Biomedical subjects

Martin Wiedmann

Publications and source records attributed to Martin Wiedmann.

At least 19 recordsLinked to original sources

PubMed2026

Proposal to validate Listeria swaminathanii sp. nov. and reassign the type strain to UTK S2-0008.

Listeria swaminathanii UTK S2-0008, isolated from soil collected in the Nantahala National Forest in North Carolina, USA, is the only L. swaminathanii strain eligible to serve as the type strain, which is needed to achieve valid status. The previously effectively published type, L. swaminathanii FSL L7-0020T, and previously described strains UTK C1-0015 and UTK C1-0024 do not conform to the International Code of Nomenclature of Prokaryotes' rules for type strains. Additionally, the currently designated type strain (FSL L7-0020T = ATCC TSD-239T) is an atypical representative of L. swaminathanii as it is the only strain lacking catalase activity. Therefore, it is proposed to reassign the type to L. swaminathanii (UTK S2-0008T = CCUG 77280T = LMG 33255T). Whole-genome sequence-based average nucleotide identity (ANI) showed that this strain clustered with the three previously described L. swaminathanii strains (FSL L7-0020 = ATCC TSD-239, UTK C1-0015, and UTK C1-0024; pairwise ANI ranged from 98.71% to 98.83%). All four strains, including the one described here, could not be classified as any validly published Listeria species and showed the highest similarity to Listeria marthii (maximum ANI of 93.92%, in silico DNA-DNA hybridization of 56.2%). L. swaminathanii exhibits the phenotypic characteristics that are currently expected of the Listeria sensu stricto species. This species lacks phenotypic characteristics associated with Listeria pathogenicity (non-hemolytic and negative for phosphatidylinositol-specific phospholipase C activity); the genomes lack genes associated with virulence (all genes found on the Listeria pathogenicity island 1 [LIPI-1], as well as the internalin genes inlA and inlB), which support L. swaminathanii is nonpathogenic.IMPORTANCEThe genus Listeria includes species of significant relevance to food safety, environmental microbiology, and public health. Accurate species identification is critical because misidentification of nonpathogenic species as pathogenic ones can lead to unnecessary recalls and regulatory complications. The validation of Listeria swaminathanii sp. nov. will ensure that this species is formally recognized and has a type strain (UTK S2-0008T) that is representative of the species. This work strengthens diagnostic accuracy by enabling the inclusion of this species in reference databases and inclusivity studies, reducing the risk of false identification. Furthermore, the identification and characterization of Listeria swaminathanii sp. nov. expands our understanding of the genetic and ecological diversity within the genus Listeria, particularly among soil-dwelling strains.

Listeria↗

PubMed2026

Motile and non-motile Listeria species adopt distinct ecological and evolutionary strategies to achieve broad geographic ranges across soil ecosystems.

Broad geographic ranges often reflect ecological versatility and are associated with lower extinction risk. Motility is a key physiological and ecological trait in bacteria. However, how some motile and non-motile bacteria achieve broad geographic ranges remains poorly understood. Here, we analyzed the genomes of 141 Listeria welshimeri and 90 Listeria booriae isolates systematically obtained from soils, representing widespread motile and non-motile species, respectively. We show that L. welshimeri lacks clear phylogeographic structure, suggesting minimal geographic barriers to dispersal. Its wide distribution is likely associated with enhanced motility and effective host colonization that facilitate wildlife-driven dispersal, particularly by regional-terrestrial birds. This pattern is supported by positive selection on flagellar and chemotaxis genes, strong associations with wildlife movement patterns, and close genomic relatedness between soil and wild bird isolates. In contrast, L. booriae displays clade endemism and a strong distance-decay relationship, suggesting dispersal limitation. Despite lacking a dispersal advantage, L. booriae's wide distribution appears to be linked to genomic flexibility and metabolic versatility that support adaptation to diverse environmental conditions, especially those shaped by iron concentration and precipitation. This is evidenced by its large, open pangenome characterized by abundant and diverse metabolic pathways and broad substrates utilization capacity; pronounced positive selection on genes involved in inorganic ion, amino acid, and coenzyme transport and metabolism; and strong associations between gene richness and abiotic factors as well as bacterial community composition. These findings suggest distinct genomic foundations and ecological and evolutionary mechanisms underlying the success of motile and non-motile cosmopolitan bacteria in soil ecosystems.

