Amostras de açúcar mascavo disponíveis em diferentes estabelecimentos comerciais no município de Araras - SP foram adquiridas para análises. No total foram coletados 49 pacotes de açúcar durante um período de quatro meses. As análises foram realizadas com o objetivo de avaliar a qualidade desse produto oferecido ao consumidor. As análises microbiológicas foram realizadas considerando a pesquisa dos seguintes microrganismos: bactérias mesófilas, bolores e leveduras, esporos de bactérias termófilas flat-sour, esporos de bactérias termófilas anaeróbias produtoras de H2S, esporos de bactérias termófilas anaeróbias não produtoras de H2S, Salmonella, coliformes totais e termotolerantes. Ainda foram determinadas a umidade (%) e atividade de água das amostras. Os resultados das análises apresentaram variações nas concentrações dos microrganismos estudados, cabendo às bactérias mesófilas e aos esporos de termófilas flat-sour as contagens mais elevadas, seguidos por bolores e leveduras. Todas as marcas apresentaram ausência de Salmonella, coliformes totais e termotolerantes, atendendo a legislação brasileira. Em relação ao padrão da National Canners Association apenas as marcas A, G, H e J estavam adequadas a essas exigências, as demais estavam em desacordo. Para o padrão da International Commission for Uniform Methods of Sugar Analysis (ICUMSA), nenhuma marca obteve resultado satisfatório em relação à qualidade microbiológica. Os teores de umidade variaram de 1,94 a 3,63% e os valores de atividade de água variaram de 0,55 a 0,64, havendo diferença significativa ao nível de 5% de significância. Pelos resultados obtidos considera-se que as marcas comercializadas estão de acordo com a legislação vigente no país, porém a maioria das marcas não atende aos padrões internacionais. / Samples of brown sugar available in different establishments in the city of Araras - SP were acquired for analysis. In the total 49 packages of sugar were collected during a period of four months. The analyses were performed to evaluate the quality of that product for the consumer. Microbiological tests were performed to search the following microorganisms: mesophilic bacteria, yeasts and molds, thermophilic flat-sour spores, thermophilic anaerobic H2S-producing spores, thermophilic anaerobic non-producing H2S spores, Salmonella and total and thermotolerant coliforms. Also humidity (%) and water activity were determined for the samples. The analysis results presented variations in the concentrations of microorganisms studied, being the mesophilic bacteria and thermophilic \"flat-sour\" spores the ones with highest scores, followed by yeasts and molds. All brands showed absence of Salmonella and total and thermotolerant coliforms, in accordance with the Brazilian legislation. Regarding the National Canners Association standards, only brands A, G, H and J were suited to these demands, the others being in disagreement. For the International Commission for Uniform Methods of Sugar Analysis (ICUMSA), standards no brands obtained satisfactory results in relation to microbiological quality. The moisture content ranged from 1.94 to 3.63% and the values of water activity ranged from 0.55 to 0.64, significant difference at 5% significance level. By the results it is considered that the brands are marketed under the law of the country, but most brands do not meet international standards. Humidity rates ranged from 1.94 to 3.63% and water activity ranged from 0.55 to 0.64, with significant difference at 5% significance level. By the results obtained it is considered that all brands sold are in accordance to the laws of the country, but most brands do not meet the international standards.

