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Listeria monocytogenes is a common post-processing contaminant in ready-to-eat vacuum packaged (VP) cold smoked salmon. Since this psychrotrophic pathogen can grow at refrigerated temperatures (~4°C), other safety barriers in addition to temperature are needed to ensure the continued safety of VP cold smoked salmon. One such novel barrier could be the pulsed light (PL) treatment of the product prior to packaging or treating the product through a transparent package. / Pulsed light destruction kinetics of L. monocytogenes were evaluated while dispensed into a liquid media, on the surface of a general purpose agar and on the surface of cold smoked salmon. Results showed that PL technology was an effective surface sanitation method (a decimal reduction time or D-value of 0.91, 1.37 and 2.25 s exposure of PL at 800, 700 and 600 V, respectively, and a resulting z value of 500 V) on the agar plate. However, it had only a limited success when applied to liquid samples as well as directly on the surface of cold smoked salmon (D-value ranged from 93 s to 24 min). / Sensory quality of VP cold smoked salmon subjected to selected PL treatments was monitored during storage for 14 days at 4°C. Both color and odor scores remained within acceptable limits over the 14 day storage period. Subsequent challenge studies were carried out with L. monocytogenes applied on VP cold smoked salmon. An overall reduction in counts was observed in samples stored at 4°C over 28 days; however, after PL treatment (day 0), there was no significant reduction in counts. Color and odor scores maintained acceptable values over 14 days. Additional experiments were carried out to determine the effects of (1) 1.5% salt, (2) 6% oil, (3) a representative salmon media and (4) background microflora (lactic acid bacteria) on the PL inactivation of L. monocytogenes. All of these factors significantly affected the destruction of L. monocytogenes by increasing the D-value (adding resistance to pulsed light destruction). / Overall, these studies have shown that PL treatment in combination with low temperature storage (4°C) has the potential to extend the shelf-life of VP cold smoked salmon products without compromising sensory quality. However further investigation into higher treatment voltages is necessary in order to achieve a higher target kill of L. monocytogenes.

Increasing consumer demand for fresh fishery products with minimized loss of their nutritional properties is forcing food industry to look for alternative technologies to maintain the fresh attributes, stability and safety of foods. Demand for fresh tuna fish is no exception, being a valuable source of nutrients with immense health benefits. However, this product is highly perishable and has been commonly implicated in scombroid (histamine) poisoning caused by microbial decarboxylation of histidine contained in high levels in the tissues of scombroid fishes. Current techniques are inadequate for the prevention of histamine formation in fresh fishery products and high pressure processing is a potential alternative for it can inactivate microorganisms and enzymes, without affecting (or only minimally altering) the quality characteristics of foodstuffs. Previous studies have shown a decrease in histamine formation after a high pressure treatment and this study focuses on the effect of high pressure on the histidine decarboxylase enzyme and selected histamine forming microorganisms involved in histamine formation. / Commercial histidine decarboxylase suspended in different media (buffer solution and fish slurry with and without added histidine) was submitted to different high pressure treatments (200--400 MPa) with distinct time durations (0--60 min) at room temperature (20°C--25°C). Enzymatic activity of pressure treated and control samples were then compared by measuring histamine formation. Results were similar in all media; a 200 MPa treatment increased the enzymatic activity a little more than 20% as time increased; a 300 MPa treatment increased activity over 20% at first, followed by a decrease in activity as time increased only to reach a level of residual activity similar or only slightly lower than control samples; and a 400 MPa treatment reduced enzyme activity as time increased to a level of 55% residual activity in a buffer solution where the greatest inactivation was observed. / Enzyme activation and inactivation were affected by a dual effect attributed to a pulse effect, which caused a shift in activity and was independent of the length of the treatment, and a pressure-hold effect, during which activation or inactivation followed first order kinetics. The enzyme appeared highly resistant to pressure in all media as observed from D-values (>2700 min) and pressure sensitivity of destruction rate (zp) values (>500 MPa). / Inactivation of non-pathogen histamine forming bacteria (HFB) Escherichia coli K12 and Bacillus megaterium was evaluated by inoculating cultures in a fish tissue homogenate. Surviving colonies were enumerated after the treatments observing inactivation described by the same dual effect described earlier. Pressures above 300 MPa achieved a significant destruction of E. coli K12 (> 4 log-cycles) while B. megaterium appeared highly resistant for only a 2 log-cycle reduction was observed after at the highest pressure treatment conditions (400 MPa, 20 min). / D-values for both microorganisms decreased as pressure increased being significantly smaller for E. coli K 12, which also appeared to be more sensitive to pressure changes as observed from the zp values (zp = 151.51 MPa and zp = 909.10 MPa for E. coli and B. megaterium respectively. Inactivation caused by the pulse effect appeared very effective for both microorganisms as pressure increased, particularly at 400 MPa (PE > 1.25).

