Tyrophagus putrescentiae, (ham mite) is difficult for commercial dry cured ham producers to control. This research was conducted to test the efficacy of C8C9C10 fatty acids combined with and without food grade coatings to control mite infestations on dry cured hams. Ham cubes were coated directly or wrapped in nets saturated with combinations of xanthan gum (XG) or carrageenan (CG), propylene glycol alginate (PGA), and either propylene glycol or C8C9C10 fatty acid. The use of 10% C8C9C10 with XG and CG + PGA in direct coatings and 1% C8C9C10 with XG or 10% with both XG and CG/PGA in saturated nets inhibited mite population growth. Unexpectedly, the soybean oil solvent effectively controlled mite infestation. Sensory evaluation indicated that 10% C8C9C10 mixed with soybean oil and 100% soybean oil did not impart sensory differences to ham when used as a coating but did impact sensory attributes when used with nets.

Mechanical performance characteristics were compared for 100% polyester treated and untreated weft-knitted mesh fabric that are used to contain dry-ageing hams, commonly referred to as ham nets. The treated net was coated with a patent-pending food-grade chemical solution (40% Propylene Glycol + 1% Propylene Glycol Alginate + 1% Carrageenan) to control ham mites. Both treated and untreated ham nets were compared for mechanical performance characteristics based on the following standards: abrasion resistance (ASTM D4966), elastic recovery (BS EN 14704-1:2005), breaking strength (ASTM D5034-09), and bursting strength (ASTM D3786). Results indicate that the coating had minimal to no impact on the mechanical performance characteristics of ham nets. SEM images also showed no negative effect on the fiber morphology due to the applied chemical solution. Findings support the use of treated ham nets to increase the end-use functionality and provide stakeholders an option for integrated pest management without compromising performance needs.

The objective of this research was to evaluate if chitosan and another food-grade compound can be incorporated into food-grade coatings in conjunction with propylene glycol and polysaccharides to control Tyrophagus putrescentiae growth without affecting sensory attributes, water activity, and moisture content of dry-cured hams. Each coating solution was coated on ham cubes (2.5 × 2.5 × 2.5 cm3, n = 5/treatment) or infused in ham nets, and dry-cured ham cubes were wrapped in the ham nets prior to inoculation with 20 mixed-sex adult mites. Randomized complete block designs with two replications were utilized to evaluate the efficacy of treatments at controlling mite growth on dry-cured ham cubes. When CH was mixed with XG and infused into a net, fewer mites (15.7 and 21.0 mites) were on ham cubes (P < 0.05) in comparison to the control (211.2 mites). Results indicated that CH contributed to controlling mite growth since 1% XG alone did not control mite growth. Difference from control test results indicated that no sensory differences existed (NS) between CH-treated and control ham slices. The addition of a food-grade compound (SP or 24P) to the XG coatings enhanced mite control efficacy. When 1% SP was added to 1% XG, it effectively controlled mite growth in both coating and netting treatments. Increasing 1% SP to 2% SP did not significantly (NS) control mite growth in the coating solution but did control mite growth when infused in the net. Both coating and netting treatments with 2% 24P + 1% XG controlled mite growth. When 24P was the only ingredient in the treatment, both 1% and 2% 24P controlled mites when infused in the net. The addition of SP did not (NS) impact the sensory attributes of the dry-cured ham. The 2% 24P + 1% XG treatment differed in moistness (P < 0.05) when compared to the blind control, but the rating was still less than 2 (moderately different). Overall, results from these studies indicate that chitosan can potentially be added in coatings or ham nets as tools to control mites in an integrated pest management program for dry-cured hams.