A mussarela de leite de búfala, principal queijo obtido a partir desse leite no Brasil, é um produto praticamente novo no mercado, com alta aceitação pelos consumidores e excelentes perspectivas. Seguindo tecnologia de produção tradicional italiana, caracteriza-se pela intensa manipulação durante a sua elaboração. No presente trabalho, avaliou-se a qualidade microbiológica de duas marcas comerciais de queijo mussarela de leite de búfala, sendo uma das marcas comercializada em embalagem com soro (A) e a outra em embalagem sem soro e a vácuo (B), adquiridas no comércio varejista da cidade de Piracicaba/SP. As análises microbiológicas compreenderam a determinação do NMP de coliformes totais e fecais, a pesquisa de Listeria spp., a contagem de Staphylococcus coagulase-positiva e a pesquisa de Salmonella spp. Com base nos resultados obtidos, pode-se afirmar que as duas marcas analisadas encontram-se em acordo com os padrões microbiológicos legais vigentes. No entanto, pôde-se notar que a qualidade microbiológica dos queijos comercializados em embalagem com soro mostrou-se inferior à dos oferecidos ao consumo em embalagem sem soro e a vácuo. / Buffalo mozzarella cheese, main cheese obtained from buffalo milk in Brazil, is practically a recent product in the market, showing high acceptance by consumers and excellent perspectives. Following traditional italian production tecnology, this cheese is intensely manipulated during its manufacture. In this study, the microbiology quality of two commercial brands of buffalo mozzarella cheese was evaluated, being one of the brands presented in bag with whey (A) while the other one is presented in bag without whey and under vacuum (B). The samples were acquired in the Piracicaba city commerce. Microbiology analysis comprehended the determination of the MPN of total and fecal coliforms, the Listeria spp. presence / absence, the coagulase-positive Staphylococcus accounting and the Salmonella spp. presence / absence. Based on the analysis results, both brands are according to current legal microbiology standards specifications. However, the microbiology quality of the cheeses packed in bag with whey was lower than the microbiology quality of those offered in bag without whey and under vacuum.

246 samples of bulk and packaged spices from retail stores in the western, southeastern, southern, midwestern, and northeastern areas of the U.S. were examined for the presence of Clostridium -perfringens. Isolates were checked for the presence of the lecithinase gene (cpa) and enterotoxin genes (cpe) by PCR. Enterotoxin formation during sporulation was investigated using the Oxoid Toxin Detection Kit. Forty-three confirmed isolates (from 17% of total samples) were cpa-positive. Of those, 27 were cpe-positive. Together, levels of C. perfringens spores ranged from 3.6-2400/gm. The amount of enterotoxin in cell extracts ranged from 2-16 ng/ml. Some of the SEM images of isolated spore (# 78) and one plasmid-borne ent control (FD-153) showed an organized surface structure termed “candy-wrapper”. This extracellular structure remained after treatment with 0.1 % SDS for 1 hr, suggesting it was not composed of membrane debris from the mother cell. The D values of spores ranged from 1.19- 3.31 min. The addition of lysozyme in the plating medium elevated the recovery rate of heat-treated spores. The growth rate of a cocktail of spores from spices (# 31, # 32, # 45) between 4 to 5 hr after inoculation was determined with a doubling time of 6.82 min in hamburger. A cocktail of spores of plasmid-borne ent control showed an optimum growth rate between 5 to 8 hr after inoculation with doubling time of 15.98 min. However; spice isolate cocktail, plasmid-borne ent control cocktail (FD-5603 and FD-153), and a chromosome-borne ent control (NTCT 8239) were unable to germinate and outgrowth at 20oC. Inoculation in laboratory medium FTG indicated the same result as hamburger at 20oC. The ability of C. perfringens spores in spices to potentially survive cooking procedures can be followed by germination and growth of vegetative cells during improper cooling to levels associated with foodborne illness caused by this organism. Our results suggest that retail spices are potential vehicles of transmission of enterotoxin-positive C. perfringens.

Human norovirus (HuNoVs) are the most common enteric pathogen around the world that cause ~50% of foodborne illness of disease outbreaks annually. HuNoVs are the member of the Caliciviridae family, which consist of small (38 nm), unenveloped, single stranded RNA (ssRNA) viruses. Norovirus are divided into 5 genogroup (GI, GII, GIII, GIV, GV, GVI and GVII). The GI, GII, and GIV cause human illness, in addition, GII.4 genotype cause the most human disease. Due to HuNoVs are difficult cultured in vitro, the cultivable HuNoVs surrogates have been widely studied. Recently, some studies have been conducted with HuNoVs surrogates, for example bacteriophage MS2. MS2 is conservative surrogate for nonenveloped viruses which there is a close relationship to the behavior of HuNoVs, thus we can examine the infection control measures for HuNoVs. Despite plenty of treatment method been done on testing antiviral effect on bacteriophage MS2, for example UV inactivation, steam ultrasound and antimicrobial etc., plant-based nanoemulsion treatment has yet to be explored. Carvacrol is a major component of oregano essential oil and is responsible for their antimicrobial activity on the growth of various microorganism. In this study, carvacrol nanoemulsions were formed by using the spontaneous emulsification for testing the nanoemulsion stability (14 days shelf life study on its droplet size and particle charge) and antimicrobial activity. In carvacrol nanoemulsion 14 days shelf life test, the droplet size and particle charge stay stable at three different treatment environments (4°C, 20°C and 37°C). The results proved that nanoemulsion (was formed with surfactant agents and medium-chain triglycerides) is stable system that gives consistent droplet size and charge. Although, the low antimicrobial activity was investigated at carvacrol nanoemulsion, the strong antimicrobial effects have been found when carvacrol or carvacrol combined with ionic surfactant of treatment on MS2 and Escherichia coli. Taken together, in the wake of growing consumer demand for different “natural” products in a number of industries, our study broadly informs the development and study of functionalized carvacrol active compound that can not only provide beneficial health for human but can also examine antimicrobial efficacy of control measures for public health.

