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Listeria Monocytogenes (listeria + monocytogene)
Kinds of Listeria Monocytogenes Terms modified by Listeria Monocytogenes Selected AbstractsEtiologic spectrum and pattern of antimicrobial drug susceptibility in bacterial meningitis in Sokoto, NigeriaACTA PAEDIATRICA, Issue 8 2000FE EmeleArticle first published online: 2 JAN 200 Etiologic agents of meningitis were prospectively investigated among patients admitted to Usman Danfodio University Teaching Hospital, Sokoto. Of 1097 cerebrospinal fluid (CSF) samples submitted to the microbiology laboratory from various wards of the hospital, 289 (26%) were microscopically, culturally and/or serologically proven to be bacterial meningitis. The etiologic spectrum was as follows: Neisseria meningitidis (61%), Streptococcus pneumoniae (18%), Haemophilus influenzae (10%), Staphylococcus aureus (6%), Coliform bacilli (3%), Escherichia coli (0.7%), Mycobacterium tuberculosis (0.7%), Listeria monocytogenes (0.4%), Flavobacterium meningosepticum (0.4%) and Pseudomonas putrifasciens (0.4%). Bacterial meningitis was most prevalent (195 or 68%) among children aged 1-9 y, while adults and neonates were least affected. Coliform bacilli caused five of eight neonatal cases. Males were more frequently affected than females (x2=12.50;p < 0.05). Culture and microscopy were comparatively less efficient than the search for bacterial antigens, especially in the diagnosis of Haemophilus meningitis. Antimicrobial susceptibility of N. meningitidis to ampicillin and benzyl penicillin reduced progressively over the years (F = 406.98;p < 0.001). Nineteen (11%) of the isolates (5 Meningococci, 7 Staph. aureus, 1 Haem. influenza and 6 others) showed simultaneous resistance to chloramphenicol, ampicillin and benzyl penicillin. [source] IL-15 is critical for the maintenance and innate functions of self-specific CD8+ T cellsEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 7 2009Momoe Itsumi Abstract IL-15 is a pleiotropic cytokine involved in host defense as well as autoimmunity. IL-15-deficient mice show a decrease of memory phenotype (MP) CD8+ T cells, which develop naturally in naďve mice and whose origin is unclear. It has been shown that self-specific CD8+ T cells developed in male H-Y antigen-specific TCR transgenic mice share many similarities with naturally occurring MP CD8+ T cells in normal mice. In this study, we found that H-Y antigen-specific CD8+ T cells in male but not female mice decreased when they were crossed with IL-15-deficient mice, mainly due to impaired peripheral maintenance. The self-specific TCR transgenic CD8+ T cells developed in IL-15-deficient mice showed altered surface phenotypes and reduced effector functions ex vivo. Bystander activation of the self-specific CD8+ T cells was induced in vivo during infection with Listeria monocytogenes, in which proliferation but not IFN-, production was IL-15-dependent. These results indicated important roles for IL-15 in the maintenance and functions of self-specific CD8+ T cells, which may be included in the naturally occurring MP CD8+ T-cell population in naďve normal mice and participate in innate host defense responses. [source] Priming of CD8+ T cell responses by pathogens typically depends on CD70-mediated interactions with dendritic cellsEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 3 2007Anita Schildknecht Abstract The CD27/CD70-interaction has been shown to provide a costimulatory and survival signal for T cells in vitro and in vivo. Recently, CD70 expression by DC was found to be important for the priming of CD8+ T cells. We show here that blocking CD70 interactions has a significant impact on priming of CD8+ T cell responses by vaccinia virus (VV), Listeria monocytogenes and vesicular stomatitis virus (VSV) in mice. However, the priming of specific CD8+ T cells upon infection with lymphocytic choriomeningitis virus (LCMV) was only marginally reduced by CD70-blockade. Blocking of CD70 prevented CD8+ T cell priming in DIETER mice, a model in which presentation of LCMV-derived epitopes can be induced selectively in dendritic cells (DC). In contrast, CD70-CD27 interactions were not important for the priming of VSV-specific CD4+ T cells or class switch of neutralizing antibodies. As we show that priming of CD8+ T cells by the pathogens used here is dependent on antigen presentation by DC and that infection results in up-regulation of CD70 on DC, we conclude that CD70 expression on DC plays an important role in the priming of CD8+ T cells by pathogens. Moreover, the lack of CD70 cannot be completely compensated for by other costimulatory molecules. [source] Virulence factor p60 of Listeria monocytogenes modulates innate immunity by inducing tumor necrosis factor ,FEMS IMMUNOLOGY & MEDICAL MICROBIOLOGY, Issue 1 2010Hiroshi Sashinami Abstract We investigated the effect of p60, a virulence factor of Listeria monocytogenes, on host immune response in vitro and in vivo. Administration of p60 before a sublethal infection with L. monocytogenes enhanced innate host resistance in naďve mice. Mouse macrophage RAW264.7 cells produced tumor necrosis factor (TNF)-, in response to stimulation with recombinant p60. Toll-like receptor 4 may be involved in TNF-, production from RAW264.7 cells and enhanced host resistance induced by p60 administration. Our findings demonstrated that p60 modulates innate immune responses against L. monocytogenes infection. [source] Listeria monocytogenes: epidemiology, human disease, and mechanisms of brain invasionFEMS IMMUNOLOGY & MEDICAL MICROBIOLOGY, Issue 2 2008Douglas A. Drevets Abstract Listeria monocytogenes is a facultative