The principal aims of this thesis were to examine whether there were hydraulic links between individual on-site wastewater systems in sandy soils at Salt Ash and the Tilgerry Creek estuary near Port Stephens, New South Wales, and whether the chemical and microbiological contaminants from on-site systems could reach surface and groundwaters, and possibly lead to impacts on estuarine oyster growing waters. The research outcomes are contained within the thesis and in four refereed papers presented at conferences, and which have been subsequently published, or are in press. Copies of each of these papers are contained within the thesis Appendices. The presence of faecal contamination from domestic systems in the estuary, and surface drains in particular, has been confirmed by work contained in this thesis. The potential level of risk to human health from the consumption of contaminated oysters is, however, regarded as very low, although an assessment of health risk using established microbial assessment models has not been undertaken / Doctor of Philosophy (PhD)

A study was undertaken to determine the food safety of the fermented Māori delicacy, Toroi. Ten batches of Toroi were prepared by a commonly used traditional method that consisted of boiling the vegetable component, either watercress or puha, and combining it with chopped mussel flesh. The mixture was cooled and then stored in a refrigerator for up to eight months to allow natural fermentation to take place. All ingredients were sourced from retail outlets. The Toroi was examined at intervals over eight months for a range of pathogens (seven in all) that have been related to incidents of food poisoning in ready-to-eat foods in New Zealand. The survival of a faecal contamination indicator, the laboratory grown strain Escherichia coli NZRM 916, was mapped over eight months. Two strategies to prevent the growth of Listeria monocytogenes in Toroi were also investigated. Only one of the seven pathogens sought was recovered from any sample. This pathogen was Bacillus cereus, a spore-former known to be associated with vegetables. All batches contained B. cereus on the day of preparation but after two weeks refrigerated storage there was no further recovery from any sample. There was a very low incidence of natural E. coli in the Toroi, consistent with levels permitted in mussels sold in retail outlets. The laboratory grown strain, E. coli declined substantially over two months and was not recovered from any samples at eight months. A laboratory grown strain of Listeria monocytogenes, (L70) was added to Toroi and grew well with an increase in concentration of about seven-fold, over 19 days storage in a refrigerator. A bacteriocin producing lactic acid bacterium, Lactobacillus sake Lb706, was added in combined culture with L. monocytogenes to Toroi. It was found that at least 5 x108 L. sake cells were required as an inoculum to ensure elimination of L. monocytogenes from the Toroi. When a purified bacteriocin; nisin, was added, a concentration of 10 mg g-1 in the Toroi was required to eliminate L. monocytogenes. The inhibition study results suggest that unacceptably high inocula or purified bacteriocin would be required to prevent the growth of L. monocytogenes in Toroi. The results of this suggest that Toroi be prepared from mussels either purchased from a retail outlet or harvested from sites known to be free from contamination. Toroi should be safe to eat if prepared carefully, chilled promptly and thoroughly and allowed to ferment for at least two weeks. In addition, care should be taken to maintain Toroi at refrigerated temperatures until it is eaten.

