Dissertations / Theses on the topic 'Food preservation/microbiology'

Lactic acid fermented fruit and vegetables are normally obtained following a natural spontaneous fermentation in which no starter cultures are added. It could be expected that a suitable starter culture would help standardise production. Several lactic acid bacteria were selected for a series of physiological studies, in a defined medium (MRS broth) and in carrot juices, under varying conditions of growth temperature, salt concentration and carbohydrate source. Based on these, the homofermenter Lactobacillus pentosus and the heterofermenter Leuconostoc mesenteroides were tested as potential starters, in single and mixed cultures, for the fermentation of carrots (Daucus carota), as a novel fermentable substrate, and cabbage (Brassica oleracea) into sauerkraut. Fermentations were performed in the presence of the natural microflora. Sugar catabolism and acid production were monitored through H.P.L.C. In the fermentation of carrots Leuconostoc mesenteroides played a major role, with no homofermenters present. For sauerkraut, the mixed starter culture composed of Leuconostoc mesenteroides and Lactobacillus pentosus gave the closest resemblance to the product normally obtained following a natural commercial fermentation. The inclusion of the heterofermenter provided the required acid balance for correct product flavour and aroma by enhancing production of acetic acid. Acetate is also a better antimicrobial than lactate. A shorter fermentation time was also obtained, reducing the time from 3-4 weeks in the natural fermentation to only 7 days with the use of the mixed starter. When reduced salt concentrations were tried, 1% NaCl (w/w) resembled the spontaneous fermentation more closely, in regard to microbial sequence, pH and total acidity. Different ratios of the two lactic acid bacteria in combination were tried, the best being that in which L, mesenteroides and L. pentosus were initially present in the same proportions. Survival of Listeria monocytogenes in fermenting sauerkraut was shorter when starter cultures were used, but no difference was detectable between mixed and single cultures.

