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1.
BMC Microbiol ; 21(1): 327, 2021 11 25.
Artigo em Inglês | MEDLINE | ID: mdl-34823476

RESUMO

BACKGROUND: Listeria monocytogenes is one of the deadliest foodborne pathogens. The bacterium can tolerate severe environments through biofilm formation and antimicrobial resistance. This study aimed to investigate the antimicrobial susceptibility, resistance genes, virulence, and molecular epidemiology about Listeria from meat processing environments. METHODS: This study evaluated the antibiotic resistance and virulence of Listeria isolates from slaughtering and processing plants. All isolates were subjected to antimicrobial susceptibility testing using a standard microbroth dilution method. The harboring of resistant genes was identified by polymerase chain reaction. The multilocus sequence typing was used to determine the subtyping of the isolates and characterize possible routes of contamination from meat processing environments. The virulence of different STs of L. monocytogenes isolates was evaluated using a Caco-2 cell invasion assay. RESULTS: A total of 59 Listeria isolates were identified from 320 samples, including 37 L. monocytogenes isolates (62.71%). This study evaluated the virulence of L. monocytogenes and the antibiotic resistance of Listeria isolates from slaughtering and processing plants. The susceptibility of these 59 isolates against 8 antibiotics was analyzed, and the resistance levels to ceftazidime, ciprofloxacin, and lincomycin were as high as 98.31% (L. m 37; L. innocua 7; L. welshimeri 14), 96.61% (L. m 36; L. innocua 7; L. welshimeri 14), and 93.22% (L. m 35; L. innocua 7; L. welshimeri 13), respectively. More than 90% of the isolates were resistant to three to six antibiotics, indicating that Listeria isolated from meat processing environments had high antimicrobial resistance. Up to 60% of the isolates harbored the tetracycline-resistance genes tetA and tetM. The frequency of ermA, ermB, ermC, and aac(6')-Ib was 16.95, 13.56, 15.25, and 6.78%, respectively. Notably, the resistant phenotype and genotype did not match exactly, suggesting that the mechanisms of antibiotic resistance of these isolates were likely related to the processing environment. Multilocus sequence typing (MLST) revealed that 59 Listeria isolates were grouped into 10 sequence types (STs). The dominant L. monocytogenes STs were ST5, ST9, and ST121 in the slaughtering and processing plant of Jiangsu province. Moreover, ST5 subtypes exhibited high invasion in Caco-2 cells compared with ST9 and ST121 cells. CONCLUSION: The dominant L. monocytogenes ST5 persisted in the slaughtering and processing plant and had high antimicrobial resistance and invasion characteristics, illustrating a potential risk in food safety and human health.


Assuntos
Antibacterianos/farmacologia , Listeria/efeitos dos fármacos , Listeria/patogenicidade , Matadouros/estatística & dados numéricos , Animais , China , Farmacorresistência Bacteriana , Inocuidade dos Alimentos , Humanos , Listeria/classificação , Listeria/genética , Carne/microbiologia , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , Suínos/microbiologia , Virulência
2.
Int J Mol Sci ; 22(15)2021 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-34360905

RESUMO

Some Listeria species are important human and animal pathogens that can be found in contaminated food and produce a variety of virulence factors involved in their pathogenicity. Listeria strains exhibiting multidrug resistance are known to be progressively increasing and that is why continuous monitoring is needed. Effective therapy against pathogenic Listeria requires identification of the bacterial strain involved, as well as determining its virulence factors, such as antibiotic resistance and sensitivity. The present study describes the use of liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) to do a global shotgun proteomics characterization for pathogenic Listeria species. This method allowed the identification of a total of 2990 non-redundant peptides, representing 2727 proteins. Furthermore, 395 of the peptides correspond to proteins that play a direct role in Listeria pathogenicity; they were identified as virulence factors, toxins and anti-toxins, or associated with either antibiotics (involved in antibiotic-related compounds production or resistance) or resistance to toxic substances. The proteomic repository obtained here can be the base for further research into pathogenic Listeria species and facilitate the development of novel therapeutics for these pathogens.


