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Thesis (M. Tech. Environmental health) -- Central University of technology, Free State, 2011 / In nature microorganisms do not exist alone, but in association with one another. These kinds of associations can also be found in food industries, where cells of the same or different species can attach to pipes (biofilm formation) and a variety of surfaces in food processing environments and in food product such as yoghurt which can contain both yeast and bacteria originating from the starter culture as well as fruit. To control food spoilage organisms and food-borne pathogens preventative measures such as good manufacturing processes, the use of sanitizers and preservatives as well as hazard analysis critical control points (HACCP) are crucial in food industries. Sanitation of the working surface, floors, pipes, containers and equipment is a stepwise application of a detergent, acid or alkali rinse, a disinfectant treatment followed by final rinsing. If rinsing of the sanitizer is not done properly it may end up in the product in sub-lethal doses. In this study the influence of Liquid Hypochlorite (LH) and Liquid Iodophore (LI) sanitizers on organism growth and toxicity was evaluated. The organisms investigated included Escherichia coli 0113, Escherichia coli 026 and Zygosaccharomyces bailii Y-1535 in yeast malt broth, which was supplemented with LH and LI at sub-lethal concentrations 0.05% LH, 0.2% LH and 0.075% LI. Subsequently, bacterial and yeast growth responses as pure cultures and in combination (E. coli + Z. bailii) were measured as colony forming units and optical density values. Incorporation of the sanitizers in the growth media resulted in different levels of growth inhibition. Z. bailii proved more robust and the growth rate was not influence significantly by the addition of sanitizers or communal growth with either E. coli strains. The growth rate of both E. coli strains decreased where grown in combination with Z. bailii as well as in the presence of sanitizers, with the most influence exerted by LH. Changes in endotoxicity following the growth of the test samples (stressed cells) and the control (unstressed) were measured by the limulus amoebocyte lysate (LAL) and porcine IL-6 ELISA methods. Where E. coli strains were cultured together with Z. bailii the toxicity of tire mixture showed a decrease over time when measured with the limulus amoebocyte assay method. Interestingly the communal growth of the E. coli strains and Z bailii produced different toxicity profiles when the IL-6 porcine method was used, hi both cases, where E. coli strains were cultured together with Z. bailii the toxicity of the mixture showed an increase over tune when measured by this assay. Other than a similar toxicity profile for E. coli 0113 grown in pure culture, the comparison between results obtained using the LAL or porcine IL-6 methods yielded no correlation in determined toxicity. It was established that LH and LI sanitizers as well as communal growth had an influence in the toxicity of LPS/EPS and the method used to determine such toxicity should be carefully considered.

Thesis (D. Tech. Environmental health) -- Central University of technology, Free State, 2011 / Apart from being the primary food source of many cultures around the world, fish contains notable amounts of essential fatty acids that are required by the human body, thus making fish a vital part of the human diet. In South Africa Cape hake is a well-known and highly consumed local fish species, which is transported from coastal areas countrywide where the fresh fish are displayed on ice in various retail stores. Fish is known to be highly susceptible to spoilage and, as a result, the maintenance of the cold-chain in related products is of particular importance. Additionally, recent trends showing a decline in natural fish resources have instigated growing concerns about the sustainability and optimal utilisation of fish as a food source. Against this backdrop, this study aimed at determining the influence of storage parameters on selected triglycerides and their possible metabolic pathways. Also applying prediction modelling of fatty acids and volatiles as instruments to assess exposure of Cape hake fillets to excessive microbial contamination and, in effect, be indicative of the environmental parameters (for example temperature) that may influence such contamination. Randomly selected juvenile hakes were filleted and stored under various simulated retail storage conditions, under either controlled or uncontrolled environmental conditions. For each hake filleted, one fillet was inoculated with an increased load of autochthonous microbiota, and the corresponding fillet was kept at similar temperature conditions. All fillets were monitored over a ten day period, during which fatty acid and volatile samples were collected and analysed. From the resulting triglycerides a selection of fatty acids were profiled and their possible metabolic pathways investigated. Fish maturity, the distribution of the fatty acids and the implication thereof in the nutritional value were also assessed. Conventional chemometric methods utilising mathematical expressions were subsequently utilised in order to predict contamination and whether the cold chain was sustained, while an artificial neural network (ANNs) were designed to predict excessive microbial contamination in the fillets. The results showed that the nutritional value of fish differs notably with its maturity and size. Mathematical equations were furthermore found to be effective assessment instruments to indicate the percentage differences in storage temperature, as well as consequent microbial influences. Thus, this approach may introduce mathematical prediction modelling as a promising mechanism to assess Cape hake spoilage. An artificial neural network (ANN) was successfully designed, that succeeded in distinguishing between Cape hake fillets displayed and stored on ice that have been exposed to excessive contamination and those that have not been exposed. In the latter case, the selected variable was a fatty acid, hexadecanoic acid, used as biochemical indicator. This modulating approach may provide a platform for future shelf-life studies on related muscle tissue. Ultimately, the study endeavoured to add to the body of knowledge regarding the biochemical and microbiological changes related to Cape hake storage, the prediction thereof via contemporary methods and contributing to the safety and effective utilization of this unique and declining South African nutritional resource.

