Esta pesquisa, para fins de determinação do comportamento de microrganismos termotolerantes em águas de aqüífero freático, foi realizada em terrenos aluviais na ETE de Vila dos Remédios, Salesópolis - SP. No local foram instalados uma vala de infiltração para esgoto doméstico bruto e 55 poços de monitoração da água subterrânea, de modo a caracterizar a pluma de contaminação em seu deslocamento ao longo da área ensaiada. Seguindo as normas técnicas de coleta e análise físico-químicabacteriológica das águas subterrâneas foram executadas, com o intuito de conhecimento específico do local de pesquisa, 55 poços com amostragem e caracterização litológica do solo local até profundidade de 2,5m, prospecção geofísica eletrorresistiva de caracterização inicial e eletromagnética (EM-31) de acompanhamento do deslocamento da pluma de contaminação, amostragem e caracterização físico-química-bacteriológica do esgoto bruto e das águas subterrâneas. Dois mil litros de esgoto bruto, coletado diretamente da rede, foi introduzido no solo através de vala de infiltração, de modo a caracterizar uma injeção pontual, sem transbordamento e de maneira contínua, com uma taxa de infiltração de aproximadamente 80L/h. O deslocamento da pluma de contaminação formada no aqüífero freático saturado foi monitorado através de amostras de água coletadas sistematicamente, ao longo do tempo, cujos resultados permitiram calcular a velocidade de deslocamento da pluma, como sendo de 8,6 x 10-4 cm/s e, um decaimento de organismos termotolerantes em ambiente de aqüífero saturado de 21dias. Estes resultados permitem o cálculo direto de perímetros de proteção aplicáveis a poços e fontes naturais de captação de água subterrânea, no caso de possibilidades de contaminação por esgoto domiciliar, neste contexto litológico. / The objective of this research was to study the behavior of thermo-tolerant microorganisms in the phreatic aquifer, in alluvial terrains where the sewage was disposed. The study area was close to the sewage treatment plant of Vila dos Remédios, Salesópolis, SP. An injection trench and 55 monitoring wells were installed in the area. The sewage was discharged in the trench and samples of the groundwater were taken to study the contamination plume migration and its physic-chemical and bacteriological characteristics through the time. The wells were constructed with 2.0 meters of depth and the samples were used to characterize the lithological setting. An initial eletroresistivity survey was performed and, together with the groundwater sampling, electromagnetic survey (EM-31) was periodically conducted to evaluate the plume migration. Two thousand liters of domestic sewage \"in natura\" was introduced in the trench to characterize a punctual injection, with a continuous infiltration rate of 80L/h. The migration of the contamination plume was continuously monitored during nine months and the results allowed calculating the migration speed as being 7,67x10-4 cm/s. The thermo-tolerant microorganisms were extinguished 21 days after the sewage injection. These results allow calculating the protection perimeter applied to groundwater wells and natural springs to avoid sewage contamination in the same lithological context.