Assuntos
Antibacterianos/farmacologia , Proteínas de Bactérias/química , Farmacorresistência Bacteriana Múltipla , Listeria/efeitos dos fármacos , Listeria/patogenicidade , Proteoma/química , Fatores de Virulência/química , Transportadores de Cassetes de Ligação de ATP/química , Cromatografia Líquida/métodos , Genes Bacterianos , Listeria/classificação , Listeria/genética , Peptídeos/química , Proteômica/métodos , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas em Tandem/métodos
3.
Int J Mol Sci ; 21(22)2020 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-33218089

RESUMO

Listeria monocytogenes is a pathogen responsible for severe cases of food poisoning. Listeria spp. strains occurring in soil and water environments may serve as a reservoir of resistance determinants for pathogenic L. monocytogenes strains. A large collection of Listeria spp. strains (155) isolated from natural, agricultural, and urban areas was screened for resistance to heavy metals and metalloids, and the presence of resistance determinants and extrachromosomal replicons. Of the tested strains, 35% were resistant to cadmium and 17% to arsenic. Sequence analysis of resistance plasmids isolated from strains of Listeria seeligeri and Listeria ivanovii, and the chromosome of L. seeligeri strain Sr73, identified a novel variant of the cadAC cadmium resistance efflux system, cadA6, that was functional in L. monocytogenes cells. The cadA6 cassette was detected in four Listeria species, including strains of L. monocytogenes, isolated from various countries and sources-environmental, food-associated, and clinical samples. This resistance cassette is harbored by four novel composite or non-composite transposons, which increases its potential for horizontal transmission. Since some cadAC cassettes may influence virulence and biofilm formation, it is important to monitor their presence in Listeria spp. strains inhabiting different environments.


Assuntos
Proteínas de Bactérias/genética , Farmacorresistência Bacteriana/efeitos dos fármacos , Genoma Bacteriano/genética , Listeria/genética , Metaloides/farmacologia , Metais Pesados/farmacologia , Farmacorresistência Bacteriana/genética , Microbiologia Ambiental , Heterozigoto , Listeria/classificação , Listeria/patogenicidade , Listeria monocytogenes/genética , Listeria monocytogenes/patogenicidade , Testes de Sensibilidade Microbiana , Plasmídeos/genética , Especificidade da Espécie , Virulência/genética
5.
PLoS One ; 15(5): e0232485, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32357157

RESUMO

Various produce including cantaloupe, caramel-coated apples, and packaged salads, have been recognized in recent years as vehicles for listeriosis, a human foodborne disease caused by intracellular pathogen Listeria monocytogenes. Our knowledge regarding the role of these foods in L. monocytogenes virulence, however, is limited. Understanding their role in modulating L. monocytogenes virulence can be useful in risk assessments and for developing control measures. In this study, we employed the Galleria mellonella larvae model to evaluate virulence potential of fifteen clinical, environmental and food isolates of L. monocytogenes, related to three major outbreaks, after growth on different foods. The non-human pathogen Listeria innocua was also included in the panel. Strains were inoculated in parallel in 5ml of brain heart infusion (BHI) broth, and on the surfaces of cantaloupe and apple fragments (5g each) at about 105 colony forming units (CFU)/ml/fragment. One set of inoculated broth and food fragments was incubated at 10°C for 5 days while the second set was kept at 25°C for 3 days. L. monocytogenes cells were recovered from the fruits and BHI, washed twice, re-suspended in saline, and used to inoculate G. mellonella larvae at final concentrations of 106 and 105 CFU/larva. The larvae were incubated at 37°C and monitored for mortality (LT50-time taken to kill 50% of the larvae) and phenotypic changes over seven days. L. monocytogenes grown on cantaloupe and apple flesh surfaces resulted in higher virulence than when grown in BHI. L. monocytogenes infection at 106 CFU/larvae resulted in an average LT50 of ≤ 30, 36 and 47 hours on cantaloupe, apples and BHI, respectively. These results represent a 2.5-4-fold increased mortality compared with an LT50 ≥120 hours in larvae infected with the same doses of L. innocua grown in corresponding matrices. Similar trends were also recorded with doses of about 105 CFU /larvae.


