Bacterial phospholipases C with dual activity: phosphatidylcholinesterase and sphingomyelinase.

Bacterial phospholipases and sphingomyelinases are lipolytic esterases that are structurally and evolutionarily heterogeneous. These enzymes play crucial roles as virulence factors in several human and animal infectious diseases. Some bacterial phospholipases C (PLCs) have both phosphatidylcholinesterase and sphingomyelinase C activities. Among them, Listeria monocytogenes PlcB, Clostridium perfringens PLC, and Pseudomonas aeruginosa PlcH are the most deeply understood. In silico predictions of substrates docking with these three bacterial enzymes provide evidence that they interact with different substrates at the same active site. This review discusses structural aspects, substrate specificity, and the mechanism of action of those bacterial enzymes on target cells and animal infection models to shed light on their roles in pathogenesis.

Anti-inflammatory activity of caffeine (1,3,7-trimethylxanthine) after experimental challenge with virulent Listeria monocytogenes in Swiss mice.

BACKGROUND: Immunomodulatory therapies are claimed to enhance antimicrobial immunity and counterbalance antimicrobial resistance mechanisms of pathogenic bacteria. PURPOSE: To investigate whether caffeine can be useful for control of inflammation derived from experimental systemic infection with Listeria monocytogenes. METHODS: Peritoneal macrophages (pMØ) from Swiss mice were cultured with caffeine in 96-well plates, and then infected with virulent L. monocytogenes 619. In another experiment, the pMØ were first infected with the bacterium and then treated with caffeine. Swiss mice were inoculated intraperitoneally with L. monocytogenes and then treated intravenously with caffeine (0.05; 0.5 or 5 mg/Kg). RESULTS: Caffeine did not exert direct antibacterial activity in vitro against L. monocytogenes. Macrophages exposed to caffeine before or after infection with L. monocytogenes had increased cell viability, although the intracellular bacterial loads were similar to the control groups. Caffeine treatments of Swiss mice reduced leukocyte infiltration into the peritoneal cavity after L. monocytogenes infection. However, the bacterial burden was reduced in the spleen and liver. The mRNA expressions of IL-1ß, IL-6 and the enzyme inducible nitric oxide synthase (iNOS) were reduced whereas IL-10 was increased. CONCLUSION: Caffeine has an anti-infectious potential and ameliorated infection-derived inflammation following experimental infection with L. monocytogenes.

Listeria monocytogenes: review of pathogenesis and virulence determinants-targeted immunological assays.

Listeria monocytogenes is one of the most invasive foodborne pathogens and is responsible for numerous outbreaks worldwide. Most of the methods to detect this bacterium in food require selective enrichment using traditional bacterial culture techniques that can be time-consuming and labour-intensive. Moreover, molecular methods are expensive and need specific technical knowledge. In contrast, immunological approaches are faster, simpler, and user-friendly alternatives and have been developed for the detection of L. monocytogenes in food, environmental, and clinical samples. These techniques are dependent on the constitutive expression of L. monocytogenes antigens and the specificity of the antibodies used. Here, updated knowledge on pathogenesis and the key immunogenic virulence determinants of L. monocytogenes that are used for the generation of monoclonal and polyclonal antibodies for the serological assay development are summarised. In addition, immunological approaches based on enzyme-linked immunosorbent assay, immunofluorescence, lateral flow immunochromatographic assays, and immunosensors with relevant improvements are highlighted. Though the sensitivity and specificity of the assays were improved significantly, methods still face many challenges that require further validation before use.

Genetic diversity of Listeria monocytogenes serotype 1/2a strains collected in Brazil by Multi-Virulence-Locus Sequence Typing.

Listeria monocytogenes is an opportunistic pathogen with the ability to adapt to different environmental conditions, resulting in safety issues for food producers. Foods contaminated by L. monocytogenes can represent a risk if consumed by susceptible individuals such as elderly, pregnant women and the immunocompromised. The aim of this study was to evaluate the genetic diversity of a collection of L. monocytogenes isolated from different matrices in Brazil during the period of 1979-2015. A total of 51 L. monocytogenes serotype 1/2a strains isolated from clinical samples (n = 3) and food samples (n = 48) were characterized by Multi-Virulence-Locus Sequence Typing (MVLST). The strains were assigned to nine virulence types (VT): VT-11 (n = 3, 5·9%), VT-45 (n = 27, 52·9%), VT-59 (n = 11, 21·6%), VT-68 (n = 3, 5·9%), VT-94 (n = 2, 3·9%), VT-107 (n = 2, 3·9%), VT-184 (n = 1, 1·9%), VT-185 (n = 1, 1·9%) and VT-186 (n = 1, 1·9%); and four of them (VT-11, VT-45, VT-59 and VT-68) have already been associated with cases of listeriosis worldwide. The VT-11, VT-59 (Epidemic Clone V) and VT-186 were identified in blood culture samples, as well as in different classes of foods. It is recommended that the epidemiological surveillance agencies evaluate the risk that foods contaminated with L. monocytogenes VTs pose to susceptible populations.