Thesis (MScVoedselwet)--University of Stellenbosch, 2004.
ENGLISH ABSTRACT: Red meat has a limited shelf-life at refrigerated temperatures, where spoilage is mainly due to the proliferation of bacteria, yeast and moulds, acquired during the dressing process. In addition, almost a fifth of food-borne disease outbreaks, caused by microorganisms such as Escherichia coli 0157:H7, Listeria monocytogenes and Staphylococcus aureus are associated with red meat. To improve the microbiological quality of red meat, systems such as HACCP, GHP and GMP are currently practiced; however, these practices are not able to extend the shelf-life of these products. At present suitable food-grade preservatives are recommended, but the use of some of these preservatives is increasingly being questioned with regard to their impact on human health. Additionally, food service customers demand high quality products that have a relatively long shelf-life, but still prefer the appearance of minimally processed food. All these factors challenge the food manufacturing industry to consider more natural means of preservation. Antimicrobial metabolites of food grade bacteria, especially lactic acid bacteria, are attracting increasing attention as food preservatives. Bacteriocins are antimicrobial peptides (3 to 10 kDa) with variable activity spectra, mode of action, molecular weight, genetic origin and biochemical properties that are bacteriostatic or bactericidal to bacteria closely related and bacteria confined within the same ecological niche. Micro-organisms were isolated from beef, lamb and pork, obtained from four commercial retailers. The number of viable cells three days after the sell-by date at 4ºC ranged from 80 cfu.g-1 to 1.4 × 108 cfu.g-1. Fifty-three percent were Gram-negative bacteria, 35% Gram-positive and 12% yeast. The microbial population of the meat was greatly influenced by the origin, i.e. the retailer. Bacteriocins produced by Enterococcus faecalis BFE 1071, Lactobacillus curvatus DF 38, Lb. plantarum 423, Lb. casei LHS, Lb. salivarius 241 and Pediococcus pentosaceus ATCC 43201 were screened for activity against bacteria isolated from the different meat samples. Sixteen to 21% of the isolates, identified as members of Klebsiella, Shigella, Staphylococcus, Lactobacillus, Lactococcus, Leuconostoc, Enterococcus, Pediococcus, Streptococcus and Bacillus were sensitive to the bacteriocins. Curvacin DF 38, plantaricin 423 and caseicin LHS (2.35 to 3.4 kDa) had the broadest activity range and inhibited species of Lactobacillus, Pediococcus, Enterococcus, Listeria, Bacillus, Clostridium and Propionibacterium. The bacteriocins remained stable at 121ºC for 20 min, in buffers with a pH ranging from 2 to 10 and in NaCl concentrations of between 0.1 and 10% (m/v). Like most peptides, they were sensitive to proteolytic enzymes. Curvacin DF 38 is sensitive to amylase, suggesting that the bacteriocin might be glycosylated. To assess the efficiency of curvacin DF 38, plantaricin 423 and caseicin LHS as meat preservatives, they were partially purified by ammonium sulphate precipitation and separation in a Sep Pak C18 cartridge. The shelf-life of pork may be extended by up to two days. Meat samples treated with bacteriocins were darker than the control (untreated) sample. Descriptive sensory evaluation by a seven-member panel indicated that there were significant differences (P ≤ 0.05) regarding the aroma, sustained juiciness, first bite and metallic taste attributes of the control and the 4 day-treated samples. The control and 2 day-treated samples and the 2 day- and 4 day treated samples did not differ significantly regarding these attributes. There were no significant differences regarding the initial juiciness, residue and pork flavour attributes. Concluded from the results obtained in this study, bacteriocins produced by Lb. curvatus DF 38, Lb. plantarum 423 and Lb. casei LHS effectively extended the shelf-life of pork loins by up to 2 d at refrigerated temperatures with no drastic changes on sensory characteristics. In edition, the stability of these bacteriocins broadens their application as preservatives in many foods.
AFRIKAANSE OPSOMMING: Die rakleeftyd van rooivleis by yskastemperature is beperk, waar bederf hoofsaaklik deur die vermenigvuldiging van bakterieë, giste en swamme veroorsaak word. Die meeste van hierdie kontaminante is afkomstig van die slagtingsproses. Byna ’n vyfde van alle uitbrake van voedselvergiftigings wat deur organismes soos Escherichia coli 0157:H7, Listeria monocytogenes en Staphylococcus aureus veroorsaak word, word met rooivleis geassosieër. Die praktyke HACCP, GMP en GHP word tans toegepas om die mikrobiologies kwaliteit van vleis te handhaaf, maar is egter nie voldoende om die rakleeftyd van rooivleis the verleng nie. Die preserveermiddels wat huidiglik aanbeveel word vir dié doel, word toenemend bevraagteken aangaande die invloed daarvan op die menslike gesondheid. Hierby is daar ’n aanvraag na hoë kwaliteit, ongeprosesseerde produkte met ’n verlengde rakleeftyd. Gevolglik word die voedsel vervaardigings industries aangemoedig om meer natuurlike vorms van preservering the oorweeg. Die aandag word tans op die anti-mikrobiese metaboliete van voedselgraad microbes, veral melksuurbakterieë, gevestig. Bakteriosiene is anti-mikrobiese peptiede (3 tot 10 kDa) met verskeie aktiwiteitsspektra, werkswyse, molekulêre massa, genetiese oorsprong en biochemiese eienskappe. Bakteriosiene is meestal bakterie-dodend of - staties teen taksonomies naby geleë organismes en organismes vanuit dieselfde ekologiese nis. Mikroörganismes is geïsoleer vanuit bees-, skaap- en varkvleis, verkry vanaf vier supermarkte. Die aantal lewensvatbare selle per gram (cfu.g-1) het drie dae na die “verkoop”-datum by 4ºC vanaf 80 cfu.g-1 tot 1.4 × 108 cfu.g-1 gevarieër. Drie en vyftig persent van die isolate is as Gram-negatief, 35% as Gram-positief en 12% as giste geïdentifiseer. Die sensitiwiteit van hierdie isolate teen bakteriosiene wat deur Enterococcus faecalis BFE 1071, Lactobacillus curvatus DF 38, Lb. plantarum 423, Lb. casei LHS, Lb. salivarius 241 en Pediococcus pentosaceus ATCC 43201 geproduseer is, is vervolgens getoets. Tussen 16% en 21% van die isolate was sensitief teen die bacteriosiene en is onder andere as Klebsiella, Shigella, Staphylococcus, Lactobacillus, Lactococcus, Leuconostoc, Enterococcus, Pediococcus, Streptococcus en Bacillus geïdentifiseer. Die bakteriosiene met die wydste aktiwiteitsspektrum, naamlik, curvacin DF 38, plantaricin 423 en caseicin LHS is verder ondersoek. Hierdie antimikrobiese peptiede (2.35 tot 3.4 kDa) toon aktiwiteit teen spesies van Lactobacillus, Pediococcus, Enterococcus, Listeria, Bacillus, Clostridium and Propionibacterium. Die bakteriosiene is stabiel by 121ºC vir 20 min, in buffers met ‘n pH-reeks van tussen 2 en 10 en soutkonsentrasies vanaf 0.1% tot 10%. Soos die geval by meeste peptiede is hierdie bakteriosiene sensitief vir proteolitiese ensieme. Curvacin DF 38 is ook sensitief vir amylase, wat daarop dui dat hierdie bakteriosien moontlik geglikosileer is. Die effektiwiteit van curvacin DF 38, plantaricin 423 en caseicin LHS as preserveermiddel in voedselsisteme is getoets deur dit te suiwer (ammonium sulfaat presipitasie en Sep Pak C18 kolom) en op vark lendestukke aan te wend. Mikrobiese analise het bewys dat die rakleeftyd van vark met sowat 2 dae verleng kan word. Volgens die vleiskleurevaluering was die bakteriosien behandelde vark donkerder as die kontrole. Die aroma-, sappigheid-, tekstuur- en metaalgeur-eienskappe van die kontrole en die 4-dag behandelde monster het volgens ‘n opgeleide sensoriese paneel betekenisvol verskil (P ≤ 0.05). Die kontrole en die 2-dag behandelde en die 2-dag behandelde en die 4-dag behandelde monsters het nie betekenisvol verskil nie. Daar was geen betekenisvolle verskil aangaande die aanvanklike sappigheid-, residu- en varkgeur-eienskappe nie. Hierdie sensoriese eienskappe is belangrik ten opsigte van die verbruiker se aanvaarding van die produk. Vervolgens kan uit hierdie resultate afgelei word dat die bakteriosiene wat deur Lb. curvatus DF 38, Lb. plantarum 423 en Lb. casei LHS geproduseer word voldoende is om die rakleeftyd van vark lendestuk by 4ºC met 2 dae te verleng met min of geen effek op die sensoriese persepsie van die vleis. Hierdie bakteriosiene is ook stabiel onder verskeie kondisies wat die toepassing as preserveermiddel aansienlik verbreed.

