Antimicrobial activity of the essential oils of Dittrichia viscosa subsp viscosa on Helicobacter pylori

Dittrichia viscosa subsp. viscosa (Compositae) is found on edges, wood clearings and in waste places of the Iberian Peninsula. Aerial parts of D. viscosa were collected at flowering phase in September-October 2001 around Lisbon, Portugal and the essential oils isolated by hydro-distillation for 4 h using a Clevenger-type apparatus. The oils were analyzed by gas chromatography and gas chromatography-mass spectrometry. Preliminary examination of the essential oils allowed the identification of 32 components. Only four components reached percentages over 5%: fokienol (11.8%), T-muurorol (7.9%), (E)-nerolidol (5.5%) and delta-cadinene (5.0%). The essential oils were tested against Helicobacterpylori and Listeria monocytogenes. Essential oils did not have antimicrobial activity against L. monocytogenes. The essential oil at 0.88 to 22.22 mu g.ml(-1) did not inhibit the growth of H. pylori, affected the growth slightly at 44.40 mu g.ml(-1), and completely inhibited the growth at 88.80 to 133.20 mu g.ml(-1) Results show that use of D. viscosa essential oil in the treatment of gastric disorders caused by H. pylori can be effective.

Microbiological quality and safety of minimally processed fruits in the marketplace of southern Portugal

The availability of fresh-cut fruit (FCF) in the marketplace has been increasing in Portugal, although reports of its microbial quality are not known. Due to the growing concerns of these commodities over their microbial safety, the objectives of this work were to study the microbiological quality and prevalence of Salmonella and Listeria monocytogenes on fresh-cut fruits sold in southern Portugal. A study to examine the changes in pH and microbial counts, before and after the expiration dates, was also made. A total of 160 samples was purchased in the local grocery stores between September 2011 and August 2014, before their sell-by date. These samples were assayed for aerobic mesophilic (AM) and psychrotrophic (AP) microorganisms, yeasts and molds (YM), lactic-acid bacteria (LAB), coliforms (TC), Escherichia coli and coagulase positive staphylococci as well as L. monocytogenes and Salmonella. The microbiological counts ranged from 3.0-9.2 lg cfu/g (AM); 2.2–10.7 lg cfu/g (AP); 2.3–10.4 lg cfu/g (YM); 1.9–9.0 lg cfu/g (LAB) and less than 1–9.1 lg cfu/g (TC). The melons and watermelon presented the highest levels of the microbial quality parameters studied. However, no E. coli, staphylococci, Salmonella and L. monocytogenes were detected in any of the samples. After the sell-by date, an increase of the AM, AP, LAB and YM values was observed in all fruits. Conversely, the differences found in TC counts before and after the best-before date had no statistical significance. A decrease in pH was observed in all fruits except pineapple whose pH slightly increased after 14 days of storage. The results highlight the importance of preventing contamination and cross contamination, selecting adequate decontamination technologies and maintaining a strict temperature control during processing, distribution and selling of FCF.

ADDITION OF VINEGAR TO EXTEND THE SHELF-LIFE OF CABEÇA DE XARA

Cabeça de xara is a ready-to-eat meat product, whose production is very characteristic in Alentejo, a particular region of Portugal. It is a galantine usually moulded into parallelepiped shape made with various meats obtained from the Alentejano pig breed reared in the same region, namely deboned pork heads, tongue and connective tissue to which a number of condiments like salt, parsley, wine and pepper, are added. This work intended to test the feasibility of adding vinegar in order to increase the shelf-life of cabeça de xara, by reducing the contaminating microbiota responsible for spoilage, as well as controlling the pathogen Listeria monocytogenes. Three independent batches were produced and proximate composition, pH, aw, microbiological parameters and biogenic amines content evaluated. A sensory analysis was also performed throughout the storage period. No significant differences between control and vinegar samples was found regarding the proximate composition of cabeça de xara. As expected, pH is lower in the vinegar samples, however no differences in aw were observed between the two treatments. L. monocytogenes was present from the first month on only in one batch in the control treatment. However, it is inhibited by the addition of vinegar until the third month of storage, where L. monocytogenes is present but below the limit established in the 2073/2005 regulation. The presence of vinegar significantly decreased the content in biogenic amines, particularly cadaverine, putrescine and tyramine, throughout the storage period. Concerning sensory evaluation, no vinegar taste was reported by the panellists in a depreciating way.