Environmental and food spoilage fungi cause billions of dollars in damage in North America alone each year, in the form of rotted wood and crops, spoiled food, and human and animal illness. Each of these threats could be drastically reduced if early and more rapid detection processes are developed to replace the serological methods that are currently in practice. The current North American protocol for establishing identification of contaminating fungi both in environment and food have a time frame of approximately one week to twenty-two days. The use of a Fourier transform infrared (FTIR) spectrometer, coupled with a focal-plan-array (FPA) detector, can theoretically shorten the time (analysis within minutes after obtaining a pure culture) it takes to identify and classify a fungal cell. FPA-FTIR spectroscopy is advantageous as little to no sample preparation is required and results are obtained in less than one minute per sample. The fungal subset chosen for this study includes representatives from five phyla, including Zygomycota (Mucor heimalis), Ascomycota (Neurospora crassa, Ophiostoma minor, Chaetomium globosporum, Alternaria brassicicola), Basidiomycota (Schizophyllum commune, Chaetomium globosporum), Deutromycota (Aspergillus niger, Penicillium notatum, Aureobasidium pullulans) and the Mycetozoa (dictyostelium discoideum, physarum polycephalum). Different variables were tested and evaluated, including variability in growth parameters, wet deposition of fungi versus dry smearing of fungi, optimal absorbance range, and spectral processing parameters as well as discrepancies from one instrument to another, as well as spectral reproducibility from one instrument to another. By following the experimental protocol developed, reproducible spectra were attained, and differentiation of the fungi within the set selected for this study was achieved. The results of this work demonstrate that FPA-FTIR spectroscopy can potentially be employed for the accurate identification of environmental and food spoilage fungi.

The objectives of this study were to determine the microbial ecology of “blown-pack” fresh beef obtained from a federally inspected facility and to use biopreservation to prevent spoilage of vacuum packaged chilled beef. Organic acids and alcohols in the purge obtained from the “blown” beef packages were detected using HPLC. PCR analysis indicated that Clostridium spp. were present in the purge of the commercial sample. Biochemical tests, RFLP and 16S rDNA sequencing were used to identify organisms isolated from the meat. Out of 66 isolated strains, 26 isolates were strict anaerobes and RFLP indicated that all were clonal isolates. Based on sequence analysis, the isolate was identified as Clostridium putrefaciens. The isolate caused “blown-pack” spoilage and produced butyric and propionic acids when inoculated onto fresh meat and Carnobacterium maltaromaticum UAL307 prevented the production gas and obvious signs of spoilage. / Food Science and Technology

Thesis (Ph. D.)--Ohio State University, 2007.