Soil Microbiology↗

PubMed2026

Clustering and Source Association of Clinical and Nonclinical Listeria monocytogenes Isolates, New York, USA, 2000-20211.

We analyzed whole-genome sequencing data for 1,046 human clinical and 1,332 nonclinical Listeria monocytogenes isolates collected across New York, USA, during 2000-2021. Several hypervirulent clonal complexes (CCs) were significantly associated with clinical isolates, and several hypovirulent CCs were associated with nonclinical isolates. Specific CCs also showed association with specific food categories (e.g., processed meat); specific genetic markers (e.g., inlA premature stop codons) were also significantly associated with processed meat isolates. Analysis of clusters that contained food isolates, as well as subsequently identified clinical isolates, showed that time of isolation between food isolates and clinical isolates was significantly shorter for produce isolates than for isolates from meat, dairy, or fish. This finding suggests unique transmission pathways for produce, which might reflect short shelf life or limited L. monocytogenes persistence (e.g., in agricultural environments). This study highlights new opportunities for use of whole-genome sequencing to improve outbreak investigations and source attribution.

Listeria monocytogenes↗

PubMed2007

Pulsed-field gel electrophoresis (PFGE) analysis of temporally matched Listeria monocytogenes isolates from human clinical cases, foods, ruminant farms, and urban and natural environments reveals source-associated as well as widely distributed PFGE types.

A total of 495 temporally and geographically matched Listeria monocytogenes isolates from human clinical cases, foods, ruminant farms, and urban and natural environments were used to investigate L. monocytogenes pulsed-field gel electrophoresis (PFGE) type diversity. Two-enzyme (AscI and ApaI) PFGE discriminated 310 PFGE types and exhibited higher overall discriminatory power (Simpson's index of discrimination [D] = 0.995) than either EcoRI ribotyping (D = 0.950) or AscI or ApaI single-enzyme PFGE (D = 0.992 for both). Seven PFGE types showed significant associations with specific sources, including one and four PFGE types, respectively, associated with human clinical cases and foods. Spatial analysis of 13 PFGE types occurring >5 times showed that two PFGE types were specific to a single processing facility each, where they appear to have persisted over time. Nine PFGE types were geographically widespread and occurred among isolates from multiple sources. For example, a PFGE type that matched isolates from listeriosis outbreaks in Los Angeles and Switzerland occurred among isolates from farms (n = 7), human clinical cases (n = 4), environmental sources (n = 3), and foods (n = 1). Our data indicate that (i) PFGE is highly discriminatory for the subtyping of L. monocytogenes, (ii) some L. monocytogenes PFGE types are associated with specific sources, and (iii) some L. monocytogenes PFGE types are widely distributed and appear to be stable and pandemic. Large PFGE type databases representing isolates from different sources are thus needed to appropriately interpret subtype data in epidemiological investigations and to identify common as well as source-specific PFGE types.

Animal Husbandry↗

PubMed2006

Markov chain approach to analyze the dynamics of pathogen fecal shedding--example of Listeria monocytogenes shedding in a herd of dairy cattle.

Fecal shedding is an important mechanism of spreading of a number of human and animal pathogens. Understanding of the dynamics of pathogen fecal shedding is critical to be able to control or prevent the spread of diseases caused by these pathogens. The objective of this study was to develop a model for analysis of the dynamics of pathogen fecal shedding. Fecal shedding of Listeria monocytogenes in dairy cattle was used as a model system. A Markov chain model (MCM) with two states, shedding and non-shedding, has been developed for overall L. monocytogenes fecal shedding (all L. monocytogenes subtypes) and fecal shedding of three L. monocytogenes subtypes (ribotypes 1058A, 1039E and 1042B) using data from one study farm. The matrices of conditional probabilities of transition between shedding and non-shedding states for different sets of covariates have been estimated by application of logistic regression. The covariate-specific matrices of conditional probabilities, describing the presence of different risk factors, were used to estimate (i) the stationary prevalence of dairy cows that shed any L. monocytogenes subtype or ribotypes 1058A, 1039E, and 1042B, (ii) the duration of overall and subtype specific fecal shedding, and (iii) the duration of periods without shedding. A non-homogeneous MCM was constructed to study how the prevalence of fecal shedders changes over time. The model was validated with data from the study farm and published literature. The results of our modeling work indicated that (i) the prevalence of L. monocytogenes fecal shedders varies over time and can be higher than 90%, (ii) L. monocytogenes subtypes exhibit different dynamics of fecal shedding, (iii) the dynamics of L. monocytogenes fecal shedding are highly associated with contamination of silage (fermented feed) and cows' exposure to stress, and (iv) the developed approach can be readily used to study the dynamics of fecal shedding in other pathogen-host-environment systems.

