The antimicrobial efficacy of carvacrol, trans-cinnamaldehyde, eugenol and thyme oil (0.5, 1, 2 and 5% v/v) was evaluated against Salmonella on turkey breast cutlets as 2 min dip treatments at 4°C. Carvacrol revealed maximum efficacy against Salmonella on turkey cutlets and was selected for 30s and 60s dip treatments at 4°C. Only 5% carvacrol was effective and exhibited 1.0 and 1.8 log CFU/g reduction of Salmonella with 30s and 60s dip treatments, respectively. However, carvacrol showed synergistic activity with carbon dioxide in modified atmosphere packaging (95% CO2/5% O2) against Salmonella and Campylobacter and caused 1.0-2.0 log CFU/g reductions with lower concentrations (0.25, 0.5 and 1.0%). This combination reduced the growth of lactic acid bacteria. In the third experiment, efficacy of lauric arginate (200 and 400ppm) against C. jejuni was tested on chicken breast fillets. Both these concentrations significantly reduced C. jejuni on chicken fillets (1.0-1.5 log CFU/g) at 4°C.

Gram-positive bacteria are part of the normal flora of fish from different aquatic environments. They are mesophilic bacteria and demonstrate optimum growth at ambient temperature. In the sub-tropics, marine fish are caught from seas at temperatures of 16 to 34C, they are usually not iced and are handled at ambient temperature. It was hypothesized that under these conditions Gram-positive bacteria will be abundant in sub-tropical marine fish and will have roles in the spoilage of fish. A review of literature showed that there is a gap in understanding the Gram-positive bacterial populations in sub-tropical marine fish. This is partly due to the fact that the selective media used for isolating Gram-positive bacteria have limitations. Ecological and speciation studies have revealed that the ecology and speciation of many Gram-positive bacteria have not been clearly elucidated. The effect of ambient storage on the individual genera and species of Gram-positive bacteria in fish has been rarely studied. The spoilage potential of Gram-positive bacteria of marine fish origin has not been clearly determined. Therefore, the main aims of this study were to isolate Gram-positive bacteria from fresh and ambient-temperature-stored sub-tropical marine fish, speciate the isolates and study the spoilage potential of the isolates. The practical components of this study were conducted in four parts. The first part dealt with validation of tryptone soya agar with 0.25% phenylethyl alcohol (PEA-TSA) to enumerate Gram-positive bacteria. The second part enumerated Gram-positive bacteria from the muscles, gills and gut of Pseudocaranx dentex (Silver Trevally), Pagrus auratus (Snapper) and Mugil cephalus (Sea Mullet) stored at 25C for 15 hours using PEA-TSA. The third part dealt with the speciation of the isolates using appropriate methods such as polymerase chain reaction, 16S rRNA gene sequence, the VITEK JR system and conventional biochemical methods. In the fourth part, the isolates were assayed qualitatively for their ability to produce volatile sulphur compounds (VSC), reduce trimethylamine oxide and decarboxylate histidine, lysine and ornithine at mesophilic temperature, 32C. Initial studies indicated that PEA-TSA significantly (P< 0.05) reduced the total aerobic bacterial count of fish whereas control Gram-positive bacteria were not affected (P> 0.05). Gram-positive aerobic bacterial counts (GABC) significantly (P< 0.05) increased in the muscles and gills during ambient storage for 15 hours. Within each species, no significant (P> 0.05) differences were found in GABC between muscles and gills. Moreover, there were no significant differences (P> 0.05) in GABC between fish species during storage. In total, 390 bacteria were isolated from the fresh and stored fish; 339 isolates (87%) were found to be Gram-positive. Two hundred and sixty-six isolates (78%) of Gram-positive bacteria were identified to fall into 13 genera, namely Staphylococcus, Micrococcus, Bacillus, Virgibacillus, Brevibacillus, Corynebacterium, Streptococcus, Enterococcus, Aerococcus, Exiguobacterium, Carnobacterium, Vagococcus and Sporosarcina and 30 species. In fresh fish, Staphylococcus epidermidis and Micrococcus luteus were the most frequent isolates. The effect of storage at 25C for 15 hours resulted in a change of Gram-positive bacterial populations; while S. epidermidis, S. xylosus and Bacillus megaterium were no longer present, S. warneri, B. sphaericus, Brevibacillus borstelensis, Enterococcus faecium and Streptococcus uberis increased. Three species, E. faecium, Str. uberis and B. sphaericus, were the most prevalent at the end of storage. Micrococcus luteus and S. warneri were the most prevalent isolates from Pseudocaranx dentex, but E. faecium and Str. uberis were the most frequently isolated from Pagrus auratus and Mugil cephalus. With respect to different parts of the fish body, E. faecium, Str. uberis and B. sphaericus were the most