Please read the abstract in the section 00front of this document / Dissertation (MSc (Food Science))--University of Pretoria, 1999. / Food Science / MSc / unrestricted

Cocoa bean fermentation is an essential but spontaneous fermentation process to obtain the necessary raw material for the production of cocoa-derived products, among which chocolate. Successful cocoa bean fermentation processes are typically dominated by three microbial groups, namely yeasts, lactic acid bacteria, and acetic acid bacteria. The use of functional starter cultures may allow to gain a better control over the fermentation process. Previously, a number of candidate functional starter cultures have been proposed for the lactic acid bacteria, namely Lactobacillus fermentum 222 and Lactobacillus plantarum 80, and for the acetic acid bacteria, namely Acetobacter pasteurianus 386B, Acetobacter ghanensis LMG 23848T, and Acetobacter senegalensis 108B. The metabolism of bacteria determines an important part of their physiology, and this is recently being investigated by using computational models. The aim of this PhD thesis was to develop such models for the candidate functional starter cultures for the cocoa bean fermentation process and to perform the related computational analysis. The computational models developed were genome-scale metabolic models, which constitute a comprehensive repertoire of metabolic enzymes with their concomitant reactions, and this at genome-scale. The reconstruction of such models requires a combination of high-quality genome re-annotation, comparative genomics, manual curation, and experimental validation. Genome-scale metabolic modeling together with the use of previously published experimental data under cocoa fermentation conditions allowed to contextualize the experimental data and to gain new insights into the metabolic properties of the candidate functional starter cultures. Simulations with the A. pasteurianus 386B genome-scale metabolic model revealed the metabolic roles of lactate and ethanol, the energetic properties of the strains’ aerobic respiratory chain, and the possible functional role of an NAD(P)+ transhydrogenase. Modeling the metabolite dynamics of A. ghanensis LMG 23848T under cocoa fermentation conditions revealed an alternative strategy for its diauxic growth, compared with A. pasteurianus 386B, which was related to a difference in lactate consumption rate and pyruvate overflow. For A. senegalensis 108B, it was shown that, next to lactic acid, also citric acid could sustain its growth in vitro as the sole carbon source. Furthermore, the absence of the glyoxylate cycle predicted from its genome did not correspond with its species description that reports growth on ethanol as the sole carbon source. For L. fermentum 222 and L. plantarum 80, core genome-scale metabolic models allowed to gain insight into the possible metabolic flux distributions as a function of environmental conditions. The modeling also indicated a current lack in knowledge; for example, concerning the presence and consumption of undefined substrates in the complex medium used.In summary, genome-scale metabolic modelling of candidate functional starter cultures for the cocoa bean fermentation process provided useful in silico tools to gain insight into their metabolic properties at a systemic level. / La fermentation du cacao est un processus essentiel pour obtenir la matière première nécessaire pour la production de produits dérivés du cacao, comme par exemple le chocolat. Une fermentation de cacao favorable est caractérisée par la domination de trois groupes de microorganismes :les levures, les bactéries lactiques, et les bactéries acétiques. L'utilisation de cultures de départ fonctionnelles permet un meilleur contrôle sur le processus de fermentation. En ce qui concerne les bactéries, de nombreuses cultures "starter" ont été proposées, à savoir Lactobacillus fermentum 222 et Lactobacillus plantarum 80 pour les bactéries lactiques et Acetobacter pasteurianus 386B, Acetobacter ghanensis LMG 23848T, et Acetobacter senegalensis 108B pour les bactéries acétiques. Le métabolisme des bactéries constitue une partie importante de leur physiologie et la recherche actuelle se concentre de plus en plus sur la modélisation du métabolisme et la simulation des flux métaboliques par ordinateur. Cette thèse de doctorat a été consacrée au développement et à l'analyse de tels modèles computationnels pour des cultures fonctionnelles "starter" proposés pour la