Assuntos
Microbiologia de Alimentos , Listeria monocytogenes/patogenicidade , Animais , Carga Bacteriana , Cucumis melo/microbiologia , Meios de Cultura , Doenças Transmitidas por Alimentos/etiologia , Humanos , Larva/microbiologia , Listeria/crescimento & desenvolvimento , Listeria/patogenicidade , Listeria monocytogenes/crescimento & desenvolvimento , Listeriose/etiologia , Malus/microbiologia , Modelos Biológicos , Mariposas/microbiologia , Medição de Risco , Virulência
6.
Probiotics Antimicrob Proteins ; 12(4): 1562-1570, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32430585

RESUMO

Chitosan films loaded with bacteriocin were examined by FTIR spectroscopy, tested for color, puncture strength, water vapor permeability, and as antimicrobials of Listeria innocua HPB13. Divergicin M35, a bacteriocin produced by Carnobacterium divergens, was incorporated into films made with chitosan of molecular mass 2 kDa, 20 kDa, or 100 kDa and de-acetylated either 87% or 95%. Only 100 kDa chitosan yielded films that could be peeled and handled easily. The higher degree of de-acetylation increased the total color factor (ΔE) of bacteriocin-loaded films, their permeability, and puncture strength. Incorporation of divergicin M35 into the films increased amide I peak intensity but otherwise did not induce significant structural change. The FTIR spectra of divergicin M35 shed from the films did not differ from those of the original free bacteriocin, except in overall peak intensity. The release of active divergicin M35 from the film was faster into the buffer than into tryptic soy broth and peaked at 10-12 h in both cases. Chitosan 95% de-acetylated and loaded with divergicin M35 was the most active, producing a six-log drop in Listeria innocua HPB13 viable count within 24 h. These results suggest that the biocompatible and biodegradable films developed here have the potential for application as antimicrobials of Listeria spp. in foods, especially ready-to-eat, minimally processed products.


Assuntos
Antibacterianos/farmacologia , Bacteriocinas/farmacologia , Carnobacterium/metabolismo , Quitosana/química , Listeria/efeitos dos fármacos , Antibacterianos/química , Antibacterianos/isolamento & purificação , Bacteriocinas/biossíntese , Bacteriocinas/isolamento & purificação , Contagem de Colônia Microbiana , Embalagem de Alimentos/métodos , Humanos , Listeria/crescimento & desenvolvimento , Listeria/patogenicidade , Membranas Artificiais , Peso Molecular , Permeabilidade , Vapor/análise
7.
Mol Microbiol ; 113(3): 627-637, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31972870

RESUMO

Teichoic acids (TAs) are the most abundant glycopolymers in the cell wall of Listeria, an opportunistic Gram-positive pathogen that causes severe foodborne infections. Two different structural classes of Listeria TA exist: the polyribitolphosphate-based wall teichoic acid (WTA) that is covalently anchored to the peptidoglycan, and the polyglycerolphosphate-based lipoteichoic acid (LTA) that is tethered to the cytoplasmic membrane. While TA polymers govern many important physiological processes, the diverse glycosylation patterns of WTA result in a high degree of surface variation across the species and serovars of Listeria, which in turn bestows varying effects on fitness, biofilm formation, bacteriophage susceptibility and virulence. We review the advances made over the past two decades, and our current understanding of the relationship between TA structure and function. We describe the various types of TA that have been structurally determined to date, and discuss the genetic determinants known to be involved in TA glycosylation. We elaborate on surface proteins functionally related to TA decoration, as well as the molecular and analytical tools used to probe TAs. We anticipate that the growing knowledge of the Listeria surface chemistry will also be exploited to develop novel diagnostic and therapeutic strategies for this pathogen.


Assuntos
Listeria/metabolismo , Relação Estrutura-Atividade , Ácidos Teicoicos/metabolismo , Membrana Celular/metabolismo , Parede Celular/metabolismo , Glicosilação , Lipopolissacarídeos/metabolismo , Listeria/patogenicidade , Proteínas de Membrana/metabolismo , Peptidoglicano/metabolismo , Virulência
8.
Sci Rep ; 10(1): 302, 2020 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-31942003