frequent isolates from the muscles, E. faecium, Str. uberis from the gills and M. luteus from the gut. Among the 228 isolates examined, Br. borstelensis 73, Br. borstelensis 291, Str. uberis 339, Vagococcus fluvialis 31 and Vag. fluvialis 132 produced VSC from sodium thiosulphate, cysteine and methionine. However, strains varied in sulphur source utilization. Exiguobacterium acetylicum 5, Exiguobacterium spp. 191, Carnobacterium spp. 338, Br. borstelensis 73, Br. borstelensis 291, Str. uberis 30, Str. uberis 339, Vag. fluvialis 31 and Vag. fluvialis 132 reduced TMAO. No histidine decarboxylase activity was found in the Gram-positive bacterial species tested. Lysine and ornithine were decarboxylated mainly by different strains of S. warneri, S. epidermidis and M. luteus. During ambient storage of fish, the frequency of lysine-decarboxylating bacteria increased and became more diverse after 5 hours of storage. Among fish species examined, the frequencies of lysine- and ornithine-decarboxylating bacteria were higher and more diverse in Pseudocaranx dentex than in Pagrus auratus and Mugil cephalus. This study found that Gram-positive bacteria were abundant and diverse in sub-tropical marine fish; however, their frequencies were affected by fish habitat and fish body part. Ambient temperature storage determined which Gram-positive bacterial species were dominant. With the exception of one isolate of S. aureus, Gram-positive bacteria isolated from sub-tropical marine fish caught from unpolluted water were not potential pathogens. The study also showed that Gram-positive bacteria had greater ability to decarboxylate lysine and ornithine than to produce VSC or reduce TMAO, and the spoilage potential of a bacterial species was a strain-dependent behaviour. This is a significant study as it is the first study on this aspect sub-tropical marine fish. It validated a selective medium that can be used to enumerate most Gram-positive bacteria from a marine environment. Most of the Gram-positive bacterial species from sub-tropical marine fish identified in this study were documented for the first time. The effects of ambient storage and the spoilage potential of Gram-positive bacteria from sub-tropical marine were clearly elucidated.

Thesis (MScAgric (Viticulture and Oenology))--Stellenbosch University, 2008. / The production of good quality wine is essential to ensure competitiveness on an international level. Wine quality is usually evaluated for the visual, olfactory and taste characteristics of that specific wine. The winemaking process starts with the grapes in the vineyard followed by oenological practises in the winery until the final wine is bottled. Factors that could influence wine quality include the grape quality from which the wine is made and different techniques used during wine production. Other factors include the presence as well as the interaction between microorganisms found in the grape juice and wine, and the biochemical effect these microorganisms have on certain chemical compounds in the wine. The different microorganisms found in grape juice and wine can either have a negative or positive contribution to the final quality of the wine. During certain stages of the winemaking process the growth and metabolic activity of certain microorganisms is a necessity to produce good wine. During other stages the presence of certain microorganisms can lead to the development of compounds that is regarded as off-flavours and therefore lead to unpalatable wines of low quality. Yeast strains that naturally present on the grapes and in the winery can also contribute to the final quality of the wine. Brettanomyces yeasts are part of the natural flora of winemaking and can drastically influence the aroma characters of a wine through the production of volatile phenols. The general aroma descriptions of volatile phenols include "smoky", "spicy", "barnyard", "animal" and "medicinal". Although some wine drinkers believe that these characters can add to the complexity of a wine, high levels of volatile phenols is mostly regarded as off-flavours and mask the natural fruity flavours of a wine. With this study we wanted to generate a better understanding of the effect of different winemaking practises on the production of volatile phenols by B. bruxellensis. We evaluated the difference in volatile phenol production when B. bruxellensis was introduced before or after alcoholic fermentation. We have shown that B. bruxellensis could grow and produce volatile phenols during alcoholic fermentation. Results obtained also showed that commercial wine yeast strains could produce the vinyl derivatives that serve as precursors for Brettanomyces yeast to produce the ethyl derivatives. The commercial yeast strains differed in their ability to produce vinyl derivatives. Different malolactic fermentation scenarios were evaluated, namely spontaneous versus inoculated, and with or without yeast lees. Results