intracellular bacterium that has predilection for causing central nervous systemic infections in humans and domesticated animals. This pathogen can be found worldwide in the food supply and most L. monocytogenes infections are acquired through ingestion of contaminated food. The main clinical syndromes caused by L. monocytogenes include febrile gastroenteritis, perinatal infection, and systemic infections marked by central nervous system infections with or without bacteremia. Experimental infection of mice has been used for over 50 years as a model system to study the pathogenesis of this organism including the mechanisms by which it invades the brain. Data from this model indicate that a specific subset of monocytes, distinguished in part by high expression of the Ly-6C antigen, become parasitized in the bone marrow and have a key role in transporting intracellular bacteria across the blood-brain barriers and into the central nervous system. This Minireview will summarize recent epidemiologic and clinical information regarding L. monocytogenes as a human pathogen and will discuss current in vitro and in vivo data relevant to the role of parasitized monocytes and the pathogenetic mechanisms that underlie its formidable ability to invade the central nervous system. [source] Isolation and characterisation of a 13.8-kDa bacteriolytic enzyme from house dust mite extracts: homology with prokaryotic proteins suggests that the enzyme could be bacterially derivedFEMS IMMUNOLOGY & MEDICAL MICROBIOLOGY, Issue 2 2002Leslie T. Mathaba Abstract Bacteriolytic activity was detected in extracts of whole mite and spent growth medium (SGM) from the clinically important Dermatophagoides pteronyssinus and Dermatophagoides farinae mites and was most abundant in whole mite extract. Gram-positive organisms Micrococcus lysodeikticus, Bacillus megaterium and Listeria monocytogenes were preferentially lysed and the lytic activity was enhanced by thiols, destroyed by mite proteases, inhibited by HgCl2 and high concentrations of NaCl but was resistant to heat and acid treatment. Substrate SDS,PAGE analysis indicated the presence of several lytic enzymes, two of which were isolated from D. pteronyssinus spent growth medium extract by hydroxyapatite chromatography. The N-terminal amino acid sequence of one of them was then used in PCR-based cloning studies. The complete amino acid sequence of this protein was determined and cDNA found to encode a 130-amino acid residue mature protein with a 20-amino acid leader sequence. The deduced protein demonstrated sequence similarity with the C-terminal regions of a group of bacterial proteins belonging to the P60 superfamily. These data suggest that the enzyme is derived from bacteria within the mites rather than from mites per se. [source] Viability of Listeria monocytogenes in co-culture with Acanthamoeba spp.FEMS MICROBIOLOGY ECOLOGY, Issue 1 2009Alisha Akya Abstract Listeria monocytogenes is a human pathogen, ubiquitous in the environment, and can grow and survive under a wide range of environmental conditions. It contaminates foods via raw materials or food-processing environments. However, the current knowledge of its ecology and, in particular, the mode of environmental survival and transmission of this intracellular pathogen remains limited. Research has shown that several intracellular pathogens are able to survive or replicate within free-living amoebae. To examine the viability of L. monocytogenes in interaction with Acanthamoeba spp., bacteria were co-cultured with three freshly isolated amoebae, namely Acanthamoeba polyphaga, Acanthamoeba castellanii and Acanthamoeba lenticulata. The survival of bacteria and amoebae was determined using culture techniques and microscopy. Under the experimental conditions used, all amoebae were able to eliminate bacteria irrespective of the hly gene. Bacteria did not survive or replicate within amoeba cells. However, extra-amoebic bacteria grew saprophytically on materials released from amoebae, which may play an important role in the survival of bacteria under extreme environmental conditions. [source] Listeriolysin O: a key protein of Listeria monocytogenes with multiple functionsFEMS MICROBIOLOGY REVIEWS, Issue 4 2006Samer Kayal Abstract Cholesterol-dependent cytolysins (CDCs) are produced by a large number of pathogenic Gram-positive bacteria. Most of these single-chain proteins are secreted in the extracellular medium. Among the species producing CDCs, only two species belonging to the genus Listeria (Listeria monocytogenes and Listeria ivanovii) are able to multiply intracellularly and release their toxins in the phagosomal compartment of the infected host cell. This review provides an updated overview on the importance of listeriolysin O (LLO) in the pathogenicity of L. monocytogenes, focusing mainly on two aspects: (1) the structure,function relationship of LLO and (2) its role in intra- and extracellular signalling. We first examine the specific sequence determinants, or protein domains, that make this cytolysin so well adapted to the intracellular lifestyle of L. monocytogenes. The roles that LLO has in cellular signalling events in the context of relations to pathogenesis are also discussed. [source] Methods for the isolation and identification of Listeria spp. and Listeria monocytogenes: a reviewFEMS MICROBIOLOGY REVIEWS, Issue 5 2005Uta Gasanov Abstract Listeria monocytogenes is an important food-borne pathogen and is widely tested for in food, environmental and clinical samples. Identification traditionally involved culture methods based on selective enrichment and plating followed by the characterization of Listeria spp. based on