RESUMO

We have previously demonstrated that a recombinant Listeria ivanovii (LI) strain expressing the ESAT-6 or Ag85C protein of Mycobacterium tuberculosis (Mtb) as a tuberculosis (TB) vaccine candidates induced antigen-specific cellular immune responses after intravenous immunization of mice. However, whether such recombinant strains could induce desired immune responses in the lung, where TB infection occurs, is not clear. In this paper, C57BL/6 J mice were intranasally vaccinated with attenuated LIΔactAplcB-Rv3875 (Δ refers to gene deletion in the bacterial genome) or LIΔactAplcB-Rv0129c, the two vaccine candidates that utilize LI as an antigen delivery vector. Bacterial load in the target organs, histological changes in the infected organs, the percentage of specific cytokine-secreting T cells in the lung and spleen, IgG levels in the serum and secretory IgA (SIgA) levles in bronchoalveolar lavage (BAL) fluid were determined at specific days post inoculation (dpi). The results showed that both strains were mainly confined to the lung and were eliminated at 10 dpi. The histological damage caused by the infection in the lung was slight and recovered by day 5. Intranasal vaccination of the mice twice at an interval of 4 weeks notably elicited TB antigen-specific CD4+ and CD8+ T cell responses in the lung and SIgA secretion in the pulmonary mucosa, and significantly enhanced the percentage of double-functional CD8+ T cells (IFN-γ+ TNF-α+ CD8+). To our knowledge, this is the first report regarding the used of LI vector vaccines to induce promising lung-localized cellular and humoral immune responses by intranasal vaccination. These data suggest that LI could be a novel and promising live vector to construct an intranasal vaccine against respiratory diseases.


Assuntos
Antígenos de Bactérias/metabolismo , Imunidade Celular , Imunidade Humoral , Listeria/metabolismo , Pulmão/imunologia , Administração Intranasal , Animais , Antígenos de Bactérias/genética , Antígenos de Bactérias/imunologia , Carga Bacteriana , Líquido da Lavagem Broncoalveolar/química , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/citologia , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Feminino , Imunoglobulina A/metabolismo , Imunoglobulina G/sangue , Interferon gama/metabolismo , Listeria/patogenicidade , Pulmão/metabolismo , Pulmão/patologia , Camundongos , Camundongos Endogâmicos C57BL , Vacinas contra a Tuberculose/imunologia , Fator de Necrose Tumoral alfa/metabolismo , Vacinação
9.
Artigo em Inglês | MEDLINE | ID: mdl-31812649

RESUMO

Listeria spp. are a group of gram-positive bacteria consisting of 20 species. Among them, Listeria monocytogenes is one of the major species that infects humans since it contaminates raw fruits, vegetables, and many others food products. The conventional methods for the detection of Listeria spp. and L. monocytogenes are time-consuming, taking 5-7 days. Herein, a duplex lateral flow dipstick (DLFD) test combined with loop-mediated isothermal amplification (LAMP) was developed for the identification of Listeria spp. and L. monocytogenes within approximately 45 min with the optimized LAMP reaction times at 63 °C. Under the optimized conditions, the method detection limits (MDL) with reference to genomic DNA and pure culture were 900 femtograms (fg) and 20 cfu/mL, respectively. The LAMP-DLFD showed no cross-reactivity with eighteen - other pathogenic bacteria such as Salmonella spp., Staphylococcus aureus, Escherichia coli, Campylobacter coli, C. jejuni, Enterococcus faecalis, Vibrio cholerae, V. parahaemolyticus, Pseudomonas aeruginosa, Shigella dysenteriae, S. flexneri, Bacillus cereus, Lactobacillus acidophilus, L. casei and Pediococcus pentosaceus. Among 100 samples of food products, LAMP-DLFD demonstrated 100% accuracy when compared to other standard detection methods, such as ISO11290-1, enzyme-linked fluorescent assay (ELFA) technology (VIDAS) and PCR. In conclusion, LAMP-DLFD proved to be highly specific and sensitive assays for screening detection of Listeria spp. and L. monocytogenes.


Assuntos
Listeria monocytogenes/genética , Produtos da Carne/microbiologia , Técnicas de Amplificação de Ácido Nucleico/instrumentação , Técnicas de Amplificação de Ácido Nucleico/métodos , DNA Bacteriano/genética , Desenho de Equipamento , Genes Bacterianos/genética , Limite de Detecção , Listeria/genética , Listeria/isolamento & purificação , Listeria/patogenicidade , Listeria monocytogenes/isolamento & purificação , Listeria monocytogenes/patogenicidade
10.
mBio ; 10(4)2019 08 27.
Artigo em Inglês | MEDLINE | ID: mdl-31455648

RESUMO

Various bacterial pathogens display an intracellular lifestyle and spread from cell to cell through actin-based motility (ABM). ABM requires actin polymerization at the bacterial pole and is mediated by the expression of bacterial factors that hijack the host cell actin nucleation machinery or exhibit intrinsic actin nucleation properties. It is increasingly recognized that bacterial ABM factors, in addition to having a crucial task during the intracellular phase of infection, display "moonlighting" adhesin functions, such as bacterial aggregation, biofilm formation, and host cell adhesion/invasion. Here, we review our current knowledge of ABM factors and their additional functions, and we propose that intracellular ABM functions have evolved from ancestral, extracellular adhesin functions.


