Search Results (11)

Wastewater treatment plants (WWTPs) are identified as contributors to faecal pollution and the spread of antimicrobial resistance (AMR) in water ecosystems. This research examined the prevalence, profiles of antimicrobial resistance, and pathogenic types of Escherichia coli in effluent from WWTPs and nearby river systems in the Vhembe District. Between May and June 2025, 28 water samples were collected from two WWTP discharge points as well as upstream and downstream locations along the Mvudi, Luvuvhu, and Madadzhe Rivers. The enumeration of E. coli was conducted using Colilert Quanti-Tray method, with isolates obtained via membrane filtration and confirmed using API 20E and VITEK^®2 systems. Antimicrobial susceptibility was assessed using VITEK^®2, while pathotypes were detected through multiplex PCR. E. coli was found at all sampling locations; however, differences in concentrations across sampling sites and sampling periods were not statistically significant (p > 0.05). Out of 26 confirmed isolates, a significant resistance to β-lactam antibiotics was noted, especially ampicillin (100%). Pathotype analysis revealed strains such as ETEC, EAEC, and EPEC. These results underline extensive contamination by antimicrobial-resistant E. coli in rivers affected by WWTP discharge, which poses potential public health concerns and underscores the necessity for enhanced monitoring efforts. Additional research is needed to validate these findings. Full article

Quantitatively assessing fecal indicator bacteria in drinking water from limited resource settings (e.g., disasters, remote areas) can inform public health strategies for reducing waterborne illnesses. This study aimed to compare two common approaches for quantifying Escherichia coli (E. coli) density in natural water versus the ColiPlate™ kit approach. For comparing methods, 41 field samples from natural water sources in Kentucky (USA) were collected. E. coli densities were then determined by (1) membrane filtration in conjunction with modified membrane-thermotolerant E. coli (mTEC) agar, (2) Idexx Quanti-Tray^® 2000 with the Colilert^® substrate, and (3) the Bluewater Biosciences ColiPlate kit. Significant correlations were observed between E. coli density data for all three methods (p < 0.001). Paired t-test results showed no difference in E. coli densities determined by all the methods (p > 0.05). Upon assigning modified mTEC as the reference method for determining the World Health Organization-assigned “very high-risk” levels of fecal contamination (>100 E. coli CFU/100 mL), both ColiPlate and Colilert exhibited excellent discrimination for screening very high-risk levels according to the area under the receiver operating characteristic curve (~89%). These data suggest ColiPlate continues to be an effective monitoring tool for quantifying E. coli density and characterizing fecal contamination risks from water. Full article

The current study investigated the impact of chicken litter application on the abundance of multidrug-resistant Enterococcus spp. in agricultural soil. Soil samples were collected from five different strategic places on a sugarcane farm before and after manure application for four months. Chicken litter samples were also collected. Enterococci were enumerated using the Enterolert^®/Quanti-Tray 2000^® system and confirm and differentiated into species using real-time PCR. The antibiotic susceptibility profile of the isolates was determined using the disk diffusion method following the European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines. The overall mean bacterial count was significantly higher (p < 0.05) in manure-amended soil (3.87 × 10⁷ MPN/g) than unamended soil (2.89 × 10⁷ MPN/g). Eight hundred and thirty-five enterococci (680 from soil and 155 from litter) were isolated, with E. casseliflavus being the most prevalent species (469; 56.2%) and E. gallinarum being the least (16; 1.2%). Approximately 56% of all the isolates were resistant to at least one antibiotic tested, with the highest resistance observed against tetracycline (33%) and the lowest against chloramphenicol (0.1%); 17% of E. faecium were resistant to quinupristin-dalfopristin. Additionally, 27.9% (130/466) of the isolates were multidrug-resistant, with litter-amended soil harbouring more multidrug-resistant (MDR) isolates (67.7%; 88/130) than unamended soil (10.0%; 13/130). All isolates were susceptible to tigecycline, linezolid and gentamicin. About 7% of the isolates had a multiple antimicrobial resistance index > 0.2, indicative of high antibiotic exposure. Although organic fertilizers are regarded as eco-friendly compared to chemical fertilizers for improving soil fertility, the application of untreated animal manure could promote the accumulation of antibiotics and their residues and antibiotic-resistant bacteria in the soil, creating an environmental reservoir of antimicrobial resistance, with potential human and environmental health risks. Full article