colony morphology, sugar fermentation and haemolytic properties. These methods are the gold standard; but they are lengthy and may not be suitable for testing of foods with short shelf lives. As a result more rapid tests were developed based on antibodies (ELISA) or molecular techniques (PCR or DNA hybridization). While these tests possess equal sensitivity, they are rapid and allow testing to be completed within 48 h. More recently, molecular methods were developed that target RNA rather than DNA, such as RT-PCR, real time PCR or nucleic acid based sequence amplification (NASBA). These tests not only provide a measure of cell viability but they can also be used for quantitative analysis. In addition, a variety of tests are available for sub-species characterization, which are particularly useful in epidemiological investigations. Early typing methods differentiated isolates based on phenotypic markers, such as multilocus enzyme electrophoresis, phage typing and serotyping. These phenotypic typing methods are being replaced by molecular tests, which reflect genetic relationships between isolates and are more accurate. These new methods are currently mainly used in research but their considerable potential for routine testing in the future cannot be overlooked. [source] Signalling mechanisms for Toll-like receptor-activated neutrophil exocytosis: key roles for interleukin-1-receptor-associated kinase-4 and phosphatidylinositol 3-kinase but not Toll/IL-1 receptor (TIR) domain-containing adaptor inducing IFN-, (TRIF)IMMUNOLOGY, Issue 3 2009Agnieszka A. Brzezinska Summary Lipopolysaccharide (LPS) stimulates exocytosis in neutrophils. The signalling molecules involved in the regulation of this mechanism are currently unknown. Using neutrophils from interleukin-1-receptor-associated kinase (IRAK)-4- and Toll/IL-1 receptor (TIR) domain-containing adaptor inducing IFN-, (TRIF)-deficient mice, we dissected the signalling pathways that control exocytosis. We analysed exocytosis of peroxidase-negative and azurophilic granules by following the mobilization of the ,2-integrin subunit CD11b and myeloperoxidase (MPO)-containing granules, respectively. IRAK-4-null neutrophils showed marked defects in both peroxidase-negative and azurophilic granule exocytosis in response to LPS. In contrast, the exocytic response to LPS of TRIF-deficient neutrophils was not different from that of wild-type cells. No differences were observed in the exocytosis of secretory organelles between IRAK-4-null and wild-type neutrophils when they were stimulated with the phorbol ester phorbol 12-myristate 13-acetate (PMA). Electron microscopy analysis showed that no morphological abnormalities were present in the granules of IRAK-4-deficient neutrophils, suggesting that the lack of exocytic response to LPS is not attributable to developmental abnormalities. Using pharmacological inhibitors, we found that p38 mitogen-activated protein kinase (p38MAPK) is essential for the exocytosis of all neutrophil secretory organelles in response to LPS. Interestingly, we found that phosphatidylinositol 3-kinase (PI3K) is essential for azurophilic granule exocytosis but not for the mobilization of other neutrophil granules in response to LPS. Azurophilic granule exocytosis in response to Listeria monocytogenes was dependent on PI3K but not IRAK-4 activity, suggesting that alternative signalling pathways are activated in IRAK-4-deficient neutrophils exposed to whole bacteria. Our results identified IRAK-4, p38MAPK and PI3K as important regulatory components with different roles in the signalling pathways that control Toll-like receptor ligand-triggered neutrophil exocytosis. [source] Importance of murine V,1+,, T cells expressing interferon-, and interleukin-17A in innate protection against Listeria monocytogenes infectionIMMUNOLOGY, Issue 2 2008Satoru Hamada Summary Murine ,, T cells participate in the innate immune response against infection by an intracellular pathogen Listeria monocytogenes. V,1+,, T cells coexpressing V,6 are a major ,, T-cell subpopulation induced at an early stage of L. monocytogenes infection in the livers of infected mice. To investigate the protective role of the V,6/V,1+,, T cells against L. monocytogenes infection, V,1 gene-deficient (V,1,/,) mice were analysed because these mice selectively lacked a V,6/V,1+,, T-cell subpopulation in the L. monocytogenes -infected liver. The V,1,/, mice showed increased bacterial burden in the liver and spleen, and decreased survival rate at an early stage of L. monocytogenes infection when compared to wild-type mice. Histological examination showed abscess-like lesions and unorganized distribution of macrophages in the liver of the V,1,/, mice but not in the wild-type mice after L. monocytogenes infection. The V,6/V,1+,, T cells produced interferon-, and interleukin-17A. All the results suggest that murine V,6/V,1+,, T cells control the innate protective response against L. monocytogenes infection through production of the proinflammatory cytokines interferon-, and interleukin-17A in the infected liver. [source] The contribution of both oxygen and nitrogen intermediates to the intracellular killing mechanisms of C1q-opsonized Listeria monocytogenes by the macrophage-like IC-21 cell lineIMMUNOLOGY, Issue 1 2000C. Álvarez-Domínguez Summary Listeria monocytogenes is a facultative intracellular pathogen which is internalized by host mammalian cells upon binding to their surface. Further listerial growth occurs in the cytosol after escape from the phagosomal,endosomal compartment. We have previously reported that C1q is able to potentiate L. monocytogenes phagocytosis upon bacterial opsonization by