Assuntos
Actinas/metabolismo , Bactérias/patogenicidade , Fenômenos Fisiológicos Bacterianos , Biofilmes/crescimento & desenvolvimento , Aderência Bacteriana , Proteínas de Bactérias/metabolismo , Interações Hospedeiro-Patógeno , Humanos , Listeria/patogenicidade , Listeria/fisiologia , Shigella/patogenicidade , Shigella/fisiologia
11.
Hig. aliment ; 33(288/289): 2455-2459, abr.-maio 2019. tab
Artigo em Português | LILACS, VETINDEX | ID: biblio-1482239

RESUMO

Queijos tipo Minas frescal podem veicular microrganismos patogênicos. Este estudo objetivou isolar Listeria spp. e identificar as espécies L. innocua, L. seeligeri, L. ivanovii e L. monocytogenes na obtenção do leite e na elaboração de queijos tipo Minas frescal e detectar a presença de genes de virulência. Foram realizadas coletas em cinco pequenas propriedades rurais produtoras deste tipo de queijo em Jaboticabal-São Paulo. Foram coletadas amostras de suabes de fezes bovinas, amostras de mãos de ordenhador, balde de ordenha, leite, água, superfície de elaboração de queijos, mãos de manipulador do queijo, peneiras, bandejas, fôrmas e escumadeiras. O gênero Listeria spp. teve alta prevalência nas amostras, entretanto, nenhuma das espécies pesquisadas foi identificada. Assim, conclui-se que a presença de Listeria spp. em alta percentagem representa potencial risco de contaminação de Queijos tipo Minas frescal e exige uma vigilância contínua para a presença deste gênero.


Assuntos
Laticínios/análise , Laticínios/microbiologia , Leite/microbiologia , Listeria/isolamento & purificação , Listeria/patogenicidade , Manipulação de Alimentos , Microbiologia de Alimentos , Queijo/análise , Queijo/microbiologia
12.
Autophagy ; 15(9): 1620-1633, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-30909843

RESUMO

Due to the critical role played by autophagy in pathogen clearance, pathogens have developed diverse strategies to subvert it. Despite previous key findings of bacteria-autophagy interplay, asystems-level insight into selective targeting by the host and autophagy modulation by the pathogens is lacking. We predicted potential interactions between human autophagy proteins and effector proteins from 56 pathogenic bacterial species by identifying bacterial proteins predicted to have recognition motifs for selective autophagy receptors SQSTM1/p62, CALCOCO2/NDP52 and MAP1LC3/LC3. Using structure-based interaction prediction, we identified bacterial proteins capable to modify core autophagy components. Our analysis revealed that autophagy receptors in general potentially target mostly genus-specific proteins, and not those present in multiple genera. The complementarity between the predicted SQSTM1/p62 and CALCOCO2/NDP52 targets, which has been shown for Salmonella, Listeria and Shigella, could be observed across other pathogens. This complementarity potentially leaves the host more susceptible to chronic infections upon the mutation of autophagy receptors. Proteins derived from enterotoxigenic and non-toxigenic Bacillus outer membrane vesicles indicated that autophagy targets pathogenic proteins rather than non-pathogenic ones. We also observed apathogen-specific pattern as to which autophagy phase could be modulated by specific genera. We found intriguing examples of bacterial proteins that could modulate autophagy, and in turn being targeted by autophagy as ahost defense mechanism. We confirmed experimentally an interplay between a Salmonella protease, YhjJ and autophagy. Our comparative meta-analysis points out key commonalities and differences in how pathogens could affect autophagy and how autophagy potentially recognizes these pathogenic effectors. Abbreviations: ATG5: autophagy related 5; CALCOCO2/NDP52: calcium binding and coiled-coil domain 2; GST: glutathione S-transferase; LIR: MAP1LC3/LC3-interacting region; MAP1LC3/LC3: microtubule associated protein 1 light chain 3 alpha; OMV: outer membrane vesicles; SQSTM1/p62: sequestosome 1; SCV: Salmonella containing vesicle; TECPR1: tectonin beta-propeller repeat containing 1; YhjJ: hypothetical zinc-protease.