>Magister Scientiae - MSc / Informal settlements are often implicated in surface water pollution with faecal matter. In most instances faecal pollution in the associated surface waters persists despite improvements in sewage removal infrastructure. This study evaluates the importance of investigating the water quality of the Plankenbrug River before it reaches Khayamnandi settlement by comparing water quality in spring and in winter upstream (Pre-Khayamnandi) and downstream (Post- Khayamnandi) from the settlement. In this study, faecal indicator bacteria (Escherichia coli and total coliforms) were enumerated using Chromocult agar. E. coli was further characterized with analytical profiling index (API) and haemolysis assays. Both Pre- and Post-Khayamnandi were not significantly different from each other for both total coliforms and E. coli in winter. Pre-Khayamnandi had between 105 and 108 cfu/100 ml for total coliforms while Post-Khayamnandi had total coliform colony count between 106 and 107 cfu/100 ml. E. coli also exhibited a similar pattern with slightly higher counts at Post-Khayamnandi with colony counts from 104 to 107 and 105 to 107 cfu/100 ml. Spring microbial count demonstrated a significant difference to winter counts within each test site (p ≤ 0.01) and across the two sites (p ≤ 0.05). Both total coliforms and E. coli were 102 fold higher at Post-Khayamnandi than at Pre-Khayamnandi in spring. The API assay demonstrated significant difference (p ≤ 0.05) between the two test sites. Pre- Khayamnandi predominantly had two different profiles while Post-Khayamnandi had three. These profiles represented five distinct E. coli biotypes. Sorbitol and sucrose tests within the API assay demonstrated significant differences (p ≤ 0.05) between the two test sites. The prevalence of sorbitol fermenters at Pre-Khayamnandi was 100% while at Post-Khayamnandi it was 73%. Pre-Khayamnandi also demonstrated a significantly higher prevalence of sucrose fermenters than Post-Khayamnandi at 100% and 59% respectively. These differences indicated dissimilar sources of faecal contamination around these sites. Differences in the distributions of sorbitol and sucrose fermenting biotypes demonstrate different toxicity potentials across these two test sites. The haemolysis assay demonstrated that 9% of isolates were haemolytic with reference to both known α- and β-haemolyitic streptococci at Post-Khayamnandi. At Pre-Khayamnandi there was a higher percentage of α- and β-haemolyitic species, 29% and 28%, respectively. Post- Khayamnandi and Pre-Khayamnandi were significantly different from each other with reference to both α- and β-haemolysis (p ≤ 0.05). These haemolytic activities also demonstrate different toxicity potentials across the two sites. In conclusion Khayamnandi contributes to an already heavy faecal load in the Plankenbrug River. Thus remedial measures to maintain high surface water quality of Plankenbrug River should be directed upstream from the Khayamnandi settlement as well as within the settlement equally. This study recommends integration of microbial loads with programs such as the National Microbial Monitoring Program of South Africa to drive prioritization process in directing reclaiming of water quality, inter alia.

In the year 2000, 170 countries decided to stand together and solve some primary and common global problems like poor health, water, and sanitation. Obtaining access to safe drinking water is every individual’s right. The UN defines safe drinking water as “the water required for each personal or domestic use must be safe, therefore free from micro-organisms, chemical substances and radiological hazards that constitute a threat to a person's health” (UNDESA, 2014). This thesis focuses on identifying the number of people who potentially receive spring water from a piped source in rural mountainous areas. There are three significant steps and data requirements which are necessary to meet this study’s objective. These are to: 1) obtain data classifying the various sources of drinking water in each country by specifying the percentage of population served by a particular water in rural and urban areas, 2) determine the number of countries which are undeveloped or developing, and 3) identify specific countries which are defined as mountainous (and thus likely to have spring fed piped water systems) based on elevation and slope in order to estimate the number of people living in mountainous areas. Results show that 183.54 (million) people were estimated to receive piped water from the springs in mountainous areas. Approximately, 34% of the population is from the Eastern Asia and South-Eastern Asia regions, and 33% reside in the Latin America and Caribbean Islands. These were followed by Western Asia and North Africa Regions with 15% and Central Asia and Southern Asia Regions with 14% of their population estimated to receive piped spring water.