Animals↗

PubMed2006

Listeria monocytogenes subgroups IIIA, IIIB, and IIIC delineate genetically distinct populations with varied pathogenic potential.

Listeria monocytogenes lineage III strains belonging to subgroups IIIA (n = 8), IIIB (n = 5), and IIIC (n = 6) were examined along with other known serotype strains (n = 11) by PCR and Southern hybridization using several recently described species-, virulence-, and serotype-specific primers and probes. The virulence of seven representative lineage III strains was then evaluated in mice via the intraperitoneal route. The results suggest that subgroup IIIA consists of typical rhamnose-positive avirulent serotype 4a and virulent serotype 4c strains, subgroup IIIC consists of atypical rhamnose-negative virulent serotype 4c strains, and subgroup IIIB consists of atypical rhamnose-negative virulent non-serotype 4a and non-serotype 4c strains, some of which may be related to serotype 7. It is possible that subgroup IIIB (including serotype 7) may represent a novel subspecies within L. monocytogenes.

Animals↗

PubMed2006

Combined sigB allelic typing and multiplex PCR provide improved discriminatory power and reliability for Listeria monocytogenes molecular serotyping.

Conventional serotyping has traditionally been used to subtype Listeria monocytogenes, but has several limitations, including low discriminatory power and poor reproducibility. Molecular serotyping methods have been developed for L. monocytogenes, but generally show limited discriminatory power and high misclassification rates. We selected 157 Listeria isolates to evaluate a combination of a previously described multiplex PCR assay and sigB allelic typing as an alternative molecular serotyping and subtyping strategy for L. monocytogenes. While the multiplex PCR assay differentiated five L. monocytogenes subtypes (Simpson's Index of Discrimination [SID]=0.78), including classification of the most common disease-associated serotypes (1/2a, 1/2b, 1/2c, and lineage I 4b) into four distinct groups, it misclassified 3.8% of the isolates studied here. sigB allelic typing differentiated 29 subtypes (SID=0.87) and also allowed identification of lineage III L. monocytogenes, which could not be differentiated from the other Listeria spp. by the multiplex PCR assay. sigB allelic typing failed to differentiate serotype 1/2c and 1/2a isolates and one sigB allelic type included serotype 4b and 1/2b isolates. A molecular serotyping approach that combines multiplex PCR and sigB sequence data showed increased discriminatory power (SID=0.91) over either method alone as well as conventional serotyping (SID=0.87) and classifies the four major serotypes (i.e., 1/2a, 1/2b, 1/2c, and 4b) into unique subgroups with a lower misclassification rate as compared to the multiplex PCR assay. This combined approach also differentiates lineage I serotype 4b isolates from the genetically distinct serotype 4b isolates classified into lineage III.

Alleles↗

PubMed2006

Listeria monocytogenes internalins are highly diverse and evolved by recombination and positive selection.

To probe the evolution of internalins with confirmed or suspected roles in Listeria monocytogenes virulence we sequenced the full inlB, inlC2, inlC, inlD, inlE, inlF, inlG, and inlH ORFs from 40 L. monocytogenes isolated from human (n=10) and animal (n=10) clinical cases, foods (n=10), and the natural environment (n=10). inlB and inlE were present in all isolates, representing 26 and 20 alleles, respectively. inlC was found in all lineage I and II isolates and represented 21 alleles. inlC2 and inlD represented 22 and 24 alleles, respectively, and were found in all L. monocytogenes isolates, with the exception of three lineage II isolates, which carried inlH, an apparent fusion of the 5' end of inlC2 with the 3' end of inlD. inlF and inlG were absent from lineage I isolates and represented 16 and 11 alleles, respectively. Average pairwise nucleotide differences per site (pi) ranged from 0.00849 (inlF) to 0.07020 (inlE). Phylogenetic trees generally showed clustering of internalin genes into two major evolutionary lineages consistent with lineages I and II previously assigned by ribotyping. In addition to detection of recombination events within each internalin gene, inlB, inlC, inlC2, and inlF showed significant evidence for positive selection (i.e., selection for an advantageous mutant allele). Overall, our data indicated that (i) internalin genes are highly diverse, (ii) internalin gene sequences cluster consistent with the phylogenetic lineages of L. monocytogenes, (iii) both intragenic recombination and positive selection have contributed to the evolution of L. monocytogenes internalins, and (iv) L. monocytogenes internalins show distinct evolutionary histories.