Este trabalho avaliou a ocorrência e o transporte de microrganismos no aqüífero freático do cemitério de Vila Nova Cachoeirinha, localizado em terrenos pré-cambrianos, zona norte do município de São Paulo. A metodologia aplicada foi dividida em etapas de laboratório e de campo. No laboratório, foram montadas colunas de solo do cemitério. Traçadores químico e biológico foram injetados nas colunas e o seu fluxo monitorado no efluente. Um modelo numérico foi usado para simular o transporte dos traçadores nas colunas. Em campo, foram realizadas investigações a fim de caracterizar o aqüífero freático. A monitoração da qualidade das águas foi realizada para estudar a ocorrência e o transporte de elementos químicos, bactérias e vírus nas águas subterrâneas. No cemitério, o embasamento está a cerca de 9,0 m de profundidade na cota mais baixa e 20,5 m no topo. O nível freático encontra-se entre 4 e mais de 16 m. O solo do cemitério é formado pelo material de alteração das rochas graníticas, de caráter predominantemente argiloso (~43% de argila), pH =5,0, matéria orgânica entre 0,7 e 4,2% e capacidade de troca de cátions entre 10,2 e 109,0 mmolc/kg. A condutividade hidráulica do aqüífero varia de 2,90 x 10-8 a 8,41 x 10-5 m/s. O gradiente hidráulico na porção oeste do cemitério é de aproximadamente 0,07 m/m; considerando o meio homogêneo e isotrópico e uma porosidade efetiva de 2%, a velocidade linear média foi estimada em 8 cm/dia. As amostras de água do aqüífero freático do cemitério de Vila Nova Cachoeirinha apresentaram, principalmente, bactérias heterotróficas (53 x 103 UFC/mL), bactérias proteolíticas (31 NMP/100 mL) e clostrídios sulfito-redutores (45 NMP/100 mL). Também foram encontrados enterovírus e adenovírus nas amostras. As principais fontes de contaminação das águas subterrâneas no cemitério são as sepulturas com menos de um ano, localizadas nas cotas mais baixas, próximas ao nível freático. Nestes locais, é maior a ocorrência de bactérias em geral. Há um grande consumo do oxigênio existente nas águas. As sepulturas ainda provocam um acréscimo na quantidade de sais minerais, aumentando a condutividade elétrica destas águas. Parece haver um aumento na concentração dos íons maiores bicarbonato, cloreto, sódio e cálcio, e dos metais ferro, alumínio, chumbo e zinco nas águas próximas de sepulturas. As bactérias são transportadas poucos metros, diminuindo em concentração com o aumento da distância à fonte de contaminação. Os vírus parecem ter uma mobilidade maior que as bactérias, podendo atingir algumas dezenas de metros no aqüífero freático do cemitério de Vila Nova Cachoeirinha. Os vírus foram transportados, no mínimo, 3,2 m na zona não saturada até alcançar o aqüífero. / This work assessed occurrence and transport of microorganisms in the unconfined aquifer of Vila Nova Cachoeirinha cemetery, located on pre-cambrian terrains at the northern zone of the city of São Paulo The applied methodology was divided in laboratory and field stages. In the lab, cemetery soil columns were designed; chemical and biological tracers were injected in the columns and the effluent was monitored. A numerical model was used to simulate the tracers’ transport through the columns. In the field, several investigations were done to characterize the unconfined aquifer; water quality was monitored to study occurrence and transport of chemicals, bacteria and viruses in groundwater. In Vila Nova Cachoeirinha cemetery, the depth to the bedrock is about 9.0 m at small elevation areas and 20.5 m at the top of the hill. The depth to the water table varies from 4 to over 16 m. The soil is formed by the weathered material of the granite rocks, clay content of 43%, pH = 5,0, cation exchange capacity between 10.2 and 109.0 mmolc/kg. The hydraulic conductivity of the aquifer varies from 2.90 x 10-8 to 8.41 x 10-5 m/s. The hydraulic gradient at the western part of the study area is about 0.07 m/m; considering a homogeneous and isotropic medium and an effective porosity of 2%, the average linear velocity was estimated in 8 cm/day. The water samples of the unconfined aquifer of Vila Nova Cachoeirinha cemetery presented, mainly, heterotrophic bacteria (53 x 103 UFC/mL), proteolitic bacteria (31 NMP/100 mL) and clostridium perfringes (45 NMP/100 mL). We have also found enterovirus and adenovirus in groundwater. The main sources of contamination are the within-a-year-graves located at the low elevation areas, close to the water table. At these sites, the occurrence of bacteria is greater and there is a greater consumption of oxygen in the water due to oxidation of organic matter. Moreover, the graves cause an increase in salts and electrical conductivity of the groundwater. There seems to be an increase in major ions: hydrogen carbonate, chloride, sodium, calcium; and metals: iron, aluminium, lead and zinc, next to the graves. The bacteria traveled a distance of a few meters, decreasing in concentration with increasing distance to the graves. The viruses seem to be more mobile than bacteria, they traveled distances of tens of meters at the Vila Nova Cachoeirinha cemetery. The viruses were transported at least 3.2 m through the unsaturated zone before reaching the unconfined aquifer.