Growth kinetics of Salmonella Typhimurium and Listeria monocytogenes in buffalo milk under different processing and storage conditions / Cinética de crescimento de Salmonella Typhimurium e Listeria monocytogenes em leite bubalino sob diferentes condições de processamento e armazenamento

Buffalo milk is rich in nutrients and can serve as a substrate for the proliferation of microorganisms. Thus, the objective of the present study was to evaluate the growth kinetics of Salmonella Typhimurium and Listeria monocytogenes in buffalo milk under different processing and storage conditions. Samples of raw and pasteurized milk were inoculated with 1 CFU of each bacterium, separately and together, per 25 mL. After contamination, samples were stored at 8 °C or 37 °C, and bacterial counts were performed at 24, 48, and 168 h. In addition, the accompanying microbiota growth, pH, and the effect of these variables on the growth kinetics of microorganisms were monitored. The pathogens tested were able to proliferate under most conditions tested, reaching high titers throughout the experimental period. At 37 °C, there was a decrease in pH and an increase in the accompanying microbiota that interfered with the microbial growth curve. It was also observed that pasteurized milk subjected to 8 °C provided better conditions for the multiplication of bacteria. Therefore, it was concluded that care throughout the production chain, storage, and commercialization of milk must be adopted to guarantee the microbiological safety of this food.

O leite bubalino é rico em nutrientes e pode servir de substrato para a proliferação de micro-organismos. Assim, o objetivo do presente estudo foi avaliar a cinética de crescimento de Salmonella Typhimurium e Listeria monocytogenes em leite bubalino, em diferentes condições de processamento e armazenamento. Para isso, utilizaram-se amostras de leite cru e pasteurizado, que foram inoculadas com 1 UFC/25 mL de cada bactéria separadamente e em conjunto. Após a contaminação, as amostras foram armazenadas nas temperaturas de 8 ºC e 37 ºC e realizadas contagens bacterianas em 24, 48 e 168h. Além disso, foi acompanhado o crescimento da microbiota acompanhante, o pH e o efeito dessas variáveis sobre a cinética de crescimento dos micro-organismos. Os patógenos testados conseguiram se proliferar na maioria das condições testadas, atingindo altos títulos durante todo o período experimental. Na temperatura de 37 ºC, houve uma diminuição do pH e um aumento da microbiota acompanhante, o que interferiu na curva de crescimento microbiana. Observou-se também que o leite pasteurizado e submetido a 8 ºC possibilitou melhores condições para a multiplicação das bactérias. Sendo assim, concluiu-se que cuidados durante toda a cadeia de produção, armazenamento e comercialização do leite devem ser adotados para garantir a segurança microbiológica desse alimento.

Growth kinetics of Salmonella Typhimurium and Listeria monocytogenes in buffalo milk under different processing and storage conditions / Cinética de crescimento de Salmonella Typhimurium e Listeria monocytogenes em leite bubalino sob diferentes condições de processamento e armazenamento

Buffalo milk is rich in nutrients and can serve as a substrate for the proliferation of microorganisms. Thus, the objective of the present study was to evaluate the growth kinetics of Salmonella Typhimurium and Listeria monocytogenes in buffalo milk under different processing and storage conditions. Samples of raw and pasteurized milk were inoculated with 1 CFU of each bacterium, separately and together, per 25 mL. After contamination, samples were stored at 8 °C or 37 °C, and bacterial counts were performed at 24, 48, and 168 h. In addition, the accompanying microbiota growth, pH, and the effect of these variables on the growth kinetics of microorganisms were monitored. The pathogens tested were able to proliferate under most conditions tested, reaching high titers throughout the experimental period. At 37 °C, there was a decrease in pH and an increase in the accompanying microbiota that interfered with the microbial growth curve. It was also observed that pasteurized milk subjected to 8 °C provided better conditions for the multiplication of bacteria. Therefore, it was concluded that care throughout the production chain, storage, and commercialization of milk must be adopted to guarantee the microbiological safety of this food.(AU)

