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Abstract Background Virulence determinants are crucial to the risk assessment of pathogens in an environment. This study investigated the presence of eleven key virulence-associated genes in Vibrio cholerae (n = 111) and Vibrio mimicus (n = 22) and eight virulence determinants in Vibrio alginolyticus (n = 65) and Vibrio parahaemolyticus (n = 17) isolated from six important water resources in Eastern Cape, South Africa, using PCR techniques. The multiple virulence gene indexes (MVGI) for sampling sites and isolates as well as hotspots for potential vibriosis outbreaks among sampling sites were determined statistically based on the comparison of MVGI. Result The PCR assay showed that all the V. cholerae isolates belong to non-O1/non-O139 serogroups. Of the isolates, Vibrio Cholera (84%), V. mimicus (73%), V. alginolyticus (91%) and V. parahaemolyticus (100%) isolates harboured at least one of the virulence-associated genes investigated. The virulence gene combinations detected in isolates varied at sampling site and across sites. Typical virulence-associated determinants of V. cholerae were detected in V. mimicus while that of V. parahaemolyticus were detected in V. alginolyticus. The isolates with the highest MVGI were recovered from three estuaries (Sunday river, Swartkopps river, buffalo river) and a freshwater resource (Lashinton river). The cumulative MVGI for V. cholerae, V. mimicus, V. alginolyticus and V. parahaemolyticus isolates were 0.34, 0.20, 0.45, and 0.40 respectively. The targeted Vibrio spp. in increasing order of the public health risk posed in our study areas based on the MVGI is V. alginolyticus > V. parahaemolyticus > V. cholerae > V. mimicus. Five (sites SR, PA5, PA6, EL4 and EL6) out of the seventeen sampling sites were detected as the hotspots for potential cholera-like infection and vibriosis outbreaks. Conclusions Our findings suggest that humans having contact with water resources in our study areas are exposed to potential public health risks owing to the detection of virulent determinants in human pathogenic Vibrio spp. recovered from the water resources. The study affirms the relevancy of environmental Vibrio species to the epidemiology of vibriosis, cholera and cholera-like infections. Hence we suggest a monitoring program for human pathogenic Vibrio spp. in the environment most especially surface water that humans have contact with regularly.

The current study determines the density of Vibrio spp. and isolates V. cholerae and Vibrio mimicus from fish-anatomical-sites, prawn, crab and mussel samples recovered from fish markets, freshwater and brackish water. Virulence and antibiotic resistance profiling of isolates were carried out using standard molecular and microbiology techniques. Vibrio spp. was detected in more than 90% of samples [134/144] and its density was significantly more in fish than in other samples. Vibrio. cholerae and V. mimicus were isolated in at least one sample of each sample type with higher isolation frequency in fish samples. All the V. cholerae isolates belong to non-O1/non-O139 serogroup. One or more V. cholerae isolates exhibited intermediate or resistance against each of the eighteen panels of antibiotics used but 100% of the V. mimicus were susceptible to amikacin, gentamycin and chloramphenicol. Vibrio cholerae exhibited relatively high resistance against polymyxin, ampicillin and amoxicillin/clavulanate while V. mimicus isolates exhibited relatively high resistance against nitrofurantoin, ampicillin and polymixin. The multiple-antibiotic-resistance-index [MARI] for isolates ranges between 0 and 0.67 and 48% of the isolates have MARI that is >0.2 while 55% of the isolates exhibit MultiDrug Resistance Phenotypes. The percentage detection of acc, ant, drf18, sul1, mcr-1, blasvh, blaoxa, blatem, blaoxa48, gyrA, gyrB and parC resistance-associated genes were 2%, 9%, 14%, 7%, 2%, 25%, 7%, 2%, 2%, 32%, 25% and 27% respectively while that for virulence-associated genes in increasing other was ace [2%], tcp [11%], vpi [16%], ompU [34%], toxR [43%], rtxC [70%], rtxA [73%] and hyla [77%]. The study confirmed the potential of environmental non-O1/non-O139 V. cholerae and V. mimicus to cause cholera-like infection and other vibriosis which could be difficult to manage with commonly recommended antibiotics. Thus, regular monitoring of the environment to create necessary awareness for this kind of pathogens is important in the interest of public health.