Foodborne pathogens, including antibiotic-resistant species, constitute a severe menace to food safety globally, especially food animals. Identifying points of concern that need immediate mitigation measures to prevent these bacteria from reaching households requires a broad understanding of these pathogens’ spread along the food production chain. We investigated the distribution, antibiotic susceptibility, molecular characterization and clonality of Enterococcus spp. in an intensive pig production continuum in South Africa, using the farm-to-fork approach. Enterococcus spp. were isolated from 452 samples obtained along the pig farm-to-fork continuum (farm, transport, abattoir, and retail meat) using the IDEXX Enterolert^®/Quanti-Tray^® 2000 system. Pure colonies were obtained on selective media and confirmed by real-time PCR, targeting genus- and species-specific genes. The susceptibility to antibiotics was determined by the Kirby–Bauer disk diffusion method against 16 antibiotics recommended by the WHO-AGISAR using EUCAST guidelines. Selected antibiotic resistance and virulence genes were detected by real-time PCR. Clonal relatedness between isolates across the continuum was evaluated by REP-PCR. A total of 284 isolates, consisting of 79.2% E. faecalis, 6.7% E. faecium, 2.5% E. casseliflavus, 0.4% E. gallinarum, and 11.2% other Enterococcus spp., were collected along the farm-to-fork continuum. The isolates were most resistant to sulfamethoxazole-trimethoprim (78.8%) and least resistant to levofloxacin (5.6%). No resistance was observed to vancomycin, teicoplanin, tigecycline and linezolid. E. faecium displayed 44.4% resistance to quinupristin-dalfopristin. Also, 78% of the isolates were multidrug-resistant. Phenotypic resistance to tetracycline, aminoglycosides, and macrolides was corroborated by the presence of the tetM, aph(3′)-IIIa, and ermB genes in 99.1%, 96.1%, and 88.3% of the isolates, respectively. The most detected virulence gene was gelE. Clonality revealed that E. faecalis isolates belonged to diverse clones along the continuum with major REP-types, mainly isolates from the same sampling source but different sampling rounds (on the farm). E. faecium isolates revealed a less diverse profile. The results suggest that intensive pig farming could serve as a reservoir of antibiotic-resistant bacteria that could be transmitted to occupationally exposed workers via direct contact with animals or consumers through animal products/food. This highlights the need for more robust guidelines for antibiotic use in intensive farming practices and the necessity of including Enterococcus spp. as an indicator in antibiotic resistance surveillance systems in food animals. Full article

Antibiotic resistance profiles of Escherichia coli were investigated in an intensive pig production system in the uMgungundlovu District, South Africa, using the ‘farm-to-fork’ approach. Four hundred seventeen (417) samples were collected from pig and pig products at different points (farm, transport, and abattoir). E. coli was isolated and enumerated using the Colilert^® 18/Quanti-Tray^® 2000 system. Ten isolates from each Quanti-tray were selected randomly and putatively identified on eosin methylene blue agar. Real-time PCR targeting the uidA gene was used to confirm isolates to the genus level. The Kirby–Bauer disc diffusion method was used to determine the isolates’ antibiotic susceptibility profiles against 20 antibiotics. A total of 1044 confirmed E. coli isolates were obtained across the three critical points in the food chain. Resistance was observed to all the antibiotics tested with the highest and lowest rates obtained against tetracycline (88.5%) and meropenem (0.2%), respectively. Resistance was also observed to chloramphenicol (71.4%), ampicillin (71.1%), trimethoprim-sulfamethoxazole (61.3%), amoxicillin-clavulanate (43.8%), cephalexin (34.3%), azithromycin (23.9%), nalidixic acid (22.1%), cefoxitin (21.1%), ceftriaxone (18.9%), ciprofloxacin (17.3%), cefotaxime (16.9%), gentamicin (15.5%), cefepime (13.8%), ceftazidime (9.8%), amikacin (3.4%), piperacillin-tazobactam (1.2%), tigecycline (0.9%), and imipenem (0.3%). Multidrug resistance (MDR) was observed in 71.2% of the resistant isolates with an overall multiple antibiotic resistance (MAR) index of 0.25, indicating exposure to high antibiotic use environments at the farm level. A high percentage of resistance was observed to growth promoters and antibiotics approved for veterinary medicine in South Africa. Of concern was resistance to critically important antibiotics for animal and human use and the watch and reserve categories of antibiotics. This could have adverse animal and human health consequences from a food safety perspective, necessitating efficient antibiotic stewardship and guidelines to streamline antibiotic use in the food-animal production chain. Full article