ingestion through C1q-binding structures. In this report, we analysed the post-phagocytic events upon internalization of C1q-opsonized L. monocytogenes and found an induction of macrophage (M,)-like IC-21 cell bactericidal mechanisms displayed by the production of oxygen and nitrogen metabolites. Both types of molecules are effective in L. monocytogenes killing. Further analysis of the cellular responses promoted by interaction of C1q with its surface binding structures, leads us to consider C1q as a collaborative molecule involved in M, activation. Upon interaction with surface binding structures, C1q was able to trigger and/or amplify the production of reactive oxygen and nitrogen intermediates induced by stimuli such as interferon-, and L. monocytogenes phagocytosis. [source] Early bacterial dependent induction of inducible nitric oxide synthase (iNOS) in epithelial cells upon transfer of CD45RBhigh CD4+ T cells in a model for experimental colitisINFLAMMATORY BOWEL DISEASES, Issue 12 2007Gerard Dijkstra MD Abstract Background: Both the role of inducible nitric oxide synthase (iNOS) in the development of inflammatory bowel disease (IBD) as well as the molecular details governing its mucosal induction remain unclear. Methods: In the present study we evaluated the role of the residing intestinal microflora in the induction of epithelial iNOS upon transfer of CD45RBhigh CD4+ T cells to SCID mice. CB-17 SCID mice were reared with conventional flora (CNV) or germfree CB-17 SCID mice were monoassociated with Helicobacter muridarum, act A(,) mutant Listeria monocytogenes, segmented filamentous bacteria (SFB), or Ochrobactrum anthropi. Results: Within 2 weeks CNV SCID mice injected with CD45RBhigh CD4+ T cells showed a focal, epithelial iNOS expression on the apical site of villi that preceded the infiltration of CD4+ T cells and cytokine production followed by extension of this expression to the entire surface along the whole crypt axis as the colitis progressed. SCID mice monoassociated with H. muridarum developed a severe colitis and showed high epithelial iNOS expression. CNV-SCID mice without T cells and SCID mice monoassociated with SFB did not show any iNOS expression, whereas SCID mice monoassociated with act A(,) mutant L. monocytogenes and O. anthropi showed some scattered epithelial iNOS staining on the apical site of a few villi, but none of these mice developed colitis. Conclusions: These findings demonstrate that the expression of epithelial iNOS is highly bacterium-specific and correlates with the severity of disease, suggesting an important role for this enzyme in the development of IBD. (Inflamm Bowel Dis 2007) [source] Incidence and antibiotic susceptibility of bacteriocin-producing lactic acid bacteria from dairy productsINTERNATIONAL JOURNAL OF DAIRY TECHNOLOGY, Issue 4 2008MAHBOUBEH MIRHOSSEINI Screening for bacteriocin production by strains of lactic acid bacteria (LAB) from local dairy products in Iran resulted in the detection of 10 bacteriocin-producing strains. Among 105 isolated, 10 bacteriocin producers were phenotypically and genotypically identified as Enterococcus spp. The antimicrobial compounds produced by these novel strains were inactivated by trypsin, proteinase k. These bacteriocins also were active in a wide range of pH and temperature values, and inhibited not only the closely related LAB, but also Listeria monocytogenes. [source] Evaluation of pre-heating and extraction solvents in antioxidant and antimicrobial activities of garlic, and their application in fresh pork pattiesINTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, Issue 2 2010Sung Y. Park Summary The objectives of this study were to screen the optimum conditions for antioxidant and antimicrobial activities of garlic as affected by pre-heating and different extraction solvents, and to evaluate the antioxidant and antimicrobial effects of these extracts in ground meat during refrigerated storage. Methanol extracted garlic had a greater total phenolic content, 1,1-diphenyl-2-picrylhydrazyl (DPPH)-radical scavenging activity and reducing power than water extracted one (P < 0.05), whereas the latter had a greater yield and iron chelating ability than the former (P < 0.05). Moreover, water extract from fresh garlic (WEFG) and methanol extract from heated garlic (MEHG) produced an inhibition zone against Escherichia coli O157:H7 and Listeria monocytogenes. The addition of garlic extracts (WEFG, MEHG and their combinations WEFMEHG)) to pork patties decreased the pH, hunter a values (redness), thiobarbituric acid substances values and the number of total plate count and Enterobacteriaceae (P < 0.05), while the hunter b values (yellowness) increased (P < 0.05). Results of this study indicated that the use of the garlic extracts was able to control lipid oxidation and microbial growth in pork patties. [source] Chemical composition and inhibitory effect of essential oil and organic extracts of Cestrum nocturnum L. on food-borne pathogensINTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, Issue 6 2009Sharif M. Al-Reza Summary In this study, we examined the chemical compositions of essential oil and tested the efficacy of oil and organic extracts of Cestrum nocturnum L. against food-borne pathogens. The chemical compositions of the oil was analysed by gas chromatography-mass spectrometry (GC-MS). Forty-seven compounds representing 93.28% of the total oil were identified. The oil [5 ,L of 1:5 (v/v) dilution of oil with methanol] and organic extracts of hexane, chloroform, ethyl acetate and methanol (300 ,g per disc) of C. nocturnum displayed a great potential of antibacterial activity against Staphylococcus