fermentation du cacao.Les modèles qui ont été développés dans cette thèse sont des modèles métaboliques à l’échelle du génome. La reconstruction du réseau métabolique a entraîné la ré-annotation du génome, une étude de génomique comparative, la curation manuelle des annotations et la validation du modèle par des expériences in vitro. La modélisation nous a permis de contextualiser des données expérimentales déjà publiées pour en obtenir de nouvelles informations concernant les propriétés métaboliques des cultures starter. Des simulations utilisant le modèle métabolique de A. pasteurianus 386B ont clarifié les rôles métaboliques de l’acide lactique et de l’éthanol, les propriétés énergétiques de sa chaîne respiratoire, et ont permis d'assigner un rôle possible à une NAD(P)+ transhydrogénase. La modélisation de la dynamique des métabolites provenant d’un milieu de croissance de A. ghanensis LMG 23848T dans des conditions simulant la fermentation du cacao, a mis en évidence une stratégie alternative de croissance biphasique comparé à A. pasteurianus 386B. Ceci est dû à une différence dans le taux de consommation de l’acide lactique et à l’éventuelle production de pyruvate. Pour A. senegalensis 108B, les expériences ont démontré, tant pour l’acide lactique que pour l’acide citrique, que ces sources de carbone permettaient, à elles seules, la croissance de cette bactérie. L’absence du cycle du glyoxylate chez A. senegalensis 108B ne correspondait pas à la description de cette espèce, laquelle pouvant croître sur l’éthanol comme seule source de carbone. Pour L. fermentum 222 et L. plantarum 80, la modélisation de leur métabolisme du carbone a permis d’explorer les distributions de flux métaboliques en fonction des substrats consommés. Les simulations ont aussi révélé le manque de connaissance que nous avons sur ces bactéries lactiques, telle que la consommation de substrats non identifiés venant du milieu de croissance et qui pourrait influencer leur dynamique de croissance.En résumé, la modélisation métabolique à l’échelle du génome des cultures starter proposées pour la fermentation du cacao a permis le développement d’outils in silico qui peuvent être utilisés pour mieux comprendre le métabolisme global de ces souches. / Het cacaoboonfermentatieproces is een essentieel maar spontaan proces dat nodig is om de noodzakelijke grondstof, met name de gefermenteerde cacaobonen, voor de productie van cacao-afgeleide producten, waaronder chocolade, te bekomen. Succesvolle cacaoboonfermentatieprocessen worden typisch gedomineerd door drie microbiële groepen, met name gisten, melkzuurbacteriën en azijnzuurbacteriën. Om meer controle te verkrijgen over het fermentatieproces is het gebruik van functionele starterculturen aangewezen. In vorige studies werd reeds een reeks kandidaat-functionele starterculturen voorgesteld. Voor de melkzuurbacteriën zijn dit Lactobacillus fermentum 222 en Lactobacillus plantarum 80 en voor de azijnzuurbacteriën zijn dit Acetobacter pasteurianus 386B, Acetobacter ghanensis LMG 23848T en Acetobacter senegalensis 108B. Het metabolisme van bacteriën bepaalt in grote mate hun fysiologie, en dit wordt recent onderzocht door middel van computationele modellen. Het ontwikkelen en analyseren van zulke modellen voor de voorgestelde kandidaat-functionele starterculturen vormde het onderwerp van deze doctoraatsthesis.De computationele modellen waarvan sprake waren genoomwijde metabole modellen (GEMs), dewelke het repertoire aan metabole enzymen en de biochemische reacties die zij katalyseren in de bacteriële cellen omvat. De reconstructie van het metabole netwerk op genoomschaal vraagt om een gecombineerde aanpak van hoge-kwaliteit genoomherannotatie, comparatieve genomica en experimentele validatie. De GEMs werden gebruikt om reeds gepubliceerde experimentele data onder cacaofermentatiecondities te contextualiseren en nieuwe inzichten te verkrijgen in de metabole karakteristieken van de kandidaat-functionele starterculturen. Door middel van simulaties met het A. pasteurianus 386B GEM kon de metabole rol van melkzuur en ethanol, en de energetische karakteristieken van de aerobe respiratieketen van deze stam aangetoond worden, alsook de mogelijke metabole functie van een NAD(P)+ transhydrogenase. Het modelleren van de microbiële dynamica van A. ghanensis LMG 23848T onder cacaofermentatiecondities wees op een alternatieve strategie voor de tweevoudige groei van deze stam ten opzichte van de tweevoudige groei van A. pasteurianus 386B onder