M. Tech. / Gold (Au) mining in South Africa resulted in vast volumes of hazardous waste being generated. Poor management of most of the tailings dams has resulted in the release of acid mine drainage, which caused stream water and soil contamination with their run-offs. The consequence of mine closure has not only been observed in large-scale land degradation, but also in widespread pollution of surface water and groundwater in the Wonderfonteinspruit Catchment Area (WCA). Thus, clean-up methods must be developed in order to remove heavy metals from contaminated water bodies in this area. The efficacy of algae, zeolite and zeolite functionalized with humic acid in reducing the concentration of the heavy metals iron (Fe3+), zinc (Zn2+), manganese (Mn2+) and nickel (Ni2+) to acceptable levels in WCA was investigated in this study. It is also envisaged that the heavy metals to be removed from contaminated water can be useful in various industries. A sampling exercise was undertaken with the aim of identifying the heavy metals that contaminate the water in the catchment, as well as identify the priority heavy metals for laboratory sorption tests. Batch experiments were conducted to study the adsorption behavior of natural zeolite clinoptilolite and algae Desmodesmus sp. with respect to Fe3+, Mn2+, Ni2+, and Zn2+. The data was analysed using the Langmuir and Freundlich isotherms. Two kinetic models namely, pseudo-first order and pseudo second order were also tested to fit the data. It was found that the concentration of Fe3+, Mn2+, Ni2+ and Zn2+ was 115 mg/L, 121 mg/L, 26.5 mg/L and 6.9 mg/L from the sampled water bodies in the WCA, respectively. The Langmuir isotherm was found to correlate the adsorption of Fe3+, Mn2+, Ni2+, and Zn2+ better, with the adsorption capacity of 11.9 mg/g, 1.2 mg/g, 1.3 mg/g, and 14.7 mg/g, for the functionalized zeolite (FZ), respectively. The algae system gave adsorption capacities of 1.523 mg/g, 144 mg/g and 71.94 mg/g for Fe3+, Mn2+ and Ni2+; respectively. Pseudo second-order equation was found to be the best fit for the adsorption of heavy metals by unfunctionalized zeolite (UFZ) and the algae system. Zeolite functionalization with humic acid increased its uptake ability. The best results for kinetic study was obtained in concentration 120 ppm for Fe3+ and Mn2+, whilst for Ni2+ was at 20 mg/L , which is about the same concentrations found in contaminated water in the WCA (Fe3+ 115 mg/L, Mn2+121 mg/L and Ni2+ 26.5 mg/L).

M.Tech. (Biomedical Technology) / Malawi is located in the south eastern Africa. It is a landlocked country covering an area of 118,484 Square Kilometres (45,747Sq.Miles) of which 20.6% is water. It is bordered with Zambia to the north-west, Tanzania to the north and Mozambique surrounding it on the south, east and west (refer map of Malawi on page 2). Malawi is one of the sub-Sahara heavily populated countries, having a population of about 13 million people of which about 80% live in the rural areas relying on small scale farming. The country is divided into three regions, south, centre and north and these regions are further divided into districts. The southern region has most districts and is the most heavily populated. Malawi’s economy heavily depends on agriculture. Cash crops such as tobacco, tea and sugar canes are grown by commercial farmers while smallholder farmers grow a variety of crops such as tobacco, coffee, beans, fava beans, cotton, rice, sorghum, soybeans, groundnuts, cassava, potatoes, wheat, millet, peas, bananas, citrus fruits, sugar canes, vegetables and maize which is the main staple food. Malawi has a subtropical climate with the rainy season starting from mid October up to April and the dry season is from May to October. During the dry season it is hot and humid in most parts of the country especially along the lake and the lower Shire valley, while the rest of the country can be cold at night with temperatures between 5ºC and 14ºC. The southern region has the highest annual rainfall and is humid most part of the year. This is where many tea plantations are and the climate allows growth of almost all types of crops and fruits. Most Malawians consume home grown unprocessed food commodities. There are only commodities are imported, mainly from South Africa. Malawi is self reliant on maize but when drought strikes, Malawi is forced to import from neighbouring countries or from South Africa and other countries as was the case in 2004 and 2005

M. Tech. / Fusarium species are common contaminants of maize and are also capable of producing mycotoxins, in particular the fumonisin. These are implicated in animal and human mycotoxins fumonisin B1 (FB1) for example, has been associated in the aetiology of oesophageal cancer in South Africa and other parts of the world, i.e., China and Iran. Because maize is the staple diet of the South African rural population, this study was designed with the aim of monitoring Fusarium spp. and FB1 in the food of rural people of Venda, Limpopo province of South Africa, during the course of processing maize into porridge which gave a means of estimating dietary exposure to this mycotoxin. Measurement of fumonisin in the excreta of these people allowed a determine of the extent to which FB1 the body is actually exposed to the mycotoxin.Fumonisin B1 has been identified as a major fungal contaminant on maize, especially in the home grown crops intended for human consumption. Thus the rural population of Limpopo Province is at high risk from FB1 exposure and it is therefore of importance to assess this exposure by the analysis of suitable samples.It can be seen that levels of FB1 in maize from Venda are quite high, as several of these samples had exceeded levels above 1750 μg/kg as recommended as maximum tolerance levels by theEuropean Commission. It is equally seen that a much higher proportion of this mycotoxin was destroyed by processing maize to porridge. And because porridge and other maize-based products are usually consumed on a daily basis, the low levels found in the present study must not be under-estimated, as such levels may accumulate over time and cause more severe chronic effects in humans. When setting daily tolerable levels of FB1 in foods in South Africa, it is imperative to take into account the food habits, especially those within the rural communities