aureus (ATCC 6538 and KCTC 1916), Listeria monocytogenes (ATCC 19166 and ATCC 15313), Bacillus subtilis ATCC 6633, Pseudomonas aeruginosa KCTC 2004, Salmonella typhimurium KCTC 2515 and Escherichia coli ATCC 8739. Also the oil had strong detrimental effect on the viable count of the tested bacteria. The results obtained from this study may contribute to the development of new antimicrobial agents with potential applications in food industries as natural preservatives to control food-borne pathogens. [source] Antimicrobial activity of nisin incorporated in pectin and polylactic acid composite films against Listeria monocytogenesINTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, Issue 2 2009Tony Jin Summary An extruded composite food packaging film containing pectin, polylactic acids (PLAs) and nisin was developed to inhibit Listeria monocytogenes. The mechanical properties and surface structure of the film were also examined. Cells of L. monocytogenes were reduced by 2.1, 4.5 and 3.7 log units mL,1 by the pectin plus PLA (pectin/PLA) film containing nisin (1000 IU mL,1 of tested liquid) in Brain Heart Infusion (BHI) broth, liquid egg white and orange juice, respectively, after 48 h at 24 °C. Pectin played an important roll in embedding nisin into the film. The pectin/PLA film had a similar stiffness but lower tensile strength, elongation and fracture energy than the pure PLA film. These data suggested that nisin incorporated into the pectin/PLA film was an effective approach to reducing L. monocytogenes in a typical growth medium (e.g. BHI broth) as well as in foods (e.g. orange juice and liquid egg). [source] Adhesion of Listeria monocytogenes to materials commonly found in domestic kitchensINTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, Issue 7 2008Pilar Teixeira Summary The aim of this work was to investigate the adhesion of Listeria monocytogenes ATCC 15313 to glass, granite, marble, polypropylene from a bowl (PPb), polypropylene from a cutting board (PPcb) and stainless steel (SS), which are materials commonly used in kitchens. Marble and granite were chosen because they are applied as kitchen bench covers and pavements in many countries and there are no literature reports on their behaviour in terms of microbial adhesion. The effect of surface hydrophobicity and roughness on the adhesion process was also analysed. The results showed that the highest extent of adhesion of L. monocytogenes occurred to stainless steel, followed by glass and in less extent to the other materials studied. However, it was not possible to establish a correlation between surface hydrophobicity or roughness and the extent of adhesion of L. monocytogenes. The adherence of L. monocytogenes should be dependent on other factors, like the presence of exopolymers and surface charge. [source] Incidence and sources of Listeria monocytogenes in a traditional hot-smoked rainbow trout processing plant in Turkey,INTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, Issue 11 2007Duygu K Summary In recent years, microbial fish safety is getting a close attention from regulatory agencies and consumers. Therefore, fish farm raising rainbow trout and affiliated slaughterhouse and smoking plants were evaluated for the occurrence of Listeria monocytogenes. Samples including raw fish, swabbings of equipment or other surfaces, as well as processing water, salt, fish feed and fish samples taken after various stages of processing were collected from thirty different locations in the plant. For the detection of L. monocytogenes, both conventional and Listeria Rapid Test (LRT) were used. L. monocytogenes was detected in thirty out of sixty samples (50%) by LRT, while it was detected in thirty-four out of sixty samples (57%) by conventional method. No L. monocytogenes was detected from raw fish, smoked fish (before handling) and processing water, but it was detected in all environmental samples including swabbings of equipment or other surfaces and smoked fish samples after filleting. [source] smcL as a novel diagnostic marker for quantitative detection of Listeria ivanovii in biological samplesJOURNAL OF APPLIED MICROBIOLOGY, Issue 3 2010D. Rodríguez-Lázaro Abstract Aims:, To develop a novel molecular tool for the quantitative detection of the ruminant pathogen Listeria ivanovii in different biological matrices. Methods and Results:, A real-time PCR (RTi-PCR) for the quantitative and species-specific identification of L. ivanovii was designed to target the region of the smcL gene. The assay includes an internal amplification control (IAC) to avoid false-negative results. The smcL -IAC RTi-PCR assay was 100% selective and allowed the detection of as little as one genome equivalent in 45% of reactions. The quantification accuracy was excellent, as demonstrated by its high linearity (R2 > 0ˇ9989) and PCR efficiency (E > 0ˇ984) over a 6-log dynamic range, down to 10 genome equivalents. Finally, the applicability of this assay was evaluated with artificially contaminated biological matrices implicated in the transmission of this bacterium such as sheep raw milk, blood and amniotic fluid. The smcL -IAC RTi-PCR assay allowed the detection of as few as 50 colony forming unit numbers (CFUs) per 25 ml of raw milk, 43 CFUs per 1 ml of blood or 50 CFUs per 1 ml of amniotic fluid. Conclusions:, This method can be an adequate alternative for the identification of L. ivanovii and for complete diagnosis of animal and human listeriosis. Significance and Impact of the Study:, We present an alternative for the detection of another pathogenic member of Listeria genus, which can help to distinguish from Listeria monocytogenes and therefore facilitates