Listeria monocytogenes septic arthritis.
Listeria monocytogenes was isolated from both the blood and the synovial fluid of a diabetic patient with a septic arthritis involving the ankle.
Aged↗
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Comparison of cold enrichment and U.S. Department of Agriculture methods for isolating Listeria monocytogenes from naturally contaminated foods. The Listeria Study Group.
We compared the cold enrichment (CE) and U.S. Department of Agriculture (USDA) methods for isolating Listeria monocytogenes by examining 402 food samples. The food samples were collected from refrigerators of listeriosis patients as part of a multistate active surveillance project to determine the role of foods in sporadic listeriosis in the United States. L. monocytogenes was isolated from 51 food samples (13%). The USDA method was significantly better (P less than 0.001) than the CE method. The isolation efficiencies of the USDA and CE methods were 96 and 59%, respectively. Quantitation of L. monocytogenes in the food samples revealed that many food samples containing less than 0.3 CFU/g were negative as determined by the CE method but positive as determined by the USDA method.
Cold Temperature↗
[Prokaryotic expression and bioactivity of the Listeria monocytogenes listeriolysin O].
To express the Listeria monocytogenes hly gene in Escherichia coli and study its primary biological characteristics, hly gene without signal peptide sequence was amplified from Listeria monocytogenes serotype 4b by PCR and inserted into T-easy vector. Sequencing showed this hly gene was 1518 bp and nucleotide homology was more than 99.8% compared with three Listeria monocytogenes hly genes in GenBank. The cloned hly gene was then inserted into prokaryotic expression vector pGEX-6P-1 and transformed into E. coli BL21. The predicted fusion protein was detected by SDS-PAGE after IPTG inducion, which had molecular weight approximately 82 kD. Hemolytic experiment demonstrated the expressed fusion protein can lyse human red cell and its hemolytic titer attained 2.26 x 10(4) HU/mg. Conclusively, the Listeria monocytogenes hly gene was successfully cloned and expressed in E. coli. The successful expression of LLO in E. coli BL21 constituted a solid foundation for further researches such as pathogenesis and immune mechanism, MAb and vaccine development.
Animals↗
Comparison of selective and non-selective enrichment media in the detection of Listeria monocytogenes from meat containing Listeria innocua.
AIMS: This study investigated whether the higher incidence of recovery from meat of Listeria innocua compared with L. monocytogenes could be due to the laboratory media used, leading to an artificially lower detection of the pathogenic species, L. monocytogenes. METHODS AND RESULTS: Minced beef was inoculated with L. monocytogenes, L. innocua, or a mixture of these species, and stored at 0 or 10 degrees C under vacuum or aerobic conditions for up to 28 days. Listeria were recovered from the minced beef using selective (University of Vermont Medium, UVM) and non-selective (Buffered Peptone Water, BPW) enrichment broths after 0, 14, and 28 days of storage. In general, there were no significant differences (P < 0.05) between the numbers of L. monocytogenes recovered from minced beef samples after 24 h enrichment in BPW and the numbers recovered using UVM. In addition, the presence of L. innocua in meat samples containing L. monocytogenes did not significantly (P < 0.05) affect the numbers of L. monocytogenes recovered using either enrichment broth. In most cases there were no significant differences (P < 0.05) between the numbers of L. innocua recovered from minced beef samples after 24 h enrichment in BPW compared with numbers recovered using UVM. CONCLUSION: Listeria innocua was found to have no significant competitive advantage over L. monocytogenes in selective or non-selective enrichment media. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggest that, in some instances, the use of a selective enrichment broth offers no advantage over a non-selective enrichment broth for the recovery of Listeria species from minced beef.
Bacterial Typing Techniques↗
Listeria monocytogenes endophthalmitis.
Listeria endophthalmitis developed in an otherwise healthy woman. Early identification of the microorganism after anterior chamber paracentesis and aggressive treatment with specific antibiotics through the intravitreal route resulted in complete recovery of visual acuity.
Anti-Bacterial Agents↗
Proteomic and microscopic analysis of biofilms formed by Listeria monocytogenes 568.
Biofilm formation may be important in the colonization of the food-processing environment by the food-borne pathogen Listeria monocytogenes. Listeria monocytogenes 568 formed adherent multicellular layers on a variety of test surfaces following growth at 37 degrees C with multiple transfers of the test surface into fresh medium. Microscopic examination of these adherent layers suggest that the cells were surrounded by extracellular material. The presence of a carbohydrate containing extracellular polymeric matrix was confirmed by labelling hydrated adherent layers with fluorescein-conjugated concanavalin A, indicating that these adherent layers are biofilms. To gain insight into the physiological state of cells in these biofilms, the proteomes from biofilm- and planktonic-grown cells from the same cultures were compared using 2-dimensional polyacrylamide gel electrophoresis. Nineteen proteins, which exhibited higher levels of expression in biofilm-grown cells, were successfully identified from the 2-D gels using a combination of MALDI-TOF and MS/MS. Proteins that were found to be more highly expressed in biofilm-grown cells were involved in stress response, envelope and protein synthesis, biosynthesis, energy generation, and regulatory functions. In biofilm-grown cells, many proteins in the pH range 4-6 ran as multiple spots arranged horizontally across the 2-D gels.