showed that spontaneous malolactic fermentation had higher volatile phenol levels in the wine than inoculated malolactic fermentation. The treatment with lees reduced the level of volatile phenols, probably due to absorption by yeast cells. The presence of the phenyl acrylic decarboxylase (PAD1) gene and the production of volatile phenols by S. cerevisiae commercial yeast strains were evaluated in Shiraz grape juice and in synthetic grape juice. The results indicated that the yeast strains differ in their ability to produce 4-vinylphenol and 4-vinylguaiacol. All the yeast strains tested had the PAD1 gene. We also evaluated the presence of the phenolic acid decarboxylase (padA) gene and the ability of different lactic acid bacteria strains to produce volatile phenols in synthetic wine media. Although some of these strains tested positive for the phenolic acid decarboxylase gene most of them only produced very low levels of volatile phenols. This study made a valuable contribution on the knowledge about the effect of Brettanomyces yeast on the volatile phenol content of red wines during different stages of the winemaking process and when applying different winemaking practices. It also showed the effect between Brettanomyces yeast and other wine microorganisms and the possible influence it could have on the final quality of wine. Research such as this can therefore aid the winemaker in making certain decisions when trying to manage Brettanomyces yeast spoilage of wines.

The ability of yeasts to grow and adapt under extreme environmental conditions including within the presence of weak organic acid preservatives has led to substantial economic losses through manufactured food and beverage spoilage. The food industry has employed the use of various weak organic acids such as sorbic, benzoic and acetic acid as preservatives to help prevent spoilage by yeasts and moulds. The mechanisms by which S. cerevisiae is able to adapt to these weak organic acids have been extensively studied. A lesser studied weak organic acid preservative is citric acid. The aim of this study was to gain further information on the mechanisms of citric acid adaptation and through this identify potential targets for new preservation strategies. Current knowledge indicates the involvement of the HOG pathway in citric acid adaptation. A citric acid sensitivity screen from a previous study also isolated a SR protein kinase Sky1p, involved in polyamine metabolism, which has been connected with other crucial cellular processes including modulation of ion homeostasis and osmotic shock. In this study we have undertaken a systematic screen for genes that confer increased sensitivity to citric acid paying particular attention to those involved in polyamine metabolism and those known to encode proteins which have evidence of interactions with Sky1p. Many of the deletion strains tested exhibited hypersensitivity to citric acid including Δsky1. Protein-protein interaction maps for Sky1p highlighted an interesting secondary interacting protein Nmd5p, an importin crucial for the nuclear localization of Hog1p. This information suggested there may be the possibility of linkage between Sky1p and Hog1p and their roles in citric acid tolerance, perhaps through Nmd5p. This provided an incentive to perform a range of experiments to test this theory. Proteomic and phosphoproteomic analyses were carried out to study protein expression and phosphorylation changes in response to citric acid stress. Comparative proteomic analyses for Δsky1, Δhog1 and BY4741a with and without citric acid identified four instances of analogous protein expression responses in both Δsky1 and Δhog1, suggesting functional overlap upon exposure to citric acid. Epistasis studies of Δhog1Δsky1 suggested that the two protein kinases do not function on the same pathway. However, overexpression analyses did suggest some functional interaction between Hog1p and Sky1p in mediating citric acid resistance since overexpression of Sky1p in Δhog1 resulted in partial rescue of growth. Further supporting evidence for some functional interaction or linkage was provided by Hog1p phosphorylation and localisation studies. Δsky1 exhibited dual phosphorylation of Hog1p in the absence of citric acid stress; implying that loss of SKY1 results in dual phosphorylation of Hog1p by either prompting phosphorylation or perhaps by interfering with dephosphorylation of Hog1p. Localisation studies of Hog1p proved that like osmotic stress, citric acid stress results in nuclear translocation of Hog1p and deletion of SKY1 seemed to interfere with this localisation to some extent. In light of the results attained in this study we believe we have evidence to propose a novel role for Sky1p in mediating resistance to citric acid and that there is also substantial evidence to suggest that Sky1p shares some functional redundancy and perhaps functional linkage with Hog1p in citric acid adaptation.