the establishment of preventive and prophylactic measures in food and farm environments. [source] Microbiological analysis of composts produced on South Carolina poultry farmsJOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2010M.W. Shepherd Jr Abstract Aims:, The purpose of this study was to determine whether the methods used in compost operations of small and medium-sized poultry forms resulted in the production of an amendment free of foodborne pathogens. Methods and Results:, Nine compost heaps on five South Carolina poultry farms were surveyed at different stages of the composting process. Compost samples were analysed for coliforms and enriched for Escherichia coli O157:H7, Salmonella spp. and Listeria monocytogenes. The waste materials and composting practices differed among the surveyed farms. On two farms, new materials were added to heaps that had previously completed the active composting phase. Five compost heaps did not reach an internal temperature of 55°C, and c. 62% of all internal samples in the first composting phase contained moisture contents <40%. Escherichia coli was detected in 63% of the surface samples (n = 38) and 9ˇ8% of the internal samples (n = 82) from the first composting phase, as compared with 16ˇ7% of the surface samples (n = 12) and 0% internal samples (n = 24) from the second composting phase. Salmonella was detected in 26 and 6ˇ1% of all surface and internal samples collected from heaps in the first composting phase, respectively, but was absent in all compost samples undergoing a second composting phase. The predominant Salmonella serotypes were Thompson, Montevideo and Anatum. Neither E. coli O157:H7 nor L. monocytogenes was detected in any of the samples. Conclusions:, Our results indicate that the conditions at the compost surface are suitable for pathogen survival, and the complete composting process can result in the elimination of pathogens in poultry wastes. Significance and Impact of the Study:, This research provides information regarding the effectiveness of the composting practices and microbiological quality of poultry compost produced by small- and medium-sized farms. Ensuring the safety of compost that may be applied to soils should be an integral part of preharvest food safety programme. [source] Assessment of survival of Listeria monocytogenes, Salmonella Infantis and Enterococcus faecalis artificially inoculated into experimental waste or compostJOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2010N. Paniel Abstract Aims:, To evaluate survival of pathogenic strains, Listeria monocytogenes and Salmonella Infantis and a sanitation indicator Enterococcus faecalis in composts at different stages of the composting process and during storage. Methods and Results:, The studied pathogenic and indicator strains, originally isolated from compost, were inoculated into compost samples from the various stages of the composting process. During incubation, indigenous microflora diversity was monitored with DGGE analysis. After 90 days of incubation, strain survival was observed in compost sampled before the beginning of the cooling phase, and DGGE analysis demonstrated an increase of microbial diversity up to the cooling phase. However, inoculated strains were not detected in composts after 30, 60 or 90 days of incubation in compost sampled after the start of the cooling phase. Microbial diversity also became stable, and DGGE profiles reached a maximum number of bands at this stage. Conclusions:, Strain survival was not observed in stabilized composts. The cooling phase seems to be the turning point for pathogen survival and at this stage the indigenous microflora appeared to play a significant role in suppression. Significance and Impact of the Study:, The importance of indigenous microflora in the survival of pathogens in four different composts was demonstrated. Stabilized composts were recommended for spreading on land. [source] Cloning and comparison of phylogenetically related chitinases from Listeria monocytogenes EGD and Enterococcus faecalis V583JOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2009J.J. Leisner Abstract Aims:, To compare enzymatic activities of two related chitinases, ChiA and EF0361, encoded by Listeria monocytogenes and Enterococcus faecalis, respectively. Methods and Results:, The chiA and EF0361 genes were amplified by PCR, cloned and expressed with histidine tags, allowing easy purification of the gene products. ChiA had a molecular weight as predicted from the amino acid sequence, whereas EF0361 was 1840 Da lower than expected because of C-terminal truncation. The ChiA and EF0361 enzymes showed activity towards 4-nitrophenyl N,N,-diacetyl-,- d -chitobioside with Km values of 1ˇ6 and 2ˇ1 mmol l,1, respectively, and kcat values of 21ˇ6 and 6ˇ5 s,1. The enzymes also showed activity towards 4-nitrophenyl ,- d - N, N,, N,-triacetylchitotriose and carboxy-methyl-chitin-Remazol Brilliant Violet but not towards 4-nitrophenyl N- acetyl-,- d -glucosaminide. Chitinolytic specificities of the enzymes were supported by their inactivity towards the substrates 4-nitrophenyl ,- d -cellobioside and peptidoglycan. The pH and temperature profiles for catalytic activities were relatively similar for both the enzymes. Conclusion:, The ChiA and EF0361 enzymes show a high degree of similarity in their catalytic activities although their hosts share environmental preferences only to some extent. Significance and Impact of the Study:, This study contributes to an understanding of the chitinolytic activities by L. monocytogenes and Ent. faecalis. Detailed information on their chitinolytic systems will help define potential reservoirs