Na cidade de São Paulo, cerca de 50 mil pessoas são portadoras do HTLV-1. O HTLV-1 é o agente causador da paraparesia espástica tropical/mielopatia associada com o HTLV-1 (TSP/HAM) e os mecanismos desta patogênese são obscuros. A TSP/HAM é considerada uma doença imuno-mediada e algumas citocinas, podem estar associadas, com a desmielinização da membrana de mielina da coluna espinhal, provavelmente, estimuladas pela presença de antígenos virais. A PCR em tempo real é uma técnica utilizada para a detecção de citocinas e apresenta maior sensibilidade na expressão de mRNA e carga proviral do HTLV-1. Objetivos: Detecção de citocinas pelas técnicas sorológicas e de biologia molecular (PCR em tempo real) que auxiliaram no aconselhamento e avaliação de pacientes infectados pelo HTLV-1, além de uma avaliação da carga proviral dos pacientes com infecção pelo HTLV-1. Casuística: Grupo I: indivíduos soronegativos para HCV, HIV-1 e HTLV-1 (Controle); Grupo II: pacientes HTLV-1 assintomáticos; Grupo III: pacientes com TSP/HAM. Observamos que o INF-<font face=\"symbol\">g apresenta-se alterado na infecção HTLV-1, sendo um dos fatores mais importantes da evolução para TSP/HAM, ambos os ensaios apresentaram o mesmo resultado. A carga proviral pode ser um marcador de progressão para a TSP/HAM. A interação complexa existente entre o HTLV-1 e as células responsáveis pela liberação de citocinas e <font face=\"symbol\">b-quimiocinas pró-inflamatórias, assim como elementos de ativação celular, resultam na ativação imunológica. Isto, lentamente, pode levar ao processo inflamatório crônico que atua diretamente no micro ambiente neuronal e/ou membrana de mielina da coluna espinhal, em alguns portadores de HTLV-1. Com a persistente ativação do sistema imunológico, este processo provocará destruição das células gliais e dos neurônios, dando início ao processo de desmielinização. / In São Paulo, about 50 thousand people are HTLV-1 carriers. HTLV-1 is the agent that causes tropical spastic paraparesis and HTLV-1 associated myelopathy (TSP/HAM) despite the mechanisms of this disease are still unclear. TSP/HAM is considered an immune mediated illness and some cytokines might be involved with axonal damage and demyelination, most pronounced in the midthoracic spinal cord, probably stimulated by the presence of viral antigens. Real Time PCR is a used technique to cytokine detection and shows higher sensitivity on mRNA expression and HTLV-1 proviral load. Objectives: cytokine detection through molecular biology (Real time PCR) and serologic techniques that helped on advising and assessment of HTLV-1 infected patients and also on their proviral load. Casuistic: Group I: seronegative individuals for HCV, HIV-1 and HTLV-1 (control); Group II: asymptomatic HTLV-1 infected patients; Group III: TSP/HAM patients. We\'ve been observed that <font face=\"symbol\">g-interferon presents changing on the HTLV-1 infection, being one of the most important factors to TSP/HAM progression, both essays showed the same results. The proviral load may be an important marker for TSP/HAM development.

Na cidade de São Paulo, cerca de 50 mil pessoas são portadoras do HTLV-1. O HTLV-1 é o agente causador da paraparesia espástica tropical/mielopatia associada com o HTLV-1 (TSP/HAM) e os mecanismos desta patogênese são obscuros. A TSP/HAM é considerada uma doença imuno-mediada e algumas citocinas, podem estar associadas, com a desmielinização da membrana de mielina da coluna espinhal, provavelmente, estimuladas pela presença de antígenos virais. A PCR em tempo real é uma técnica utilizada para a detecção de citocinas e apresenta maior sensibilidade na expressão de mRNA e carga proviral do HTLV-1. Objetivos: Detecção de citocinas pelas técnicas sorológicas e de biologia molecular (PCR em tempo real) que auxiliaram no aconselhamento e avaliação de pacientes infectados pelo HTLV-1, além de uma avaliação da carga proviral dos pacientes com infecção pelo HTLV-1. Casuística: Grupo I: indivíduos soronegativos para HCV, HIV-1 e HTLV-1 (Controle); Grupo II: pacientes HTLV-1 assintomáticos; Grupo III: pacientes com TSP/HAM. Observamos que o INF-<font face=\"symbol\">g apresenta-se alterado na infecção HTLV-1, sendo um dos fatores mais importantes da evolução para TSP/HAM, ambos os ensaios apresentaram o mesmo resultado. A carga proviral pode ser um marcador de progressão para a TSP/HAM. A interação complexa existente entre o HTLV-1 e as células responsáveis pela liberação de citocinas e <font face=\"symbol\">b-quimiocinas pró-inflamatórias, assim como elementos de ativação celular, resultam na ativação imunológica. Isto, lentamente, pode levar ao processo inflamatório crônico que atua diretamente no micro ambiente neuronal e/ou membrana de mielina da coluna espinhal, em alguns portadores de HTLV-1. Com a persistente ativação do sistema imunológico, este processo provocará destruição das células gliais e dos neurônios, dando início ao processo de desmielinização. / In São Paulo, about 50 thousand people are HTLV-1 carriers. HTLV-1 is the agent that causes tropical spastic paraparesis and HTLV-1 associated myelopathy (TSP/HAM) despite the mechanisms of this disease are still unclear. TSP/HAM is considered an immune mediated illness and some cytokines might be involved with axonal damage and demyelination, most pronounced in the midthoracic spinal cord, probably stimulated by the presence of viral antigens. Real Time PCR is a used technique to cytokine detection and shows higher sensitivity on mRNA expression and HTLV-1 proviral load. Objectives: cytokine detection through molecular biology (Real time PCR) and serologic techniques that helped on advising and assessment of HTLV-1 infected patients and also on their proviral load. Casuistic: Group I: seronegative individuals for HCV, HIV-1 and HTLV-1 (control); Group II: asymptomatic HTLV-1 infected patients; Group III: TSP/HAM patients. We\'ve been observed that <font face=\"symbol\">g-interferon presents changing on the HTLV-1 infection, being one of the most important factors to TSP/HAM progression, both essays showed the same results. The proviral load may be an important marker for TSP/HAM development.