dezelfde condities, en dit omwille van een verschil in melkzuurconsumptiesnelheid en pyruvaatsecretie. Voor A. senegalensis 108B werd aangetoond dat deze stam, naast melkzuur, ook op citroenzuur als enige koolstofbron kon groeien. De afwezigheid van de glyoxylaatcyclus, voorspeld op basis van het genoom, bij A. senegalensis 108B is in tegenstelling tot de soortbeschrijving, dewelke stipuleert dat deze azijnzuurbacteriesoort in staat is tot groei op ethanol als enige koolstofbron. Voor L. fermentum 222 en L. plantarum 80 leidde de ontwikkeling van GEMs tot nieuwe inzichten in de mogelijke metabole fluxverdelingen, voornamelijk ten aanzien van substraatverbruik. Het modelleren van de microbiële dynamica wees ook op een tekortkoming aan huidige kennis over deze stammen, bijvoorbeeld met betrekking tot het gebruik van ongedefinieerde substraten in een rijk groeimedium.Samenvattend werden door middel van de ontwikkelde GEMs van de kandidaat-functionele starterculturen voor cacaoboonfermentatieprocessen nieuwe inzichten verkregen in hun metabolisme en dit op systeemniveau. / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

To assess the effects of gamma radiation at low doses, spores of Penicillium expansum NRRL 971 and Penicillium patulum NRRL 989 were exposed to doses of 0, 50, 100, 150, and 200 krads. The amount of patulin for each culture was quantitatively determined and growth at each dose was evaluated. The results showed a 31% increase in the amount of patulin synthesis after a dose of 150 krads for Penicillium expansum At 200 krads, both fungi showed low levels of patulin production and inhibited growth. Eight color mutants of P. expansum and eight color mutants of P. patulum were produced by exposing spores of each fungus to doses of 150 and 200 krads of gamma radiation. An evaluation of patulin production and growth for each mutant showed that color mutation could result in mutants with an enhanced ability to grow or produce increased levels of patulin. When grown at a subatmospheric pressure of 320 mm Hg, P. expansum produced 30% more patulin per flask, when compared to control cultures. At 160 mm Hg, P. expansum showed a 75% decrease over the control cultures. P. patulum was greatly affected by the temperature of 15℃ and showed decreases both in growth and patulin production. Effective control of growth and patulin synthesis was found for these fungi at 160 mm Hg.

Amino acids derived from the degradation of casein in cheese serve as precursors for the generation of key flavor compounds. Microbial degradation of tryptophan (Trp) is thought to promote formation of aromatic compounds that impart putrid fecal or unclean flavors in cheese, but pathways for their production have not been established. This study investigated tryptophan catabolism by Lactobacillus casei LC301 and LC202 and Lactobacillus helveticus CNRZ32 and LH212 cheese flavor adjuncts in carbohydrate starvation (pH 6.5, 30 or 37°C, no sugar) and cheese-like conditions (pH 5.2, 4% NaCl, 15°C, no sugar). Enzyme assays of cell-free extracts revealed both species of Lactobacillus catabolized tryptophan to indole lactic acid via indole pyruvic acid through transamination followed by dehydrogenation. Micellar electrokinetic capillary chromatography of culture supernatants showed these enzymes also catalyzed the reverse reactions, i.e., conversion of indole lactic acid to tryptophan. Tryptophan decarboxylase activity was detected in Lactobacillus cell-free extracts, but tryptamine was not detected in culture supernatants. Analysis of culture supernatants showed that tryptophan metabolism in Lactobacillus casei did not differ between the two conditions of incubation as it did in Lactobacillus helveticus LH212 and CNRZ32. Lactobacillus helveticus LH212, for example, did not catabolize Trp in carbohydrate starvation but did in cheese-like conditions. While cells of L. helveticus CNRZ32 did not catabolize Trp in either condition, they catabolized indole pyruvic acid to only Trp in carbohydrate starvation and to both Trp and indole lactic acid in cheese-like conditions. Micellar electrokinetic capillary chromatography of culture supernatants incubated under either starvation or cheese-like conditions showed Lactobacillus casei strains produced more indole lactic acid, and Lactobacillus helveticus strains favored tryptophan anabolic reactions. Based on the results obtained in this study, a putative pathway for the catabolism of tryptophan by lactobacilli in cheese is proposed.

