Fresh produce is increasingly implicated in outbreaks of foodborne illness. Internalization of bacterial pathogens into produce is of particular concern as internalized pathogens are unlikely to be removed by surface sanitizers. It is therefore necessary to develop treatments that will reduce their prevalence and numbers on fresh produce. Irradiation is a penetrating nonthermal treatment that effectively eliminates bacteria. Irradiated baby spinach leaves up to 1.0 kGy showed negligible (P>0.05) changes in color, texture, vitamin C, total carotenoids, and chlorophyll content compared to non-irradiated controls throughout storage (15 days at 4oC). This research also shows that irradiation effectively reduces viable Escherichia coli cells internalized in lettuce, and that decontamination is not influenced (P>0.05) by lettuce variety. Irradiation effectively reduced the population of internalized pathogens in a dose-dependent manner (3-4 log reduction at 1.0 kGy). Microscopy images suggest that the contamination sites of pathogens in leafy vegetables are mainly localized on crevices and in the stomata. A careful design of the treatment (understanding dose distribution) will effectively eliminate pathogens while maintaining produce quality. The use of modified atmosphere packaging increased (P<0.05) the sensitivity of pathogens (Salmonella spp. and Listeria spp.) to irradiation in baby spinach leaves (up to 25%). Increasing concentration of oxygen increased (P<0.05) sensitivity of both microorganisms. Radiosensitization could be affected (P<0.05) by production of ozone, which increases with increasing dose-rate and oxygen concentration, and reducing temperatures. Antimicrobial effectiveness of various active compounds was determined against Salmonella spp. and Listeria spp. Inclusion complexes were prepared with antimicrobial compounds and -cyclodextrin. The effectiveness of the microencapsulated compounds was tested by spraying them on the surface of baby spinach leaves inoculated with Salmonella spp. The increase in radiosensitivity (up to 40%) varied with the antimicrobial compound. Spherical poly (DL-lactide-co-glycolide) (PLGA) nanoparticles with entrapped eugenol and trans-cinnamaldehyde were synthesized for future antimicrobial delivery applications. All loaded nanoparticles proved to be efficient in inhibiting growth of Salmonella spp. and Listeria spp. The entrapment efficiency for eugenol and trans-cinnamaldehyde was 98% and 92%, respectively. Controlled release experiments (in vitro at 37oC for 72 hrs) showed an initial burst followed by a slower release rate of the antimicrobial entrapped inside the PLGA matrix.

This thesis is an investigation of the potential of Allyl Isothiocyanate (AITC) derived from oriental mustard meal (Brassica juncea meal) as a natural preservative suppression moulds growth on bread. Currently, clean labels and natural antimicrobial agents are interests of alternative preservatives. In this study, an antimicrobial sachet/patch containing B. juncea meal was developed to produce AITC vapour in situ; the efficacy of gaseous AITC/B. juncea meal on suppression of Penicillium spp. and other mould growth was investigated. The growth was completely inhibited for 28 days at 23˚C in the presence of 0.7-1.3 ppm AITC in the headspace (released from 50-100 mg B. juncea meal). Fifty mg mustard meal showed fungistatic activity, and ≥100 mg were fungicidal. The shelf life of sliced white bread (600 g) was prolonged for 14 days using 3g of B. juncea meal at 23˚C thereby illustrating the potential of AITC as an alternative to chemical preservatives. / Developing Innovation Agricultural Products (DIAP) program of AAFC and Mustard 21 (RBPI 2109)

