T. G. Barnard

Fungi in housefly (Musca domestica L.) as a disease risk indicator—A case study in South Africa

Houseflies are the commonest insects which have increasingly overcrowded human dwellings, particularly in rural areas and constitute a health hazard. In the environment they move back and forth by feeding and breeding on food commodities and filth. This may lead to the spread of diseases and also mycotoxin-producing fungi. Thus frequent exposure to the activity of such houseflies will have an impact on the welfare of humans. The study investigated the natural occurrence of fungal contamination in housefly samples captured from different households and pit toilets from a rural community in South Africa. Fungal contamination data were based on the prevalence, contamination level and morphological characteristics of the different identified species. A total of 497 fungal isolates of 15 genera including Aspergillus (37%), Fusarium (17%), Penicillium (21%), Alternaria (1.4%), Chrysosporium (2%), Cladosporium (0.2%), Curvularia (0.4%), Epicoccum (1%), Eupenicillium (1%), Moniliella (9%), Mucor (2%), Nigrospora (1%), Rhizopus (2%), Scopulariopsis (2%) and Yeasts (3%) were identified from the external surfaces of both female and male houseflies. The range of fungal contamination per total fungal count isolated from female and male houseflies were recorded with mean fungal load of 4.1×10(6), 8.4×10(6), 4.4×10(6), 3.3×10(5), 9.8×10(6), 2.2×10(4), 5.6×10(4), 2.9×10(6), 5.2×10(6), 4.7×10(6), 4.5×10(7), 4.6×10(6), 2.3×10(6), 4.9×10(7) and 6.4×10(6)CFU/ml, respectively. However, the range from The most dominant fungal isolates of the female housefly samples were Aspergillus flavus, Fusarium verticillioides, Penicillium verrucosum and Moniliella suaveolens, while A. flavus, Aspergillus parasiticus, F. verticillioides, Fusarium proliferatum and Penicillium aurantiogriseum were most prevalent in male samples. The study proves that housefly is a vector for fungal spores. Therefore, it is crucial to implement housefly-control measures to curb the spread of diseases.

Pathogenic Escherichia coli in rural household container waters.

Plastic containers in the range of 5-20 L are widely used - especially in rural African settings - to collect, transport and store water for domestic use, including drinking, bathing and hygiene. The pathogen content of the waters in these containers has not been adequately characterized as yet. This paper presents the primary findings of a synoptic survey of drinking water quality samples from these containers and involved collection of bacterial indicator and pathogenicity gene data. In total, 571 samples of a variety of waters were taken in rural communities in South Africa and the Escherichia coli numbers measured. Of the E. coli positive samples, 46% (n = 148) were screened for the presence of E. coli pathogen gene markers. Though synoptic, the survey provided many insights into the issues that drove the study. Container use markedly degraded water quality as judged by indicator counts, even where improved water supply services were in place. Household container use also appeared to promote regrowth or contamination of containers with pathogenic E. coli strains. Polymerase chain reaction (PCR) analysis also showed that the diversity of potential pathogenic E. coli carrying virulence genes was great. All seven genes screened for (Ial, Stx1, Stx2, EaeA, Eagg, ST, LT) were found in the waters, alone or as mixtures (number of different combinations = 31) including those characteristic of the more dangerous invasive and haemorrhagic E. coli strains. Given the central role of containers in the management of water supply to rural communities, it is clear the microbiology of these waters requires much further characterization.

Fungal dissemination by housefly (Musca domestica L.) and contamination of food commodities in rural areas of South Africa

Several insects that act as vectors, including houseflies (Musca domestica L.), are often considered to be an important source of fungal contamination in human foods. Houseflies are also involved in the transmission of bacterial pathogens that may pose a serious hazard to human health. Thus, the rural population of South Africa, as typified by that in the Gauteng Province investigated in this study, is at high risk from fungal exposure disseminated by houseflies and it is therefore important to assess the role of flies in contaminating various food commodities. Eighty four samples of houseflies (captured from households and pit toilets) were studied for their potential to carry fungal spores into food commodities. The fungi occurring in samples of raw maize (15) and porridge (19) were also assessed. Fungal isolates were identified based on morphological characteristics by conventional identification methods. Fifteen genera of fungi were isolated and identified, of which Aspergillus, Fusarium, Penicillium, Cladosporium, Moniliella and Mucor were the most prevalent in all three sample types analysed. The incidence rates of fungal contamination per total fungal count isolated in houseflies, maize and porridge were recorded with mean fungal load of 2×10(8) CFU/ml, 1×10(7)CFU/g and 2×10(7)CFU/g respectively. Additionally, A. flavus, A. parasiticus, F. verticillioides, F. proliferatum, P. verrucosum, P. aurantiogriseum and M. suaveolens were the most frequent fungal isolates in houseflies with incidence rate of 34%, 11%, 27%, 21%, 22%, 17% and 32% respectively. F. verticillioides, A. flavus, A. niger and P. oslonii were the most prevalent species contaminating porridge and maize with incidence rate of 23%, 32%, 16% and 28% in maize samples, while incidence rates of 59%, 15% and 29% were recorded in porridge samples with the exception of F. verticillioides. The prevalence of these genera of fungi may pose serious health risks.

