin (86 %), ciprofloxacin (81 %) and chloramphenicol. Contrariwise, high resistances observed to other antibiotics were in the order; ampicillin (95 %), penicillin (93 %), erythromycin (90 %) and clarithromycin (83 %) among wastewater isolates, ciprofloxacin and norfloxacin (75 %) in soil isolates; penicillin, vancomycin and erythromycin (98 %), rifampicin and clarithromycin (93 %), sulphamethoxazole (83 %), ampicillin (81 %), tetracycline and imipenem (76 %), trimethoprim (72 %) and amoxicillin (71 %) among vegetable isolates, with multidrug resistance patterns ranging from three to seventeen. CONCLUSIONS: Our results reveal the treated wastewater as a reservoir of enterotoxigenic E. coli as well as multidrug resistance that may pose a health hazard for humans and animals when released to the natural environment. Hence, there is need to develop management strategies and ensure compliance in order to prevent water-borne diarrhoea caused by ETEC and reduce the menace of antibiotic resistance in the environment.

Determination of levels of some metal contaminants in the freshwater environments of Osun State, Southwest Nigeria: A risk assessment approach to predict health threat.

This study evaluated levels of heavy metals and macro-elements in ten major rivers in Osun State, Southwest Nigeria. Triplicate water samples collected from selected rivers were analyzed for metal pollutants by atomic absorption spectrophotometry. Concentrations were obtained as follows: iron: 60-960 µg L-1; manganese: ND - 3 µg L-1; chromium 0-2100 µg L-1; aluminium: 0-800 µg L-1; copper: 0-1350 µg L-1; zinc: 10-650 µg L-1; calcium: 6400-232000 µg L-1 and magnesium: 2000-71000 µg L-1 but lead was not detected. While most of the parameters fell within the threshold values for drinking water, iron, chromium, aluminium and calcium exceeded at some locations. The pollution order of the rivers, especially with heavy metals had the order: R8 > R3 > R2 > R5 > R10 > R6 > R4 > R1 > R7 > R9; which implies that R8 and R3 are the most polluted. The health risk assessment results revealed that hazard quotient and hazard index were below the acceptable limit of 1.0, suggesting no substantial non-carcinogenic effect would arise from individual exposure to the elements. However, the carcinogenic risks were 3.18 × 10-6 and 4.43 × 10-6 for adults and children, respectively at R8, implying that 1 person in 314,206 adults and 225,836 children drinking from the water may be at risk of having cancer correspondingly. Hence, there is need to moderate the levels of these pollutants in these waterbodies to safeguard public health.

Molecular identification of some wild Nigerian mushrooms using internal transcribed spacer: polymerase chain reaction.

Identification of fungal species based on morphological characteristics is tedious, complex, prone to errors, and thus cannot be completely relied upon. In this study, internal transcribed spacers (ITS 1 and 4)-polymerase chain reaction was employed to amplify DNA of 19 mushroom isolates collected at Environmental Pollution Science and Technology farm, Ilesa, Southwest Nigeria. The PCR amplification of ITS1 and 4 of the mushrooms isolates yielded approximately 850 bp. Amplicons obtained were sequenced and identified using BLASTn in the NCBI. The BLASTn results revealed that Termitomyces aurantiacus (3), Tricholoma matsutake (8), Tricholoma robustum (2), P. ostreatus (4), Schizophyllum commune (1) and Pleurotus pulmonarius (1) were fully represented. Only Tricholoma matsutake (KT273371), Pleurotus pulmonarius (KY962469) and Tricholoma matsutake (AF438605) had 100% similarity with reference strain. However, the phylogenetic analysis of the isolates showed low genetic relatedness with reference strains. This study revealed the novelty of the mushroom strains and thus advocating the need for strict conservation measures and further investigations on their potential benefits to mankind.

Antimicrobial resistance determinants of Escherichia coli isolates recovered from some rivers in Osun State, South-Western Nigeria: Implications for public health.

The inevitable development of resistance has sunk the great success achieved in the discovery of antimicrobial agents and dashed the hope of man in the recovery from infections and illnesses, as diseases and disease agents that were once thought to be controlled by antimicrobials are now re-emerging in new leagues resistance to therapy. A total of 300 PCR confirmed Escherichia coli isolates recovered from different river sources in Osun State, Nigeria were evaluated for their antibiogram profiling by the disc diffusion method and the resistant isolates were further profiled for their genotypic antimicrobial resistance determinants by polymerase chain reaction assays. Among the 20 antimicrobials selected from 10 families, resistance among sulfonamides, ß-lactams and tetracyclines were found to be most frequent than phenicols and aminoglycosides with a noticeable increase in the number of multi-drug resistance ranging from three to nine antimicrobials. A total of 19 resistance determinants were assessed with their prevalence and distributions obtained as follows; [sulfonamides sulI (8%), sulII (41%)], [ß-lactams; ampC 22%; blaTEM, (21%), and blaZ (18%),], [tetracyclines tetA (24%), tetB (23%), tetC (18%), tetD (78%), tetK (15%), and tetM, (10%)], [phenicols; catI (37%), catII (28%), and cmIA1 (19%)] and [aminoglycosides; aacC2 (8%), aphA1 (80%), aphA2 (80%), aadA (79%) and strA (38%)]. The Pearson chi-square exact test revealed many strong significant associations among ampC, blaTEM, blaZ and tetA genes with some determinants screened. The findings signify high increase in the prevalence of multidrug resistant E. coli isolates and resistance determinants indicating increased public health risks associated with the ingestion of waters from untreated sources. Hence, a necessity for safe water supply, provision of proper sanitation facilities and good surveillance programmes to monitor antimicrobial resistance patterns in water bodies.

Multiple antibiotic resistance indexing of Escherichia coli to identify high-risk sources of faecal contamination of water.

We evaluated the antibiogram profile of Escherichia coli (n = 300) isolated from selected rivers in Osun State, Nigeria. The identities of the E. coli isolates were confirmed by polymerase chain reaction (PCR) technique. Susceptibility of the isolates to 20 antibiotics conventionally used in clinical cases was assessed in vitro by the standardized agar disc-diffusion method. All the isolates were susceptible to imipenem, meropenem, amikacin and gatilofloxacin. The isolates were variously susceptible to the other antibiotics as follows: ciprofloxacin (96 %), kanamycin (95 %), neomycin (92 %), streptomycin (84 %), chloramphenicol (73 %), nalidixic acid (66 %), nitrofurantoin (64 %), gentamycin (63 %), doxycycline (58 %), cefepime (57 %), tetracycline (49 %) and cephalothin (42 %). The multiple antibiotic resistance indexing ranged from 0.50 to 0.80 for all the sampling locations and exceeded the threshold value of 0.2, suggesting the origin of the isolates to be of high antimicrobial usage. Our findings signify an increase in the incidence of antimicrobial resistance of E. coli towards conventionally used antibiotics necessitating proper surveillance programmes towards the monitoring of antimicrobial resistance determinants in water bodies.