Alleles↗

PubMed2006

Ribotype diversity of Listeria monocytogenes isolates from two salmon processing plants in Norway.

The purpose of this study was to use automated ribotyping procedure to track Listeria monocytogenes transmission in the cold smoked fish production chain and to characterize L. monocytogenes subtypes associated with the salmon processing industry. A total of 104 isolates, which had previously been obtained from a raw fish slaughter and processing plant (plant B) and an adjacent, downstream, salmon smoking operation (plant A), were characterized. These isolates had been obtained through a longitudinal study on Listeria presence, which covered a 31-week period, in both plants. Isolates had been obtained from samples taken from different machinery used throughout the production process. In addition, six isolates obtained from products produced in plant A two years after the initial study were included, so that a total of 110 isolates were characterized. Automated ribotyping was performed using both the restriction enzymes EcoRI and PvuII to increase the discriminatory power. The 110 L. monocytogenes isolates could be divided into 11 EcoRI ribotypes; PvuII ribotype data yielded multiple subtypes within 7 EcoRI ribotypes for a total of 21 subtypes based on both EcoRI and PvuII ribotyping. A total of three EcoRI ribotypes (DUP-1023C, DUP-1045B, and DUP-1053E) were isolated at multiple sampling times from both plants. In addition, one subtype (DUP-1053B) was isolated at multiple sampling times in only plant A, the salmon smoking operation. These data not only support that L. monocytogenes can persist throughout the salmon production system, but also showed that L. monocytogenes may be transmitted between slaughter and smoking operations or may be unique to smoking operations. While the majority of subtypes isolated have been rarely or never linked to human listeriosis cases, some subtypes have previously caused human listeriosis outbreaks and cases. Molecular subtyping thus is critical to identify L. monocytogenes transmission and niches in order to allow design and implementation of control strategies at the appropriate stage of production and in order to reduce the prevalence of L. monocytogenes linked to human disease.

Animals↗

PubMed2006

Listeria monocytogenes in multiple habitats and host populations: review of available data for mathematical modeling.

Listeria monocytogenes has the ability to survive and multiply in diverse habitats and to cause infection in a variety of animal species and humans. We evaluated the literature on survival and multiplication within and transmission among multiple host populations and habitats, including man, sewage, general environment (soil, water, and vegetation), silage (fermented plant material), animals (including wild and domestic animals), and food processing plants. The available knowledge on L. monocytogenes transmission dynamics was translated into the key process nodes of interrelated host- and habitat-specific mathematical models, providing a starting framework for future modeling work and the ultimate development of a system-wide model for evaluation of its transmission, and strategies to reduce human exposure. Because of the ability of L. monocytogenes to survive and multiply in many habitats and hosts, and the number of possible transmission routes, it is highly unlikely that it could be eradicated from any habitat or host, including man. However, L. monocytogenes load within and transmission among habitats and host populations could probably be reduced. Based on the published information, we hypothesize that three recent anthropogenic practices increase the load within and transmission among reviewed habitats and host populations: extended refrigerated storage of ready-to-eat foods allowing L. monocytogenes growth in foods that are contaminated during production or subsequent handling; feeding domestic ruminants with silage often contaminated with L. monocytogenes; and dispersal of contaminated products of sewage treatment to agricultural fields and waters. Future mathematical modeling work could test how much the reduction of L. monocytogenes load and transmission in hosts and habitats associated with these anthropogenic practices would reduce human exposure and consequently human listeriosis.

Animal Feed↗

PubMed2006

Genetic and phenotypic characterization of Listeria monocytogenes lineage III.