Thesis (MTech (Chemical Engineering))--Cape Peninsula University of Technology, 2016.
Microbial spoilage has been reported in various food products and this has led to increased food, fruit and beverage losses, thereby threatening economic growth, food safety and security. Furthermore, statistics have shown that more than 30% of agricultural produce in developing countries, mostly in Africa, is lost owing to microbial spoilage. Beverages, food and fruits are predominant contributors to the South African export market. In recent years, contamination of these products resulting in spoilage has been a problem, although partial spoilage control has been achieved using chemical preservatives such as dimethyl dicarbonate, sodium benzoate, potassium sorbate, and sulphur dioxide (SO2). However, prolonged exposure to these chemical preservatives can cause human health problems such as skin and/or eyesight damage, muscle and stomach pain, cardiovascular disease and the impairment of brain function. To mitigate such health concerns, biologically benign alternatives are deemed suitable, providing the rationale for this study.

High pressure (HP) kinetics of the microbial destruction and changes in the physicochemical characteristics of milk and pork were studied. Raw milk samples containing indigenous microflora of approximately 106 CFU/mL were heat sealed in dual peel sterilization pouches and subjected to HP treatment from 150--400 MPa with holding times ranging 5--120 min. The kinetic parameters (rate constant, k and decimal reduction time, D) for the microorganisms, alkaline phosphatase, color and viscosity were evaluated, based on first order kinetics and the pressure dependence of kinetic parameters was evaluated using pressure destruction time (PDT) and Arrhenius models. Kinetic data was well described by the first order model (R 2 > 0.90).
The application of pressure pulse was explored for pressure destruction of microorganisms as well as changes in physical-chemical characteristics of pork chops. Pork chops (2 days post-rigor) were subjected to HP treatment from 200--350 MPa for 0--120 min. Results showed that pressure changes of pork variables followed a dual effect consisting of an instantaneous pressure kill (IPK) with the application of pressure pulse (no holding) and a subsequent first order rate of destruction during the pressure hold time. The IPK values were pressure dependent and increased with pressure level. Parameters k and D indicated a higher rate of pressure destruction of microorganisms compared to quality attributes.
Kinetics of pressure destruction of Listeria monocytogenes Scott A were studied in relation to those of indigenous microorganism of milk and pork. The IPK was more pronounced with L. monocytogenes than with indigenous microflora. However, the kinetic parameters (k and D values) indicated a larger pressure resistance for L. monoctyogenes. HP processes were developed based on the standard plate count (SPC) kinetic data for indigenous microflora of milk as well as L. monocytogenes in milk and pork. The results showed that SPC kinetics permitted good estimation of microbial destruction in low pressure-lethality processes of milk and pork but its application at higher pressure-lethality levels were inaccurate. On the other hand, processes established based an destruction of L. monocytogenes were more predictable. Pressure pulse application to microbial lethality was also well predicted.
The shelf-life of milk and pork increased with the level of applied pressure lethality, but Q10 values suggested that low storage temperature was nevertheless required to control microbial growth and maintain quality. Storage of HP treated park offered some improvement in the texture but resulted in large color changes and drip losses. L. monocytogenes were not detected in any of the stored milk samples HP treated to achieve a lethality ≥10D.

Thesis (M. Tech. Environmental health) -- Central University of technology, Free State, 2011
Although the application of low pH is common practice in food preservation, the emergence of acid tolerance has been reported world-wide amidst a growing concern that preservation with weak acids, such as organic acids may be influenced as a result of food-borne bacteria becoming acid tolerant or acid resistant. The present study was conducted to assess the acid tolerance of a wide range of bacterial species and consequently the sustainable application of organic acids as food preservatives in particularly acidic foodstuffs. Acid tolerance was determined in 19 bacterial strains predominantly associated with food spoilage and food poisoning. After exposure to hydrochloric acid 16% of the isolates were found to be intrinsically tolerant to low pH and included amongst others the enteric bacteria Escherichia coli and Salmonella spp. The latter organisms are known causative agents in food spoilage and poisoning, and the results highlight the predicaments related to their ability to survive in acidic foodstuffs as well as the human gastric environment. Bacterial strains were further exposed to increasing concentrations of various acidic foodstuffs in order to determine the development of acid tolerance by gradual decrease in pH, as opposed to exposure to acid shock. After induction, the protein profiles of resulting acid tolerant isolates were compared with those of the original un-induced strains. Exposure to acidic foodstuffs resulted in various survival profiles, where not only pH but also the type of acidulant (foodstuff or inorganic acid) were found to be contributing factors in acid tolerance development. Bacterial protein composition after exposure to acidic foodstuffs showed considerable variation which may be indicative of acid tolerance development whereas the mechanisms involved may be the result of multiple modifications in bacterial composition. After the induction of acid tolerance, susceptibility of induced strains to various organic acids were determined at various pH values. This was done to investigate whether acid tolerance would influence the inhibitory activity of organic acids as antimicrobial agents in acidic food. Decreased susceptibility was not significantly demonstrated with the exception of only selected isolates, the latter including E. coli and S. typhimurium. Organic acid activity was found to be much more effective at lower pH values and it would be necessary to elucidate whether this inhibition is the result of a lower pH or more specifically the activity of the organic acids. The effect of exposure to an acidic environment on phenotypic characteristics of Gram-negative bacteria, and more specifically psychrotrophic organisms was evaluated in order to show the combined effect of organic acids and low temperature preservation. The characteristic yellow pigment of various Chryseobacterium species was found to be not as apparent after acid exposure while in some cases the colonies were observed as white. In Pseudomonas aeruginosa the characteristic green pigment was much more prominent after acid exposure. These morphological alterations may be important factors that should be considered in identification procedures employed in food safety laboratories. Finally, the influence of acidic exposure via acidic foodstuffs and also organic acids on the protein composition and outer membrane protein structure of various bacterial cells was investigated. No specific relationships with the MICs (Minimum Inhibitory Concentrations) of organic acids after induction with the selected acidic foodstuffs could be established, although various differences were found in protein expression. From the results, it may be suggested that the outer membrane of various pathogenic bacteria is involved in acid tolerance development and this supports the reports on the importance of membrane integrity in the protection against low pH. In conclusion, the study endeavoured to add to the body of knowledge with regard to alternative food preservation regimes utilising organic acids, either solely or in combination with selected extrinsic and intrinsic parameters.