in the natural environment and possible transmission routes into food-manufacturing plants. [source] The growth limits of a large number of Listeria monocytogenes strains at combinations of stresses show serotype- and niche-specific traitsJOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2008S. Van Der Veen Abstract Aims:, The aim of this study was to associate the growth limits of Listeria monocytogenes during exposure to combined stresses with specific serotypes or origins of isolation, and identify potential genetic markers. Methods and Results:, The growth of 138 strains was assessed at different temperatures using combinations of low pH, sodium lactate, and high salt concentrations in brain heart infusion broth. None of the strains was able to grow at pH , 4ˇ4, aw , 0ˇ92, or pH , 5ˇ0 combined with aw , 0ˇ94. In addition, none of the strains grew at pH , 5ˇ2 and NaLac , 2%. At 30°C, the serotype 4b strains showed the highest tolerance to low pH and high NaCl concentrations at both pH neutral (pH 7ˇ4) and mild acidic conditions (pH 5ˇ5). At 7°C, the serotype 1/2b strains showed the highest tolerance to high NaCl concentrations at both pH 7ˇ4 and 5ˇ5. Serotype 1/2b meat isolates showed the highest tolerance to low pH in the presence of 2% sodium lactate at 7°C. ORF2110 and gadD1T1 were identified as potential biomarkers for phenotypic differences. Conclusions:, Differences in growth limits were identified between specific L. monocytogenes strains and serotypes, which could in some cases be associated with specific genetic markers. Significance and Impact of the Study:, Our data confirm the growth limits of L. monocytogenes as set out by the European Union for ready-to-eat foods and provides an additional criterion. The association of L. monocytogenes serotypes with certain stress responses might explain the abundance of certain serotypes in retail foods while others are common in clinical cases. [source] Isolation and partial characterization of a bacteriocin produced by Pediococcus pentosaceus K23-2 isolated from KimchiJOURNAL OF APPLIED MICROBIOLOGY, Issue 2 2008M.S. Shin Abstract Aims:, Screening and partial characterization of a bacteriocin produced by Pediococcus pentosaceus K23-2 isolated from Kimchi, a traditional Korean fermented vegetable. Methods and Results:, A total of 1000 lactic acid bacteria were isolated from various Kimchi samples and screened for the production of bacteriocin. Pediocin K23-2, a bacteriocin produced by the Pediococcus pentosaceus K23-2 strain, showed strong inhibitory activity against Listeria monocytogenes. The bacteriocin activity remained unchanged after 15 min of heat treatment at 121°C or exposure to organic solvents; however, it diminished after treatment with proteolytic enzymes. The bacteriocin was maximally produced at 37°C, when the pH of the culture broth was maintained at 5ˇ0 during the fermentation, although the optimum pH for growth was 7ˇ0. The molecular weight of the bacteriocin was about 5 kDa according to a tricine SDS-PAGE analysis. Conclusions:,Pediococcus pentosaceus K23-2 isolated from Kimchi produces a bacteriocin, which shares similar characteristics to the Class IIa bacteriocins. The bacteriocin is heat stable and shows wide antimicrobial activity against Gram-positive bacteria, especially L. monocytogenes. Significance and Impact of the Study:, Pediocin K23-2 and pediocin K23-2-producing P. pentosaceus K23-2 could potentially be used in the food and feed industries as natural biopreservatives, and for probiotic application to humans or livestock. [source] Investigation into the effect of detergents on disinfectant susceptibility of attached Escherichia coli and Listeria monocytogenesJOURNAL OF APPLIED MICROBIOLOGY, Issue 1 2008J.T. Walton Abstract Aims:, Investigate the effect of detergent treatment on susceptibility of attached Escherichia coli and Listeria monocytogenes to subsequent disinfectant treatment. Methods and Results:, Plate counts show that E. coli attached to stainless steel surfaces became significantly more susceptible to benzalkonium chloride (BAC) after treatment with sodium alkyl sulfate (SAS) and fatty alcohol ethoxylate (FAE). No change in susceptibility was observed with Sodium dodecyl sulfate (SDS). L. monocytogenes became significantly less susceptible to BAC after treatment with SAS and SDS yet no change in susceptibility was observed with FAE. Flow cytometry using the fluoresceine propidium iodide revealed significant increases in cell membrane permeability of both organisms by SAS and FAE, although the effect was much greater in E. coli. No change was observed with SDS. Hydrophobic interaction chromatography showed that both organisms became less hydrophobic following treatment with SAS and SDS but FAE had no effect. Conclusions:, In E. coli, detergents that increase susceptibility to BAC increase membrane permeability. In L. monocytogenes, detergents that reduce susceptibility to BAC lower cell surface hydrophobicity. Significance and Impact of the Study:, Detergents can influence the sensitivity of pathogenic food borne micro-organisms to BAC. [source] Influence of temperature on biofilm formation by Listeria monocytogenes on various food-contact surfaces: relationship with motility and cell surface hydrophobicityJOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2008G. Di Bonaventura Abstract Aims:, To assess the ability of Listeria monocytogenes to form biofilm on different food-contact surfaces with regard to different temperatures, cellular hydrophobicity and motility. Methods and Results:, Forty-four L. monocytogenes strains from food and food environment were tested for biofilm formation by crystal