Ces travaux de thèse s’inscrivent dans le cadre du projet ANR HUMIBATex « Comment prédire les désordres causés par l’humidité ? Quelles solutions techniques pour rénover le bâti existant ? » (2012-2016). Elle traite de la modélisation numérique et expérimentale des transferts couplés hydro-thermo-aérauliques à différentes échelles : matériau, paroi et ambiance de bâtiment. Sur le plan théorique, un modèle phénoménologique des transferts couplés de chaleur, d’air et d’humidité à travers les enveloppes de bâtiments (HAM) a été élaboré. Après la phase de validation (confrontation avec des solutions analytiques et des résultats expérimentaux), ce modèle a été implémenté avec confiance dans un code de simulation thermique dynamique du bâtiment (BES). Ceci a permis, ainsi, de développer une plateforme de co-simulation HAM-BES. Grâce à l’outil mis en œuvre, les comportements hygrothermiques de la paroi et de l’ambiance habitable des bâtiments ont été prédits finement. Deux cas d’études ont été entrepris. Le premier avait pour but de mettre en évidence l’impact des transferts hygrothermiques sur la prédiction des consommations énergétiques. Le deuxième cas d’étude a été dédié à l’étude de l’efficacité de différentes stratégies de ventilation (extraction ou insufflation) sur le contrôle et la diminution des risques d’apparition de désordres liés à l’humidité au niveau des bâtiments résidentiels. Sur le plan expérimental, une campagne de caractérisation des propriétés physiques, hydriques et thermophysiques des matériaux de construction a été effectuée. Cette campagne expérimentale s’est focalisée sur l’analyse de l’impact de l’état thermique et hydrique du matériau sur les valeurs des propriétés hygrothermiques. Dans un autre travail expérimental, des dispositifs expérimentaux, à petite échelle mais également à l’échelle de la paroi, ont été conçus au laboratoire dans le but d’étudier la réponse hygrothermique des enveloppes de bâtiment ainsi que valider la plateforme de co-simulation dynamique HAM-BES. La confrontation des résultats a montré une bonne concordance entre la résolution numérique et les mesures expérimentales. Les résultats obtenus dans le cadre de ce travail de thèse ont mis en exergue l’influence d’une modélisation fine des transferts couplés de chaleur, d’air et d’humidité, à la fois sur la prédiction du comportement hygrothermique des ambiances habitables mais aussi sur le calcul des besoins énergétiques des bâtiments. / The present PhD thesis work is conducted in the framework of the National Program ANR HUMIBATex Project « How to predict the disorders caused by moisture? What technical solutions to renovate the existing buildings? ». It deals with the numerical and experimental modeling of a coupled heat, air and moisture transfers at different scales: material, envelope and building ambience. In the theoretical part, based on expression of heat and moisture (vapor, liquid and air) balances equations, a phenomenological model describing the coupled heat, air and moisture transfer (HAM) through the wall has been developed. After validation stage (comparison with experimental results and analytical solution), the model has been implemented with confidence in a building energy simulation code (BES). Using this HAM-BES dynamic co-simulation tool, the hygrothermal behavior of the wall and indoor air of buildings were predicted finely. Two cases studies have been undertaken. The aim of the first one was to highlight the impact of hydrothermal transfers on the prediction of building energy consumption. However, the second case study was devoted to study efficiency of different ventilation strategies (extracting or insufflation) on the control and reduction of disorders caused by moisture in residential buildings. In the experimental part, a characterization campaign of physical, hydric and thermophysical properties of construction materials has been carried out. This experimental campaign has been focused on analyzing of the impact of thermal and hydric state of the construction material on the hygrothermal properties values. In another work, several experimental devices, at small-scale and wall scale, were designed in the laboratory to study the hygrothermal response of different building envelopes configuration and validate the developed HAM-BES dynamic co-simulation platform. Results of confrontation have showed good agreement between the numerical solution and experimental measurements.The obtained results in the framework of this PhD thesis have highlighted the influence of a detailed modeling of coupled heat air and moisture transfer through the wall on the hygrothermal behavior prediction of the indoor air, on assessment of pathology indicators and on the evaluation of the buildings energy loads.