Le sel joue un rôle essentiel dans la conservation des produits de charcuterie puisqu’il inhibe le développement bactérien. Or les recommandations nutritionnelles visent à réduire les teneurs en sel dans les denrées alimentaires. Les objectifs de ce projet de thèse étaient (1) de caractériser la communauté bactérienne des chipolatas et le phénomène d’altération sous l’effet d’une réduction de sel et (2) de corréler la dégradation organoleptique des produits aux modifications de la communauté bactérienne.Nous avons tout d’abord caractérisé l’altération des chipolatas par des mesures sensorielles et physico-chimiques. Les travaux démontrent que l’intensité de l’altération est plus importante quand la teneur en sel est réduite et plus particulièrement lorsque les chipolatas sont conditionnées sous atmosphère modifiée. L’altération est caractérisée par la production d’odeurs soufrées, aigres et rances, une diminution du pH des chipolatas et une augmentation de la production d’exsudat.Parallèlement, nous avons décrit la diversité bactérienne des chipolatas altérées par analyse des ARNr 16S bactériens et l’abondance des espèces bactériennes a été quantifiée par qPCR. Par ces méthodes, nous avons pu distinguer la population dominante de la population sous-dominante. La baisse de sel entraine un déséquilibre d’abondance entre ces deux populations et ce déséquilibre résulte non pas de l’augmentation des espèces dominantes mais d’une diminution d’abondance des espèces sous-dominantes.Pour comprendre le rôle respectif de ces populations bactériennes, nous avons analysé leurs activités métaboliques par approche RNAseq. Les travaux montrent une forte activité métabolique des espèces sous-dominantes. Lorsque le sel est en plus faible concentration, l’expression des gènes de Serratia spp. impliqués dans la fermentation du pyruvate pour produire de l’éthanol, du CO2 et de l’acétate est plus importante. La production d’acétate par cette espèce bactérienne peut être reliée aux défauts d’altération observés (baisse de pH, production d’exsudat, odeur aigre).Ces travaux démontrent que le sel impacte la communauté bactérienne des chipolatas (abondance et activités métaboliques) et que cette perturbation compromet la qualité organoleptique des produits. / Salt content plays a key role in meat product preservation since it inhibits bacterial growth. However, dietary guidelines aim to reduce salt content in food. The objectives of this study were (1) to characterize the bacterial community of raw pork sausages and the spoilage phenomenon of these products under salt reduction conditions and (2) to correlate the organoleptic deterioration of the products to modifications in bacterial community.We first characterized the raw pork sausages spoilage by sensory and physicochemical analysis. The work demonstrates that spoilage intensity is greater under a reduced salt content, particularly when sausages are packaged under modified atmosphere. The spoilage is characterized by the production of sulfur, sour and rancid off-odors, a decrease in pH of the sausages and an increase of exudate production.At the same time, we described the bacterial diversity of spoiled sausages through 16S rRNA analysis. Abundance of bacterial species was quantified by qPCR. With these methods, we were able to distinguish the dominant population from the subdominant population. Reducing salt content causes an abundance imbalance between these two populations. This imbalance does not result from an increase of the dominant species; it results from a decrease in abundance of subdominant species.To understand the roles of these bacterial populations, we analyzed their metabolic activities by RNA-Seq approach. The works highlight a high metabolic activity of the subdominant species. When the salt concentration is lowest, the expression of Serratia sp. genes involved in the fermentation of pyruvate to produce ethanol, CO2 and acetate is most important. The acetate production may be connected to the spoilage defaults observed (decrease in pH, exudate production and sour off-odors).These studies reveal that salt reduction impacts the bacterial community of raw pork sausages (abundance and metabolic activities) and this disruption compromises the organoleptic quality of the products.