Listeria monocytogenes has been previously grouped into three evolutionary groups, termed lineages I, II and III. While lineages I and II are commonly isolated from various sources, lineage III isolates are rare and have several atypical and unique phenotypic characteristics. Relative to their prevalence in other sources, lineage III strains are overrepresented among isolates from food-production animals, and underrepresented among isolates from human clinical cases and foods. This work describes an extensive genotypic and phenotypic characterization of 46 lineage III isolates. Phylogenetic analyses of partial sigB and actA sequences showed that lineage III represents three distinct subgroups, which were termed IIIA, IIIB and IIIC. Each of these lineage III subgroups is characterized by differentiating genotypic and phenotypic characteristics. Unlike typical L. monocytogenes, all subgroup IIIB and IIIC isolates lack the ability to ferment rhamnose. While all IIIC and most IIIB isolates carry the putative virulence gene lmaA, the majority of subgroup IIIA isolates lack this gene. All three lineage III subgroups contain isolates from human clinical cases as well as isolates that are cytopathogenic in a cell culture plaque assay, indicating that lineage III isolates have the potential to cause human disease. The identification of specific genotypic and phenotypic characteristics among the three lineage III subgroups suggests that these subgroups may occupy different ecological niches and, therefore, may be transmitted by different pathways.

Animals↗

PubMed2006

Contributions of Listeria monocytogenes sigmaB and PrfA to expression of virulence and stress response genes during extra- and intracellular growth.

Listeria monocytogenes sigmaB and PrfA are pleiotropic regulators of stress response and virulence gene expression. Quantitative RT-PCR (qRT-PCR) was used to measure transcript levels of sigmaB- and PrfA-dependent genes in exponential-phase L. monocytogenes wild-type and DeltasigB strains as well as in bacteria exposed to environmental stresses (0.3 M NaCl or growth to stationary phase) or present in the vacuole or cytosol of human intestinal epithelial cells. Stationary-phase or NaCl-exposed L. monocytogenes showed sigmaB-dependent increases in opuCA (10- and 17-fold higher, respectively) and gadA transcript levels (77- and 14-fold higher, respectively) as compared to non-stressed, exponential-phase bacteria. While PrfA activity, as reflected by plcA transcript levels, was up to 95-fold higher in intracellular L. monocytogenes as compared to non-stressed bacteria, sigmaB activity was only slightly higher in intracellular than in non-stressed bacteria. Increased plcA transcript levels, which were similar in both host cell vacuole and cytosol, were associated with increases in both prfA expression and PrfA activity. qRT-PCR assays were designed to measure expression of prfA from each of its three promoter regions. Under all conditions, readthrough transcription from the upstream plcA promoter was very low. The relative contribution to total prfA transcription from the sigmaA-dependent P1prfA promoter ranged from approximately 17 % to 30 %, while the contribution of the P2prfA region, which appears to be transcribed by both sigmaA and sigmaB, ranged from approximately 70 % to 82 % of total prfA transcript levels. In summary (i) sigmaB is primarily activated during environmental stress and does not contribute to PrfA activation in intracellular L. monocytogenes and (ii) the partially sigmaB-dependent P2prfA promoter region contributes the majority of prfA transcripts in both intra- and extracellular bacteria.

Bacterial Proteins↗

PubMed2006

Allelic exchange and site-directed mutagenesis probe the contribution of ActA amino-acid variability to phosphorylation and virulence-associated phenotypes among Listeria monocytogenes strains.

To test the hypothesis that actA allelic variation contributes to virulence differences among Listeria monocytogenes strains, cell-to-cell spread and intracellular ActA phosphorylation patterns were characterized for 14 wild-type isolates and selected isogenic mutants. Our data show that (i) while actA allelic variation is not responsible for enhanced cell-to-cell spread observed in epidemic clone I strains, actA allelic variation may contribute to reduced plaque size observed in some isolates, (ii) actA sequence alone determines phosphorylation-dependent ActA banding patterns, and (iii) sequence variation at the positively selected ActA residue 498 does not contribute to ActA phosphorylation patterns or to differences in cell-to-cell spread.

Alleles↗

PubMed2006

Exposure to salt and organic acids increases the ability of Listeria monocytogenes to invade Caco-2 cells but decreases its ability to survive gastric stress.