<p>Greywater is domestic household wastewater without inputfrom the toilet, i.e. wastewater from sinks, the shower,washing machine and dishwasher in a home. Source separation ofgreywater can be a strategy to enhance recirculation of plantnutrients and/or improve water use. The risk for transmissionof disease when reusing greywater is largely dependent on thecross-contamination by faeces. High levels of faecalindicators, mainly thermotolerant coliform bacteria, have beenreported in greywater, indicating substantial faecal pollution.However, growth of indicator bacteria within the system leadsto an overestimation of thefaecal input and thus the hygienerisk. The faecal input of the greywater in Vibyåsen,Sollentuna, North of Stockholm, was estimated to be 0.04 ±0.02 g faeces person-1 day-1 from the quantification of thefaecal sterol coprostanol, compared to 65 g, 5.2 g and 0.22 gp-1 d-1 using E. coli, enterococci and cholesterolrespectively.</p><p>Prevalence of pathogens in the population and the faecalload based on coprostanol concentrations were used to form thebasis of a screening-level quantitative microbial riskassessment (QMRA) that was undertaken for rotavirus, Salmonellatyphimurium, Campylobacter jejuni, Giardia intestinalis andCryptosporidium parvum, looking at the treatment required to bebelow an acceptable level of risk (10-3) for reuse or dischargeof the greywater. The different exposure scenarios simulated–groundwater recharge, direct contact, irrigation andrecreational water–showed that a reduction of 0.7–3.7 log was needed for rotavirus, with the measured level offaecal load in Vibyåsen. The other pathogen of concern wasCampylobacter, where a 2.2 log reduction was needed forgroundwater recharge. The infectious dose of Salmonella is highand the excretion numbers of Giardia cysts and Cryptosporidiumoocysts low, resulting in no treatment requirements for theseorganisms under these circumstances. Pathogen input fromcontaminated food via the kitchen sink had a minor effect onthe microbiological quality of the greywater. Studies on virusoccurrence in greywater as well as validation of the faecalload of greywater at another site would give valuable input forfuture QMRAs.</p><p>Greywater treatment efficiency studies, especially on virusremoval, are scarce and more investigations are warranted.Active sludge may not be a suitable technique for greywater dueto the low carbon content in this flow. Chemical precipitationhas the advantage of removing phosphorus as well as virusesefficiently and it is suggested as one possible method fortreating greywater. Otherwise the most common practice forgreywater treatment in Sweden is soil infiltration. However, itis suggested that the recommendations for wastewaterinfiltration also be observed for greywater, despite the lowfaecal load, due to the simulated results on virus reductionneeded.</p><p><b>Key words:</b>greywater, greywater reuse, greywatertreatment, microbial risk assessment, groundwater recharge,irrigation, recreational water, faecal contamination, indicatorbacteria, index organisms, faecal sterols, bacteriophages,enteric pathogens, rotavirus, Salmonella, Campylobacter,Giardia, Cryptosporidium, Legionella</p>

L’objectif général de cette thèse est de caractériser la dynamique des transferts des bactéries fécales à l’aide d’une modélisation spatio-temporelle, à l’échelle du bassin versant (BV) dans une région agricole et à l’échelle événementielle. Ce projet vise à mieux comprendre l'influence des processus hydrologiques, les facteurs environnementaux et temporels impliqués dans l’explication des épisodes de contamination microbienne des eaux récréatives. Premièrement, un modèle bayésien hiérarchique a été développé pour quantifier et cartographier les niveaux de probabilité des eaux à être contaminées par des effluents agricoles, sur la base des données spectrales et des variables géomorphologiques. Par cette méthode, nous avons pu calculer les relations pondérées entre les concentrations d’Escherichia coli et la distribution de l’ensemble des paramètres agro-pédo-climatiques qui régissent sa propagation. Les résultats ont montré que le modèle bayésien développé peut être utilisé en mode prédictif de la contamination microbienne des eaux récréatives. Ce modèle avec un taux de succès de 71 % a mis en évidence le rôle significatif joué par la pluie qui est la cause principale du transport des polluants. Deuxièmement, le modèle bayésien a fait l’objet d'une analyse de sensibilité liée aux paramètres spatiaux, en utilisant les indices de Sobol. Cette démarche a permis (i) la quantification des incertitudes sur les variables pédologiques, d’occupation du sol et de la distance et (2) la propagation de ces incertitudes dans le modèle probabiliste c'est-à-dire le calcul de l’erreur induite dans la sortie par les incertitudes des entrées spatiales. Enfin, une analyse de sensibilité des simulations aux différentes sources d’incertitude a été effectuée