Orientador: Oswaldo Durival Rossi Junior / Banca: Luiz Augusto do Amaral / Banca: Ana Maria Centola Vidal Martins / Resumo: A deterioração "blown pack" é caracterizada por abundante produção de gás, induzindo a completa distensão da embalagem durante o processo de estocagem sob refrigeração. Quando a embalagem é aberta, há um odor desagradável, levemente fecal. O gás presente na embalagem é composto por dióxido de carbono e hidrogênio e por vários tipos butíricos do metabolismo fermentativo. O objetivo deste experimento foi determinar possíveis causadores deste tipo de deterioração, quantificando as populações de bactérias da família Enterobacteriaceae, e caracterizando-as nos principais gêneros e espécies encontradas, o número de bactérias ácido-lácticas, a freqüência de Clostridium estertheticum e do Clostridium gasigenes, em carnes próprias para o consumo e em carnes que apresentaram a deterioração "blown pack". Para contagem e identificação dos membros da família Enterobacteriaceae e contagem de bactérias ácido-lácticas utilizou-se de técnicas microbiológicas clássicas. Já para pesquisa do C. estertheticum e C. gasigenes fez-se uso de técnicas de biologia molecular. Os microrganismos da família Enterobacteriaceae e bactérias ácido-láticas estavam presentes em populações elevadas e em maior número nas carnes com deterioração "blown pack". A espécie mais freqüentemente encontrada foi a Hafnia alvei. As amostras com deterioração "blown pack' apresentaram maior positividade para o C. estertethicum que amostras não deterioradas. Não houve diferença estatística de positividade para a presença do C. gasigenes entre amostras com deterioração "blown pack" e carnes não deterioradas. A principal forma de controle desta deterioração é a prevenção da contaminação da carne por material fecal. / Abstract: The "blown pack" spoilage is characterised by abundant gas production, leading to complete gross distention pack during refrigerated storage. When the packaging is opened, there is an unpleasant smell, lightly fecal. The gas present in the package is composed of carbon dioxide and hydrogen and also of several butyric types of metabolism fermentation. The purpose of this experiment was to determine possible causes of this spoilage type, quantifying the populations of bacteria of the family Enterobacteriaceae, and characterizing them in the major genera and species found, the number of lactic acid bacteria, the frequency of Clostridium estertheticum and Clostridium gasigenes in meat proper for consumption and meat which showed the "blown pack" spoilage. In order to enumerate and identify the members of the Enterobacteriaceae family, and to enumerate the lactic acid bacteria the procedure was classical microbiological techniques. However to search the C. estertheticum and C. gasigenes the procedure was molecular biology techniques. The microorganisms of the family Enterobacteriaceae and lactic acid bacteria were present in large populations and in greater numbers in meat with "blown pack" spoilage. The species which were found more often was the Hafnia alvei. Samples of "blown pack" spoilage had greater positive features for C. estertethicum than samples not damaged. There was no statistical difference of positive features for the presence of C. gasigenes between samples of "blown pack" spoilage and not damaged meat. The main way to control this spoilage is the prevention of contamination of meat by fecal material. / Mestre

Dissertação(mestrado) - Universidade Federal do Rio Grande, Programa de Pós-Graduação em Engenharia e Ciência de Alimentos, Escola de Química e Alimentos, 2008. / Submitted by Caroline Silva (krol_bilhar@hotmail.com) on 2012-09-17T00:08:20Z No. of bitstreams: 1 tese mestrado liane.pdf: 1042970 bytes, checksum: bb0c0bb945531f1ebe8e3aca39dd22de (MD5) / Approved for entry into archive by Bruna Vieira(bruninha_vieira@ibest.com.br) on 2012-09-20T18:27:46Z (GMT) No. of bitstreams: 1 tese mestrado liane.pdf: 1042970 bytes, checksum: bb0c0bb945531f1ebe8e3aca39dd22de (MD5) / Made available in DSpace on 2012-09-20T18:27:46Z (GMT). No. of bitstreams: 1 tese mestrado liane.pdf: 1042970 bytes, checksum: bb0c0bb945531f1ebe8e3aca39dd22de (MD5) Previous issue date: 2008 / Durante a produção, armazenamento, transporte e embalagem de produtos alimentícios, a presença de microrganismos é inevitável. A carne de frango, especialmente, é um alimento altamente perecível e, juntamente com outros tipos de carnes provenientes de ave, apresenta grande variedade de bactérias patogênicas e deteriorantes quando comparada com outros alimentos. Assim, a avaliação do crescimento microbiano e o controle da temperatura de armazenamento são importantes para garantir a segurança e vida útil dos alimentos. O trabalho objetivou caracterizar microbianamente