Assuntos
Autofagia/genética , Proteínas de Bactérias/metabolismo , Proteínas Associadas aos Microtúbulos/metabolismo , Proteínas Nucleares/metabolismo , Proteína Sequestossoma-1/metabolismo , Motivos de Aminoácidos , Bacillus/metabolismo , Bacillus/patogenicidade , Análise por Conglomerados , Biologia Computacional , Evolução Molecular , Interações Hospedeiro-Patógeno/genética , Humanos , Listeria/metabolismo , Listeria/patogenicidade , Proteínas de Membrana/metabolismo , Proteínas Associadas aos Microtúbulos/genética , Proteínas Nucleares/genética , Ligação Proteica/genética , Mapas de Interação de Proteínas , Salmonella/metabolismo , Salmonella/patogenicidade , Proteína Sequestossoma-1/genética , Shigella/metabolismo , Shigella/patogenicidade , Fatores de Virulência/metabolismo
13.
Infect Immun ; 87(4)2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30670551

RESUMO

Listeria innocua is considered a nonpathogenic Listeria species. Natural atypical hemolytic L. innocua isolates have been reported but have not been characterized in detail. Here, we report the genomic and functional characterization of representative isolates from the two known natural hemolytic L. innocua clades. Whole-genome sequencing confirmed the presence of Listeria pathogenicity islands (LIPI) characteristic of Listeria monocytogenes species. Functional assays showed that LIPI-1 and inlA genes are transcribed, and the corresponding gene products are expressed and functional. Using in vitro and in vivo assays, we show that atypical hemolytic L. innocua is virulent, can actively cross the intestinal epithelium, and spreads systemically to the liver and spleen, albeit to a lesser degree than the reference L. monocytogenes EGDe strain. Although human exposure to hemolytic L. innocua is likely rare, these findings are important for food safety and public health. The presence of virulence traits in some L. innocua clades supports the existence of a common virulent ancestor of L. monocytogenes and L. innocua.


Assuntos
Doenças das Aves/microbiologia , Listeria monocytogenes/patogenicidade , Listeria/isolamento & purificação , Listeria/patogenicidade , Listeriose/microbiologia , Listeriose/veterinária , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Patos , Fezes/microbiologia , Galliformes , Genoma Bacteriano , Ilhas Genômicas , Humanos , Listeria/classificação , Listeria/genética , Listeria monocytogenes/classificação , Listeria monocytogenes/genética , Listeria monocytogenes/isolamento & purificação , Filogenia , Sorotipagem , Virulência , Sequenciamento Completo do Genoma
14.
Microb Pathog ; 126: 123-133, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30381253

RESUMO

The present study aimed to determine the prevalence of Listeria spp. in stray dogs in the Kayseri province of Turkey. In addition, serotyping, genotyping and virulence gene analysis of the isolated Listeria spp. were performed and their pathogenicity and antibacterial susceptibility were investigated. The study included 80 rectal swaps taken from 80 stray dogs of different ages and gender that were sheltered in the Kayseri Municipal Dog Shelter. Listeria selective broth and Listeria selective agar were used for the isolation of Listeria spp. and the isolates were identified using a Microbact 12L (Oxoid, England) identification test kit. 16S rDNA sequencing and species-specific polymerase chain reaction (PCR) were performed for molecular identification of the isolates, multiplex PCR and a serological test were performed for serotyping, and PCR was used for virulence gene analysis. For determining the pathogenicity of L. monocytogenes and L. innocua isolates, a total of 100 mice (50 pregnant and 50 non-pregnant) were used. The mice were infected intraperitoneally; the inoculation dose was 1 × 109 CFU/mL and 0.2 mL was used for each animal. Tissue samples obtained from infected mice were processed for the re-isolation of the Listeria spp. and then stained with hematoxylin eosin and Brown-Brenn Gram stain. The antibiotic susceptibilities of the isolates were determined by the disk diffusion method. Listeria spp. were isolated from 5 (6.25%) of the 80 fecal samples. While 1 of the isolates was identified as L. monocytogenes, 4 of them were identified as L. innocua. Serotyping by serological and molecular methods revealed the isolate of L. monocytogenes to be serotype 1/2a. According to the phylogenetic trees, L. innocua and L. monocytogenes strains were clustered in different groups. The L. monocytogenes isolate was positive for all virulence genes tested. All L. innocua isolates were positive for the plcB gene. While all L. innocua isolates were negative for the lin1068 gene, 3 L. innocua isolates were found to be positive for the lin0558 gene. In mice infected with L. monocytogenes, pathological findings were observed in the uterus, intestines, pancreas, and heart. In mice infected with L. innocua, pathological findings were observed in the stomach, intestines and spleen. L. monocytogenes- or L. innocua-related infections or other inflammatory reactions were not observed in the brains of infected animals. On histopathological examination with Gram stain, an abundance of Listeria spp. was observed in the lesions of the liver, spleen, uterus, and kidney. Moreover, while abortion was observed in all animals infected with L. monocytogenes, it was not observed in any of the animals infected with L. innocua. Antibiotic susceptibility testing revealed that all 5 isolates were sensitive to ampicillin, amoxicillin/clavulanic acid, erythromycin, gentamicin, penicillin G, and trimethoprim-sulfamethoxazole and were resistant to nalidixic acid, streptomycin, and cefuroxime sodium. Considering also the pathogenicity of the isolated microorganisms, it can be suggested that stray dogs as carriers of Listeria spp. are a significant risk to public health. As L. innocua isolates, which are considered apathogenic, led to the occurrence of lesions similar to those caused by L. monocytogenes, detailed studies on the pathogenesis of L. innocua infections caused by L. innocua isolates recovered from various sources are required.