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2016-04-29T15:01:00Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Priscilla_Gregorio_de_Oliveira_PPSHMA_2015_.pdf: 1709908 bytes, checksum: aa7683bc61b779c42677bd3a2f56d1cf (MD5) / Made available in DSpace on 2016-04-29T15:01:01Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Priscilla_Gregorio_de_Oliveira_PPSHMA_2015_.pdf: 1709908 bytes, checksum: aa7683bc61b779c42677bd3a2f56d1cf (MD5) Previous issue date: 2015-05-20 / FACEPE / O queijo de coalho é um produto que apresenta insegurança microbiológica, sendo apontado como um importante veículo de Listeria monocytogenes. O objetivo dessa pesquisa foi verificar a bioatividade de quitosana como cobertura comestível em queijo de coalho na inibição de Listeria monocytogenes. Para isso foram elaborados produtos com coberturas de quitosana em diferentes concentrações: T1 (15mg/mL), T2 (10mg/mL) e T3 (5mg/mL). Como controle utilizou-se amostras de queijo de coalho sem tratamento (C) e com coberturas de ácido acético 1% (CA). A cinética de inibição bacteriana da cobertura de quitosana foi avaliada através da contagem de células viáveis (curva de sobrevivência bacteriana) de Listeria monocytogenes (ATCC 3533) durante os intervalos 0,1,2,4,8,12 e 16 dias de armazenamento, após aplicação da cobertura. Os produtos foram avaliados quanto as características físico-químicas, aos padrões microbiológicos sanitários e sensorialmente. Os resultados observados no teste de inibição bacteriana contra Listeria monocytogenes apontaram o tratamento T1 como o mais eficiente, por proporcionar menores contagens da carga microbiana, em comparação aos demais tratamentos, ao longo do período de armazenamento avaliado, favorecendo reduções logarítmicas de 2,50; 2,46; 2,84; 1,41; 1,09; 1,38 e 0,81, respectivamente. Quanto às características físico-químicas e aos padrões microbiológicos sanitários, os produtos avaliados (T1, T2, T3, C e CA) se enquadraram nos requisitos exigidos pela legislação para queijo de coalho. Verificou-se nos produtos variações de umidade entre 50,07±0,2 e 50,86±0,5 e de gordura entre 46,54±0,1 e 47,77±0,2. Quanto aos coliformes termotolerantes observou-se que as amostras sob tratamento T1, T2, T3, e controles C e CA apresentaram valores de: 1,51 x 10¹ NMP/mL, 1,83 x 10¹ NMP/mL, 1,95 x 10¹ NMP/mL, 2,40 10¹ NMP/mL e 1,99 x 10¹ NMP/mL, respectivamente. Na contagem de Staphylococcus coagulase positivo as amostras apresentaram resultados < 10 UFC/g. Para L. monocytogenes e Salmonella spp. não foi encontrada a presença destes microrganismos em 25g dos produtos. Quanto as análises sensoriais, os resultados para a aceitação foram satisfatórios, e verificou-se que a amostra com maior concentração de quitosana (T1) obteve o maior índice de aceitabilidade para os atributos sabor (76,66%) e textura (78,33%), assim como melhores resultados nos testes de preferência e intenção de compra. Concluiu-se que a cobertura comestível de quitosana pode ser utilizada na conservação de queijo de coalho frente a contaminação de Listeria monocytogenes, e que o emprego dessa tecnologia não altera os parâmetros físico-químicos e aceitação sensorial do queijo de coalho. / The coalho-type cheese is a product its food security is not recommended and it has been pointed out as an important vehicle for Listeria monocytogenes. The objective of this study was to assess the bioactivity of chitosan as edible coating in coalho-type cheese in inhibiting Listeria monocytogenes. For this purpose, treatments were realized with coating of chitosan in different concentrations: T1 (15 mg.mL-1), T2 (10 mg.mL-1) and T3 (5 mg.mL-1). Untreated samples of cheese were used as a control (C) and coated samples, only, with acetic acid 1% (CA). Kinetic studies of bacterial inhibition from chitosan coating was assessed by viable cell counting (microbial survival curves) of Listeria monocytogenes (ATCC 3533) during storages intervals (0,1,2,4,8,12 e 16 days), after coating application. The products were also assessed for physical-chemical and sensory characteristics, as well as, sanitary-microbiological criteria. The results obtained from the test of inhibition against the Listeria monocytogenes showed T1 treatment as the most efficient, by providing a lower growth rate, compared to the other treatments, during the storage period, representing a log reduction of 2.50; 2.46; 2.84; 1.41; 1.09; 1.38 e 0.81. In relation to the physical-chemical characteristics and sanitary-microbiological criteria, the evaluated products (T1, T2, T3, C e CA) met the requirements by the legislation for coalho-type cheese and showed moisture variations between 50.07±0.2 and 50.86±0,5; and fat from 46.54±0.1 to 47.77±0.2.Thermotolerant