três peitos de frango (cru, temperado com NaCl e cozido), produzidos no Brasil e exportados para a Europa, bem como, estimar a vida útil de tais produtos após o descongelamento, quando armazenados em diferentes condições de temperatura. Foi simulada a cadeia de abastecimento dos peitos de frango congelados, durante 20 dias a -18 ± 0,5 C para simulação do transporte destes produtos por navio até a Europa, desde sua expedição no Brasil e, após descongelamento, durante 21 dias a 4 ± 0,5 C para simulação da vida útil em gôndola de supermercado. Os produtos foram analisados quanto a Pseudomonas spp., Salmonella spp., Listeria monocytogenes, Staphylococcus spp. e microrganismos viáveis totais (mesófilos e psicrotróficos). Em termos de contagens de número de colônias viáveis totais, durante os primeiros 20 dias (a -18°C), a presença de microrganismos se manteve estável em baixos níveis de detecção. Após o descongelamento foram observadas as curvas de crescimento dos microrganismos, onde as fases lag ocorreram durante os primeiros 6 dias. A máxima concentração microbiana foi atingida após 15-18 dias, dependendo do produto. Em nenhuma das amostras foi detectada a presença de Salmonella spp. e Listeria monocytogenes. Após este estudo inicial foi avaliado o tempo de vida útil dos três produtos em diferentes condições de temperatura de armazenamento (2, 4, 7, 10, 15 e 20ºC). O estudo foi baseado na determinação de microrganismos aeróbios psicrotróficos, Pseudomonas spp., aeróbios mesófilos e Staphylococcus spp. O aumento da temperatura fez reduzir a vida útil dos três produtos estudados, em relação a todos os microrganismos. De modo geral, os produtos apresentaram faixas de vida útil de 10 até mais de 26 dias, a 2ºC; de 9 até mais de 21 dias, a 4ºC; de 6 até 12 dias, a 7ºC; de 4 até 8 dias, a 10ºC; de 2 até 4 dias, a 15ºC; e de 1 a 2 dias, a 20ºC. Quando armazenadas em temperaturas de refrigeração (2, 4 e 7°C), as amostras apresentaram pouca variação no tempo de vida útil, especialmente a 2 e 4°C. Já quando armazenadas à temperatura ambiente (temperaturas iguais ou superiores a 10ºC), a cada 5ºC de elevação na temperatura de armazenamento, a vida útil reduziu-se à metade do tempo. / During production, storage, transport and packaging of food products, the presence of microorganisms is unavoidable. The chicken meat, especially, is a highly perishable food and, together with other types from poultry meat presents great variety of pathogenic bacteria and spoilage when compared with other foods. Like this, the evaluation of the microbial growth and the control of the storage temperature are important to guarantee the safety and shelf life of the foods. The work aimed to characterize microbially, three chicken breast (raw, tempered with NaCl and cooked), produced in Brazil and exported to Europe, as well as, to estimate the shelf life of such products after thawing, when stored in different temperature conditions. The supply chain of frozen chicken breast was simulated for 20 days at -18±0,5oC for simulation of the transport of these products by ship to Europe, from its expedition in Brazil and, after thawed, during 21 days at 4 ± 0,5oC for simulation of the supermarket shelf life. Pseudomonas spp., Salmonella spp., Listeria monocytogenes, Staphylococcus spp. and total viable microorganisms (mesophilic and psicrotrophic) were analyzed in the products. In terms of accountings of total viable colonies number, during the first 20 days (-18°C), the presence of microorganisms was stable at low levels of detection. After thawing the microorganism growth curves showed that the lag phases happened during the first 6 days. The highest microbial concentration was reached after 15-18 days, according to the product. In none of the samples the presence of Salmonella spp. and Listeria monocytogenes were detected. After this initial study it was evaluated the shelf life time of the three products in different conditions of storage temperature (2, 4, 7, 10, 15 and 20ºC). The study was based on the determination of aerobic psicrotrophic, Pseudomonas spp., aerobics mesophilic and Staphylococcus spp. The increase of temperature reduce the shelf life of the three studied products, in relation to all of the microorganisms. In general, the products presented shelf life ranges from 10 to more than 26 days, at 2ºC; from 9 to more than 21 days, at 4ºC; from 6 to 12 days, at 7ºC; from 4 to 8 days, at 10ºC; of 2 to 4 days, at 15ºC; and, from 1 to 2 days at 20ºC. When stored in refrigerating temperature (2, 4 and 7°C), the samples presented a few variation in the shelf life time, especially at 2 and 4°C. But when stored to room temperature (at 10ºC or higher temperature), to each 5ºC of elevation in the storage temperature, the shelf life was reduced to half time.