There is a need for accessible and low-cost microbiological water quality testing in contexts where diarrheal illness is a major public health concern. In most cases, the quantification of Escherichia coli and other microbial indicators by conventional culture methods requires an incubation step for processed samples at specific temperatures for bacterial growth over a prescribed time. However, incubators can be the most expensive equipment required for such microbial analyses, limiting the number and scope of water quality testing available in low-resource contexts. In this study, a low-cost incubator was developed using a locally available expanded polystyrene (EPS) foam cooler, with two water bottles filled with hot water to heat incubator to a target of 35 °C. The EPS incubator performance was validated by processing 150 water samples in duplicates using the Colilert Quanti-tray/2000 system, incubated in either the EPS incubator or a standard laboratory incubator set at 35 °C. Statistically significant correlations of results indicated that the quantification of E. coli was comparable between both methods. Risk categorizations from standard and EPS incubation results agreed for 141 of 150 (94%) samples, with zero false negatives. In addition to being reasonably mobile the EPS incubator would reduce the cost of such water quality testing, thus potentially increasing the scope of water quality testing coverage. Full article

Background: Young children exhibit a high susceptibility to several diarrhoea-causing bacterial microorganisms. In this study, the prevalence of fecal contamination on children’s toys was determined using total coliform and E. coli as bacterial fecal indicators. The prevalence of diarrhoeagenic E. coli strains were used as an indication of the potential health risks. Materials and Methods: A cross-sectional descriptive study was carried out for 3 months in rural communities in the Vhembe district, Limpopo province of South Africa. Nonporous plastic toys (n = 137) used by children under 5 years of age in households and day care centres (DCCs) from rural villages were collected for assessment. New toys (n = 109) were provided to the households and DCCs and collected again after 4 weeks. Microbiological assessment was carried out using the Colilert^® Quanti-Tray/2000 system. Diarrhoeagenic E. coli strains were identified using a published multiplex PCR protocol. Results: Water, sanitation and hygiene (WASH) conditions of the children in the households and DCCs were assessed. Statistical analysis was used to identify the relationship between fecal contamination of the existing and introduced toys. All the existing and introduced toy samples, both from DCCs and households, tested positive for total coliform counts and 61 existing and introduced toy samples tested positive for E. coli counts. Diarrhoeagenic E. coli strains identified included EHEC, ETEC, EPEC, EIEC and EAEC. Conclusions: The results indicated that water, sanitation and hygiene conditions could be responsible in the contamination of children’s toys and the transmission of diarrhoea to young children. Full article

Although many developing countries use harvested rainwater (HRW) for drinking and other household purposes, its quality is seldom monitored. Continuous assessment of the microbial quality of HRW would ensure the safety of users of such water. The current study investigated the prevalence of pathogenic Escherichia coli strains and their antimicrobial resistance patterns in HRW tanks in the Eastern Cape, South Africa. Rainwater samples were collected weekly between June and September 2016 from 11 tanks in various areas of the province. Enumeration of E. coli was performed using the Colilert^®18/Quanti-Tray^® 2000 method. E. coli isolates were obtained and screened for their virulence potentials using polymerase chain reaction (PCR), and subsequently tested for antibiotic resistance using the disc-diffusion method against 11 antibiotics. The pathotype most detected was the neonatal meningitis E. coli (NMEC) (ibeA 28%) while pathotype enteroaggregative E. coli (EAEC) was not detected. The highest resistance of the E. coli isolates was observed against Cephalothin (76%). All tested pathotypes were susceptible to Gentamicin, and 52% demonstrated multiple-antibiotic resistance (MAR). The results of the current study are of public health concern since the use of untreated harvested rainwater for potable purposes may pose a risk of transmission of pathogenic and antimicrobial-resistant E. coli. Full article