Coliforms counts were detected in T1, T2, T3, C e CA samples that proved values: 1,51 x 10¹ MPN.mL-1, 1.83 x 10¹ MPN.mL-1, 1.95 x 10¹ MPN.mL-1, 2.40 10¹ MPN.mL-1 e 1.99 x 10¹ MPN.mL-1, respectively. The counts of positive-coagulase staphylococci expressed results < 10 CFU.g-1. There was no evidence of L. monocytogenes e Salmonella spp. in 25 grams of cheese. Application of sensory test revealed that the addition of coating was well appreciated by a taste panel and the sample with the highest concentration of chitosan (T1) was better accepted for taste (76.66%) and texture (78.33%) attributes, in addition to superior performance in preference test and intention to purchase. In summary that chitosan as edible coating can be used to preserve coalho-type cheese against Listeria monocytogenes and the appropriate use this technology cannot affect sensory properties and physical chemistry of the product.

Made available in DSpace on 2016-08-17T18:39:51Z (GMT). No. of bitstreams: 1 6095.pdf: 2718900 bytes, checksum: c4d734a7b2927b9a3c31ecfb042e26bd (MD5) Previous issue date: 2014-04-16 / Many efforts have been dedicated in the searching of natural compounds which are suitable to be applied in food conservation. Among them, the chitosan has presented promising results concerning the antimicrobial activity, structural preservation and delay of fruits senescence. However, the chitosan, as found commercially, is soluble only in acidic medium solutions what is considered a hindrance for fully applications as food conservative. In this sense, chitosan derivations which are able to be solubilized in neutral pH have been developed. The N,N,N Trimethylchitosan (TMC) is one of them, characterized as a hydrosoluble derivative of chitosan, which has been considered as an efficient antimicrobial agent with positive action as fresh food conservative. The aim of the present was the evaluation of the antimicrobial activity of the TMC against Gram-positive and Gram-negative bacteria and filament fungi. The TMC antimicrobial activity was assessed by comparing with the Commercial Chitosan (CC), by means of the determination of the Minimum Inhibitory Concentration (MIC), following the Clinical and Laboratory Standards Institute (CLSI) according to M7-A6 (NCCLS, 2003) and M38-A (NCCLS, 2002) standards, both recognized by the Agência Nacional de Vigilância Sanitária (ANVISA). Additionally, the TMC and CC antifungal activity was carried out by recoding the visual fungi growth and estimated from the percentage of infected samples by the fungy on cut faces of grape-tomatoes (Solanum lycopersicum variety cerasiforme), in coated and non-coated conditions, along seven days of incubation. The results concerning antibacterial activity points to a greater action of the TMC when comparing to some analysis performed on CC coating against four of the six strains analyzed: Enterococcus faecalis, Staphylococcus aureus, Escherichia coli and Salmonella enterica. Nevetheless the CC antifungal activity was considerable superior to that observed to TMC for three different fungi tested: Penicillium sp., Aspergillus flavus standard and Aspergillus flavus wild. However, both polymers showed high antibacterial and antifungal activity and are therefore considered as natural agents with relevant antimicrobial potential for the fresh food conservation. / Muitos têm sido os esforços pela procura de substâncias naturais que possam ser aplicadas à conservação de alimentos. Dentre as substâncias em estudo, a quitosana tem apresentado resultados promissores tanto com relação à atividade antimicrobiana quanto na manutenção estrutural de alimentos e redução da senescência em frutos. No entanto, a quitosana, em sua forma comercial, é solúvel apenas em soluções ácidas diluídas, o que dificulta sua plena utilização como conservante alimentício. Dessa forma, derivados de quitosana que apresentem capacidade de solubilização em pH neutro têm sido desenvolvidos. Dentre esses derivados a N,N,N-trimetilquitosana (TMQ), um polímero hidrossolúvel derivado da quitosana, tem sido avaliado como promissor agente antimicrobiano e conservador de alimentos frescos. Assim, o presente projeto teve como objetivo avaliar a atividade antimicrobiana