Molecular epidemiology of human bocavirus infection in hospitalized children with acute gastroenteritis in South Africa, 2009-2015.

Human bocavirus (HBoV) is known to be associated with a variety of clinical manifestation including acute gastroenteritis (AGE). Despite their global prevalence, no data is available on the epidemiology of HBoV associated with AGE in South Africa (SA). Between April 2009 and April 2015, 3765 stool specimens were collected from children less than 5 years of age hospitalized with diarrhea. Specimens were screened for selected enteric viruses by enzyme immunoassay and quantitative polymerase chain reaction, bacteria by culture and parasites by staining and microscopy. HBoV was detected in 5.63% (212 of 3765) of cases, the majority of which were children ≤2 years (92%, 195 of 212), and were common in the summer and autumn months (60%; 128 of 212). Further investigations of coinfections showed that bacteria (adjusted odds ratio [aOR] = 2.20; 95% confidence interval [CI], 1.41-3.45; P = .001) and sapovirus (aOR = 2.05; 95% CI, 1.08-3.86; P = .027) were significantly associated with HBoV in multivariate analysis. HBoV genotyping was successful in 191 of the 212 samples with HBoV-1 being the most prevalent genotype observed (79.6%; 152 of 191) followed by HBoV-3 (13.6%; 26 of 191), HBoV-2 (5.2%; 10 of 191), and HBoV-4 (1.6%; 3 of 191). The high prevalence of HBoV-1, a virus known to be associated with respiratory infections, and the association between HBoV-positive specimens and already established AGE agents, suggests that HBoV may play a limited role in the observed AGE cases in SA.

Relative proportions of E. coli and Enterococcus spp. may be a good indicator of potential health risks associated with the use of roof harvested rainwater stored in tanks.

A total of 285 water samples were collected from 71 roof harvested rainwater tanks from four villages in different provinces over a two-year (2013-2014) period during the early (October to December) and late (January to March) rainy season. Water quality was evaluated based on Escherichia coli, faecal coliforms and Enterococcus spp. prevalence using the IDEXX Quanti-Tray quantification system. Real-Time PCR was used to analyse a subset of 168 samples for the presence of Shigella spp., Salmonella spp. and E. coli virulence genes (stx1, stx2 and eaeA). Escherichia coli were detected in 44.1% of the samples, Enterococcus spp. in 57.9% and faecal coliforms in 95.7%. The most prevalent E. coli concentrations in harvested rainwater were observed in 29.1% of samples and 22.5% for Enterococcus spp. and, were within 1-10 cfu/100 ml and 10-100 cfu/100 ml, respectively, whereas those for faecal coliforms (36.6%) were within 100-1000 cfu/100 ml. On average 16.8% of the samples had neither E. coli nor Enterococcus spp. detected, while 33.9% had only Enterococcus spp. and 23.7% had only E. coli. E. coli and Enterococcus spp. were detected together in 25.5% of the samples. Evaluation of samples for potential pathogenic bacteria showed all tested samples to be negative for the Shigella spp. ipaH gene, while five tested positive for Salmonella ipaB gene. None of the samples tested positive for the stx1 and stx2 genes, and only two tested positive for the eaeA gene. These findings are potentially useful in the development of a simplified risk assessment strategy based on the concentrations of indicator bacteria.

Antimicrobial Resistance Profiles of Salmonella spp. from Agricultural Environments in Fruit Production Systems.