Le coproduit de fabrication de jambon sec est issu de l'opération de désossage de la cuisse de porc parée, salée, séchée et affinée. Il est constitué à plus de 85 % d'os et de tissus associés (cartilages, ligaments, tendons). Le concassage au broyeur à marteau permet d'homogénéiser le coproduit en morceaux de taille inférieure à 8 cm (> 90 % compris entre 40 et 80 mm). La composition chimique de la matière sèche du mélange (77 ± 3 % de MS) est de 33 ± 5 % en protéines (89 % de collagène, 14 % de protéines hydrosolubles, 6 % d'acide aminés libres), 31 ± 3 % en lipides (triglycérides : 70 % ; diglycérides : 3,5 % ; acides gras libres : 11 % ; saturé/insaturé : 0,87 ; 24 % C16 :0 ; 13 % C18 :0 ; 2 % C16 :1 ; 38 % C18 :1 ; 4 % C18 :2) et 26 ± 4 % de matière minérale (phosphate de calcium 88 % ; NaCl 9 %). L'extraction aqueuse des lipides et des protéines du coproduit est étudiée en contacteur agité. Le raffinat solide est séparé par filtration à chaud sous forme de granulat et la matière grasse entraînée est séparée par décantation à froid. L'étude de l'influence des principaux facteurs de l'extraction liquide/solide (temps de contact : 30 à 90 min, température : 40 à 90°C ; ratio eau/coproduit : 4 à 10) grâce à la réalisation d'un plan d'expérience met en évidence les effets de la solubilisation et la coagulation des protéines sur l'entraînement des lipides et leur décantation sous forme de matière grasse. Mis en oeuvre à l'échelle pilote (64 kg de coproduit de jambon sec concassé, 207 kg d'eau, 30 min à 90°C en contacteur agité), le procédé de fractionnement aqueux conduit par filtration centrifuge et séchage à un granulat stable (rendement : 59 % ; matière minérale : 41 % ; protéines : 43 % ; lipides : 16 %), source de phosphate de calcium (95 % de la matière minérale) et de gélatine ou de colle d'os (88 % de protéines de nature collagénique). La fraction matière grasse décantée (rendement : 24 % ; lipides : 93 % ; triglycérides : 75 % ; diglycérides : 4 % ; acides gras libres : 7 % ; saturé/insaturé : 0,82 % ; 37 % C16 :0 ; 15 % C18 :0 ; 2 % C16 :1 ; 44 % C18 :1 ; 8 % C18 :2) présente les mêmes caractéristiques physicochimiques que le saindoux, avec une odeur proche de celle du jambon sec (19 COV aromatiques identifiés présents dans les arômes majoritaires de jambon). La fraction protéines solubilisées, obtenue sous forme de lyophilisat après concentration de la phase aqueuse (rendement : 8 % ; protéines : 52 % dont 29 % d'acides aminés libres ; matière minérale : 29 % dont 90 % NaCl, lipides : 3 %), contient aussi des glucosaminoglycanes sulfatés (GAGs : 3,4 %). Ces caractéristiques de composition, associées à ses propriétés épaississantes et gélifiantes, adhésives et stabilisantes d'émulsion, font de cette fraction minoritaire du procédé de fractionnement aqueux du coproduit de jambon sec, un extrait aux multiples applications à forte valeur ajoutée (source de peptones pour la culture de champignons et de levures, adhésif et liant naturel, ingrédient de formulation alimentaire nutracétique et cosmétique). / The ham production by-product comes from the deboning of dressed, salted, dried and refined pork leg. It consists of more than 85% of bone and associated tissues (cartilage, ligaments, tendons). Hammer mill crushing allows homogenizing the by-product into pieces smaller than 8 cm (> 90% between 40 and 80 mm).Dry matter chemical composition of the blend; (77 ± 3% DM) is 33 ± 5% protein (89% collagen, 14% of watersoluble proteins, 6% free amino acid), 31 ± 3% lipids (triglycerides: 70% diglycerides: 3.5%; free fatty acids: 11%; saturated / unsaturated: 0.87; 24% C16: 0; 13% C18: 0; 2% C16: 1; 38% C18: 1; 4% C18: 2) and 26 ± 4% mineral matter (calcium phosphate 88%, 9% NaCl). Lipids and proteins aqueous extraction of the by-product is studied in an agitated contactor reactor. The solid raffinate was separated by hot filtration to an aggregate and the fat is separated by cold decantation.The study of the influence of main factors of the liquid / solid extraction (contact time: 30 to 90 min, temperature: 40 to 90 °C; ratio water / by-product: 4 to 10) through the implementation of an experimental design, highlights the effects of proteins dissolution and coagulation on lipid output and decantation as