M.Tech. (Biotechnology) / Please refer to full text to view abstract

Several countries have enacted regulations on tolerance limits for common mycotoxins because of the hazardous nature and widespread occurrence of these fungal secondary metabolites in agricultural commodities. Screening of agricultural commodities destined for animal consumption for the presence of mycotoxins is now becoming a prerequisite in several countries as a means of minimizing ingestion of these toxins. Silage samples were analyzed for pH, % dry matter (DM) content, and the presence of total fungi, yeasts and the types of lactic acid bacteria present. The samples were also analyzed for mycotoxins that have been reported to commonly occur in silage. The pH of the samples was found to be acidic ranging from pH 3.4 to 4.7, with few samples having pH values above 6. There was a significant difference in the % DM content amongst the sampling regions. There was no significant difference in the extent of fungal contamination amongst the different regions. Aspergillus fumigatus was the predominant species from all the samples. Most of the yeast species were isolated from the Bergville region. The yeast species isolated from all samples were Trichosporon, Cryptococcus and Candida species, which are all regarded as nonlactate fermenters. Lactobacillus plantarum and Lactobacillus buchneri were the only two lactic acid producers isolated from the samples. Aflatoxins, citrinin and patulin were the most predominant toxins in the samples. Ochratoxin A and deoxynivalenol was not detected in all samples using thin layer chromatography, while the latter two toxins were only detected in two samples using VICAM fluorometry. The level of fumonisins that was found in the forage crops used for silage production was fairly low with the highest level being 9.36 ppb. Most of the mycotoxin extracts were found to reduce the % cell viability of human lymphocytes after 18 hours of incubation as determined by the MTT assay. / Professor Mike Dutton Mr. F. E. Van Zyl

M.Tech. / Please refer to full text to view abstract

M.Tech. (Biomedical Technology) / A study was conducted to determine the extent of fungi and mycotoxins contamination of Rwandan selected food commodities. A total of one-hundred food samples including maize, rice, cassava, beans and peanuts were collected from all five provinces of Rwanda and analysed. Mycological data obtained revealed a high level of contamination of common toxigenic fungi belonging mainly to the Aspergillus, Penicillium and Fusarium genera. Accordingly, Aspergillus flavus was the most prevalent fungal contaminant in maize (90%), while A. carbonarius was mainly concentrated in peanuts at an incidence rate of 70%. Aspergillus fumigatus was mostly found in cassava (85%) in combination with Penicillium decumbens at the rate of 70%, meanwhile P. citrinum was found at an incidence rate of 80% in rice. The genus Fusarium was dominantly present with F. verticillioides and F. graminearum found in all analysed commodities. A toxigenicity study was also conducted to evaluate the capacity of these fungi recovered to produce their respective mycotoxins. Certain species such as A. flavus and A. parasiticus isolated from these commodities produced the aflatoxins (AFs). Other Aspergillus spp. such as A. carbonarius produced ochratoxin A (OTA) and F. verticillioides and F. graminearum also showed their capacity in producing different mycotoxins viz: zearalenone (ZEA), fumonisins (FBs) and deoxynivalenol (DON). The analysis of mycotoxins in these commodities was performed following thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). Data obtained revealed that peanuts and maize were the most contaminated with mycotoxins at incidence rates of 85 and 80%, respectively, and at the highest contamination levels. The highest AF-contaminated commodity was maize from Western province (range: 1.3-3219.6 μg/kg; mean: 829.3 μg/kg) followed by peanut from the same region whose mean level found was 401.5 μg/kg (range: 3.2–1755.8 μg/kg). Ochratoxin A was also found in peanuts with a mean concentration of 302.6 μg/kg, while DON was found at the highest level of 419.6 μg/kg in a rice from Kigali-city. Maize was the main substrate for FBs (mean: 134 μg/kg; max: 4591 μg/kg). Zearalenone was also recovered from samples but at a low incidence rate of 40% with the highest level of 5.2 μg/kg recorded. It was also observed that 65% of samples analysed were contaminated with more than one mycotoxin.

Magister Scientiae - MSc / Listeria monocytogenes is a Gram-positive bacterium responsible for listeriosis, a food-borne disease, which may result in severe illness and possible death. The importance of L. monocytogenes as a food-borne pathogen has been recognized since the 1980's when a correlation between the cunsumption of contaminated foodstuffs and human listeriosis outbreaks was observed. Listeriosis occurs with the ingestion of contaminated foods. The aim of this study involved developing DNA based methods to aid the food industry for the fast detection of L. monocytogenes in food products. Therefore assays were developed in such a way that they will have potential applications in the food idustry. / South Africa