O leite bubalino é rico em nutrientes e pode servir de substrato para a proliferação de micro-organismos. Assim, o objetivo do presente estudo foi avaliar a cinética de crescimento de Salmonella Typhimurium e Listeria monocytogenes em leite bubalino, em diferentes condições de processamento e armazenamento. Para isso, utilizaram-se amostras de leite cru e pasteurizado, que foram inoculadas com 1 UFC/25 mL de cada bactéria separadamente e em conjunto. Após a contaminação, as amostras foram armazenadas nas temperaturas de 8 ºC e 37 ºC e realizadas contagens bacterianas em 24, 48 e 168h. Além disso, foi acompanhado o crescimento da microbiota acompanhante, o pH e o efeito dessas variáveis sobre a cinética de crescimento dos micro-organismos. Os patógenos testados conseguiram se proliferar na maioria das condições testadas, atingindo altos títulos durante todo o período experimental. Na temperatura de 37 ºC, houve uma diminuição do pH e um aumento da microbiota acompanhante, o que interferiu na curva de crescimento microbiana. Observou-se também que o leite pasteurizado e submetido a 8 ºC possibilitou melhores condições para a multiplicação das bactérias. Sendo assim, concluiu-se que cuidados durante toda a cadeia de produção, armazenamento e comercialização do leite devem ser adotados para garantir a segurança microbiológica desse alimento.(AU)

Ten years of molecular epidemiology surveillance of Listeria monocytogenes in Chile 2008-2017.

Listeria monocytogenes causes severe diseases in humans, including febrile gastroenteritis and systemic infections that has a high mortality despite antibiotic treatment. This pathogen may cause massive outbreaks associated to the consumption of contaminated food products, which highlight its importance in public health. In the last decade, L. monocytogenes has emerged as a foodborne pathogen of major importance in Chile. A previous work showed that in Chile during 2008 and 2009, L. monocytogenes serotypes 1/2a, 1/2b and 4b were the most frequently identified in food and clinical strains. Here we report the molecular characterization of L. monocytogenes strains isolated from 2008 to 2017 in the country. Our results indicate that serotypes 1/2a, 1/2b and 4b continue to be the most commonly found in food products. In addition, we identify persistent and widespread PFGE subtypes. This study reports ten years of epidemiological surveillance ofL. monocytogenes in Chile.

Experimental listeriosis: A study of purinergic and cholinergic inflammatory pathway.

The cholinergic, purinergic and oxidative stress systems were related to nervous system damage in some pathologies, as well as being involved in pro-inflammatory and anti-inflammatory pathways. The objective was to investigate changes in purinergic, cholinergic systems and oxidative stress related to the neuropathology of listeriosis. Gerbils were used as experimental models. The animals were divided in two groups: control and infected. The animals were orally infected with 5 × 108 CFU/animal of the pathogenic strain of Listeria monocytogenes. Collected of material was 6 and 12th days post-infection (PI). Infected animals showed moderate mixed inflammatory infiltrates in the liver. The spleen and brain was used for PCR analyses, confirming infection by L. monocytogenes. Increase in number of total leukocytes because of an increase in lymphocytes in infected (P < 0.001). ATP and ADP hydrolysis by NTPDase was lower at 6 and 12th days PI in infected animals than in the control group. ADA (adenosine deaminase) activity was higher on the 6th day PI (P < 0.05) and decreased on the 12th day PI (P < 0.05) in infected animals. AChE (acetylcholinesterase) activity did not differ between groups on the 6th day PI; however, activity decreased in infected group on the 12th day PI (P < 0.05). On the 12th day PI, an increase of oxygen-reactive species levels and lower catalase and superoxide dismutase activities in the infected group was observed, characterizing a situation of cerebral oxidative stress. The inflammatory and oxidative mechanisms are present in listeriosis in asymptomatic animals, and that ectonucleotidases and cholinesterase's are involved in immunomodulation.