fat matter.By pilot scale implementation (64 kg of crushed by-product of dry-cured ham, 207 kg of water, 30 min at 90 ° C in agitated contactor), the aqueous fractionation process leads, by centrifugal filtration and drying, to a stable aggregate (yield: 59%; mineral matter: 41%; protein 43%; lipids: 16%), source of calcium phosphate (95% of the mineral) and gelatin or bone glue (88% collagenous protein).The decanted fat fraction (yield: 24%; lipids: 93%; triglycerides: 75%; diglycerides: 4% free fatty acids: 7%; saturated / unsaturated: 0.82%; 37% C16: 0; 15% C18: 0; 2% C16: 1; 44% C18: 1; 8% C18: 2) has the same physicochemical characteristics as lard, with an odor similar to that of dry-cured ham (19 identified aromatic VOC part of ham main flavors). The solubilized protein fraction, obtained as a lyophilized extract after concentration of the aqueous phase (yield: 8%; protein: 52% with 29% of free amino acids; mineral matter: 29%, with 90% NaCl, lipids: 3%), also contains sulfated glycosaminoglycans (GAGs: 3.4%). These composition characteristics, associated with its thickening and gelling properties, adhesive and stabilizing for emulsion, transforms this minor fraction of the aqueous fractionation process of the dry-cured ham byproduct, in an high added value multiple applications extract (source of peptones for culture for fungi and yeasts, a natural and binding adhesive, ingredient for food nutraceutic and cosmetic formulation).

Doctor of Philosophy / Department of Entomology / Thomas W. Phillips / Robert "Jeff" J. Whitworth / Dry-cured ham is protected from infestations of Tyrophagus putrescentiae (Schrank) (Sarcoptiformes: Acaridae) with the fumigant methyl bromide. Developing feasible alternatives to methyl bromide is necessary due to the phase out of methyl bromide. The effectiveness of food-safe compounds for preventing infestations of T. putrescentiae on dry-cured hams was evaluated by dipping ham pieces in solutions of various food additives. Propylene glycol (1, 2-propanediol), lard, ethoxyquin and butylated hydroxytoluene prevented or significantly reduced mite population growth. A combination of carrageenan + propylene glycol alginate + 40% propylene glycol was effective in reducing mite numbers on treated whole-aged hams compared with untreated hams. Dose/response tests with twelve registered residual insecticides were conducted to assess contact toxicity to T. putrescentiae. Three of these insecticides were evaluated for persistence over a 2-month period on different surfaces. Commercial formulations of deltamethrin plus chlorpyrifos-methyl, chlorfenapyr, and malathion showed promising results for contact toxicity against T. putriscentiae. Chlorfenapyr applied to metal, concrete, and wood resulted in 100% mortality of treated ham mites for up to 8 weeks. The effect of high and low temperatures on mortality of T. putrescentiae was studied in the laboratory. Groups of 10 eggs and groups of a mixture of 40 adults and nymphs were separately exposed to several high and low temperatures, ranging from +35 to 45°C and from −20 to +5°C, for several time periods. Eggs were found to be more tolerant to both high and low temperatures compared with adults and nymphs. Results showed that high temperatures from 40-45°C killed all T. putrescentiae eggs, adults, and nymphs within 4-1 d, while −10°C or lower killed all the same stages in less than 1 d. Combinations of the fumigant sulfuryl fluoride (SF) applied under high temperatures ranging from 25-40ºC, were studied to determine the highest temperature, shortest exposure time, and the lowest value of a concentration-by-time product (CTP) of SF against T. putrescentiae. Results showed that complete control of all life stages of T. putrescentiae was achieved at 40ºC with SF. More than one day of exposure was required to kill adults and nymphs and eggs at 40ºC at a CTP close to the EPA labeled rate of 1500 gh.m⁻³. Results indicated that adults and nymphs were more susceptible to SF compared to eggs. This study focused on investigating different control methods for T. putrescentiae in dry-cured ham facilities and most of examined techniques are preventive, although some of them can be applied as remedial methods after mite infestations are noticed.