Foodborne disease outbreaks involving fresh produce have increased in recent years. The risk of infection from contaminated food is worsened by the increased prevalence of antibiotic-resistant strains. This study evaluated the prevalence of antibiotic resistance in Salmonella isolates (n = 263) from agricultural production systems through to the final packed product. Salmonella isolates were preliminarily identified by matrix-assisted laser desorption ionization-time-of-flight mass spectroscopy (MALDI-TOF MS) and API 20E and identities confirmed by invA gene polymerase chain reaction. Antimicrobial susceptibility was performed with 15 antimicrobial agents using the Kirby-Bauer disk diffusion test. Of the 263 Salmonella isolates assessed, 59.3% were resistant to one or more antimicrobials. The most frequently detected resistance was against chloramphenicol and kanamycin (46.7%), trimethoprim-sulfamethoxazole (28%), and streptomycin (14%), and the less frequently detected resistance was toward ampicillin (1.14%), amikacin (0.76%), and amoxicillin-clavulanic acid (0.38%). Multiple antimicrobial resistance (MAR) (resistance to ≥3 antibiotics) was found in 48.7% (76/156) isolates. The most common MAR phenotype was to chloramphenicol and trimethoprim/sulfamethoxazole-kanamycin (43.6%). Resistance to chloramphenicol, kanamycin, or trimethoprim/sulfamethoxazole was only observed in MAR phenotypes. All isolates were susceptible to ceftiofur, cefoxitin, ceftriaxone, ciprofloxacin, nalidixic acid, gentamicin, and tetracycline. This study confirms the importance of fresh produce production environments as potential reservoirs and fresh produce as carriers of antibiotic-resistant Salmonella spp. with significant clinical importance. Further studies to evaluate the actual level of health risk from these pathogens should include characterization of the antibiotic resistance determinant genes among the isolates.

Spatial and Temporal Variations in Pollution Indicator Bacteria in the Lower Vaal River, South Africa.

The aim of this study was to evaluate the spatial and temporal variation of microbiological parameters of the Lower Vaal River, with emphasis on the Staphylococci population. River water concentrations of Staphylococci, heterotrophic, total coliforms, fecal coliforms and Streptococci groups of sanitary indicator bacteria were monitored. Results indicated significant contamination from agricultural land use inputs and municipal waste applications. Significant temporal and spatial variation was observed in response to the varying river discharge. Higher microbial concentrations were detected during high river discharge whereas during low river discharge both low and high microbial concentrations were detected. The varying responses associated with river discharge helped to identify the importance of different sources of contamination in the catchment and the mechanisms transferring them. The overall impact of contamination on the water quality of the Vaal River could have potentially serious public health risks but also provide valuable data for integrated water resource management.

Pyrosequencing analysis of roof-harvested rainwater and river water used for domestic purposes in Luthengele village in the Eastern Cape Province of South Africa.

Pyrosequencing targeting the V1-V3 hypervariable of the 16S rDNA was used to investigate the bacterial diversity in river and roof-harvested rainwater (RHRW) used for potable purposes by rural households in Luthengele village in the Eastern Cape Province of South Africa. The phylum Proteobacteria dominated the data set (80.5 % of all reads), while 4.2 % of the reads could not be classified to any of the known phyla at a probability of 0.8 or higher (unclassified bacteria). At class level, the classes; Betaproteobacteria (50.4 % of all reads), Alphaproteobacteria (16.2 %), Verrucomicrobiae (6.6 %), Planctomycetacia (5.7 %), and Sphingobacteria (3 %) dominated the data set in all the samples. Although the class Verrucomicrobiae constituted 6.6 % of all sequences, 88.6 % of the sequences were from the river sample where the class represented 43.7 % of the observed sequences in the sample. The bacteria community structure clearly showed significant similarities between RHRW and differences with the river water control sample, suggesting different levels of contamination and environmental factors affecting the various water sources. Moreover, signatures of potential pathogens including Legionella, Acinetobacter, Pseudomonas, Clostridia, Chromobacterium, Yersinia, and Serratia were detected, and the proportions of Legionella were relatively higher suggesting a potential health risk to households using RHRW. This work provides guidance for prioritizing subsequent culturable and quantitative analysis to ensure that potentially significant pathogens are not left out of risk estimations.

Antibiotic resistance in Escherichia coli isolates from roof-harvested rainwater tanks and urban pigeon faeces as the likely source of contamination.