Efficiency of chitosan synergism with clove essential oil in the coating of intentionally contaminated Tambaqui fillets / Eficiência do sinergismo de quitosana com óleo essencial de cravo no revestimento de filés de tambaqui contaminados intencionalmente

The edible coating of chitosan with clove essential oil (CEO) was studied for its ability to reduce the microbial growth of pathogens (Escherichia coli O157:H7 CDCEDL933, Listeria monocytogenes CERELA, Salmonella Enteritidis ATCC13076, Staphylococcus aureus ATCC43300, and Pseudomonas aeruginosa ATCC27853) in Tambaqui fillets kept under refrigeration. In in vitro tests, chitosan showed higher antimicrobial activity against S. aureus and L. monocytogenes (MIC 0.5%), and CEO for L. monocytogenes (MIC 0.08%). Based on the antimicrobial activity of chitosan and CEO, Tambaqui fillets were subjected to different treatments, T1: chitosan 2%; T2: chitosan 2% + CEO 0.16%, and T3: chitosan 0.5% + CEO 0.08%, kept at 4 ºC for 72 h. The chitosan coating, incorporated with CEO, inhibited microorganisms in Tambaqui fillets and enhanced coating efficiency (p < 0.05). It was most effective against L. monocytogenes and S. aureus at the lowest CEO concentration (0.08%). Chitosan coating in combination with CEO enhanced the antimicrobial effect of pathogens on Tambaqui fillets, increased their shelf life under refrigeration, and was more effective against Gram-positive pathogens than Gram-negative pathogens.(AU)

O revestimento comestível de quitosana com óleo essencial de cravo (OEC) foi estudado por sua capacidade em reduzir o crescimento microbiano de patógenos (Escherichia coli O157:H7 CDCEDL933, Listeria monocytogenes CERELA Salmonella Enteritidis ATCC 13076, Staphylococcus aureus ATCC43300 e Pseudomonas aeruginosa ATCC27853) em filés de tambaqui mantidos sob refrigeração. Nos testes in vitro, a quitosana apresentou maior atividade antimicrobiana para S. aureus e L. monocytogenes (CIM 0,5%) e o OEC para L. monocytogenes (CIM 0,08%). Com base na atividade antimicrobiana da quitosana e OEC, os filés de Tambaqui foram submetidos a T1: quitosana a 2%; T2: quitosana 2% + OEC 0,16% e T3: quitosana 0,5% + OEC 0,08%, mantidos a 4 ºC por 72 h. O revestimento de quitosana, incorporado ao OEC, inibiu os micro-organismos nos filés de Tambaqui aumentando a eficiência do revestimento (p< 0,05); e foi mais eficaz para L. monocytogenes e S. aureus na menor concentração do OEC (0,08%). O revestimento de quitosana quando combinado ao OEC aumentou o efeito antimicrobiano de patógenos nos filés de Tambaqui, aumentando sua vida útil sob refrigeração, sendo mais eficaz contra patógenos Gram positivos do que os patógenos Gram negativos.(AU)

Distribution, adhesion, virulence and antibiotic resistance of persistent Listeria monocytogenes in a pig slaughterhouse in Brazil.

Listeria monocytogenes is a relevant pathogen usually associated with meat and ready-to-eat products. This study aimed to assess the distribution, adhesion, virulence and antibiotic resistance of L. monocytogenes in a pork production chain. Environment, carcass and food samples (n = 894) were obtained from different steps of a pork production chain over a 6-month period (10 samplings), including from farms and the slaughterhouse (reception, slaughtering, processing, storage and end products). L. monocytogenes was detected in samples from the reception (lairage floor, 1/10), slaughtering (drains, 2/20) and cutting room stages (conveyor belts in the final packing stage - 11/20, knife - 1/40, and cutting boards - 1/20). Positive results for conveyor belts were recorded in seven consecutive samplings. L. monocytogenes isolates (n = 87) were characterized as belonging to serogroup IVb and presented positive PCR results for inlA, inlB, inlC, inlJ, hlyA, plcA, actA and iap. Isolates were selected according to the original samples (n = 31) and subjected to Pulsed Field Gel Electrophoresis (PFGE), demonstrating their high clonal identity (98.4-100%). According to PFGE results and their original samples, isolates were selected (n = 16) and subjected to phenotypic assay to assess their adhesion potential and tested for resistance against 15 antibiotics; all tested isolates presented weak adhesion potential and were resistant to ampicillin. The present study demonstrated the persistence of L. monocytogenes in the pork processing facility, indicating the potential risk for cross-contamination with a potential virulent and resistant clone.