In Australia, rainwater is an important source of water for many households. Unlike municipal water, rainwater is often consumed untreated. This study investigated the potential contamination of rainwater by microorganisms. Samples from 53 rainwater tanks across the Adelaide region were collected and tested using Colilert™ IDEXX Quanti-Tray*/2000. Twenty-eight out of the 53 tanks (53%) contained Escherichia coli. Samples collected from ten tanks contained E. coli at concentrations exceeding the limit of 150 MPN/100 mL for recreational water quality. A decline in E. coli was observed in samples collected after prolonged dry periods. Rainwater microbiological values depended on the harvesting environment conditions. A relationship was found between mounted TV antenna on rooftops and hanging canopies; and E. coli abundance. Conversely, there was no relationship between seasonality and E. coli or roof and tank structure materials and E. coli. In several tanks used for drinking water, samples collected prior to and after filtration showed that the filtration systems were not always successful at completely removing E. coli. These results differed from a study undertaken in the laboratory that found that a commercially available in-bench 0.45 µm filter cartridge successfully reduced E. coli in rainwater to 0 MPN/100 mL. After running a total of 265 L of rainwater which contained high levels of E. coli through the filter (half of the advertised filter lifespan), the filter cartridge became blocked, although E. coli remained undetected in filtered water. The difference between the laboratory study and field samples could be due to improper maintenance or installation of filters or recontamination of the faucet after filtration. The presence of E. coli in water that is currently used for drinking poses a potential health concern and indicates the potential for contamination with other waterborne pathogens. Full article

Background: Water quality testing is dictated by microbial agents found at the time of sampling in reference to their acceptable risk levels. Human activities might contaminate valuable water resources and add to the microbial load present in water bodies. Therefore, the effects of human activities on the microbial quality of rivers collected from twelve catchments in the Vhembe District in South Africa were investigated, with samples analyzed for total coliform (TC) and Eschericha coli (E. coli) contents. Methods: Physical parameters and various human activities were recorded for each sampling site. The Quanti-Tray^® method was adopted for the assessment of TC and E. coli contents in the rivers over a two-year period. A multiplex polymerase chain (PCR) method was used to characterize the strains of E. coli found. Results: The microbial quality of the rivers was poor with both TC and E. coli contents found to be over acceptable limits set by the South African Department of Water and Sanitation (DWS). No significant difference (p > 0.05) was detected between TC and E. coli risks in dry and wet seasons. All six pathogenic E. coli strains were identified and Enteroaggregative E. coli (EAEC), atypical Enteropathogenic E. coli (a-EPEC) and Enterotoxigenic E. coli (ETEC) were the most prevalent E. coli strains detected (respectively, 87%, 86% and 83%). Conclusions: The study indicated that contamination in the majority of sampling sites, due to human activities such as car wash, animal grazing and farming, poses health risks to communities using the rivers for various domestic chores. It is therefore recommended that more education by the respective departments is done to avert pollution of rivers and prevent health risks to the communities in the Vhembe District. Full article

The use of contaminated surface water continues to be a pressing issue in areas of the world where people lack improved drinking water sources. In northern coastal Ecuador, many communities rely on untreated surface water as their primary source of drinking water. We undertook a study to explore how microscale river hydrodynamics affect microbial water quality at community water collection locations at three rivers with varying stream velocity and turbidity profiles. To examine how the distance from river shore and physiochemical water quality variables affect microbial contamination levels in the rivers; we collected a total of 355 water samples within six villages on three rivers; and tested for Escherichia coli concentrations using the IDEXX Quanti-tray method. We found that log₁₀ E. coli concentrations decreased with increasing distance from shore (β = −0.017; p = 0.003). Water in the main channel had E. coli concentrations on average 0.12 log₁₀ lower than within eddies along the river shore and 0.27 log₁₀ lower between the sample closest to shore and any sample >6 m from the shore. Higher E. coli concentrations were also significantly associated with increased turbidity (β = 0.003; p < 0.0001) and decreased dissolved oxygen levels (β = −0.310; p < 0.0001). The results of this study can help inform community members about the safest locations to collect drinking water and also provide information on watershed scale transport of microbial contaminants between villages. Full article