Si les flores contaminantes représentent la plupart du temps, dans les conserves, un risque industriel aujourd'hui maitrisé, la flore d'altération, de par sa résistance importante à la température, continue à constituer une cause de pertes économiques majeures. Pourtant cette dernière restait cependant peu caractérisée. En s'appuyant sur les travaux réalisés ces dernières années au sein de l'unité de microbiologie EMaiRIT'S du CTCPA (unité d'Expertise dans la Maitrise du Risque Industriel en Thermorésistants Sporulés du Centre Technique de la Conservation des Produits Agricoles), les principaux objectifs de cette thèse ont été (i) d'identifier et caractériser, en vue de sa maitrise ultérieure, la flore d'altération sporulante (ii) d'identifier l'origine de ces flores dans les conserveries et enfin (iii) de déterminer des moyens de maitrise.Pour cela, un état des lieux des bactéries sporulées d'altération des conserves a été effectué avec la collaboration de 122 conserveries sur plus de 10 ans en France. Cette caractérisation des espèces altérantes a permis l'élaboration d'un outil de biologie moléculaire (SporeTraQTM) afin d'identifier rapidement ces germes ou de pouvoir les détecter au sein d'une population complexe. En parallèle, l'amélioration de la connaissance de la thermorésistance de ces espèces, principale caractéristique de la flore sporulante, a été menée. A ce paramètre, il a été associé une relation avec la chimio résistance des spores. Identifiée, nous avons cherché à localiser cette flore d'altération au sein des usines à l'aide de plusieurs campagnes de prélèvements sur différents légumes. Au final, la flore spécifique du procédé de fabrication des conserves a été identifiée, caractérisée et localisée en vue d'améliorer la maitrise du risque microbien soit par une maitrise des contaminations et/ou un nettoyage plus performant (localisation au niveau d'étapes unitaires, efficacité de molécules sporicides) soir par un barème optimisé (en relation avec la thermorésistance). De plus, ce travail a été conduit au sein d'une approche bénéfice/risque représentant le futur de l'évolution des procédés agro-alimentaires associant amélioration de la qualité nutritionnelle et maintien de la maitrise sanitaire. Cette thèse s'appuie sur 5 publications de rang A. / Microbial contaminants of safety concern represent most of time, in canned food, an industrial risk which is well mastered. However, the spoilage flora, due to its high heat resistance, is responsible for major economic losses. Nevertheless, these bacteria remained poorly characterized. Based on the works realized during last 10 years within the EMaiRIT'S unit of microbiology of the CTCPA (expertise unit of the French Technical Center of the Preservation of Food, focused on Management of Industrial Risk liked to Heat Resistant Spores), the main objective of this thesis were: i) to identify and to characterize, with the aim of its later control, the spoilage spore forming bacteria florae ii) to identify the origin of these florae in canning factories and finally iii) to determine ways of control.For that purpose, a current inventory of spore forming bacteria in spoiled canned food was made with the cooperation of 122 canning factories over more than 10 years in France. This characterization of the spoilage species allowed the elaboration of a molecular biology tool (SporeTraQTM) for quick identification of these germs or their detection within a complex population. In parallel, the improvement of the knowledge about the heat resistance of these species, main characteristic of the spores, was led. In addition, the chemical resistance of spores was investigated. When identified, we tried to localize these spores on canning factories lines, with several sampling plans, on various vegetables. At the end, the specific spore forming bacteria related to the industrial canning process was identified, characterized and localized, allowing to improve the microbial risk control either by a more efficient cleaning, and through optimized process schedules. Furthermore, this work was driven within a benefic / risk approach representing the future of the food-processing evolution with improvement of the nutritional quality and the preservation of the sanitary control.This thesis leans on 5 publications of rank A.