Pesquisa de Salmonella spp. e Listeria monocytogenes em queijo mussarela fatiado comercializado em hipermercados de Recife-PE / Search of Salmonella spp. and Listeria monocytogenes in sliced mussarella cheese marketed in hypermarkets of Recife-PE

O queijo mussarela, tipo mais utilizado no Brasil em preparações culinárias especialmente por características como fatiamento e derretimento, está sujeito a contaminações microbiológicas durante todo o processo que o leva à mesa do consumidor. Diante disso, objetivou-se pesquisar Salmonella spp. e Listeria monocytogenes em queijo mussarela fatiado comercializado em hipermercados de Recife-PE. Foram analisadas quarenta e nove amostras de queijo mussarela fatiado e todas foram negativas para os microrganismos pesquisados. Porém, observou-se Listeria innocuaem 4,1% das amostras (2/49). Estes resultados sugerem provável falha de higienização no local de fatiamento ou embalagem do produto, sendo necessárias ações que garantam a inocuidade dos alimentos ofertados ao consumidor, de forma a não pôr em risco a saúde pública.(AU)

Isolamento de Listeria monocytogenes em salsichas comercializadas no Distrito Federal / Isolation of Listeria monocytogenesin sausages commercialized in the Distrito Federal

A bactéria Listeria monocytogenes é o patógeno causador da listeriose, importante doença transmitida por alimentos que acomete humanos e animais podendo causar morte. Foram coletadas 100 amostras de salsicha de mercados do Distrito Federal. A metodologia usada para o isolamento foi a descrita na Instrução Normativa Nº 40 do Ministério da Agricultura, Pecuária e Abastecimento. As amostras que apresentaram motilidade característica de Listeria sp p. foram submetidas a técnica de PCR para confirmação da espécie L. monocytogenes. Os resultados positivos para Listeria sp p. foram vistos em 14 amostras e em 5 confirmou-se a presença de L. monocytogenes. Através dos resultados desse experimento conclui-se que salsichas podem estar contaminadas com bactérias do gênero Listeria sp p. e L. monocytogenes, oferecendo risco de listeriose aos consumidores.(AU)

Isolamento de Listeria monocytogenes em salsichas comercializadas no Distrito Federal / Isolation of Listeria monocytogenesin sausages commercialized in the Distrito Federal

A bactéria Listeria monocytogenes é o patógeno causador da listeriose, importante doença transmitida por alimentos que acomete humanos e animais podendo causar morte. Foram coletadas 100 amostras de salsicha de mercados do Distrito Federal. A metodologia usada para o isolamento foi a descrita na Instrução Normativa Nº 40 do Ministério da Agricultura, Pecuária e Abastecimento. As amostras que apresentaram motilidade característica de Listeria sp p. foram submetidas a técnica de PCR para confirmação da espécie L. monocytogenes. Os resultados positivos para Listeria sp p. foram vistos em 14 amostras e em 5 confirmou-se a presença de L. monocytogenes. Através dos resultados desse experimento conclui-se que salsichas podem estar contaminadas com bactérias do gênero Listeria sp p. e L. monocytogenes, oferecendo risco de listeriose aos consumidores.

Pesquisa de Salmonella spp. e Listeria monocytogenes em queijo mussarela fatiado comercializado em hipermercados de Recife-PE / Search of Salmonella spp. and Listeria monocytogenes in sliced mussarella cheese marketed in hypermarkets of Recife-PE

O queijo mussarela, tipo mais utilizado no Brasil em preparações culinárias especialmente por características como fatiamento e derretimento, está sujeito a contaminações microbiológicas durante todo o processo que o leva à mesa do consumidor. Diante disso, objetivou-se pesquisar Salmonella spp. e Listeria monocytogenes em queijo mussarela fatiado comercializado em hipermercados de Recife-PE. Foram analisadas quarenta e nove amostras de queijo mussarela fatiado e todas foram negativas para os microrganismos pesquisados. Porém, observou-se Listeria innocuaem 4,1% das amostras (2/49). Estes resultados sugerem provável falha de higienização no local de fatiamento ou embalagem do produto, sendo necessárias ações que garantam a inocuidade dos alimentos ofertados ao consumidor, de forma a não pôr em risco a saúde pública.

Association of Listeria monocytogenes LIPI-1 and LIPI-3 marker llsX with invasiveness.