Além das características tecnológicas das bactérias lácticas para o processamento de alimentos fermentados, novas propriedades são requeridas, como a produção de bacteriocinas. Vários desses compostos protéicos possuem atividade antimicrobiana direcionada à patógenos e apresentam potencial de uso como bioconservadores naturais de alimentos. O objetivo desse trabalho foi avaliar a viabilidade de aplicação de cinco culturas lácticas produtoras de bacteriocinas como bioconservantes em alimentos. Avaliaram-se as propriedades de segurança e probióticas dessas culturas e determinaram-se os parâmetros de produção e purificação parcial das bacteriocinas, bem como os efeitos de fatores limitantes de crescimento (cloreto de sódio e nitrito). Os resultados demonstraram que essas linhagens não possuem atividade de enzimas indicadoras de patogenicidade (termonuclease, -hemolisinas e gelatinases). A tolerância às condições ácidas variou dependendo da linhagem. Lactobacillus plantarum CTC 368 e Enterococcus avium CTC 469 apresentaram maior tolerância a valores de pH 2,0 e 3,0. Lactococcus lactis subsp. cremoris CTC 204 e Enterococcus avium CTC 483 também apresentaram 100% de sobrevivência a pH 3,0 e, mesmo com uma diminuição da tolerância a pH 2,0, a sobrevivência foi cerca de 80%. Com relação à susceptibilidade aos sais de bile, observou-se que a viabilidade variou entre as linhagens em função da concentração e tempo de incubação. Enterococcus avium CTC 469 e Lactococcus lactis subsp. hordinae CTC 484 foram as mais tolerantes e apresentaram sobrevivência de 57% e 58%, respectivamente. As linhagens mostraram excelente sobrevivência tanto a baixos valores de pH, quanto às concentrações de sais biliares indicando potencial probiótico. As características de crescimento e de produção de bacteriocinas foram semelhantes para todas as linhagens e meios avaliados. A viabilidade aumentou durante o período de incubação e a produção de bacteriocinas mostrou uma cinética de metabólito primário. A suplementação do meio MRS com 1,0% de NaCl e 0,1% de NaNO2, não afetou o crescimento de Lc. lactis subsp. cremoris CTC 204, mas promoveu a redução da atividade da bacteriocina de 42,5% e 17,5%, respectivamente. O crescimento de Lactococcus lactis subsp. hordinae CTC 484 foi afetado com a adição de NaCl e NaNO2 ao meio APT, mas ainda reteve 50% da atividade bacteriocigênica. O espectro de atividade das bacteriocinas variou de acordo com a susceptibilidade da linhagem indicadora, a concentração ou atividade da bacteriocina, a viabilidade e/ou presença das células produtoras e os procedimentos de extração das bacteriocinas. Os resultados revelaram que as bacteriocinas de Lactococcus lactis subsp. cremoris CTC 204 e Lactococcus. lactis subsp. hordinae CTC 484 promoveram um efeito bactericida contra Staphylococcus aureus. Concluiu-se que essas duas culturas produtoras de bacteriocinas apresentam propriedades para utilização como bioconservantes em alimentos e que estudos mais aprofundados, relacionados com a purificação da bacteriocina e avaliação de seu uso em alimentos, devem ser realizados.
In addition to the technological characteristics of lactic bacteria for the processing of fermented foods, other properties need to be developed and explored, such as bacteriocin production. Several of these protein-based compounds have antimicrobial activities targeted towards pathogens and have a significant potential for use as natural biopreservatives in processed foods. The objective of this study was to evaluate the viability of using five bacteriocin-producing lactic cultures as biopreservatives in foods. The safety and probiotic properties of these cultures were evaluated, followed by the determination of bacteriocin production and purification parameters and the effect of growth-limiting factors (sodium chloride and nitrite). The results demonstrated that these strains do not show activity of marker enzymes of pathogenicity (thermonuclease, - hemolysins and gelatinases). Tolerance to acidic conditions varied depending on the specific strain. Lactobacillus plantarum CTC 368 and Enterococcus avium CTC 469 exhibited greater tolerance to pH values 2,0 and 3,0. Lactococcus lactis subsp. cremoris CTC 204 and En. avium CTC 483 also presented 100% survival at pH 3,0, and even with reduced tolerance at pH 2,0, the survival rate was 80%. Variation in susceptibility to bile salts was observed between the strains as a function of concentration and incubation time. En. avium CTC 469 and Lactococcus lactis subsp. hordinae CTC 484 were found to be the most tolerant, exhibiting survival rates of 57% and 58%, respectively. The strains demonstrated excellent survival not only at low pH values, but also at high bile salt concentrations, thereby indicating probiotic potential. The growth and bacteriocin-producing characteristics were similar for all the strains and culture media evaluated. Bacterial multiplication increased during the incubation period, while bacteriocin production displayed primary metabolite kinetics. The activity spectrum of the bacteriocins varied with the susceptibility of the indicator strain, the concentration or activity of the bacteriocin, the viability and/or presence of bacteriocinproducing cells and the bacteriocin extraction procedures. The results reveal that the bacteriocins produced by Lc. lactis subsp. cremoris CTC 204 and Lc. lactis subsp. hordinae CTC 484 exerted a bactericidal effect against Staphylococcus aureus. Supplementation of the MRS medium with 1,0% NaCl and 0,1% NaNO2, did not affect the growth of Lc. lactis subsp. cremoris CTC 204, but did reduce by 42,5% and 17,5% the activity of the bacteriocin, respectively. Growth of Lc. lactis subsp. hordinae CTC 484 was affected by the addition of NaCl and NaNO2 to the APT medium, but even so still exerted 50% of its bacteriocinogenic activity. It was concluded that Lc. lactis subsp. cremoris CTC 204 and Lc. lactis subsp. hordinae CTC 484 exhibited adequate properties for use as biopreservatives in foods and that more extensive studies related to the purification of bacteriocins and assessment of their use in foods should be conducted.