pour évaluer la contribution de chaque facteur sur l’incertitude globale en prenant en compte leurs interactions. Il apparaît que sur l’ensemble des scénarios, l’incertitude de la contamination microbienne dépend directement de la variabilité des sols argileux. Les indices de premier ordre de l’analyse de Sobol ont montré que parmi les facteurs les plus susceptibles d’influer la contamination microbienne, la superficie des zones agricoles est le premier facteur important dans l'évaluation du taux de coliformes. C’est donc sur ce paramètre que l’attention devra se porter dans le contexte de prévision d'une contamination microbienne. Ensuite, la deuxième variable la plus importante est la zone urbaine avec des parts de sensibilité d’environ 30 %. Par ailleurs, les estimations des indices totaux sont meilleures que celles des indices de premier ordre, ce qui signifie que l’impact des interactions paramétriques est nettement significatif pour la modélisation de la contamination microbienne Enfin, troisièmement, nous proposons de mettre en œuvre une modélisation de la variabilité temporelle de la contamination microbiologique du bassin versant du lac Massawippi, à partir du modèle AVSWAT. Il s'agit d'une modélisation couplant les composantes temporelles et spatiales qui caractérisent la dynamique des coliformes. La synthèse des principaux résultats démontrent que les concentrations de coliformes dans différents sous-bassins versants se révèlent influencées par l’intensité de pluie. La recherche a également permis de conclure que les meilleures performances en calage sont obtenues au niveau de l'optimisation multi-objective. Les résultats de ces travaux ouvrent des perspectives encourageantes sur le plan opérationnel en fournissant une compréhension globale de la dynamique de la contamination microbienne des eaux de surface. / Abstract : The aim of this study was to predict water faecal contamination from a bayesian probabilistic model, on a watershed scale in a farming area and on a factual scale. This project aims to better understand the influence of hydrological, environmental and temporal factors involved in the explanation of microbial contamination episodes of recreational waters. First, a bayesian probabilistic model: Weight of Evidence was developed to identify and map the probability of water levels to be contaminated by agricultural effluents, on the basis of spectrals data and geomorphologic variables. By this method, we were able to calculate weighted relationships between concentrations of Escherichia coli and distribution of key agronomic, pedologic and climatic parameters that influence the spread of these microorganisms. The results showed that the Bayesian model that was developed can be used as a prediction of microbial contamination of recreational waters. This model, with a success rate of 71%, highlighted the significant role played by the rain, which is the main cause of pollution transport. Secondly, the Bayesian probabilistic model has been the subject of a sensitivity analysis related to spatial parameters, using Sobol indications. This allowed (1) quantification of uncertainties on soil variables, land use and distance and (2) the spread of these uncertainties in the probabilistic model that is to say, the calculation of induced error in the output by the uncertainties of spatial inputs. Lastly, simulation sensitivity analysis to the various sources of uncertainty was performed to assess the contribution of each factor on the overall uncertainty taking into account their interactions. It appears that of all the scenarios, the uncertainty of the microbial contamination is directly dependent on the variability of clay soils. Sobol prime indications analysis showed that among the most likely to influence the microbial factors, the area of farmland is the first important factor in assessing the coliforms. Importance must be given on this parameter in the context of preparation for microbial contamination. Then, the second most important variable is the urban area with sensitivity shares of approximately 30%. Furthermore, estimates of the total indications are better than those of the first order, which means that the impact of parametric interaction is clearly significant for the modeling of microbial contamination. Thirdly, we propose to implement a temporal variability model of microbiological contamination on the watershed of Lake Massawippi, based on the AVSWAT model. This is a model that couples the temporal and spatial components that characterize the dynamics of coliforms. The synthesis of the main results shows that concentrations of Escherichia coli in different sub-watersheds are influenced by rain intensity. Research also concluded that best performance is obtained by multi-objective optimization. The results of these studies show the prospective of operationally providing a comprehensive understanding of the dynamics of microbial contamination of surface water.