The effects of environmental stress exposure on Listeria monocytogenes growth and virulence-associated characteristics were investigated. Specifically, we measured the effects of temperature (7 or 37 degrees C), pH (5.5 or 7.4), the presence of salt and organic acids (375 mM NaCl, 8.45 mM sodium diacetate [SD], 275 mM sodium lactate [SL], or a combination of NaCl, SD, and SL), and deletion of sigB, which encodes a key stress response regulator, on the ability of L. monocytogenes to grow, invade Caco-2 cells, and survive exposure to synthetic gastric fluid (pH 2.5 or 4.5). Our results indicate that (i) L. monocytogenes log-phase generation times and maximum cell numbers are not dependent on the alternative sigma factor sigmaB in the presence of NaCl and organic acids at concentrations typically found in foods; (ii) growth inhibition of L. monocytogenes through the addition of organic acids is pH dependent; (iii) the ability of L. monocytogenes to invade Caco-2 cells is affected by growth phase, temperature, and the presence of salt and organic acids, with the highest relative invasion capabilities observed for cells grown with SL or NaCl at 37 degrees C and pH 7.4; (iv) growth of L. monocytogenes in the presence of NaCl, SD, or SL reduces its ability to survive exposure to gastric fluid; and (v) exposure of L. monocytogenes to gastric fluid reduces the enhanced invasiveness caused by growth in the presence of NaCl or SL. These findings suggest that virulence-associated characteristics that determine the L. monocytogenes infectious dose are likely to be affected by food-specific properties (e.g., pH or the presence of salt or organic acid).

Bacterial Proteins↗

PubMed2006

Associations among Listeria monocytogenes genotypes and distinct clinical manifestations of listeriosis in cattle.

OBJECTIVE: To determine whether specific strains of Listeria monocytogenes, as determined by genetic characteristics and virulence phenotypes, were associated with distinct clinical manifestations of listeriosis in cattle and thus may potentially have tissue specificity. ANIMALS: 32 cattle. PROCEDURE: DNA sequence data for the virulence genes actAand inlAwere used to infer the phylogeny of L. monocytogenes and to test for positive selection. Isolates were screened for the presence or absence of internalin genes and assigned an internalin profile. Plaquing assays were performed to determine the relative cytopathogenicity of each isolate. Categorical data analyses were performed to describe associations among L. monocytogenes genotypes, virulence phenotypes, and clinical manifestations of listeriosis. RESULTS: Results confirmed that L. monocytogenes represents 2 deeply separated evolutionary lineages. Genes actA and inlA contained amino acid sites under positive selection, and specific residues at some sites were associated with lineage and manifestation of listeriosis. Whereas lineage I was clonal and predominantly composed of isolates from cases of encephalitis, lineage II was more genetically diverse and equally represented by isolates from cases of encephalitis versus septicemia and fetal infection. Lineage I isolates also had greater cytopathogenicity in vitro, compared with lineage II isolates. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that L. monocytogenes virulence genes underwent positive selection that is consistent with the diversification of 2 evolutionary lineages: lineage I is clonal and associated with encephalitis, and lineage II is more genetically diverse and equally likely to cause both major forms of listeriosis in cattle.

Animals↗

PubMed2006

Molecular characterization of Listeria monocytogenes from natural and urban environments.

Characterization of 80 Listeria monocytogenes isolates from urban and natural environments differentiated 7 and 26 EcoRI ribotypes, respectively. Whereas the majority of isolates from the natural environment represented L. monocytogenes lineage II (12 of 13 isolates), urban isolates grouped evenly into lineages I and II (32 and 33 isolates, respectively) and included two lineage III isolates. Multilocus sequence typing of all natural isolates and a randomly selected subset of 30 urban isolates showed a higher overall diversity (Simpson index of discrimination [D] of 0.987 and 0.920, respectively) than did EcoRI ribotyping (D = 0.872 and 0.911, respectively). Combined analysis with ribotype and lineage data for 414 isolates from farm sources, 165 isolates from foods and food-processing environments, and 342 human clinical isolates revealed that lineage I was significantly more common among human (P < 0.0001) isolates, whereas lineage II was more common among isolates from the natural environment, farms, and foods (P < or = 0.05). Among a total of 92 ribotypes, 31 showed significant associations with specific isolate sources. One ribotype (DUP-1039C) was significantly associated with both natural environments and farms. A spatial analysis showed a marginal association between locations in the natural environment positive for L. monocytogenes and a proximity to farms. Our data indicate that (i) L. monocytogenes strains from different sources show a high level of diversity; (ii) L. monocytogenes subtypes differ significantly in their associations with different environments, even though populations overlap; and (iii) a higher proportion of isolates from environmental sources than from human clinical cases can be classified into L. monocytogenes lineage II, which supports the classification of this lineage as an environmentally adapted subgroup.