A nisina é uma bacteriocina produzida por Lactococcus factis subsp. lactis e seu uso é permitido como conservador de alimentos em diversos países. Em 1998, o Ministério da Agricultura e Abastecimento do Brasil, em atitude pioneira, autorizou a aplicação de 200 ppm de Nisaplin® em solução de ácido fosfórico grau alimentício na superfície de produtos cárneos embutidos, tais como salsichas, como alternativa tecnológica para aumentar a segurança microbiológica e a vida-de-prateleira desses produtos. Este trabalho avaliou a eficiência deste tratamento na inibição da deterioração microbiana e controle da multiplicação de Listeria monocytogenes em salsichas tratadas com nisina conforme preconizado, e armazenadas sob refrigeração (8°C) e abuso de temperatura (12°C). Salsichas foram coletadas em uma planta processadora da cidade de São Paulo e submetidas a tratamento por imersão em solução de 200 ppm de Nisaplin® em ácido fosfórico grau alimentício a 0,1 % durante 1 minuto. Para o estudo da eficiência do tratamento sobre Listeria monocytogenes, as salsichas foram contaminadas experimentalmente após o tratamento através de imersão em suspensões contendo L. monocytogenes Scott A, resultando em dois níveis de contaminação superficial denominados baixo (103-104UFC/cm2) e alto (106-107 UFOcm2). Controles não tratados e não inoculados foram realizados concomitantemente. As amostras foram embaladas a vácuo e armazenadas a 8°C e 12°C durante 42 dias. A enumeração das populações de microrganismos aeróbios mesófilos, bactérias láticas, microrganismos psicrotróficos e L. monocytogenes foi realizada semanalmente até o final do período de estocagem. Os resultados indicam que não há diferença estatisticamente significativa entre as populações de bactérias aeróbias mesófilas, psicrotróficas e bactérias láticas em produtos tratados e não tratados com solução de Nisaplin® e armazenados a 8°C ou a 12°C. A redução das populações de L. monocytogenes em salsichas tratadas com Nisaplin® quando a contaminação superficial foi de 103 a 104UFC/cm2 e o produto foi armazenado a 8°e foi pequena (<1 log UFC/cm2), porém estatisticamente significativa. Esses resultados indicam serem necessárias medidas adicionais para garantir a segurança do produto. / Nisin is a bacteriocin produced by Lactococcus lactis subsp lactis and its use is permitted as a food preservative in severaI countries. In 1998, the Brazilian Ministry of Agriculture authorized the application of 200 ppm nisin (Nisaplin®) on the surface of meat products by immersion or spraying, as a technological alternative to increase safety and shelf-life of cooked frankfurters. This work intended to evaluate the effectiveness of this treatment on inhibition of microbial spoilage and control of growth of Listeria monocytogenes in frankfurters during storage under refrigeration (8°C) and temperature abuse (12°C). Frankfurters from a local meat industry were submerged for 1 min in nisin solution (200 ppm) in 0.1% phosphoric acid and experimentally contaminated with low (103-104 CFU/cm2) and high (106-107CFU/cm2) levels of L. monocytogenes. Non-treated and non-inoculated controls were also prepared. Counts of lactic acid bacteria, aerobes (mesophilic and psychrotrophic) and L. monocytogenes were carried out weekly up to 42 days, in four genuine repetitions. Results available indicate that the differences of counts of mesophilic and psychrotrophic aerobes, lactic acid bacteria in nisin-treated and non-treated controls are not significant. Reduction of L. monocytogenes populations in nisin-treated frankfurters experimentally contaminated with low (103-104CFU/cm2) leveis of L. monocytogenes stored under refrigeration (8°C) was low (<1 log UFC/cm<SUP2), but statistically significant. Results indicate that additional measures are necessary to ensure the safety of these products.