A cor vermelha brilhante característica do presunto Parma é resultante, principalmente, do pigmento Zinco-protoporfirina IX (ZnPP). A ZnPP é formada a partir da mioglobina por uma reação de transmetalação, catalisada pela enzima ferroquelatase (FECH), em que o íon de Fe(II) coordenado ao grupo heme é substituído pelo íon Zn(II). O presunto Parma apresenta uma maior estabilidade oxidativa em relação aos demais produtos cárneos curados além de não conter nitrito e nitrato, portanto, são considerados mais saudáveis. A utilização da FECH no processamento de carnes curadas pode permitir a produção de produtos cárneos curados mais saudáveis e em menor tempo. No presente trabalho a proteína ferroquelatase de Bacillus subtilis (BsFECH) foi expressa em células de E. coli BL21(DE3), purificada por cromatografia de afinidade ao níquel e exclusão por tamanho e caracterizada por dicroísmo circular, emissão de fluorescência do triptofano e cromatografia de exclusão por tamanho analítico. Em termos de estabilidade foi encontrado que altas concentrações de sal aumentam a estabilidade da proteína frente aos agentes denaturantes ureia e temperatura. A BsFECH produzida é capaz de ligar-se ao substrato modelo de porfirina (TPPS), conforme verificado por espectroscopia de UV-Vis, com uma Ka = 3,8x105 M-1 e é capaz de se associar à metamioglobina, conforme verificado por reação de cross-linking com dissuccinimidil suberato e avaliado por SDS-PAGE. A BsFECH aumenta significativamente a taxa de inserção de íons de zinco na TPPS e mostra uma cinética de saturação com uma constante de ligação aparente de Zn(II) ao complexo [BsFECH-TPPS] de 1,3x104 M e uma constante de primeira ordem de 6,6x10-1 h-1 para a dissociação do complexo ternário. A reação de troca ferro/zinco na mioglobina catalisada pela BsFECH é facilitada pela proteólise limitada da mioglobina com pepsina que abre um caminho para a reação de troca metálica com base na interação proteína-proteína entre o fragmento globina da mioglobina e a BsFECH. / The bright red color, characteristic of the Parma ham, results mainly of the pigment Zinc-Protoporphyrin IX (ZnPP). The ZnPP is formed from myoglobin by the reaction, catalyzed by ferrochelatase enzyme (FECH), in which Fe(II) ions coordinated to the heme group is replaced by Zn(II) ions. Parma ham shows greater oxidative stability when compared to others cured meat products besides do not contain nitrite and nitrate and, therefore, is considered healthier. The use of FECH in the processing of cured meats may allow the production of healthier cured meat products in a shorter period of time. In this work, the ferrochelatase protein from Bacillus subtilis was expressed in E. coli BL21(DE3) cells, purified by nickel affinity chromatography and size exclusion, and characterized by circular dichroism, fluorescence emission of tryptophan and analytical size exclusion chromatography. In terms of stability, it was found that the high salt content enhances the protein stability against the denaturation agents urea and temperature. The BsFECH produced is able to bind to the porphyrin model substrate (TPPS), as verified by UV-Vis spectroscopy, with Ka = 3.8x105 M-1 and is capable to associate to metamyoglobin as verified by cross-linking reaction with dissuccinimidil suberato, as observed by SDS-PAGE. The BsFECH increases, significantly, the zinc ions insertion rate in TPPS and shows a saturation kinetics behavior with an apparent biding constant of Zn(II) to the [BsFECH-TPPS] complex of 1.3x104 M and a first order rate constant for the dissociation of ternary complex of 6.6x10-1 h-1. The Fe/Zn exchange reaction in the myoglobin as catalyzed by BsFECH is facilitated by myoglobin-limited proteolysis with pepsin that opens a reaction channel for the metallic exchange based on protein-protein interaction between the globin moiety of myoglobin and BsFECH.