Water resources contaminated with pathogenic Vibrio species are usually a source of devastating infection outbreaks that have been a public health concern in both developed and developing countries over the decades. The present study assessed the prevalence of six medically significant Vibrio species in some water resources in Eastern Cape Province, South Africa for 12 months. We detected vibrios in all the 194 water samples analyzed using polymerase chain reaction (PCR). The prevalence of Vibrio cholerae, Vibrio mimicus, Vibrio fluvialis, Vibrio vulnificus, Vibrio alginolyticus, and Vibrio parahaemolyticus in freshwater samples was 34, 19, 9, 2, 3, and 2%, and that in brackish water samples was 44, 28, 10, 7, 46, and 51%, respectively. The population of the presumptive Vibrio spp. isolated from freshwater (628) and brackish water (342) samples that were confirmed by PCR was 79% (497/628) and 85% (291/342), respectively. Twenty-two percent of the PCR-confirmed Vibrio isolates from freshwater (n = 497) samples and 41% of the PCR-confirmed Vibrio isolates from brackish water samples (n = 291) fall among the Vibrio species of interest. The incidences of V. cholerae, V. mimicus, V. fluvialis, V. vulnificus, V. alginolyticus, and V. parahaemolyticus amidst these Vibrio spp. of interest that were recovered from freshwater samples were 75, 14, 4, 6, 1, and 1%, whereas those from brackish water samples were 24, 7, 3, 3, 47, and 18%, respectively. Our observation during the study suggests pollution as the reason for the unusual isolation of medically important vibrios in winter. Correlation analysis revealed that temperature drives the frequency of isolation, whereas salinity drives the composition of the targeted Vibrio species at our sampling sites. The finding of the study is of public health importance going by the usefulness of the water resources investigated. Although controlling and preventing most of the factors that contribute to the prevalence of medically important bacteria, such as Vibrio species, at the sampling points might be difficult, regular monitoring for creating health risk awareness will go a long way to prevent possible Vibrio-related infection outbreaks at the sampling sites and their immediate environment.

Treated sewage harbours pathogenic microbes, such as enteric bacteria and protozoa, are capable of causing several diseases. Some of these are emerging pathogens sometimes recovered in the absence of common water quality indicator organisms. The possibility of selected treatments plants serving as momentary reservoirs of Vibrio pathogens during a non-outbreak period was assessed. The occurrence and diversity of Vibrio pathogens were monitored for one year (December 2016 to November 2017) in the treated effluents and upstream and downstream areas of the receiving water bodies of two wastewater treatment plants (WWTPs), designated AL and TS. Physicochemical parameters of TS and AL WWTPs’ water samples were analysed using a multi-parameter meter (Hanna, model HI 9828, Padova, Italy) and a turbidimeter (HACH, model 2100P, Johannesburg, South Africa). Water samples were augmented with alkaline peptone water and cultured on thiosulfate citrate bile salts sucrose agar at 37 °C for 24 h. The recovered probable pathogens were confirmed via PCR amplification, using primers specific for Vibrio species of public health significance. The distribution of Vibrio species positively and significantly (p < 0.01) correlated with turbidity (r = 0.630), temperature (r = 0.615), dissolved oxygen (r = 0.615), pH (r = 0.607), biological oxygen demand (r = 0.573), total dissolved solid (r = 0.543), total suspended solid (r = 0.511), electrical conductivity (r = 0.499), residual chlorine (r = 0.463) and salinity (r = 0.459). The densities of Vibrio species were found to be significantly higher (p < 0.05) in effluents from both AL and TS WWTPs than upstream and downstream of the receiving rivers across the sampling regime. Furthermore, the maximum Vibrio species density across the sampling regime were observed during the warmer Summer and Spring season. Moreover, six medically important Vibrio species were detected in the water samples, indicating that the methods employed were efficient in revealing that WWTPs are potential reservoirs of Vibrio pathogens, which could pose a substantial public health risk if the receiving water is used for domestic purposes. Our findings further strengthen existing calls for the inclusion of emerging bacterial pathogens, including Vibrio species, as water quality indicators by the South African Department of Water Affairs. Hence, we recommend regular monitoring of treated effluents and receiving water bodies to ensure early control of potential outbreaks of vibriosis and cholera.

Limpet (Scutellastra cochlear) serves as seafood recipe and an important member of the aquatic food chain. It is an abundant mollusc in some aquatic environment in South Africa. In this study, we investigated the potential of the molluscs harvested from the Buffalo, Swartkops, and Kowie estuaries in the Eastern Cape Province, South Africa to serve as transient or maintenance reservoir of Vibrio species. The mollusc and source water samples were collected monthly from the rivers between December 2016 and November 2017. The reservoir category of the limpet samples recovered was determined by employing the combination of MPN-PCR method and statistical analysis (comparison of mean and proportion tests). The densities of Vibrio spp. in limpet and their source water samples were determined using MPN-PCR methods. Presumptive isolates were recovered by processing the samples with thiosulfate-citrate-bile salts-sucrose agar and where necessary, samples were enriched with alkaline peptone water. The presumptive isolates were identified using PCR methods with emphasis on six Vibrio species of public health importance. Vibrio spp. were detected in all the limpet samples but not in all the water samples. The densities of Vibrio spp. in the limpet samples were more than the densities of Vibrio species in their source water and these were significant at P < 0.05. In like manner, five out of the six key Vibrio pathogens targeted in this study were more prevalent in limpet samples than in source water samples. Based on our findings, we concluded that our method though could be improved on, is efficient for the determination of reservoir types of bacterial-carrying organisms. We also concluded that the limpet found in the estuaries are not just a transient but a maintenance reservoir of Vibrio spp. which could cause vibrio-related infections.