da N,N,N-trimetilquitosana sobre o crescimento de bactérias Gram-positivas e Gram-negativas e de fungos filamentosos. A atividade antimicrobiana da TMQ foi avaliada mediante comparação com a Quitosana Comercial (QC) pela determinação das Concentrações Inibitórias Mínimas (CIM) a partir de testes padronizados pela Clinical and Laboratory Standards Institute (CLSI), segundo suas normas M7-A6 (NCCLS, 2003) e M38-A (NCCLS, 2002), ambas reconhecidas pela Agência Nacional de Vigilância Sanitária (ANVISA). Além disso, a avaliação da atividade antifúngica das substâncias foi realizada mediante leitura visual e estimada a partir do percentual de amostras infectadas pelos fungos sobre fatias de tomatecereja (Solanum lycopersicum variedade cerasiforme) revestidas e não revestidas, ao longo de sete dias de incubação. Os resultados indicaram uma maior atividade antibacteriana da TMQ em relação à QC para quatro das seis cepas bacterianas analisadas: Enterococcus faecalis, Staphylococcus aureus, Escherichia coli e Salmonella enterica. No entanto, a atividade antifúngica da QC superou a da TMQ para os três fungos testados: Penicillium sp., Aspergillus flavus padrão e Aspergillus flavus selvagem. Entretanto, ambos os polímeros avaliados apresentaram atividade antibacteriana e antifúngica satisfatória sendo, portanto, considerados como agentes naturais com potencial antimicrobiano relevante para a conservação de alimentos frescos.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The edible coatings are defined as a thin layer of edible material usually applied in liquid form on the product by immersing the same. Several studies have shown that the addition of bioactive substances such as essential oils have increased antimicrobial packaging potential of these, contributing to the increased food shelf life. Thus, this study aimed to develop and evaluate in vitro and in situ microbiological efficiency of edible coatings to chitosan base incorporated with essential oils of Myrcia ovata Cambess (MYRO). The antimicrobial potential of 9 essential oils (EOs) was evaluated in vitro by diffusion technique in discs against foodborne bacteria (Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus cereus, Bacillus subtilis, Serratia marcescens, Escherichia coli, Enterococcus faecalis and Salmonella enteritidis). Due to greater availability of MYRO-174 and MYRO-175 oils and being gram-positive bacteria very sensitive to extremely sensitive to them, these were selected for incorporation into edible eatable coatings of chitosan base.The edible coatings formulations were prepared ranging cassava starch, chitosan and essential oil concentrations according the central composite rotational design (23 + 6 axial points and 3 central points). The bacteria were sensitive to edible coatings with diameters of inhibition zone between 9.0 -12.5 mm. The edible coatings formulations containing MYRO-175 oil were optimized for the B. Cereus and B. subtilis inhibition using the response surface methodology, a predictive mathematical model for the inhibition of them was obtained. The essential oil concentration was the more significant parameter to antimicrobial activity of coatings. The edible coatings incorporated with MYRO-174 oil or MYRO-175 (1.00% cassava starch, 1.00% chitosan and 1.25% essential oil) were applied in mangabas and the antimicrobial potential against natural microflora and Bacillus cereus artificially contaminated in the fruits was studied. The coated mangabas showed total mesophilic aerobic bacteria counts the order of 102 CFU/g, molds and yeasts <10 CFU/g (estimated value) and inhibition of growth of B. cereus for 12 days of storage at 10°C. On the other hand, uncoated mangabas (control) rotted from the 8th day of refrigerated storage. The edible coatings based in chitosan incorporated with M. ovata Cambess. EOs demonstrated potential to be used as an alternative in vitro microbiological control of foodborne bacteria and in situ on mangaba preservation. / As coberturas comestíveis são definidas como uma camada fina de material comestível, geralmente aplicada na forma líquida sobre o produto, por imersão do mesmo. Várias pesquisas têm demonstrado que a adição de substâncias bioativas