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With the objective of evaluating the physical/chemical, microbiological characteristics and the changes that occur in UHT milk (UAT) processed in the State of Goias during its storage for 135 days, a study was done between August and December 2008 with all the companies that produced UHT milk (UAT) processed in the State. Samples were collected from refrigerated raw milk stored in milk storage silos of industry for analysis of their physical/chemical and microbiological characteristics. From each company, twelve one liter cartons of milk were collected for analysis of their physical/chemical and microbiological properties as well as nitrogen compounds and the amount of fat separation from the milk and sedimentation within each sample. The samples were sent to the laboratories of Microbiology and Dairy of the School of Agronomy and Food Engineering at the Federal University of Goias (UFG). The physico-chemical properties of raw milk were in accordance with the parameters established by legislation, except for three brands of milk, with values of freezing point, nonfat milk solids and total solids. The Alizarol test results were normal for UHT milk (UHT) of all brands, except the milk of the mark "E", which has had worst results established by legislation. All milk samples were within the standards for titratable acidity and pH. The density indices were normal, but decreased during the stock. The freezing point values were within the standards, but increased during the storage and the values of total solids and nonfat solids were low and at variance with the laws in UHT milk (UHT) of all brands. The fat found below the minimum content of 3% recommended by the legislation. All results are suggestive of water being added accidentally during processing. Scores were found higher than those established by law for psychotropic and mesophilic aerobic refrigerated raw milk. In UHT milk (UHT) milk from only one mark was at odds as the parameters established by law for heat resistant spores of aerobic mesophiles. It was verified a progressive increase in sedimentation and fat separated from milk during storage and observed age gelation in milk from one brand to 135 days of storage. From the results obtained can be verified that the physicochemical characteristics of UHT milk (UAT), when compared to raw milk source, changed negatively and that the raw material, the type of thermal processing and storage time at room temperature are possible causes for the changes that occurred throughout the shelf-life UHT milk (UAT).
Com o objetivo de avaliar as características físico-químicas, microbiológicas e alterações que ocorrem em leite UHT (UAT) processado no Estado de Goiás, durante seu armazenamento foi feito um estudo entre agosto e dezembro de 2008, por 135 dias, com leite UHT (UAT) de todas as marcas processadas no Estado. Foram coletadas amostras de leite cru refrigerado nos silos de estocagem de leite das indústrias, para análises físico-químicas e microbiológicas. Foi feito o acompanhamento do processamento do leite desse mesmo silo pelo sistema UHT (UAT). De cada indústria foi coletado um pacote embalado, contendo doze caixas cartonadas de um litro de leite, para análises físico-químicas, microbiológicas, teor de compostos nitrogenados e para verificação de ocorrência de sedimentação e de gordura separada do leite. As amostras de leite cru refrigerado foram colocadas em caixas isotérmicas contendo gelo e as amostras do leite UHT envasado foram encaminhadas aos laboratórios de Microbiologia e de Laticínios da Escola de Agronomia e Engenharia de Alimentos da Universidade Federal de Goiás (UFG). As análises físico-químicas do leite cru se apresentaram de acordo com os parâmetros preconizados pela legislação, com exceção do leite de três marcas, com valores de crioscopia, extrato seco desengordurado e extrato seco total. Ao longo do período de estocagem os resultados das análises de Alizarol apresentaram-se normais para o leite UHT (UAT) de todas as marcas, com exceção do leite da marca E , sendo esse leite o que teve mais resultados fora dos padrões estabelecidos pela legislação. Todas as amostras de leite estavam dentro dos padrões para acidez Dornic e pH. Os índices de densidade estavam normais, mas decresceram durante o estocamento. Os valores de crioscopia estavam dentro dos padrões, mas altos e os valores de extrato seco total e extrato seco desengordurado apresentaram-se baixos e em desacordo com a legislação no leite UHT (UAT) de todas as marcas. Os teores de gordura se apresentaram abaixo do teor mínimo de 3% preconizado pela legislação. Os resultados em conjunto são sugestivos de adição de água durante o processamento. Foram encontradas contagens superiores aos estabelecidos pela legislação para bactérias psicrotróficas e Aeróbios Mesófilos no leite cru refrigerado. No leite UHT (UAT) apenas o leite de uma marca estava em desacordo com os parâmetros preconizados pela legislação para esporos termorresistentes de Aeróbios Mesófilos. Foi verificado aumento progressivo de sedimentação e de gordura separada do leite durante a estocagem e observada gelificação no leite de uma marca aos 135 dias de estocagem. Conclui-se que as características físico-químicas, do leite UHT (UAT), quando comparadas ao leite cru de origem, se alteraram negativamente e que a matéria-prima, o tipo de processamento térmico e o tempo de estocagem em temperatura ambiente são possíveis indicativos da origem das modificações ocorridas ao longo da vida-de-prateleira do leite UHT (UAT).