Poor water quality is a global health concern affecting one billion people around the world. It is important to monitor water sources in order to maintain the quality of our drinking water and to avoid disease outbreaks. Targeting Escherichia coli as a faecal indicator is a widely used procedure, but the current methods are time consuming and not adequate to prevent spreading of faecal influence.   This Master thesis demonstrates the development of a near infrared fluorescence flow cytometer sensor system targeting Escherichia coli, using fluorescently labeled chicken IgY antibodies. The near infrared light was chosen to avoid fluorescence from blue-green algae that are present in the water source.   The hardware was developed with a 785  nm laser line to detect Alexa Fluor 790 labeled antibodies, using a photomultiplier tube or two different CMOS cameras. The antibodies were labeled using a commercial labeling kit, and evaluated using antibody binding assays and the developed hardware.   The IgY antibodies were successfully labeled with Alexa Fluor 790 and the function was maintained after the labeling process. The result demonstrates the principles of the sensor system and how it solved to the problem with fluorescence from blue-green algae. An aperture was used to overcome the suboptimal laser and filter setup, and to increase the sensitivity of the system. However, only a small fraction of the cells could be detected, due to challenges with the focal depth and loss of sensitivity in the photomultiplier tube at near infrared wavelengths. Further development is required to create a working product.

Greywater is domestic household wastewater without inputfrom the toilet, i.e. wastewater from sinks, the shower,washing machine and dishwasher in a home. Source separation ofgreywater can be a strategy to enhance recirculation of plantnutrients and/or improve water use. The risk for transmissionof disease when reusing greywater is largely dependent on thecross-contamination by faeces. High levels of faecalindicators, mainly thermotolerant coliform bacteria, have beenreported in greywater, indicating substantial faecal pollution.However, growth of indicator bacteria within the system leadsto an overestimation of thefaecal input and thus the hygienerisk. The faecal input of the greywater in Vibyåsen,Sollentuna, North of Stockholm, was estimated to be 0.04 ±0.02 g faeces person-1 day-1 from the quantification of thefaecal sterol coprostanol, compared to 65 g, 5.2 g and 0.22 gp-1 d-1 using E. coli, enterococci and cholesterolrespectively. Prevalence of pathogens in the population and the faecalload based on coprostanol concentrations were used to form thebasis of a screening-level quantitative microbial riskassessment (QMRA) that was undertaken for rotavirus, Salmonellatyphimurium, Campylobacter jejuni, Giardia intestinalis andCryptosporidium parvum, looking at the treatment required to bebelow an acceptable level of risk (10-3) for reuse or dischargeof the greywater. The different exposure scenarios simulated–groundwater recharge, direct contact, irrigation andrecreational water–showed that a reduction of 0.7–3.7 log was needed for rotavirus, with the measured level offaecal load in Vibyåsen. The other pathogen of concern wasCampylobacter, where a 2.2 log reduction was needed forgroundwater recharge. The infectious dose of Salmonella is highand the excretion numbers of Giardia cysts and Cryptosporidiumoocysts low, resulting in no treatment requirements for theseorganisms under these circumstances. Pathogen input fromcontaminated food via the kitchen sink had a minor effect onthe microbiological quality of the greywater. Studies on virusoccurrence in greywater as well as validation of the faecalload of greywater at another site would give valuable input forfuture QMRAs. Greywater treatment efficiency studies, especially on virusremoval, are scarce and more investigations are warranted.Active sludge may not be a suitable technique for greywater dueto the low carbon content in this flow. Chemical precipitationhas the advantage of removing phosphorus as well as virusesefficiently and it is suggested as one possible method fortreating greywater. Otherwise the most common practice forgreywater treatment in Sweden is soil infiltration. However, itis suggested that the recommendations for wastewaterinfiltration also be observed for greywater, despite the lowfaecal load, due to the simulated results on virus reductionneeded. <b>Key words:</b>greywater, greywater reuse, greywatertreatment, microbial risk assessment, groundwater recharge,irrigation, recreational water, faecal contamination, indicatorbacteria, index organisms, faecal sterols, bacteriophages,enteric pathogens, rotavirus, Salmonella, Campylobacter,Giardia, Cryptosporidium, Legionella / NR 20140805