como os óleos essenciais têm aumentado o potencial antimicrobiano destas embalagens contribuindo para o aumento da vida de prateleira de alimentos. Diante do exposto, o presente estudo teve como objetivo desenvolver e avaliar in vitro e in situ a eficiência microbiológica de coberturas comestíveis à base de quitosana incorporadas com óleos essenciais de Myrcia ovata Cambessedes (MYRO). O potencial antimicrobiano de 9 óleos essenciais foi avaliado in vitro através da técnica de difusão em discos frente à bactérias patogênicas de alimentos (Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus cereus, Bacillus subtilis, Serratia marcescens, Escherichia coli, Enterococcus faecalis e Salmonella enteritidis). Devido a maior disponibilidade dos óleos MYRO-174 e MYRO-175 e sendo as bactérias Gram-positivas de muito sensíveis a extremamente sensíveis aos mesmos, estes foram selecionados para a incorporação nas coberturas comestíveis a base de quitosana. As formulações de coberturas comestíveis foram elaboradas variando-se as concentrações de fécula de mandioca, quitosana e óleo essencial conforme o delineamento composto central rotacional (23 + 6 pontos axiais e 3 pontos centrais). As bactérias foram sensíveis às coberturas com halos de inbição entre 9,0 –12,5 mm. As formulações contendo o óleo MYRO-175 foram otimizadas para a inibição de B. cereus e B. subtilis utilizando a metodologia de superfície de resposta, sendo possível obter um modelo matemático preditivo para a inibição das mesmas. A concentração de óleo essencial foi o parâmetro significativo para a atividade antimicrobiana das coberturas. As coberturas comestíveis incorporadas com o óleo MYRO 174 ou MYRO-175 (1,0% de fécula de mandioca, 1,0% de quitosana e 1,25% de óleo essencial) foram aplicadas em mangabas e o potencial antimicrobiano foi avaliado frente a microbiota e ao Bacillus cereus artificialmente contaminado nos frutos. As mangabas revestidas com cobertura comestível apresentaram contagens de bactérias aeróbias mesófilas totais da ordem de 102 UFC/g, bolores e leveduras < 10 UFC/g (valor estimado) e inibição do crescimento de B. cereus por 12 dias de armazenamento a 10ºC. Já as mangabas sem revestimento (controle) apodreceram a partir do 8º dia de estocagem refrigerada. As coberturas comestíveis à base de quitosana incorporadas com óleos essenciais de M. ovata Cambessedes demonstraram potencial para serem utilizadas como alternativa no controle microbiológico in vitro de bactérias patogênicas de alimentos e in situ na conservação de mangabas.

Present diploma thesis is focused on natural antimicrobial substances. The aim of the experimental part of thesis was testing of antimicrobial activity of selected natural substances in free and encapsulated forms, preparation of particles and monitoring of their long-term stability. Various teas extracts (aqueous, ethanoic and DMSO) and aqueous herbs extracts were used as plant antimicrobial substances. Liposomes and chitosan particles with encapsulated aqueous teas extracts were prepared by ultrasonication. Encapsulation efficiency, size, stability and antioxidant activity for all particles were specified. Consequently, within a two months period, long-term stability for these particles was monitored. Spectrophotometric method for determining total polyphenols was used for evaluating encapsulation efficiency as well as for monitoring of long-term stability. The DLS method was used for determination of size and stability of particles. Antioxidation activity was monitored by using spectrophotometric method using ABTS. For testing antimicrobial activity of various types of teas, herbs extracts and their mixtures and combianitons with lysozyme and antibiotics, broth dilution methods were used while using both Gram positive and Gram negative bacterial test strains. Antimicrobial activity of prepared particles was also monitored, the antimicrobial activity was especially significant with the chitosan particles. Suitable combinations of natural substances whatever with antimicrobial or antioxidation effect either in free or encapsulated form could be used for applications for example in cosmetics industry, food processing or in medicine.

No description available.