Bacterial Adhesion↗
Cholesterol oxidase from Rhodococcus equi is likely the major factor involved in the cooperative lytic process (CAMP reaction) with Listeria monocytogenes.
The CAMP reaction between Listeria monocytogenes and Rhodococcus equi was studied by a diffusion assay. Listeria monocytogenes displayed identical cooperative haemolytic effect with supernatant cultures of R. equi or with commercial cholesterol oxidase (COX). This result, even with enzymes of different sources (commercial COX is obtained from Pseudomonas spp.) suggests that this enzyme secreted by R. equi has a crucial role in the synergistic haemolytic (CAMP) reaction with L. monocytogenes. The mechanism of the cooperative lytic process between L. monocytogenes and R. equi may represent a different and novel mechanism reaction, in which the COX may not act as a conventional second-step factor, and a reaction different to the direct interaction with the cholesterol of the erythrocyte membrane may be involved.
Cholesterol Oxidase↗
Genus- and species-specific detection of Listeria monocytogenes using polymerase chain reaction assays targeting the 16S/23S intergenic spacer region of the rRNA operon.
In this study, the 16S/23S rRNA intergenic spacer (IGS) regions of six Listeria species were examined. DNA bands of 590 and 340 bp were observed following polymerase chain reaction (PCR) amplification of DNA from Listeria monocytogenes, Listeria innocua, Listeria seeligeri, Listeria welshimeri, and Listeria ivanovii strains with generic rRNA IGS oligonucleotide primers. For strains of Listeria grayi subspp. grayi and murrayi, DNA band sizes of 550 and 340 bp were observed with this primer pair. DNA bands of these sizes were not observed for other Gram-negative or- positive bacteria in this PCR assay. Four RsaI digestion profiles were noted for the Listeria PCR products. Listeria monocytogenes strains had one profile; L. innocua strains had a second; L seeligeri, L. welshimeri, and L ivanovii strains had a third; and L. grayi strains had a fourth. The small and large 16S/23S rRNA IGSs of L. monocytogenes ATCC 15313 were identical in the first 58 5' and the last 169 3' nucleotides. However, the large rRNA IGS contained a central 267-bp region with tRNA(Ile) and tRNA(Ala) genes. Large rRNA 16S/23S IGS nucleotide sequence data has not been previously reported. This data was used to develop novel Listeria genus-specific and L.monocytogenes species-specific PCR assays.
Base Sequence↗
Occurrence of Listeria monocytogenes in soft and semi-soft cheeses in retail outlets in Sweden.
Samples of 333 retail cheeses produced in or imported into Sweden were examined for the presence of Listeria monocytogenes. Listeria monocytogenes was isolated from 6% of the cheese samples. Cheeses made from raw milk were more frequently contaminated with L. monocytogenes (42%) than cheeses made from heat-treated milk (2%). The incidence of the organism in whole cheeses and pre-cut wedges was similar (6%). L. monocytogenes was only found in imported cheeses (18 from France, and one from Germany and Italy, respectively). The numbers of L. monocytogenes varied from < 1 x 10(2) to 1 x 10(5) cfu/g. All L. monocytogenes strains belonged to serogroup 1/2, except isolates from two samples that belonged to serogroup 4.
Cheese↗
Alteration of pulmonary immunity to Listeria monocytogenes by diesel exhaust particles (DEPs). II. Effects of DEPs on T-cell-mediated immune responses in rats.