La sicurezza e la qualità degli alimenti sono tutt’ora un problema critico per i paesi in via di sviluppo. Le diete a basso contenuto di acido folico, per esempio, possono causare gravi problemi di salute, soprattutto nei bambini. Gravi disturbi legati al tubo neurale (DTN) nei neonati possono derivare infatti da madri che hanno insufficiente apporto di acido folico (400-600 g / giorno) durante il periodo di gravidanza. Inoltre, se non adeguatamente protetti o trattati, I prodotti alimentari possono essere vettori di funghi e batteri patogeni rappresentando una fonte potenziale di malattie per l’uomo e una perdita economica per le industrie agro-alimentari. Nella seguente tesi si è quindi quindi studiato il ruolo di batteri lattici selezionati (LAB) in grado di aumentare il valore nutrizionale del latte attraverso la produzione di acido folico durante il processo di fermentazione. Inoltre, ci si è concentrati sul loro uso come "bio-conservanti" contro funghi e batteri, attraverso la sintesi di composti antimicrobici (batteriocine) in grado di inibire la crescita di funghi filamentosi e/o batteri patogeni.
The safety and quality of food are still a critical issue in developing countries. Diets with a low content of folic acid, for example, may cause serious health problems, especially in children. Severe disorders related to neural tube (NTD) in infants may arise from mothers having inadequate intakes of folic acid (400-600 g/dia) during the mother pregnancy period. Moreover foods, when not properly protected or treated, can be vectors of pathogenic fungi and bacteria thereby representing a potential source of human diseases and an economical loss for the food industry. In the following thesis we have therefore investigated the role of selected lactic acid bacteria (LAB) in increasing the nutritional value of milk through the production of folic acid during the fermentation process. In addition, we focused on their use as “bio-preservatives” against fungal and bacterial spoilage, through the synthesis of antimicrobial compounds (bacteriocins) able to inhibit the growth of filamentous fungi and /or pathogenic bacteria.

Os peitos de galinhas matrizes pesadas de corte, poedeiras comerciais brancas e frangos foram desossados manualmente, com o objetivo de avaliar e comparar as propriedades tecnológicas – capacidade de retenção de água e de emulsificação – bem como pH e composição centesimal, além de elaborar e avaliar sensorialmente um produto reestruturado empanado, tipo nugget. Os resultados das análises indicaram que não houve diferença significativa entre as carnes analisadas, nem entre produtos processados, implicando utilização regular desse tipo de matéria-prima na indústria de alimentos. O restante das carcaças das galinhas foi utilizado para extração de carne mecanicamente separada (CMS) para avaliação da estabilidade microbiológica. As CMSs foram submetidas a dois tratamentos: I) com 150ppm de nitrito, embaladas em saco de polietileno; e II) com 150ppm de nitrito e 500ppm de eritorbato, embaladas em saco de polietileno. Como controle, as amostras de CMS foram apenas acondicionadas em sacos de polietileno. Todas as embalagens foram estocadas e congeladas a –18ºC por 99 dias. Os resultados da análise de caracterização microbiológica indicaram que tanto as amostras de CMS de galinha matriz quanto de galinha poedeira atenderam os padrões de contagem requeridos pela legislação Brasileira. A avaliação da estabilidade microbiológica das CMSs no primeiro e no último dia para mesófilos, Escherichia coli, Staphylococcus aureus, Clostridium perfringens e Pseudomonas, bem como as análises quinzenais de microrganismos psicrotróficos, indicaram que não houve uma redução significativa nas contagens, independentemente do tratamento aplicado. Não foi detectada Salmonella em nenhuma das amostras analisadas. Tais resultados apontaram o tempo de estocagem como o principal fator na redução da carga bacteriana.
The breasts of heavy fowls, white commercial egg-layers and chickens were manually deboned in order to evaluate and compare technological properties—water hold and emulsifying capacity—and pH and centesimal composition, as well as to prepare and assess a restructured coated product. The results of the analyses indicated that there was not a significant difference between the meats analyzed, nor among the processed products, implying the regular use of such meat in the food industry. The remaining parts of the hens’ carcasses were mechanically deboned for the evaluation of microbiological stability. The mechanically deboned meat (MDM) was submitted to two treatments: I) 150ppm of nitrite, packed in polyethylene bags; II) 150ppm of nitrite and 500ppm of erythorbate, packed in polyethylene bags. The control meats were just packed in polyethylene bags. All samples were frozen-stored at –18ºC for 99 days. The results of microbiological characterization indicated that both samples from heavy fowls and egg-layers were in accordance with the Brazilian legislation. MDM microbiological stability evaluation on the first and last days for fecal coliforms, mesophiles, Staphylococcus aureus, Clostridium perfringens and Pseudomonas, as well as the analyses carried out every fifteen days for psychrotrophics bacteria indicated that there was not a significant reduction in the presence of the above, irrespective of the treatment applied. Salmonella was not detected in any of the samples. Such results indicated that freezing storage period was the main factor responsible for the reduction of the bacterial count in MDM.