A thermophilic fungus was isolated from composted horse manure. The organism was as a Chaetomium sp. by sequencing the highly conserved ITS region of the fungus and comparing to known regions in a genomic database and was referred to as TM-417. TM-417 was found to have an optimal growth temperature of 45 oC and an optimal pH of 7.0. An extracellular DNase and RNase was found to be produced by the isolate and were purified 145.58-fold and 127.6-fold respectively using a combination of size exclusion chromatography and a novel affinity membrane purification system. The extent of purification was determined electrophoretically using 4-15% gradient polyacrylamide gels. Both DNase and RNase were dependent on metal co-factors for activity. The metal ion Mg2+ was the preferred ion for the DNase, whereas for the RNase, Zn2+ and Mn2+ yielded an increase in enzyme activity over that with Mg2+. The purified DNase demonstrated maximum activity at pH 6.0 with no activity at pH 2.0 or 10.0. The RNase exhibited two peaks of maximum activity, on at pH 3.0 and the other at pH 7.0 with no activity at pH 2.0 or 10.0. The optimal temperature for the purified DNase was 65oC. The optimal temperature for the RNase was 70oC. The molecular of the DNase and RNase were determined to be 56 kDa and 69kDa respectively using a Sephadex G-75 column. A standard curve was generated using several standard proteins of known molecular weight.

Within the last decade, pork consumption has steadily increased and continues to be the most consumed meat globally. However, pathogenic bacterial strains resistant to antibiotics have also been increasingly found in pig farms, animals, and the environment. Bacterial food poisoning cases due to Salmonella spp. and Escherichia coli (E. coli) O157:H7 appear to be linked with a variety of pork products. The meat industry has recognized that research is needed to combat the multi-drug resistance in foodborne pathogens with alternative methods of control. This study evaluated the effects of both E. coli- and Salmonella-specific lytic bacteriophages and lactic acid (LA) on E. coli O157:H7, Salmonella Enteritidis, Salmonella Montevideo, and Salmonella Heidelberg growth in raw pork loins ready for marination. The efficacy of the treatments was determined after 1 h of application and marination. Lytic bacteriophage 5% significantly (PSalmonella spp. population by 2.30 log CFU/cm2 when compared with the initial surface attachment. Moreover, the combined treatment of LA 2.5% + phage 5% significantly (PSalmonella population by 2.35 log CFU/cm2 after 1 hour of attachment. In the post-tenderization surface samples, the combination of both phage and LA showed (PP>0.05) when analyzing the translocation of Salmonella spp. on pork loins. Similar treatment efficacy results were observed in the application of E. coli O157:H7 on pork. Following antimicrobial treatments, both control and treated loin samples were enumerated after 1 h at 4°C. Both the lytic bacteriophage 5% and the combination of lytic bacteriophage 5% with lactic acid 2.5% had a significant reduction of E. coli O157:H7 on surface attachment after 1 h of treatment application. Lytic bacteriophage 5% and Lactic acid 2.5% significantly (P < 0.05) reduced the surface bacterial population by 1.89 log CFU/cm2. Lytic bacteriophage 5% alone significantly (P < 0.05) reduced the surface bacterial population by 1.90 log CFU/cm2 when compared with the initial surface attachment groups. Moreover, in the post-tenderization surface samples, lytic bacteriophage 5% and the combination of lytic bacteriophage 5% with lactic acid 2.5% were the only treatments that had a significant reduction (P < 0.05) when compared with the control group. Interestingly, lactic acid 2.5% was the only treatment that had a significant reduction (P < 0.05) of 0.76 log CFU/cm2 when analyzing the translocation of pathogens on pork chops.