Previously, we showed that diesel exhaust particles (DEPs) suppressed pulmonary clearance of Listeria monocytogenes (Listeria) and inhibited the phagocytosis of alveolar macrophages and their response to Listeria in the secretion of interleukin (IL)-1 beta, tumor necrosis factor alpha, and IL-12. In this report we examined the effects of DEPs and/or Listeria on T-cell development and secretion of IL-2, IL-6, and interferon (IFN)-gamma. We exposed Brown Norway rats to clean air or DEPs at 50 or 100 mg/m3 for 4 hr by nose-only inhalation and inoculated with 100,000 Listeria. Lymphocytes in the lung-draining lymph nodes were isolated at 3 and 7 days postexposure, analyzed for CD4+ and CD8+ cells, and measured for cytokine production in response to concanavalin A or heat-killed L. monocytogenes. Listeria infection induced lymphocyte production of IL-6. At 7 days postinfection, lymphocytes from Listeria-infected rats showed significant increases in CD4+ and CD8+ cell counts and the CD8+/CD4+ ratio and exhibited increased production of IFN-gamma and IL-2 receptor expression compared with the noninfected control. These results suggest an immune response that involves the action of IL-6 on T-cell activation, yielding Listeria-specific CD8+ cells. DEP exposure alone enhanced lymphocyte production of both IL-2 and IL-6 but inhibited lymphocyte secretion of IFN-gamma. In rats exposed to 100 mg/m3 DEPs and Listeria, a 10-fold increase occurred in pulmonary bacterial count at 3 days postinfection when compared with the Listeria-only exposure group. The isolated lymphocytes showed a significant increase in the CD4+ and CD8+ cell counts and the CD8+/CD4+ ratio and exhibited increased IL-2 responsiveness and increased capacity in the secretion of IL-2, IL-6, and IFN-gamma. This T-cell immune response was sufficient to allow the Brown Norway rats to clear the bacteria at 7 days postinfection and overcome the down-regulation of the innate immunity by the acute DEP exposure.
Air Pollutants↗
In vitro antimicrobial susceptibility of Listeria monocytogenes isolated in the UK and other Listeria species.
The MICs and MBCs of 21 antimicrobial agents were determined for 103 strains of Listeria monocytogenes isolated in the UK and 27 strains of other Listeria species. Ampicillin, penicillin, azlocillin, imipenem, gentamicin, netilmicin, amikacin, erythromycin, rifampicin, trimethoprim, clindamycin and vancomycin had good activity, while cephalothin, chloramphenicol, ciprofloxacin and ofloxacin were less active, and cefuroxime, enoxacin, norfloxacin and fosfomycin were the least active. Tetracycline had good activity against many strains, but the MIC was high for some. Unlike the other Listeria species tested, Listeria ivanovii was susceptible to fosfomycin. Inoculum size and media employed were shown to affect the MBC, tryptose phosphate broth yielding higher MBCs than Mueller-Hinton or Isosensitest broths.
4-Quinolones↗
Physical and genetic map of the Listeria monocytogenes EGD serotype 1/2a chromosome.
Listeria monocytogenes is a facultative intracellular pathogen responsible for both invasive and non-invasive food-borne illness in animals and humans. In this study, macrorestriction analysis following pulsed-field gel electrophoresis was used to show that Listeria monocytogenes serovar 1/2a strain EGD has a single chromosome containing eight NotI fragments of 1100, 850, 365, 320, 275, 40, 30 and 20 kb in size and 11 AscI fragments of 860, 470, 410, 360, 320, 250, 110, 80, 50, 30 and 20 kb. The total genome therefore comprises 3000 +/- 50 kb. The creation of a physical and genetic map of the Listeria genome was achieved by generating NotI linking clones and their use in subsequent hybridisation analysis. Using isogenic mutants harbouring additional artificial NotI restriction sites, we were able to precisely map the positions of all currently known virulence genes on the chromosome.
Chromosome Mapping↗
Listeria monocytogenes septicemia in a foal.
Listeria monocytogenes is rarely reported as a cause of septicemia in foals. In this case, the foal had diarrhea 2 weeks prior to the onset of signs of lethargy, high rectal temperature, and leukopenia with a left shift. Listeria monocytogenes was isolated from the blood culture. The most commonly isolated organism causing septicemia in foals is Escherichia coli. Without the blood cultures, a definitive diagnosis would not have been possible.
Animals↗
Involvement of Listeria monocytogenes in the abortive disease.
Listeria monocytogenes, an intracellular facultative germ that causes the invasion, sometimes fatal, in susceptible hosts is a food borne pathogen with ubiquitary spread that has generated a public health problem for such risk groups as: pregnant women, foetuses, new borns. 504 women with abortive disease were serologically investigated in 1999 for serotype 1a circulating in Romania. The most affected age group proved to be that in the range of 20-30 yrs: 378 (75%) cases. 107 (21.23%) female patients had the diagnostic titer (> or = 1/320): among these, 38 (7.53%) had miscarriages in the IVth-VIIIth month and 18 (3.57%) gave birth to dead foetuses; during pregnancy, 10 (1.98%) female patients received treatment with Ampicillin and 2 (0.39%) treatment with Erythromycin. In the age group > 31 yrs, the 1/320 titer was noticed in 21 (4.16%) female patients but among these only 4 (0.79%) had a history of miscarriage in the final pregnancy months; they were administered Ampicillin during pregnancy. Although there is no clear-cut evidence, our results point to the conclusion that these female patients were contaminated with Listeria monocytogenes.