violet staining. Biofilm levels were significantly higher on glass at 4, 12 and 22°C, as compared with polystyrene and stainless steel. At 37°C, L. monocytogenes produced biofilm at significantly higher levels on glass and stainless steel, as compared with polystyrene. Hydrophobicity was significantly (P < 0ˇ05) higher at 37°C than at 4, 12 and 22°C. Thirty (68ˇ2%) of 44 strains tested showed swimming at 22°C and 4 (9ˇ1%) of those were also motile at 12°C. No correlation was observed between swimming and biofilm production. Conclusions:,L. monocytogenes can adhere to and form biofilms on food-processing surfaces. Biofilm formation is significantly influenced by temperature, probably modifying cell surface hydrophobicity. Significance and Impacts of the Study:, Biofilm formation creates major problems in the food industry because it may represent an important source of food contamination. Our results are therefore important in finding ways to prevent contamination because they contribute to a better understanding on how L. monocytogenes can establish biofilms in food industry and therefore survive in the processing environment. [source] Response of Listeria monocytogenes to liquid smokeJOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2008M. Guilbaud Abstract Aims:, To investigate the effect of liquid smoke on growth, survival, proteomic pattern and haemolytic potential of Listeria monocytogenes. Methods and Results:, Growth and survival curves were recorded in brain,heart infusion broth supplemented with three concentrations of liquid smoke. L. monocytogenes growth was inhibited in the presence of 15 ,g ml,1 phenol while a rapid decrease in cell viability occurred in the presence of 30 ,g ml,1 phenol. The proteome of L. monocytogenes cytosoluble proteins was slightly modified after 2-h incubation with 30 ,g ml,1 phenol but no protein already characterized in response to other known stresses was induced, except the protease ClpP. Liquid smoke inhibited the haemolytic potential without affecting hly gene expression, showing a potential inhibition of protein activity or stability. Conclusions:, The presence of liquid smoke in a rich medium strongly affected growth and survival of L. monocytogenes. Brief smoke stress affected the metabolic pathways and inhibited the haemolytic activity of L. monocytogenes. Significance and Impact of Study:, This study is a first step in the investigation of the influence of a smoked product on L. monocytogenes strains. [source] Listeria monocytogenes in spontaneous abortions in humans and its detection by multiplex PCRJOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2007S. Kaur Abstract Aim: To assess the extent of Listeria monocytogenes in causation of human spontaneous abortions by isolation methods and PCR analysis for the presence of virulence-associated genes. Methods and Results: A total of 305 samples comprising blood, urine, placental bits, faecal and vaginal swabs were collected from 61 patients with spontaneous abortions. Listeria spp. were isolated from 10 samples collected from nine (14ˇ8%) patients. Confirmation of these isolates was based on biochemical tests, haemolysis on blood agar, CAMP test, phosphatidylinositol-specific phospholipase C (PI-PLC) assay followed by in vivo pathogenicity tests and multiplex PCR to detect virulence-associated genes (prfA, plcA, hlyA, actA and iap). Three isolates were confirmed as L. monocytogenes. Of these, two isolates turned out to be pathogenic and found to posses all five genes. However, the remaining two haemolytic L. monocytogenes isolates lacking the plcA gene and activity in the PI-PLC assay were found to be nonpathogenic by in vivo tests. Conclusions: The occurrence of pathogenic L. monocytogenes in cases of spontaneous abortions was 3ˇ3%. It seems that the plcA gene and its expression have an important role as essential virulence determinants in pathogenic Listeria spp. Significance and Impact of the Study: The recovery of pathogenic L. monocytogenes isolates from cases of spontaneous abortion indicates the significance of listeric infection in pregnant women. [source] Viable but non-culturable Listeria monocytogenes on parsley leaves and absence of recovery to a culturable stateJOURNAL OF APPLIED MICROBIOLOGY, Issue 4 2007N. Dreux Abstract Aims:, To investigate the presence of viable but non-culturable Listeria monocytogenes during survival on parsley leaves under low relative humidity (RH) and to evaluate the ability of L. monocytogenes to recover from VBNC to culturable state under satured humidity. Methods and Results:, Under low RH (47,69%) on parsley leaves, the initial number of L. monocytogenes populations counted on non selective media (109 L. monocytogenes per leaf on TSA) was reduced by 6 log10 scales in 15 days, whereas number of viable L. monocytogenes counted under the microscope was reduced by 3,4 log10 scales, indicating the presence of VBNC cells. This was demonstrated on three L. monocytogenes strains (EGDe, Bug 1995 and LmP60). Changing from low to 100% RH permitted an increase of the culturable counts of L. monocytogenes and this growth was observed only when residual culturable cells were present. Moreover, VBNC L. monocytogenes inoculated on parsley leaves did not become culturable after incubation under 100% RH. Conclusions:, Dry conditions induced VBNC L. monocytogenes on parsley leaves but these VBNC were likely unable to recover culturability after transfer to satured humidity. Significance and Impact of Study:, Enumeration on culture media presumably under-estimates the number of viable L. monocytogenes on fresh produce after exposure to low RH. [source] | ||||||||||||||||