Les eaux florales sont des matières premières aromatiques issues de la distillation, contenant généralement moins de 1 g/L de composés volatils leur conférant leurs propriétés organoleptiques. Elles sont utilisées principalement en industrie agroalimentaire et cosmétique. Elles sont sujettes à des problèmes d’instabilité microbienne incompatibles avec leurs applications. Ces microorganismes, leurs dynamiques, ainsi que les nutriments disponibles nécessaires à leur croissance restent mal connus. Les eaux florales sont actuellement stabilisées par ajout de conservateurs dont certains sont controversés et visent à être retirés du marché. De plus, leur efficacité dans les eaux florales n’a pas été évaluée.L’objectif de cette thèse est d’apporter une meilleure connaissance de la composition des eaux florales et de ses contaminants afin de proposer une méthode de stabilisation adaptée.La composition en huile essentielle et le microbiote de 22 échantillons d’eau de fleur d’oranger (Citrus aurantium L. ssp. amara L.) et de rose (Rosa damascena Miller et Rosa centifolia L.), provenant de différents producteurs autour du bassin méditerranéen, ont été analysés afin de déterminer les facteurs responsables de leur altération. Bien que les composés volatils soient connus pour leurs propriétés antimicrobiennes, leurs concentrations dans les hydrolats ne sont pas suffisantes pour assurer la stabilité microbiologique. En plus des composés volatils, les hydrolats contiennent des composés non-volatils tels que des sucres, entraînés vers le distillat par effet de primage ou de moussage pendant la distillation, et pouvant être utilisés comme substrat de croissance par les microorganismes. La population microbienne peut atteindre 106 à 107 UFC/mL en quelques jours à température ambiante et jusqu’à 3 mois à 5°C. Des bactéries environnementales, oligotrophes, et acido-tolérantes, appartenant principalement aux genres Pseudomonas sp. et Burkholderia sp. ont été isolées et identifiées. Parmi ces bactéries, B. vietnamiensis et Novosphingobium capsulatum ont été capables de métaboliser des composés volatiles tels que le géraniol ou l’acétate de 2-phényléthyle pour produire la 6-méthyl-5-heptèn-2-one ou le 2-phényléthanol, et modifier ainsi les propriétés organoleptiques des hydrolats. Enfin, la capacité de croissance de bactéries pathogènes et d’altération dans les hydrolats a été évaluée, et différents conservateurs ont été testés sur les souches capables de se multiplier dans les hydrolats.Une distillation aseptique et un conditionnement stérile permettrait d’assurer la stabilité des hydrolats sans ajout de conservateurs. En l’absence de conditions aseptiques, l’ajout de conservateurs est nécessaire pour assurer la stabilité des hydrolats. / Hydrosols are hydrodistillation products mainly used as food flavoring agents or ingredient in cosmetics. They contain less than 1 g/L of dispersed essential oils giving the organoleptic properties. These are subjected to microbial proliferation that can prevent use due to non-compliance to professional microbiological standards. The microorganisms, their growth dynamics, and the available nutrients in hydrosols remain unknown. Hydrosols can contain few preservatives, but there is no data about their efficiency in hydrosols.The aim of this study was to have a better knowledge on hydrosols composition, their microbiota, and spoilage conditions, in order to propose an adapted stabilization method.The composition in volatile compounds and the microbiota of 22 hydrosol samples of Citrus aurantium L. ssp. amara L. (orange blossom), Rosa damascena Miller (rose D.), and Rosa centifolia L. (rose C.) flowers were analyzed to determine factors responsible for decay. Some non-volatile compounds were likely carried over during distillation by a priming and foaming effect, and could be used as nutrients by microorganisms. Concentrations of volatile compounds in hydrosols are not high enough to prevent microbial proliferation, and bacteria concentrations can reach up to 106 CFU/mL in both hydrosols. The isolated microbial population was composed of oligotrophic Gram-negative bacteria, arranged in four major genera: Pseudomonas sp., Burkholderia cepacia complex, and presumably two new genera belonging to Acetobacteraceae and Rhodospirillaceae. Among those bacteria, Burkholderia vietnamiensis and Novosphingobium capsulatum were able to metabolize volatile compounds, such as geraniol to produce 6-methyl-5-hepten-2-one or geranic acid, or 2-phenylethyl acetate to produce 2-phenylethanol. Finally, the growth potential of a range of bacteria isolated from hydrosols and of pathogenic micro-organisms was evaluated, then the anti-microbial activity in nutrient broth and/or in hydrosols of a range of chemical preservatives authorized for food and cosmetic applications was tested.Additional hurdles such as chemical preservatives or aseptic packaging will be necessary to insure microbial stability.