Abortion, Spontaneous↗
[A case of Listeria monocytogenes meningitis in an immunocompetent infant].
Listeria Monocytogenes meningitis is a rare affection after the neonatal period, but in immunocompromised patients. Listeria Monocytogenes is a Gram-positive, facultative intracellular bacterium frequently causing infection in pregnant women, in patients with cell-mediated immunity deficit and in the early and late stages of life. We present a case of Listeria Monocytogenes meningitis in an immunocompetent nomad 8-month-child, preceded by gastroenteritis. Although gastrointestinal symptoms may be due to intestinal infection by Listeria, the concomitant presence of other bacteric or viral enteric pathogens may have promoted bacterium intestinal translocation and generated disseminated disease. The main transmission route of infection after the neonatal period is ingestion of contaminated food. A diet history was taken after isolation of the bacterium in liquor and showed that the child was an eater of undercooked hot-dogs. Despite the frequency of clinical complication in such affection, the outcome in this patient was a complete recovery. Although the infection is extremely infrequent in healthy children, physicians should always consider Listeria as a possible etiologic agent of meningitis in pediatric patients, regardless of their age or immunological status, especially in patients living in precarious sanitary conditions, where weaning times and conditions are not respected and a suitable food cooking is not assured.
Humans↗
Otitis media/interna and suppurative meningoencephalomyelitis associated with Listeria monocytogenes infection in a llama.
Listeria monocytogenes was found to be the cause of fatal suppurative meningoencephalomyelitis in a 3.5-month-old cria. The cria initially had clinical signs of unilateral peripheral vestibular disease, but on the following day, the cria developed progressive signs of encephalitis. Treatment with antibiotics, flunixin meglumine, and anticonvulsant drugs failed to stop progression of the disease, and the cria was euthanatized. Post-mortem examination revealed otitis media-interna and diffuse suppurative meningoencephalomyelitis. Listeria monocytogenes was isolated from CSF and brain tissue.
Animals↗
[Misleading presentation of rhombencephalitis due to Listeria monocytogenes].
INTRODUCTION: Rhombencephalitis due to Listeria monocytogenes may present in a variety of forms. CASE REPORT: We describe the case of a 76 years old male presenting with an acute symmetrical palsy of fifth, seventh, ninth-tenth cranial nerves, ophthalmoparesia and miosis, without evident signs of long tract lesion, mimicking a peripheral nervous system or neuromuscular junction disease. The results of the blood cultures and the magnetic resonance imaging gave the diagnosis of rhombencephalitis due to Listeria. CONCLUSIONS: Rhombencephalitis due to Listeria monocytogenes may present as a symmetric palsy of cranial nerves, therefore it can mimic a cranial nerves multineuritis syndrome.
Aged↗
Redundant roles for Met docking site tyrosines and the Gab1 pleckstrin homology domain in InlB-mediated entry of Listeria monocytogenes.
The bacterial pathogen Listeria monocytogenes causes food-borne illnesses leading to gastroenteritis, meningitis, or abortion. Listeria induces its internalization into some mammalian cells through interaction of the bacterial surface protein InlB with host Met receptor tyrosine kinase. Binding of InlB leads to phosphorylation of Met and the adapter Gab1 and to activation of host phosphoinositide (PI) 3-kinase. The mammalian ligand of Met, hepatocyte growth factor, promotes cell motility and morphogenesis in a manner dependent on phosphorylation of two docking site tyrosines at positions 1349 and 1356 in the receptor's cytoplasmic tail. Here we determined if these tyrosines were essential for Listeria entry. A derivative of the human cell line T47D stably expressing a truncated Met lacking most of its cytoplasmic domain was unable to support InlB-mediated signaling or entry. Surprisingly, cells expressing mutant Met containing phenylalanine substitutions in both tyrosines 1349 and 1356 (MetYF) allowed entry and InlB-induced Gab1 phosphorylation. However, in contrast to the situation in cells expressing wild-type Met, Gab1 phosphorylation in MetYF cells required PI 3-kinase activity. The Gab1 pleckstrin homology (PH) domain was constitutively associated with the plasma membrane of cells in a PI 3-kinase-dependent manner. Overexpression of the PH domain blocked entry of Listeria into cells expressing MetYF but not into cells expressing wild-type Met. Taken together, these results indicate that the docking site tyrosines are dispensable for internalization when membrane localization of Gab1 is constitutive. Distinct pathways of recruitment by phosphorylated tyrosines in Met and PH domain ligands in the membrane are redundant for bacterial entry.
Adaptor Proteins, Signal Transducing↗