Applicability of Bio-wipes for the collection of human faecal specimens for detection and characterisation of enteric viruses

To determine whether gastroenteritis viruses and other enteric viruses could be detected in faecal specimens collected with Bio‐wipes.

Detection of Vibrio cholerae and Vibrio parahaemolyticus by molecular and culture based methods from source water to household container-stored water at the point-of-use in South African rural communities

Detection methods for Vibrio cholerae and Vibrio parahaemolyticus which included the culture based approach with polymerase chain reaction (PCR) confirmation, PCR detection without enrichment and PCR with a pre-enrichment were developed and their performance evaluated. PCR assays targeted the SodB (V. cholerae species), Flae (V. parahaemolyticus species), 16S rRNA (Vibrio and Enterobacteriacea species) genes (Multiplex 1) and V. cholerae O1 and V. cholerae O139 rfb genes, ctxA (cholera toxin) gene and 16S rRNA gene (Multiplex 2). These methods were used to determine the occurrence of selected Vibrios in source water as well as in household container-stored water. The combination of filtration, enrichment and PCR method provided a sensitive and specific method for the detection of selected Vibrios in water samples. The PCR with a pre-enrichment method detected as few as 4-10 cfu/100 mL of selected Vibrios and PCR detection without the enrichment method detected as few as 40-100 cfu/100 mL of selected Vibrios. The inclusion of an enrichment period allows detection of culturable bacteria. As an application of the developed methods, V. cholerae and V. parahaemolyticus were detected in the source water used by the population and in the water-storage containers. The results indicate that Vibrio species in the containers could have originated from the source water and survive in biofilms inside the containers.

Mycotoxigenic potentials of the genera: Aspergillus, Fusarium and Penicillium isolated from houseflies (Musca domestica L.)

A study on the potential of houseflies (Musca domestica L.) to spread fungal spores in Gauteng Province, South Africa proved that houseflies are vectors for fungal spores. Therefore, there is a need to determine the toxigenic potentials and to identify the mycotoxins produced by fungal isolates derived from this study. In total 377 potentially toxigenic isolates of Aspergillus (186), Fusarium (85) and Penicillium (106) species (spp.) were isolated. These isolates were further tested for their ability to produce aflatoxins (AFs) [aflatoxin B1, B2, G1 and G2], deoxynivalenol (DON), fumonisin B1 (FB1) ochratoxin A (OTA), and zearalenone (ZEA) by high-performance liquid chromatography (HPLC) respectively. Strains of A. flavus and A. parasiticus belonging to the genera of Aspergillus were found to be the main producers of AFB1, AFB2, AFG1, and AFG2, while A. carbonarius, A. niger and A. ochraceus produced OTA. Fumonisin B1 was produced by F. verticillioides and F. proliferatum with concentrations ranging from 20 to 1834μg/kg and 79 to 262μg/kg respectively. Deoxynivalenol produced mainly by F. culmorum (2-6μg/kg), F. graminearum (1-4μg/kg), F. poae (1-3μg/kg), and F. sporotrichioides (2-3μg/kg) species was the least detected toxin in this study. The high mycotoxins levels produced in isolates from houseflies in this study are regarded as unsafe, especially when international legislated tolerance levels for mycotoxins are considered. Thus, possible human exposure to mycotoxins may pose concerns with respect to human health and demands constant and consistent investigation.

Use of a bio-wipe kit to detect fumonisin B1 in faecal materials

Fusarium toxins with reference to fumonisin B1 (FB1) have long been regarded as contaminants of maize and maize-based related products. However, when consumed they can cause intoxication, especially in humans. Therefore, effective quantitative methods for assessing the dietary exposure of this toxic fungal metabolite are required. The objective of this investigation was to evaluate the effect on the use of a bio-wipe kit, which is a faecal material collection kit, to detect the presence of FB1. Faecal materials were collected from a rural farming community in Gauteng Province, South Africa. In total, 200 samples of faecal material were analysed for Fusarium species using a serial dilution method, while FB1 was further analysed and quantified by reversed-phase TLC and HPLC. The study showed the presence of 11 different Fusarium species grown on potato dextrose agar culture medium of which F. verticillioides and F. proliferatum, producers of FB1, and F. oxysporum were the dominant species. Fumonisin B1 was recorded at an incidence rate of 65% of the total using TLC. Results from HPLC showed that 84% were positive at different ranges of concentration for FB1. This study supports the use of a bio-wipe as a rapid method to determine human exposure to FB1.