Thesis (M. Tech. (Environmental health: Food safety )) - Central university of Technology, Free State, 2013 / Food processing plants and agricultural environments have a long-standing history of being known to provide a conducive environment for the prevalence and distribution of microorganisms which emanate as a consequence of activities undertaken in such premises. Microorganisms in the aforementioned environments may be found in the atmosphere (airborne), and/or on food contact surfaces. Airborne microorganisms from food handlers and in food products and raw materials (as part of bioaerosols) have in the past been implicated as having a potential to cause adverse health effects (especially in indoor environments) and therefore also to have economic implications. Recently their effect on food safety has received increased interest. The recent international interest in bioaerosols in the food industry has played a role in rapidly providing increased understanding of bioaerosols and their effects in different food processing environments. However, there is still a lack of research on the actual impact of bioaerosols over time in most of the food premises especially in Southern Africa and other developing countries. The overall purpose of this dissertation was to assess possible microbial contaminants and the role of selected environmental parameters on these microbes at a dairy farm plant in central South Africa. In relation to the purpose of the study, the objectives of this dissertation were to investigate and establish the food handler’s food safety knowledge, attitude, behaviour and practices. The sub-objective was to investigate the prevalence and distribution of microbial contaminants (both airborne and food contact surface populations), and concomitant environmental parameters. The microbe isolates from both investigations (i.e. air samples and food contact surfaces) were identified to strain level using matrix-assisted laser desorption ionization – time of flight mass spectrometry (MALDI-TOF MS). The findings of this study in relation to food handlers’ food safety knowledge, attitude, behaviour and practices indicated a dire need for training of employees as well as improved health and hygiene measures as emphasised by some of the identified strains. The environmental parameters (both indoor and outdoor) were similar, with no relationship established between airborne microbes’ prevalence and environmental parameters. The samples of the airborne microbial populations in both indoor and outdoor environments were similar. Airborne microbial counts at the dairy farm plant over the entire duration of the study ranged between 1.50 x 101cfu.m-3and 1.62 x 102cfu.m-3. Microbial counts on food contact surfaces ranged between 2.50 x 102 cfu.cm-2 and 1.10 x 105 cfu.cm-2 over the entire duration of the study. A wide variety of microorganisms (from air and food contact surfaces) such as the Gram-positive bacteria, Gram-negative bacteria, as well as fungi were present at the dairy farm plant. A number of the isolated genera have previously been associated with agricultural environments whilst others are associated with hospital environments. The positively identified strains were from genera such as Aeromonas, Arthrobacter, Candida, Pseudomonas, Pantoea, Citrobacter, Staphylococcus, Bacillus, Escherichia, Rhodococcus and Rhodotorula, amongst others. The isolation of microorganisms associated with food spoilage and foodborne disease outbreaks, which are known as indicator organisms such as Escherichia coli, Staphylococcus and Bacillus from both air and surface samples, signified possible faecal contamination and could be attributed to poor health and hygiene practices at the dairy farm plant. Despite the isolation of microorganisms associated with food spoilage and foodborne disease outbreaks, the isolation of microorganisms not usually associated with the food processing industry (usually associated with hospital environments) was an enormous and serious concern which suggested a need for further investigations at dairy farm plants as the implications of these pathogenic microorganisms in food is not known. The isolation of similar microorganisms from both the air samples and surface swabs suggests that airborne microbes have a potential of settling on food contact surfaces, therefore having a potential to contaminate dairy products which are known to be more prone to contamination and which, because of their nutritional status, serve as a good substrate for the growth of microorganisms.