Colony Count, Microbial↗

PubMed2006

Growth and stress resistance variation in culture broth among Listeria monocytogenes strains of various serotypes and origins.

Twenty-five Listeria monocytogenes strains of various serotypes and sources, including clinical and food isolates associated with the same outbreaks, were characterized and compared based on growth rates and heat and acid death rates. Growth was monitored in tryptic soy broth supplemented with 0.6% yeast extract (TSBYE) at 4 and 30 degrees C for 32 days and 20 h, respectively. Heat and acid stress responses in TSBYE heated to 55 degrees C or acidified to pH 3.0 with lactic acid were evaluated for 240 or 120 min, respectively. Extensive variation in growth and stress resistance was observed among the tested strains. Growth rate differences were less evident at 30 than at 4 degrees C, where growth rates (log CFU per milliliter per day) ranged from 0.28 to 0.43. Thermal and acid death rates (log CFU per milliliter per minute) ranged from -0.023 to -0.052 and from -0.012 to -0.134, respectively. Serotype appeared to play a significant role (P < 0.05) only with respect to the heat resistance of the organism. Serotype 4b isolates as a group had lower heat resistance than did isolates representing all other serotypes combined. Although no clear origin-related (food versus clinical) trends were observed under the tested conditions, outbreak-related isolates of serotype 4b had lower acid death rates (higher acid resistance) (P < 0.05) than did the rest of the strains belonging to this serotype. Strain Scott A exhibited slow growth at 4 degrees C and low acid resistance, behavior that was distinct among both clinical and serotype 4b isolates. The results of this study highlight the risks associated with extrapolation to other strains of findings obtained with only one strain of L. monocytogenes. This information should be useful when test strains are to be selected for the evaluation of antimicrobial alternatives in ready-to-eat meat and other food products and when risk assessments are to be conducted.

Adaptation, Physiological↗

PubMed2006

International Life Sciences Institute North America Listeria monocytogenes strain collection: development of standard Listeria monocytogenes strain sets for research and validation studies.

Research and development efforts on bacterial foodborne pathogens, including the development of novel detection and subtyping methods, as well as validation studies for intervention strategies can greatly be enhanced through the availability and use of standardized strain collections. These types of strain collections are available for some foodborne pathogens, such as Salmonella and Escherichia coli. We have developed a standard Listeria monocytogenes strain collection that has not been previously available. The strain collection includes (i) a diversity set of 25 isolates chosen to represent a genetically diverse set of L. monocytogenes isolates as well as a single hemolytic Listeria innocua strain and (ii) an outbreak set, which includes 21 human and food isolates from nine major human listeriosis outbreaks that occurred between 1981 and 2002. The diversity set represents all three genetic L. monocytogenes lineages (I, n = 9; II, n = 9; and III, n = 6) as well as nine different serotypes. Molecular subtyping by EcoRI automated ribotyping and pulsed-field gel electrophoresis (PFGE) with AscI and ApaI separated the 25 isolates in the diversity set into 23 ribotypes and 25 PFGE types, confirming that this isolate set represents considerable genetic diversity. Molecular subtyping of isolates in the outbreak set confirmed that human and food isolates were identical by ribotype and PFGE, except for human and food isolates for two outbreaks, which displayed related but distinct PFGE patterns. Subtype and source data for all isolates in this strain collection are available on the Internet and are linked to the PathogenTracker database (www.pathogentracker.com), which allows the addition of new, relevant information on these isolates, including links to publications that have used isolates from this collection. We have thus developed a core L. monocytogenes strain collection, which will provide a resource for L. monocytogenes research and development efforts with centralized Internet-based data curation and integration.

Animals↗