MA RESEARCH REPORT Prepared for the Department of Development Studies, University of the Witwatersrand, Johannesburg June 2016
This research argues that food sovereignty offers a plausible alternative to the current unjust, unsafe and unsustainable food system in South Africa. In addition, it argues that food sovereignty provides important solutions to hunger and the brutalities of the food system which current policies and interventions fail to address. Food sovereignty is an ideal that originated amongst a peasant movement in the global South. This ideal and framework to address hunger has since evolved and spread to international movements, and is making great strides in advocating for change in the current broken food system. Food sovereignty has lately been adapted in South Africa as a grassroots led initiative promoted by the nascent South African Food Sovereignty Campaign (SAFSC). This research uses the SAFSC as a case study to explore food sovereignty alternatives in South Africa. It does this by using in-depth interviews and participant observation in the campaign to draw out understandings of food sovereignty particular to South African activists. It further assesses tactics and strategies the SAFSC uses, and compares these to current state, business and civil society organisations’ solutions to show how a more grassroots-led approach, using the food sovereignty framework, has the potential to address the roots of hunger. These roots of hunger are shown to be at the corporate food regime level, as has been indicated by the literature and confirmed in this research. As food sovereignty is pursued by various actors in South Africa it provides important examples of approaches by which power in the food system can be reclaimed to benefit the majority instead of a few elites, as is currently the case.

Rapid detection of live pathogens is of paramount importance to ensure food safety. At present, nucleic acid-based polymerase chain reaction and antibody-based lateral flow assays are the primary methods of choice for rapid detection, but these are prone to interference from inhibitors, and resident microbes. Moreover, the positive results may neither assure virulence potential nor viability of the analyte. In contrast, the mammalian cell-based assay detects pathogen interaction with the host cells and is responsive to only live pathogens, but the short shelf-life of the mammalian cells is the major impediment for its widespread application. An innovative approach to prolong the shelf-life of mammalian cells by using formalin was undertaken. Formalin (4% formaldehyde)-fixed human ileocecal adenocarcinoma cell line, HCT-8 on 24-well tissue culture plates was used for the capture of viable pathogens while an antibody was used for specific detection. The specificity of the Mammalian Cell-based ImmunoAssay (MaCIA) was validated with Salmonella enterica serovar Enteritidis and Typhimurium as model pathogens and further confirmed against a panel of 15 S. Enteritidis strains, 8 S. Typhimurium,11 other Salmonella serovars, and 14 non-Salmonella spp. The total detection time (sample-to-result) of MaCIA with artificially inoculated ground chicken, eggs, milk, and cake mix at 1-10 CFU/25 g was 16-21 h using a traditional enrichment set up but the detection time was shortened to 10-12 h using direct on-cell (MaCIA) enrichment. Formalin-fixed stable cell monolayers in MaCIA provide longer shelf-life (at least 14 weeks) for possible point-of-need deployment and multi-sample testing on a single plate.

The effects of various pressure treatments (OK, 30K, 60K, 75K psig) and packing medium (water or cocktail sauce) on shucked oysters were investigated. The pH, moisture content, microbiological tests (including aerobic plate count (APC) and anaerobic plate count (ANPC)), enzyme assays (i.e. α-amylase, β-amylase, lipase and peroxidase activities) were conducted to determine the quality of pressure treated oysters during a 6 week shelf-life study. The moisture content in water-packed oysters under OK, 30K, 60K and 75K psig pressure treatments was slightly increased during storage, while that in cocktail sauce-packed samples was significantly lower than in water-packed samples. Addition of cocktail sauce lowered the pH in oysters, which effectively inhibited the microbial growth, but altered the appearance. The microbial shelf-life of water-packed oysters with pressure treatment of 60K and 75K psig was extended several weeks compared with the controls while 30K psig had less of an effect. Pressure treatments did not inhibit enzyme activities in oysters, however, the addition of cocktail sauce was significant in inhibiting the enzyme activities in this study.
Graduation date: 2000

The validation and potential use of mathematical models to estimate the shelf-life of refrigerated food exposed to temperature abuse and basing such estimations on microbial growth was analyzed. Combined heat transfer, microbial growth models, and non-parametric statistical procedures formed a computer-based predictive tool to assess shelf-life and estimate the accuracy of the prediction. Experiments were carried out to assess the precision of the combined model parameters. The different situations analyzed considered stepwise fluctuations in environmental temperature and a change in package characteristic (size and packaging material). Computer simulations showed that even when the temperature abuse period constitutes a small fraction of the total exposure time (2%-3%), shelf-life can be highly affected (20%-30%). To analyze the precision of the combined model response, two sources of variation were considered, microbial growth and heat transfer parameters. First order, pseudo-zero order kinetics and Arrhenius model formed the basis for the microbial model. The accuracy of lag and exponential phase of microbial growth for a mixture of three microorganisms (P. fluorescens, S. aureus, and A. Iwoffi) was assessed using a nonparametric statistical procedure based on the bootstrap method. The activation energy (E [subscript a]) and the logarithm of the frequency factor (InK₀) were found to be 109±3.4 J/mole and 48.3±1.5 for the exponential phase of this microbial mixture. The values for the exponential phase were 152±4 J/mole and 64.0±1.7, respectively. These parameters together with experimental values for the overall heat transfer coefficient were used to analyze the precision of the model response. This precision was not affected by a change in environmental temperature and packaging characteristics and remained constant at ±1 day. Two different temperature abuse situations yield estimated shelf-life of 4.8±1 and 8.9±1 day, respectively. This result can not be generalized as it depends on the particular examples analyzed.
Graduation date: 1993