Thesis (MSc Food Sc (Food Science))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: Agriculture in the Western Cape is not only one of the most important economic sectors but also provides many job opportunities. Over the last few years the sustainability of this successful industry has become threatened by the faecal pollution of rivers used to irrigate produce that will be consumed raw or after minimal processing. This situation not only poses an enormous risk to the health of the consumer but also to farmers who stand to lose their export licenses. The purpose of this study was to determine the microbial types and loads in river water, irrigation water and on irrigated produce. A baseline study was done on four sites in two Western Cape rivers. These sites were chosen to allow for the sampling of river water, irrigation water and irrigated produce so as to determine whether a link between the use of contaminated irrigation water and the microbial population found on irrigated produce exists. The physico-chemical analyses used in the study consisted of: pH, alkalinity, water temperature, conductivity and chemical oxygen demand. The microbial monitoring included the aerobic colony counts (ACC) and the enumeration of the total coliforms, faecal coliforms, staphylococci, enterococci, and aerobic and anaerobic sporeformers present in the water samples. The presence or absence of the potential pathogens like E. coli, Listeria and Salmonella, was also determined. During the baseline study faecal coliform counts as high as 160 000 organisms.100 mL-1 were noted in the Plankenburg River, while counts as high as 460 000 organisms.100 mL-1 were found in the Mosselbank River. Apart from this, high numbers of staphylococci and intestinal enterococci were often found, while E. coli, Listeria and Salmonella were present in samples from both of these rivers. Based on the results of the baseline study on the two rivers it was decided to do a more intensive study on the microbial load of the river and irrigation water as well as irrigated produce from the Mosselbank site. Lettuce and cabbages from a commercial farmer’s fields were chosen as the irrigated produce. During the warmer summer months, ACC counts in the river samples peaked at 12 8000 000 cfu.mL-1, while faecal coliform counts of 1 600 000 organisms.100 mL-1 were found. The three potential pathogens (E. coli, Listeria and Salmonella) were present in all the river samples taken during this period. While the counts of indicator bacteria in the irrigation water was often lower, faecal coliform counts as high as 1 600 000 organisms.100 mL-1 and several other potential pathogens were found on the irrigated lettuce and cabbage. This could indicate a possible “build-up” of contamination on the produce with the repeated application of the tainted irrigation water. According to guidelines published by DWAF in 2008, water to be used for irrigation should not contain more than 4 000 organisms.100 mL-1 faecal coliforms if it is used for the irrigation of crops that are to be consumed raw or after a minimal processing step, as this would increase the health risk to the consumer. Guidelines published by the South African Department of Health are even stricter and state that raw vegetables and fruit should not contain more than 200 coliform organisms per gram, while E. coli and L. monocytogenes should be absent in one gram, and Salmonella spp. in 25 grams of the produce, respectively. From the data obtained during this study it was evident that the two rivers monitored regularly contained faecal indicators at levels much higher than those proposed in national and international guidelines for safe irrigation, making them unfit for the irrigation of MPF’s. It could be concluded that the rivers investigated during this study contained high levels of faecal contamination. Since some of the pathogens isolated from the river and irrigation water and the irrigated produce, it suggests a carry-over of microbial contamination from the river water to the irrigated produce. This was, however, only done using the traditional international