Effect of autochthonous starter cultures in the production of "Paio", a traditional Portuguese dry- cured sausage

Traditional dry-cured sausages are highly appreciated in Mediterranean countries. The aim of the present study was to evaluate the effect of different starter cultures in the sausages Alentejano pig meat was used to prepare drycured sausages in a local factory. Staphylococcus xylosus, Lactobacillus sakei and a yeast strain were inoculated at a concentration of 106 cfu/g meat batter both in separate and in mixed culture. Three independent batches with two replicates per treatment were produced. Samples were collected throughout the ripening process. pH and aw were determined according to the ISO standards. Microbiological counts of total mesophiles, total psycrotrophs, anaerobes, coagulase-negative staphylococci (CNS), lactic acid bacteria (LAB), enterobacteria, yeasts and moulds and Listeria monocytogenes were done according to the respective ISO standards, as well as detection of Salmonella spp. Biogenic amines quantification was performed by HPLC as described by Roseiro et al. (1). The treatment with L. sakei alone was the most effective in reducing the contamination level both with Salmonella spp. and L. monocytogenes, however this effect seems to be lost in the mixed cultures. The presence of the yeast strain seems to increase the levels of phenylethylamine and histamine. The contents in cadaverine, putrescine and tyramine were generally lower in the inoculated sausages. Regarding tyramine, the treatments with L. sakei showed significantly lower values. No significant differences between treatments were observed for both spermine and spermidine.

Effetto di starter autoctoni sulla qualità e la sicurezza di salami tradizionali spagnoli

Il lavoro svolto in questa tesi aveva l’obiettivo di valutare il potenziale tecnologico e bioprotettivo di ceppi di batteri lattici (LAB) isolati da salami tradizionali spagnoli. In particolare due ceppi (Lactiplantibacillus paraplantarum BPF2 e Pediococcus acidilactici ST6) che avevano dimostrato buone performance in vitro sono stati utilizzati, da soli o in miscela, come colture starter per la produzione salami e i prodotti ottenuti sono stati confrontati con un controllo a fermentazione spontanea ed un prodotto addizionato di uno starter commerciale contenente LAB e stafilococchi. Per quanto riguarda gli aspetti tecnologici, il pH ha mostrato cinetiche di acidificazione simili in tutti i prodotti, mentre il calo peso era più lento nel controllo. A livello microbiologico, i campioni addizionati di colture starter hanno mostrato carichi di LAB molto più elevati già al tempo zero, senza differenze significative in relazione al ceppo utilizzato. Enterobatteri e lieviti hanno mostrato andamenti simili in tutti i campioni. L’utilizzo di colture starter ha invece avuto un impatto rilevante sul contenuto di ammine biogene, con valori totali doppi nel campione ottenuto con fermentazione spontanea, e sul profilo in metaboliti volatili (soprattutto a carico di composti derivanti dall’acido piruvico). L’aspetto più rilevante di questa tesi è stato ottenuto nel challenge test, utilizzando come microrganismo target Listeria monocytogenes (inoculo 3 log ufc/g): infatti, nel controllo e nei campioni contenenti starter commerciale, L. monocytogenes era in grado di crescere fino a valori superiori a 5.7 log ufc/g, mentre i ceppi BPF2 e ST6 hanno determinato una riduzione del suo carico cellulare (2.4 log ufc/g). Questo conferma quindi le grandi potenzialità anti-listeria dei due ceppi testati e la loro attitudine ad essere utilizzati, oltre che come starter per i salami, anche come colture bioprotettive con lo specifico compito di contrastare lo sviluppo di L. monocytogenes.

Atividade antimicrobiana de nisina e óleo essencial de orégano (Origanum vulgare L.) sobre Listeria monocytogenes isolada de alimentos

Não consta resumo na publicação.

Infecção por listeria monocytogenes pós-transplante renal: relato de seis casos

São relatados 6 casos de listeriose em pacientes transplantados renais, que ocorreram entre 13 dias e 57 meses após iniciarem imunossupressão clássica, sendo 3 casos classificados como meningite, 2 casos como meningoencefalite e 1 caso como bacteremia. Houve um óbito e os 5 pacientes restantes apresentaram boa evolução. Com dados adicionais da literatura, são discutidos a epidemiologia, o diagnóstico e tratamento, da listeriose pós-transplante renal.

Envasado activo biodegradable para el control de Listeria monocytogenes en jamón cocido

El envasado activo consiste en la adición de una sustancia activa al material del envasado, mejorando de esta forma la funcionalidad del envase. En el caso del envasado antimicrobiano se consigue minimizar o evitar la contaminación superficial de los alimentos, de ahí el interés de su aplicación en los productos cárnicos listos para el consumo.

Detección de Listeria monocytogenes y Listeria spp en puntos críticos establecidos en una planta procesadora de productos congelados para rápido consumo

Tesis ( Maestro en Ciencias con Especialidad en Microbiología) U.A.N.L.

Padronização da técnica de imunoistoquímica para identificação de Listéria monocytogenes em placentas humanas e tecido nervoso central de ruminantes.