Ready to eat (RTE) meat products are subject to recontamination after industrial processing, mainly by Listeria monocytogenes, a pathogenic microorganism that can persist for a long time in the environment. A RTE meat product that is contaminated with L. monocytogenes due to cross contamination during some stage after industrial processing, such as weighing, slicing or wrapping, can be an important causer of disease, due to absence of a kill step before consumption. The objective of this project was to measure the transfer of L. monocytogenes during slicing of cooked ham (cross contamination) at retail, simulating in the laboratory the practices in commercial slicing, and to develop a predictive model capable of describing this transfer. It was observed that in the first slices obtained after the experimental contamination of the slicer, the counts and the transfer rates of L. monocytogenes were higher than in the subsequent slices, and the counting curves presented a long tail as the slices were obtained. The data demonstrate that the slicer may be a relevant source of cross contamination of L. monocytogenes for RTE meat products, regardless of the level of contamination of the slicer. With the data obtained, a new transfer model was proposed called 4p-2se, as it contained four parameters (4p) and two environments (2se), and was independent of the quantification of the pathogen transferred to the slicer. The proposed model was compared to two pathogen transfer models previously described, and the predicted data presented lower RMSE (Root Mean Sum of squared errors) values than the other models. The 4p-2se model was able to satisfactorily predict the pathogen transfer data during slicing of cooked ham, which could assist the food retail establishments and regulatory agencies in the evaluation and control of cross contamination of RTE foods and in the design of proper risk management strategies. / Os produtos derivados da carne que são prontos para consumo estão sujeitos à recontaminação após o processamento industrial, principalmente por Listeria monocytogenes, um microrganismo patogênico capaz de persistir por longo tempo no ambiente. Um produto cárneo pronto para consumo que se contamina com L. monocytogenes devido à contaminação cruzada durante alguma etapa após o processamento industrial, tal como pesagem, fatiamento ou acondicionamento, pode ser um importante causador de enfermidade, pois não há uma etapa de eliminação do patógeno antes do consumo. Este projeto teve por objetivo mensurar a transferência de L. monocytogenes durante o fatiamento de presunto cozido (contaminação cruzada), simulando em laboratório práticas adotadas nos estabelecimentos comerciais de fatiamento de produtos prontos para o consumo, e desenvolver um modelo preditivo capaz de descrever esta transferência. Foi observado que nas primeiras fatias obtidas após a contaminação experimental do fatiador, as contagens e as taxas de transferência de L. monocytogenes eram mais altas que nas subsequentes, observando-se que as curvas de contagem apresentavam uma longa cauda ao longo do fatiamento. Os dados demonstram que o fatiador pode ser uma fonte importante de contaminação cruzada de L. monocytogenes para produtos cárneos prontos para o consumo fatiados, independentemente do nível de contaminação do fatiador. Com os dados obtidos, foi possível sugerir um novo modelo de transferência, denominado 4p-2se, formado por uma equação com apenas quatro parâmetros (4p) e dois ambientes (2se,) sendo esse modelo independente da quantificação do patógeno transferido para o fatiador. O modelo sugerido foi comparado a outros dois modelos de transferência previamente descritos, observando os dados preditos no modelo 4p-2se apresentavam valores de RMSE (Root Mean Sum of squared erros) mais baixos que os demais modelos. O modelo proposto mostrou-se capaz de predizer satisfatoriamente os dados de transferência de patógeno durante o fatiamento de presunto cozido, podendo auxiliar os estabelecimentos comerciais de alimentos e as agências reguladoras na avaliação e controle da contaminação cruzada de alimento prontos para consumo e na concepção de estratégias adequadas de gestão de risco.