Listeria monocytogenes is an opportunistic pathogen that is widely distributed in the environment. The evolution of its genome has exhibited differences in virulence among strains of the same species. Listeria monocytogenes LIPI-3 (Listeria Pathogenicity Island 3) and LIPI-1 (Listeria Pathogenicity Island 1) are considered responsible for the increased virulence in some strains. The aim of this study was to detect LIPI-1 genes and the llsX gene belonging to LIPI-3 in invasive strains of L. monocytogenes and to establish whether there is a relationship among the invasiveness, presence of the llsX and LIPI-1 genes, and the source of the strains. The results showed that 70% of the strains were invasive, and all these strains except one possessed LIPI-1, which suggests that although there is a correlation between LIPI-1 and invasiveness, the independent mechanisms of LIPI-1 may contribute to invasiveness. In contrast, 35% of the total strains were positive for llsX and were invasive; thus, the results revealed that there is a strong association between llsX and the invasiveness of L. monocytogenes in HEp-2 cells (HeLa contaminant/epithelial in origin). In addition, there is no other association with any other variable in this study. Moreover, the authors found that LIPI-1 and llsX are more frequently found in fresh than in frozen vegetables. Together, the findings provide an approximation for the better understanding of Listeriolysin S (LLS) and its role in the pathogenesis of L. monocytogenes, and a possible relation between virulence factors and food-storage temperature.

Lactic Acid Bacteria (LAB) and Their Bacteriocins as Alternative Biotechnological Tools to Control Listeria monocytogenes Biofilms in Food Processing Facilities.

Bacteriocins are antimicrobial peptides produced by bacteria Gram-negative and Gram-positive, including lactic acid bacteria (LAB), organisms that are traditionally used in food preservation practices. Bacteriocins have been shown to have an aptitude as biofilm controlling agents in Listeria monocytogenes biofilms, a major risk for consumers and the food industry. Biofilms protect pathogens from sanitization procedures, allowing them to survive and persist in processing facilities, resulting in the cross-contamination of the end products. Studies have been undertaken on bacteriocinogenic LAB, their bacteriocins, and bioengineered bacteriocin derivatives for controlling L. monocytogenes biofilms on different surfaces through inhibition, competition, exclusion, and displacement. These alternative strategies can be considered promising in preventing the development of resistance to conventional sanitizers and disinfectants. Bacteriocins are "friendly" antimicrobial agents, and with high prevalence in nature, they do not have any known associated public health risk. Most trials have been carried out in vitro, on food contact materials such as polystyrene and stainless steel, while there have been few studies performed in situ to consolidate the results observed in vitro. There are strategies that can be employed for prevention and eradication of L. monocytogenes biofilms (such as the establishment of standard cleaning procedures using the available agents at proper concentrations). However, commercial cocktails using alternatives compounds recognized as safe and environmental friendly can be an alternative approach to be applied by the industries in the future.

Lactobacillus spp. impair the ability of Listeria monocytogenes FBUNT to adhere to and invade Caco-2 cells.

OBJECTIVES: The objective of this study was to evaluate the ability of Lactobacillus curvatus CRL705, CRL1532, and CRL1533 and Lactobacillus sakei CRL1613 to survive under simulated gastrointestinal conditions. Moreover, a microencapsulation approach was proposed to improve gastrointestinal survival. Finally, experiments were performed to demonstrate that Lactobacillus spp. can modulate the ability of Listeria monocytogenes FBUNT to adhere to and invade Caco-2 cells. RESULTS: Lactobacillus strains were encapsulated in alginate beads to enhance the survival of bacteria under in vitro gastrointestinal conditions. All strains hydrolyzed bile salts using chenodeoxycholic acid as a substrate and adhered to Caco-2 cells. Cell-free supernatants (CFSs) showed antimicrobial activity against L. monocytogenes as demonstrated by agar diffusion assays. The average percentages of L. monocytogenes adhesion decreased from 67.74 to 41.75 and 38.7% in the presence of 50 and 90% (v/v), respectively, for all CFSs tested. The highest concentrations of CFSs completely inhibited the L. monocytogenes invasion of Caco-2 cells. CONCLUSIONS: The studied Lactobacillus strains have protective effects against the adhesion and invasion of L. monocytogenes FBUNT. Alginate encapsulation of these bacteria improved gastrointestinal tolerance such that they could be further studied as potential probiotics against intestinal pathogenic bacteria.