Thesis (M. Tech. (Environmental Health)) - Central University of Technology, Free State, 2014 / In the battle to sustain and produce quality food that is safe and affordable, the limited legislative and regulatory environment continues to allow opportunities for food to become contaminated during processing. The degree of contamination distributed over the final food product (including meat products) depends upon several factors that include knowledge and behaviour of the food handlers, equipment, the hygiene habits of personnel, and the monitoring that takes place at food processing plants (including butcheries). The current study was conducted in five selected butcheries (forming 15% of the registered butcheries at the time the study was conducted) in the Mangaung Metropolitan municipal area, purposely targeting the ones registered with the municipality. The hygiene practices of meat handlers were assessed (through self-administered questionnaires) because meat is a perishable product that requires labour intensive processing for production of quality products. Thus, mishandling by food handlers may create and maintain conditions favourable to microbial contamination. Furthermore, the study assessed and characterised microbial contamination on working surfaces and utensils through swabs as well as bioluminescence instrument [Adenosine Tri-phosphate (ATP) Hygiena] for cleanness of the working environment. Concomitant to the above, meat handlers’ hands and aprons were also assessed for possible microbial contamination as well as their characterisation. Lastly, aerosolised microbes [through an air sampler (Surface Air System) SAS Super 90] were also collected for iv quantification and identification during working hours as airborne microbes can settle on working surfaces and/or utensils as a result of movement of workers and other related working processes. Statistical points such as correlations, standard deviations, group standard deviations as well as significant differences were captured per respective chapter where necessary. Data reported in this study is over 3 month period with two weeks intervals during sampling and thus reported as either weekly or rounds between sampling periods. The results of the current study indicate that the food safety objectives are negligibly achieved, indicating a need for proper food safety training which is audit based. On administration of a questionnaire, food handlers showed poor knowledge of food safety awareness coupled with poor attitude and behaviour in terms of food safety. The five butchery premises were further examined regarding the airborne and surface microbial loads, as well as that of the food handlers’ hands, during processing. The microbial loads in the air appeared to comply with the suggested limits at all the sampled butcheries. Microbial loads on meat contact surfaces showed levels conforming to the South African standard or guideline of 1 × 102 cfu.m-2. Total Coliforms on hands and on aprons were compared to the general microbial target value of <2.5 cfu.m-2 as suggested by literature. In this study, Matrix Laser Desorption Time of Flight Mass Spectrophotometer (MALDI-TOF MS) was found to be an accurate, rapid and cost effective method towards v identifying of foodborne pathogens and spoilage bacteria including yeast. Moreover, in recent years South Africa’s meat scandals have increased consumer awareness and the demand for food safety. Section 11 of the Meat Safety Act (Act no. 40 of 2000) stipulates that every abattoir must utilize an independent inspection service appointed by the department of agriculture to ensure that meat of high quality and wholesomeness is produced. However, once the meat and meat products leave the abattoir, they are under the jurisdiction of the local authorities who rely only on visual assessment as opposed to microbiological inspection in the maintenance of their hygiene and quality. Despite the high incidence of foodborne illnesses in both developed and developing countries; South African data on foodborne illness incidents is still insufficient. This could be attributed to the fact that in South Africa, legislation governing the acceptable standards of the levels of microbiota in the air and on food handlers’ hands is still inadequate. Additionally, lack of obligatory usage of Hazard Analysis Critical Control Point (HACCP) procedures in the meat premises poses a risk for economic productivity. In conclusion, the identification of airborne bacteria in the butcheries strongly suggests that in the planning of the existing establishments, the building layout, control of the traffic flow of personnel, the durability and imperviousness of floors, the ventilation system and the placement of the equipment were not carefully considered. This may play a role in the prevalence and proliferation of airborne microbes as the resulting establishments provide an environment conducive to the breeding of microbes. vi In regard to swabs, it was concluded that floors may present a high point of contamination possibly through aerosolization of microbial communities. Moreover, cleaning materials and hygiene practices need to be reviewed. The results of the administered questionnaire showed that food handlers should be sufficiently trained with regard to food quality management tools such as Good Manufacturing Practices (GMP), Hazard Analysis and Critical Control Point (HACCP) systems and food safety. The evaluation of meat contact surfaces for organic soils to determine their cleanliness using the rapid ATP bioluminescence testing can be convenient for everyone involved in the food chain since visual and touch inspection cannot be conclusive enough to meet regulatory requirements in terms of microbial counts.

