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, 2013 / Introduction Dairy farms in central South Africa produce a substantial amount of milk, which is sold in Bloemfontein, Free State. Large volumes of unpasteurized (raw) milk is collected on the dairy farms, which undergoes further processing before it reaches the consumer at the end of the production line. There is a large proportion of the population that, in most cases unknowingly, consumes raw milk that has bacterial counts substantially higher than legal standards. Poor quality unpasteurized milk is either sold as fresh milk in the informal market, or as dairy products, such as cheese, manufactured from unpasteurized milk. Consumers are therefore, in most cases, unaware of the poor quality dairy products they consume. Milk quality is usually assessed in terms of bacterial content, which include Escherichia coli, coliforms and total bacterial count. The bacterial quality of milk is influenced by a number of factors, including farming practices, structural design of the milking shed, herd health and quality of water used in the dairy. If the highest level of hygiene practices is maintained, contamination of the milk by pathogenic microorganisms will be controlled, however, any drop in the vigilance of hygiene practices could result in unacceptable high levels of pathogenic microorganisms resulting in poor quality raw milk. Poor quality raw milk will inevitably result in poor quality pasteurized milk, containing unacceptably high levels of pathogenic organisms, which will eventually reach the consumer. Objectives The objectives of this study were to assess the quality of milk and influencing factors of milk produced on 83 dairy farms that supply milk intended for further processing to the greater Mangaung region, Central South Africa. Influencing factors investigated included, water quality and hygiene of milk contact surfaces, namely pulsator surfaces and milk pipeline surfaces. Methods Standard sampling procedures were followed when milk was sampled from bulk milk tanks, water at the point of use in the dairy, as well as collection of surface swabs. Escherichia coli, coliforms, total bacterial counts and somatic cell counts in milk were determined in terms of the regulations relating to milk and dairy products, and for water in terms of drinking water standards. These data were analysed and the factors that directly influence bacterial quality of milk were identified. Results 93% of the dairy farms displayed E. coli in their bulk milk containers, which did not comply with the legal standard. For coliforms, 86% of the milk samples did not comply with the legal standard. The total bacterial count of 85% of the milk samples did comply with the legal standard. The somatic cell count of 42% of the milk samples did not comply with the legal standard. The pulsator surfaces as well as the milk pipeline surfaces of 13% of the dairy farms displayed the presence of E. coli. 80% of the pulsator surfaces and 78% of the milk pipeline surfaces did comply with the legal standard pertaining to coliforms. The total bacterial count of pulsator surfaces revealed that 19% complied, whereas 29% of the milk pipeline surfaces complied with the legal standard. The water data further revealed that 31% of the dairy farms contained E. coli in the water used in the dairies. 63% of the dairy farms contained more than the allowable number of coliforms in their water. Chi-square tests revealed significant differences (p > 0.05) between the presence or absence of E. coli in milk and water; the presence or absence of E. coli in milk and milk pipeline surfaces; the presence or absence of E. coli in milk and pulsator surfaces and the presence or absence of E. coli in milk and the positioning of the cows in the milking shed. When milk quality indexes were calculated for all the farms, only four farms were classified with excellent milk, the remainder were all classified as producing poor quality milk. The hygiene quality indexes revealed that the hygiene practices on all the farms were not up to standard. Discussion and conclusion The study revealed that the milk produced for commercial processing and distribution in the greater Mangaung region of central South Africa was of poor quality. It is often mistakenly believed that the pasteurization process will remove all microorganisms from milk. As this is not the case, it is of major concern that milk delivered commercially is not of acceptable quality. Furthermore, it could be concluded that the quality of milk products from raw milk were also probably not of acceptable quality. The results further revealed that the possible contributing factors to the poor quality milk produced by the 83 commercial dairy farms were; poor quality water used in dairy sheds and contaminated milk contact surfaces. From this study it could be concluded that the overall status of milk production on the 83 commercial dairy farms studied, did not meet the standards required for milk quality, water quality and hygiene practices.

A cross-sectional study was conducted from February 2011 to August 2012 to investigate the status and identify risk factors for Bovine Tuberculosis (BTB) and cattle Brucellosis. A total of 1277 cattle from 6 selected dairy farms were tested. The farms were purposively selected on suspected risk. Samples were analysed using Rose Bengal Test (RBPT) and Complement Fixation Test (CFT) for Brucellosis and Comparative Intradermal Tuberculin Test (CITT) for BTB. The results revealed the overall prevalence of 1.6% for Brucellosis and no BTB was found. Logistic regression analysis revealed that age of cattle, herd size and number of calves a cow has had significant association with Brucellosis with p-values 0.0071, 0.0490 and 0.000 respectively. The role of location (p>0.05), gender (p>0.05) and breed (p>0.05) in the occurrence of the disease was not statistically significant at α=0.05, although higher rates were obtained in females and Jersey breed. Questionnaires were also administered to 47 individuals a total of 12 females and 35 males. Most respondents were not aware about cattle brucellosis and cattle tuberculosis. Consumption of raw or unpasteurized milk still occurs with 74.75% of respondents revealed.

Tuberculosis (TB) is an ancient infectious disease that has been infecting different populations around the globe and it has also been considered as one of the most successful human and animal disease. TB found in animals such as cattle and other known bovids is known as bovine tuberculosis. Bovine tuberculosis (BTB) is an infectious disease found in cattle mainly caused by Mycobacterium bovis. M. bovis is a member of the Mycobacterium tuberculosis complex (MTC) together with M. tuberculosis, M. africanum, and M. canetti where the natural host is humans; whereas M. caprae, M. microti and M. pinnipedii usually have animals as their natural host. In this study the molecular characterization of the MTC from cow milk in the Eastern Cape was investigated. One hundred and twenty samples (40 ml each) were collected from three dairy farms in the Eastern Cape, South Africa. These samples were processed using a modified Petroff decontamination method. Sample processing was followed by DNA isolation using a Zymo Bacterial/Fungal DNA Kit and the amplification and detection of the MTC was done using the Seeplex MTB Nested ACE assay. The drug susceptibility tests were done using GenoTypeMTBDRplus assay which detects mutations and resistance to INH (isoniazid) and RMP (rifampicin). The milk isolates were further analyzed using a spoligotyping method which is based on the PCR amplification of a highly polymorphic direct repeat locus in the M. tuberculosis genome which detects and types the MTC. A percentage of 20.8 % samples were found to be positive for MTC using the Seeplex MTB Nested ACE assay. There were 42.1 % samples that were resistant to both INH and RMP with the rest sensitive to either INH or RMP. The spoligotyping method showed that 78.3 % samples resembled Family 33 strains and the rest (21.7 %) resembled a spoligotyping signature known to be that of M.africanum. Both these strains belong to the Ancestral lineage with Indo-Oceanic and West Africa 2 lineage. The outcomes of our study showed that molecular methods for detection of MTC can be applied directly on milk samples without the need for culturing.