Action of ginger essential oil (Zingiber officinale) encapsulated in proteins ultrafine fibers on the antimicrobial control in situ.

The ultrafine fibers were produced using a polymeric blend of soy protein isolate (SPI), polyethylene oxide (PEO), and zein at a ratio of 1:1:1 (v/v/v) by electrospinning. The ginger essential oil (GEO) was encapsulated in the ultrafine fibers and the morphology, Fourier-Transform Infrared Spectroscopy (FTIR) analysis, thermal properties and relative crystallinity were evaluated. The antimicrobial activity of ginger essential oil was evaluated against five bacteria (Listeria monocytogenes, Staphylococcus aureus, Escherichia coli 0157:H7, Salmonella typhimurium, and Pseudomonas aeruginosa). Based on the preliminary tests, the concentration of GEO selected to add in the polymer solution was 12% (v/v; GEO/polymer solution). The fiber produced with 12% (v/v) GEO was used for antimicrobial analysis and in situ application (in fresh Minas cheese) against L. monocytogenes by micro-atmosphere. The ultrafine fibers produced, regardless the concentration of the essential oil, presented homogeneous morphology with cylindrical shape without the presence of beads. The application of the active fibers containing 12% GEO showed high potential to be applied in food packaging to reduce microbial contamination.

Exploiting antagonistic activity of fruit-derived Lactobacillus to control pathogenic bacteria in fresh cheese and chicken meat.

This study assessed the antagonistic activity of fruit-derived lactic acid bacteria (LAB) strains against food-related bacteria and the effects of the highest organic acids LAB producers on the survival of Listeria monocytogenes and Salmonella Enteritidis PT4 in cheese and chicken meat, respectively. The production of organic acids by the Lactobacillus strains in the tested food matrices was also monitored. All tested LAB strains showed antagonistic activity in vitro on the growth of pathogenic or spoiling food-related bacteria, particularly on L. monocytogenes and/or S. Enteritidis PT4, through the action of non-proteinaceous substances. The highest amounts of acetic and lactic acid were detected in cell free culture supernatants of L. paracasei 108 and L. plantarum 201. In "Minas Frescal" cheese, L. plantarum 49 and L. paracasei 108 decreased the counts of L. monocytogenes, and L. plantarum 201 showed bacteriostatic effects on this pathogen over time. L. paracasei 108 decreased the counts of S. Enteritidis PT4 in ground chicken breast; L. plantarum 49 and L. plantarum 201 failed to decrease the counts of this pathogen. Decreases in counts of L. monocytogenes or S. Enteritidis in "Minas Frescal" cheese and ground chicken breast, respectively, were related with increases in lactic and acetic acid contents and decreases in pH values. L. plantarum 49 and L. paracasei 108 could be used as biopreservation tools in cheese and chicken breast meat, respectively.

Changes on the activity of cholinesterase's in an immunomodulatory response of cattle infected by Listeria monocytogenes.

The aim of this study was to evaluate whether Listeria monocytogenes infection alters the activity of cholinesterases in cattle to module their inflammatory response and neurotransmission. Thus, ten male bovines (Holstein) were divided into two groups of five animals each: uninfected (control) and L. monocytogenes infected. Blood samples were collected on days 0, 7 and 14 post-infection (PI) to evaluate the activity of acetylcholinesterase (AChE) in the blood, and seric butyrylcholinesterase (BChE) activity, as well as total protein, albumin, globulin and C-reactive protein (CPR) levels in serum. The AChE activity and acetylcholine (ACh) levels were measured in the central nervous system on day 14 PI, and histopathological analyses were also performed. The infected animals did not show apparent clinical signs of listeriosis, however histopathological alterations were seen in the intestines and spleens. On days 7 and 14 PI, AChE activity in the blood was lower in infected animals, as well the seric BChE activity on day 7 PI. In the cerebral cortex and cerebellum, AChE activity was lower in infected animals compared to the control group, while the ACh levels were higher in the cerebral cortex compared to uninfected animals. Moreover, seric levels of total protein, globulin and CRP were higher in infected animals on days 7 and 14 PI compared to the control group. Therefore, we concluded that acute infection by L. monocytogenes alters the cholinergic system through the reduction of cholinesterase enzymes in the blood, serum and cerebral tissues as an adaptive response to an anti-inflammatory effect in order to increase the ACh levels, an anti-inflammatory molecule with an important role in the host immunomodulation.