Thesis (MScMedSc (Pathology. Medical Microbiology))--University of Stellenbosch, 2009. / Water is used for various purposes and contamination can have severe implications if untreated. One of the most common and cost effective water disinfectants, especially used in developing countries, is chlorine. However, microorganisms have developed different mechanisms in response to environmental stress conditions, such as the viable but nonculturable (VBNC) effects possibly displayed in this study, enabling them to survive. Chlorine may also exert several effects on microorganisms, such as the expression of multi-substrate efflux pumps, decreased membrane permeability and transport inhibition that may lead to chlorine tolerance and antimicrobial resistance. In a descriptive and comparative study, the molecular characteristics of E. coli strains isolated from environmental waters in the Western Cape and the possible relationship between chlorination and antimicrobial resistance were investigated. Water and biofilm samples were exposed to chlorine, as well as efflux pump inhibitor (EPI) concentrations, and surviving E. coli strains were tested for their phenotypic characteristics including antimicrobial susceptibility profiles and morphological types. Candidate genes possibly involved in resistance to antimicrobials, disinfection and efflux pumps were detected with polymerase chain reaction (PCR) and sequenced. Sequencing analysis and homology searches were done and E. coli strains were typed as either Enteropathogenic E. coli strains (EPEC) or Enterotoxigenic E. coli strains (ETEC) on the presence of virulence genes. All water and biofilm sources examined were heavily polluted with E. coli, and a high enumeration level of this indicator organism of faecal contamination was recorded. Chlorine tolerance was found to be associated with antimicrobial resistance. Addition of EPI with exposure to chlorine decreased enumeration levels of these organisms, suggesting that efflux pumps may play a role in tolerance to chlorine. Several morphological patterns were described amongst the E. coli strains and a change in this was recorded after exposure to chlorine. Highly resistant antibiograms displayed by the isolated strains included ampC β-lactamase producing E. coli strains and extended spectrum β-lactamases (ESBLs). Amplification of the candidate genes selected for heatshock, oxidative stress genes and efflux pump were most frequently detected while the structural genes involved in fluoroquinolones (FQs) resistance were detected less frequently in the selected strains. Sequencing of these amplified candidate genes demonstrated various changes in amino acid sequences, including one common mutational pathway taken by E. coli when exposed to stress conditions. Further homology searches of the sequenced candidate genes illustrated similarities in 19 pathogenic and 14 non-pathogenic E. coli as well as 3 Shigella strains. Detection of virulence genes found three EPEC strains (bfpA, eaeA), two EPEC (eaeA), ten EPEC (bfpA) and one ETEC strain (st) amongst the isolates. This study underlines the need for monitoring our water sources, which poses a public health risk due to incomplete chlorination, antimicrobial resistance and the spread of clinically relevant pathogenic strains.