In this study we investigate the synergistic effect of low frequency ultrasonication (14, 22, and 47 kHz) on antimicrobial activity of Cecropin P1 against Escherichia coli. The hypothesis was tested by comparing three different treatments (1) ultrasonication only (2) Cecropin P1 only (3) combination of both. The results showed that the combined treatment deactivate E. coli more efficiently by six order of magnitude. The mechanism of membrane permeabilization due to Cecropin P1 is also investigated using dye leakage experiment. The result indicated pore formation and carpet mechanism. Finally, a mathematical modeling is proposed to explain the synergistic effect, allowing us to make better prediction for cell deactivation.

The current trend of increasing demand for minimally processed food requires more effective preservation technologies than are presently used. In this study, an investigation has been made into a novel strategy to control some common foodborne pathogens, and therefore, to provide an alternative means for enhancing the safety and extending the shelf lives of food products. Modified atmosphere is able to extend the shelf life of seafood and meat products. In this study, a simulated controlled atmosphere (CA) broth system was used to investigate the potential of a modified atmosphere rich in CO2 at a concentration of 40%, supplemented with N2, to control common foodborne pathogens, such as Listeria monocytogenes, Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, Staphylococcus aureus and Vibrio parahaemolyticus. Controlled atmosphere significantly reduced the exponential growth rates of all tested pathogens, while the effects on other growth parameters (eg. lag phase duration and maximum population density) depended on the individual species and the specific growth conditions. The CA significantly extended the lag phase durations of S. aureus and V. parahaemolyticus at 20 degrees C at both pH 6.3 and 6.8, and that of L. monocytogenes at both 7 degrees C and 20 degrees C, and at both pH 6.3 and 6.8. The CA also significantly lowered the maximum population densities of S. aureus and V. parahaemolyticus at 20 degrees C, at pH 6.3 and 6.8, S. Typhimurium at pH 6.8, and L. monocytogenes at pH 6.3 and 7 degrees C. E. coli O157:H7 and S. Typhimurium were more resistant to the inhibitory effect of the CA, while S. aureus and V. parahaemolyticus were most sensitive. The inhibitory effect of CA was due mainly to the extensions of the lag phase duration and the reduction of the exponential growth rates of the test pathogens. This study confirms other studies that CA as a means for food preservation provides potential to control foodborne pathogens and therefore enhance the safety of a food product. The use of lactic acid bacteria (LAB) in controlling spoilage microorganisms and pathogens in foods has been a popular research theme worldwide. In this study, the antimicrobial effects of 18 lactic acid bacteria strains were evaluated in vitro, with emphasis on the most effective strain, the newly characterised Lactobacillus reuteri DPC16. The results demonstrated antagonistic effects of many strains against L. monocytogenes, E. coli O157:H7, S. Typhimurium and S. aureus. L. reuteri DPC16 showed the strongest antimicrobial activity against the tested pathogens including both Gram-positive and Gram-negative bacteria. Co-cultivation of L. reuteri DPC16, and co-incubation of its spent culture supernatant (DPC16-SCS), with the pathogens have demonstrated that the antimicrobial effect is bactericidal and valid at pH 4 - 6.5 and at a temperature as low as 10 degrees C. Further characterisation of the antimicrobial effect of L. reuteri DPC16 showed it to be mainly due to the presence of reuterin (ß-hydroxypropionaldehyde), although lactic acid may have also played a role. These characteristics of L. reuteri DPC16 and its metabolite reuterin make it an unique and potent candidate as a biopreservative to control both Gram-positive and Gram-negative bacteria in foods. The combination of L. reuteri DPC16 and CA was assessed for its inhibitory effect on L. monocytogenes using DPC16-SCS and the fermentative supernatant of L. reuteri DPC16 from a glycerol-water solution (DPC16-GFS). The results showed that both of these supernatants, at 25 AU/mL, in combination with CA (60% CO2:40% N2) had a combined inhibitory effect on L. monocytogenes which could not be achieved by any one of the individual factors alone. Analysis of the levels of expression of some stress response genes of L. monocytogenes, after growth in the presence of L. reuteri DPC16 supernatant and/or CA, showed that the expression of some genes was affected including genes betL, gbuA and opuCA responsible for osmosis adaptation and genes gadA, gadB and gadC responsible for acid tolerance. Induction of gbuA, gadB and gadC by the culture supernatant suggests activation of osmotic and acid adaptation and that these genes play a major role in the culture supernatant-induced stresses. An investigation was also carried out to determine if the changes in gene expression conferred a cross-protection to heat. The result showed that the survival of L. monocytogenes grown in the presence of the culture supernatant and CA was significantly increased after exposure to heat treatment at 56oC, suggesting that a cross-protection to thermal stress had been induced. Based on these findings it is proposed that a comprehensive novel strategy incorporating both L. reuteri DPC16 or its fermentative products and a modified atmosphere rich in CO2 could be developed to potentially control foodborne pathogens in food products.