methods and the presence of specific pathogens should in future be confirmed by means of molecular techniques. / AFRIKAANSE OPSOMMING: Landbou is nie net die een van die belangrikste ekonomiese sektore in die Wes-Kaap nie, maar verskaf ook vele werksgeleenthede. Oor die afgelope paar jaar word die volhoubaarheid van hierdie suksesvolle industrie egter bedreig deur die fekale kontaminasie van riviere wat gebruik word vir die besproeiing van voedsel wat rou of na ‘n minimale prosesserings stap ingeneem word. Hierdie situasie hou nie net ‘n groot gevaar vir die gesondheid van verbruikers in nie, maar ook vir boere wat hul uitvoerlisensies hierdeur kan verloor. Die doel van hierdie studie was om die ladings en tipes mikrobes in rivier water, besproeiingswater en op besproeide produkte vas te stel. ‘n Basiese studie van vier liggings in twee Wes-Kaapse riviere is gedoen. Hierdie liggings is só gekies dat dit moontlik was om die rivier water, besproeiingswater en die besproeide produkte te monitor, en daar sodoende vasgestel kon word of daar ‘n verhouding is tussen die gebruik van gekontamineerde besproeiingswater en die mikrobe populasie wat op die besproeide produkte aanwesig was. Die fisiko-chemiese analises wat gedurende die studie gedoen is, het pH, alkaliniteit, water temperatuur, geleidingsvermoë en die chemiese suurstof vereiste (COD) ingesluit. Die mikrobiese analises het die aërobe kolonie tellings (ACC) en die enumerasie van die totale kolivorme, fekale kolivorme, staphylococci, enterococci en die aërobe en anaërobe spoorvormers ingesluit. Daar is ook vir die aanwesigheid van potensiële patogene soos E. coli, Listeria en Salmonella getoets. Gedurende die basiese studie is fekale kolovorme tellings van so hoog as 160 000 organismes.100mL-1 in die Plankenburg Rivier aangeteken, terwyl tellings van so hoog as 460 000 organismes.100mL-1 in die Mosselbank Rivier gevind is. Hoë tellings stafielokokki en intestinale enterokokki is gereeld genoteer, terwyl E.coli, Listeria en Salmonella uit die waters van beide hierdie riviere geïsoleer is. Gebaseer op hierdie resultate is daar besluit om ‘n meer intensiewe studie van die rivier, besproeiingswater en die besproeide produkte van die Mosselbank Rivier te doen. Blaarslaai en kool van ‘n kommersiële boer se lande is vir hierdie doel gekies. Gedurende die warmer somer maande het die aërobe kolonie tellings in die rivier ‘n piek van 12 800 000 kve.mL-1 bereik, terwyl fekale kolivorme tellings van 1 600 000 organismes.100mL-1 genoteer is. Die drie potensiële patogene (E. coli, Listeria en Salmonella) was aanwesig in al die monsters wat gedurende hierdie tydperk van die rivierwater geneem is. Alhoewel die tellings indikator bakterieë in die besproeiingswater meestal laag was, is tellings fekale kolivorme van so hoog as 1 600 000 kve.100mL-1 en verskeie ander potensiële patogene op die besproeide blaarslaai en kool gevind. Dit kan dui op ‘n moontlike opbou van kontaminasie op die produkte met die herhaalde besproeiing met gekontamineerde besproeiingswater. Volgens die riglyne wat in 2008 deur DWAF gepubliseer is, mag water wat vir die besproeiing van minimaal geprosesseerdevoedsels gebruik word nie meer as 4 000 organismes.100mL-1 bevat nie, aangesien dit die gesondheid van die gebruiker in gevaar mag stel. Die riglyne van die Suid-Afrikaanse Departement van Gesondheid is selfs strenger en beveel aan dat rou vrugte en groente nie meer as 200 kolivorme en geen L. monocytogenes per gram, en geen Salmonella spp. in 25 g van die produk mag bevat nie. Vanuit die data wat tydens hierdie studie ingesamel is, is dit duidelik dat die twee riviere gereeld fekale indikators bevat het teen vlakke baie hoër as wat in die nasionale en internasionale riglyne aanbeveel word. Hierdie water is dus nie geskik vir die besproeiing van minimaal geprosesseerde produkte nie. Die afleiding kan gemaak word dat die riviere wat tydens hierdie studie gemonitor is, hoë vlakke van fekale kontaminasie bevat het. Aangesien sommige van die patogene vanuit beide die rivier- en besproeiingswater, en vanaf die besproeide produkte geïsoleer is, kan dit dui op ‘n moontlike oordrag van mikrobiese kontaminasie vanuit die rivierwater na die besproeide produkte. Tydens hierdie studie is daar egter net van die tradisionele internasionale metodes gebruik gemaak. Vir toekomstige navorsing word dit aanbeveel dat die aanwesigheid van die spesifieke patogene deur die gebruik van molekulêre metodes bevestig word.