Made available in DSpace on 2017-07-21T20:42:41Z (GMT). No. of bitstreams: 1 Gabriele Scheidt.pdf: 1730831 bytes, checksum: 7a5558c7626aa9889fcd9887e31fdb85 (MD5) Previous issue date: 2013-02-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This work consisted of modifying the surface of commercial glass using copper/copper oxide thin films to obtain bactericidal properties, that is, a thin film that showed a decrease in bacterial contamination either by bactericidal characteristics or inhibition of biofilm adhesion. The methodology used for film synthesis requires a certain time at elevated pressures at a temperature below the glass transition temperature (Tg). The conditions for thin film synthesis were the temperature of 320° C at constant pressure of 3 MPa during 32 h. The thin films were characterized by the techniques of X-ray diffraction and scanning electron microscopy. For biological assays, the bacteria Acinetobacter sp. was used by a standard method of immersion in Luria Broth with stages of dilution and cultured in a Petri dish, by a process of dripping broth on the surface of that blades, followed by Gram staining. The methodology of growth in broth did not allowed to evaluate the efficiency of thin film copper/copper oxide as bacteriological agent. This result is probably due to the dilution of biocidal particles in relation to the volume of the liquid culture media; consequently, a reduction in the bioactivity was observed. On the other hand, tests on slides showed that bacterial films did not adhere to the surface with copper thin films, as compared to slides without, showing effectiveness in combating biofilm adhesion that are considered one of the major means of bacterial contamination. / Este trabalho consistiu em modificar a superfície de vidros comerciais por meio da deposição de filmes finos de cobre / óxido de cobre para fins antibacterianos. Ou seja, que apresentesse redução nos índices de contaminação bacteriana, por características bactericidas ou até mesmo pela inibição da adesão de biofilmes. A metodologia utilizada para a síntese dos filmes finos utiliza o efeito da elevada pressão durante determinado tempo em temperatura abaixo da temperatura de transição vítrea (Tg). As condições de síntese dos filmes finos foram: a temperatura de 320 °C, a pressão constante de 3 MPa durante o período de 32 horas. Os filmes finos foram caracterizados pelas técnicas de difração de raios X e de microscopia eletrônica de varredura. Para os testes antibacterianos, utilizou-se como modelo experimental à bactéria Acinetobacter sp pela sua capacidade de formar biofilme. Na qual foi cultivada em caldo Luria e em laminas planas após contato com vidros com e sem filmes. A metodologia de crescimento em caldo não foi eficiente para avaliar a eficiência do filme fino de cobre/óxido de cobre como agente antibacteriano. Estes resultados provavelmente são devidos as diluição das partículas biocidas em relação ao volume do meio de cultura líquido, fazendo com que a bioatividade seja diminuída. No entanto, nos testes em lâminas planas, as fotos por microscopia óptica mostram que os biofilmes não aderiram na superfície com filmes finos de cobre quando comparados com o controle, lâminas de vidro sem filmes, sendo portanto, eficiênte no combate da adesão de biofilme, considerados um dos principais meios de contaminação bacteriana.

Thèse numérisée par la Division de la gestion de documents et des archives de l'Université de Montréal

Reliable access to safe drinking water is one necessity for humans to live without concern for major health risks. The overall goal of this research is to improve the public health, through improved drinking water, for communities in the Rakai District in Uganda, directly, and other communities in the world, indirectly, via dissemination of knowledge. This study specifically assessed the knowledge of drinking water quality in regards to public health, their sanitation measures, and water treatment methods for users of Brick by Brick rainwater harvesting tanks in the Rakai District (N = 28) by using a knowledge, attitudes, and practice survey and a sanitary inspection; tested the water quality of the Brick by Brick rainwater harvesting tanks (N = 33) in the Rakai District for physical, chemical, and microbial parameters; and piloted a sustainable treatment technology called the chulli system that uses excess heat from a cookstove to treat water. Twenty of the participants identified contaminated water as a cause of diarrheal disease (N = 28). Participants perceived boiling (1), chlorine (2), and filtering (3) as the best three methods of treating water. The average score for the sanitary inspection was 2.27±2.31, which falls between the low and medium expected risk score categories. Fourteen of the thirty-three samples showed detectable levels of colony forming units for coliforms, and two of the thirty-three samples showed detectable levels of colony forming units for E. coli. A demonstration chulli system was constructed for St. Andrew’s Primary School in Rakai District and operated successfully. The research supports that the chulli system along with proper sanitation measures identified in the sanitary inspections can be a sustainable option for users of Brick by Brick rainwater harvesting tanks in the Rakai District.