The sensing needs for the fresh produce industry can be split into two primary stages: during maturation in the field, also referred to as Precision Farming, and during storage and transport of the produce, or Postharvest Storage. This work seeks to improve the accuracy and reliability of commercially available electrochemical and spectroscopic sensors tailored to the sensing needs of the fresh produce industry. For electrochemical sensing, this study proposes the use of an inline filter to remove polar organic compounds, which can interfere with the readings of a platinum-based electrochemical sensor. A 50% improvement in measurement accuracy was achieved when monitoring the storage headspace of a container of apples. For portable spectroscopy instruments, this study suggests improvements for the alignment of the optical bench and the spectral collect protocol. Methods to reduce the influence of environmental noise, such as variability of background light (sunlight in the field) and thermal effects on hardware performance, are presented. This study also presents the first report of the calibration transfer of spectral regressions developed with Karl Norris's Derivative Quotient Method. The motivation for this aspect of research was to develop methods to collect stable and accurate data in the field, which can be used to improve the quality of fresh produce reaching the customer and reduce premature food spoilage.

No description available.

Eight samples of varying qualities and ages of surimi were used in the development of a new method for testing the quality of surimi. The effects of salt, pH, concentration, heating temperatures and times, and cooling times were observed. These results were used in the development of the McRae-Manning Test. By employing this method, it was possible to determine the difference between fresh high quality surimi, fresh low quality surimi, and old surimi. For this test, surimi was mixed at a 15% concentration, heated at 90 C for 20 minutes in plastic syringes, cooled and evaluated. The surimi was evaluated by emptying the samples onto prepared transparent sheets and measuring how far the sample spreads with time. The transparent sheets had circular measurements which indicated the amount of spread. Best results were obtained when the sheets were elevated at one end.

Growth of bacteria in beer leads to turbidity and off-flavors, resulting in a spoiled and unpalatable product and thus economic loss. The most common beer-spoilage organisms (BSOs) are lactic acid bacteria (LAB), with Lactobacillus and Pediococcus species being the most problematic. Because of the harsh environment (low nutrients, antimicrobial compounds ethanol and hops, anaerobic), only select isolates are able to sustain growth in and spoil beer. To begin understanding the phenomenon of LAB adapting to overcome stresses in beer, ethanol tolerance, hop resistance, and nutrient acquisition mechanisms were investigated. First, ethanol tolerance was analyzed in the context of beer-spoilage ability, and it was found that it is intrinsically high in LAB, thus leading to the conclusion that LAB ability to spoil beer is not dependent on ethanol resistance levels. This was then followed by genome sequencing of the BSO Pediococcus claussenii ATCC BAA-344T (Pc344) to elucidate mechanisms being used to resist hops and acquire low abundance or alternative nutrients. Subsequent analysis of Pc344 and Lactobacillus brevis BSO 464 via reverse transcription quantitative PCR demonstrated the variability found among BSOs in the presence of beer-spoilage-related genes and their use during growth in beer. Further analysis of Pc344 was performed via RNA-sequencing to get a global view of gene expression during mid-logarithmic growth in beer. It was found that several alternative nutrients were being used by Pc344 to sustain growth, and that hop resistance was enabled by a variety of mechanisms including oxidative stress response and pH control. Finally, genomic comparison of BSOs determined that conservation is only present for closely related organisms and that no specific genes/proteins are indicative of an isolate’s beer-spoilage potential. It is more likely that horizontal gene transfer plays a major role in LAB adaption for growth in beer, and that plasmids are very important for this evolution, as was demonstrated by plasmid-variants of Pc344. The main conclusions of this thesis are therefore that hop resistance is the main factor determining ability to grow in beer, and that transfer of genetic elements is the driving force behind LAB evolving into BSOs.