The objective of this study was to investigate the risks associated with the use of roof-harvested rainwater (RHRW) and the implication of pigeons as the most likely source of contamination by testing for antibiotic resistance profiles of Escherichia coli. A total of 239 E. coli were isolated from thirty fresh pigeon faecal samples (130 isolates), 11 RHRW tanks from three sites in Pretoria (78) and two in Johannesburg (31). E. coli isolates were tested against a panel of 12 antibiotics which included ampicillin, amoxicillin, amikacin, cefoxitin, ceftriaxone, chloramphenicol, ciprofloxacin, cotrimoxazole, enrofloxacin, gentamicin, nalidixic acid and tetracycline. In all samples, resistance to ampicillin (22.7.9%), gentamicin (23.6%), amikacin (24%), tetracycline (17.4) and amoxicillin (16.9%) were the most frequently encountered form of resistance. However, a relatively higher proportion of isolates from pigeon faeces (67.3%) were antibiotic resistant than those from RHRW (53.3%). The highest number of phenotypes was observed for single antibiotics, and no single antibiotic resistance was observed for chloramphenicol, ceftriaxone, gentamicin, cefoxitin, cotrimoxazole, although they were detected in multiple antibiotic resistance (MAR) phenotypes. The highest multiple antibiotic resistance (MAR) phenotypes were observed for a combination of four antibiotics, on isolates from JHB (18.8%), pigeon faeces (15.2%) and Pretoria (5.1%). The most abundant resistance phenotype to four antibiotics, Ak-Gm-Cip-T was dominated by isolates from pigeon faeces (6.8%) with Pretoria and Johannesburg isolates having low proportions of 1.3 and 3.1%, respectively. Future studies should target isolates from various environmental settings in which rainwater harvesting is practiced and the characterisation of the antibiotic resistance determinant genes among the isolates.

Microbial Load and Prevalence of Escherichia coli and Salmonella spp. in Macadamia Nut Production Systems.

ABSTRACT: This study evaluated the potential impact of environmental factors and harvesting practices on the microbial load of macadamia nuts. Three farms located in primary macadamia nut production regions, the Mbombela (A), Barberton (B) and White River (C) areas in Mpumalanga Province, South Africa, were sampled over a 2-year period. A total of 264 irrigation water (54), soil (30), and macadamia nut (180) samples were collected and evaluated for microbial load. All water samples had mean Escherichia coli loads below 1,000 MPN/100 mL, which is the standard regulatory requirement for agricultural water considered fit for irrigation in South Africa. Mean total aerobic plate counts of nut-in-husk on-tree samples (3.91 log CFU/g; n = 60) were higher after harvesting (5.98 log CFU/g; n = 60) but were lower after dehusking (to 4.89 log CFU/g; n = 60) on nut-in-shell samples. Salmonella spp. were only detected in water samples from farm B (67%; n = 18) and farm C (15%; n = 18). Neither Listeria monocytogenes nor Salmonella spp. were detected in the soil samples. E. coli was only detected in 20% (n = 10) of soil samples collected from two farms (farms A and B). None of the E. coli isolated in this study was positive for the eae, stx1, and stx2 enterohemorrhagic E. coli virulence genes. This study provides basic data that can be used in the development of macadamia nut-specific hazard assessment tools within primary production environments.

Monitoring Pesticide Residues in Fruits and Vegetables at Two of the Biggest Fresh Produce Markets in Africa.

In most countries, fresh produce sold at local markets is usually not analyzed for agricultural chemical residues as export products are, which raises concerns about the perceived safety levels of local food supplies in contrast with exported products. The aim of this study was to determine pesticide residue levels in fruits and vegetables sold at two of the biggest fresh produce markets in Africa. A total of 199 fruit and vegetable samples were collected between 2012 and 2014 and analyzed for 74 pesticides commonly used in the horticultural sector. Of the samples analyzed, 91% were compliant with set maximum residue levels (MRLs). The remaining samples either contained unregistered chemicals (8%) or exceeded set MRL values (1%). Products containing more than one pesticide residue constituted 4.02% of all samples tested. Imazalil and iprodione were found to be the most frequently detected pesticides (12 samples each). Boscalid, endosulfan, profenofos, and procymidone were associated with the most noncompliance, including exceeding MRL values or being unregistered for the specific crop. The establishment of a national pesticide monitoring program is essential for the country and would ensure that pesticides are used in accordance with good agricultural practices.