Este projeto foi desenvolvido no Laboratório de Patologia Experimental do Hospital de Clínicas de Porto Alegre (HCPA), com a aprovação da Comitê de Ética em Pesquisa do HCPA e com apoio financeiro parcial do Fundo de Incentivo à Pesquisa e Eventos do HCPA (FIPE). O experimento 1, chamado de projeto piloto, teve como objetivo implementar a técnica de IHQ para identificar a Listeria monocytogenes (L.m.), utilizando anticorpo policlonal antilisteria monocytogenes (Biodesig ). Vários testes foram realizados para acertar a diluição (1:1000) que foi diferente da preconizada pelo fabricante. Os blocos de parafina, de dez placentas provenientes de parto prematuro ou aborto foram utilizados para os cortes histológicos e a preparação das lâminas para a coloração Hematoxilina e Eosina (HE) e imunoistoquímica (IHQ). As lâminas foram identificadas por números para resguardar a identidade das pacientes. O resultado do HE mostrou alterações inflamatórias em oito placentas e L. m. foi identificada pelo IHQ em cinco dessas placentas. O objetivo do 2º experimento foi identificar a L. m. em tecido nervoso cerebral de ruminantes, utilizando a técnica implementada no projeto piloto. O material utilizado neste trabalho foi cedido pelo Setor de Patologia Veterinária do Departamento de Patologia da Universidade Federal de Santa Maria. Os casos estudados (2 ovinos, 1 caprino e 2 bovinos) tinham suspeitas clínicas diversas e as necropsias dos animais evidenciaram aspectos sugestivos da doença. Os cinco casos foram confirmados pelo IHQ, comprovando a importância da utilização desta técnica para o diagnóstico da listeriose no SNC de ruminantes. O 3º experimento objetivou identificar a L. m. em placentas encaminhadas ao Serviço de Patologia do HCPA no ano 2000. Da mesma forma que no experimento 1, as lâminas foram identificadas por números. Após o levantamento realizado nos registros dos exames anatomopatológicos (AP) deste setor, observou-se que 714 AP eram de placentas provenientes de aborto, parto prematuro e nascimento a termo examinados naquele período. Foram sorteados 254 AP para análise através de HE, revelando que 148 desses AP apresentavam alterações inflamatórias (corioamnionite, vilite e deciduite). Os blocos destas placentas foram utilizados para fazer as lâminas e realizar IHQ. A consulta aos prontuários dos casos com alterações inflamatórias permitiu observar que um deles tinha a confirmação bacteriológica de L. m. na placenta, tornando-se este o controle positivo. O controle negativo foi selecionado entre aqueles sorteados que não apresentavam alterações inflamatórias. A presença de L. m. foi identificada em 33,78% das placentas analisadas pela técnica IHQ. Corioamnionite e vilite foram as alterações inflamatórias que mostraram diferença estatística significativa nas placentas positivas. L. m. estava presente nas placentas de 1º, 2º e 3º trimestres gestacionais. A idade das gestantes, casos de aborto e/ou parto prematuro não mostraram diferença estatística significativa com a presença ou ausência de L. m. nas placentas. Abortos habituais ocorreram em pacientes com ou sem L. m. no tecido placentário. Conclusão: a técnica de imunohistoquímica pode ser utilizada para confirmar o diagnóstico histopatológico de listeriose em placentas e tecido nervoso central de ruminantes.

Listeria monocytogenes, E. coli O157, Salmonella spp. e microrganismos indicadores em carcaças bovinas para exportação

Samples were collected from 100 carcasses in a slaughterhouse exporter, located within the State of São Paulo, sampled over a year through the sponge method, applied to the chest of the animal. Samples were taken at three points, denominated A, B and C, each carcass sampled at three points located in the following steps: after bleeding (A) after skinning (B) and after washing (C). Research was conducted for Listeria sp., E. coli O157, Salmonella spp. and Micro-organism (Petrifilms ® AC, EC and EB). Listeria or E. coli O157 were not isolated in any of the 300 samples. Salmonella spp. was isolated in nine, eight at point A and one at point B. For Mesophiles, scores ranged from 0 to 6.8 log UFC/cm²; for Total coliforms, 0 to 4.57 log UFC/cm² and E. coli from 0 to 4.38 log UFC/cm². With the results obtained and compared with the literature, it is concluded that the establishment in this study has both sanitary quality (due to the low prevalence of pathogens) and hygienic quality (due to the sharp decrease in the microbial load of indicators along the line.

Types of Listeria monocytogenes predicted by the positions of EcoRI cleavage sites relative to ribosomal RNA sequences.

By using taxonomic characters derived from EcoRI restriction endonuclease digestion of genomic DNA and hybridization with a labeled rRNA operon from Escherichia coli, a polymorphic structure of Listeria monocytogenes, characterized by fragments with different frequencies of occurrence, was observed. This structure was expanded by creating predicted patterns through a recursive process of observation, expectation, prediction, and assessment of completeness. This process was applied, in turn, to normalized strain patterns, fragment bands, and positions of EcoRI recognition sites relative to rRNA regions. Analysis of 1346 strains provided observed patterns, fragment sizes, and their frequencies of occurrence in the patterns. Fragment size statistics led to the creation of unobserved combinations of bands, predicted pattern types. The observed fragment bands revealed positions of EcoRI sites relative to rRNA sequences. Each EcoRI site had a frequency of occurrence, and unobserved fragment sizes were postulated on the basis of knowing the restriction site locations. The result of the recursion process applied to the components of the strain data was an extended classification with observed and predicted members.