Assuntos
Antibacterianos/farmacologia , Genótipo , Listeria/efeitos dos fármacos , Listeria/genética , Listeria/patogenicidade , Listeriose/microbiologia , Animais , Técnicas de Tipagem Bacteriana/métodos , DNA Bacteriano/genética , Modelos Animais de Doenças , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Doenças do Cão/microbiologia , Cães , Fezes/microbiologia , Feminino , Genes Bacterianos/genética , Listeria/isolamento & purificação , Listeriose/diagnóstico , Listeriose/patologia , Camundongos , Camundongos Endogâmicos BALB C , Filogenia , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/genética , Sorotipagem , Especificidade da Espécie , Turquia , Virulência/genética
15.
J Infect Dis ; 219(1): 145-153, 2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-29733369

RESUMO

Background: Listeria generate actin-rich tubular protrusions at the plasma membrane that propel the bacteria into neighboring cells. The precise molecular mechanisms governing the formation of these protrusions remain poorly defined. Methods: In this study, we demonstrate that the prolyl cis-trans isomerase (PPIase) cyclophilin A (CypA) is hijacked by Listeria at membrane protrusions used for cell-to-cell spreading. Results: Cyclophilin A localizes within the F-actin of these structures and is crucial for their proper formation, as cells depleted of CypA have extended actin-rich structures that are misshaped and are collapsed due to changes within the F-actin network. The lack of structural integrity within the Listeria membrane protrusions hampers the microbes from spreading from CypA null cells. Conclusions: Our results demonstrate a crucial role for CypA during Listeria infections.


Assuntos
Extensões da Superfície Celular/metabolismo , Extensões da Superfície Celular/microbiologia , Ciclofilina A/metabolismo , Listeria/metabolismo , Listeriose/metabolismo , Células A549 , Citoesqueleto de Actina/metabolismo , Actinas/metabolismo , Actinas/ultraestrutura , Membrana Celular/metabolismo , Membrana Celular/microbiologia , Extensões da Superfície Celular/ultraestrutura , Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Células HeLa , Interações Hospedeiro-Patógeno/fisiologia , Humanos , Listeria/patogenicidade , Listeria monocytogenes/metabolismo , Listeria monocytogenes/patogenicidade , Peptidilprolil Isomerase/metabolismo
16.
Arq. Inst. Biol ; 86: e0072019, 2019. tab
Artigo em Inglês | VETINDEX, LILACS | ID: biblio-1024598

RESUMO

Salami is a ready-to-eat (RTE) product frequently purchased at street fairs in Porto Alegre. Salmonella enterica, Listeria monocytogenes, and coagulase-positive Staphylococcus (CPS) are important causes of foodborne disease and can be transmitted through the consumption of RTE foods. The aim of this study was to evaluate the presence of these pathogens in salami sold at street fairs. Ninety salami samples from three commercial brands available at street fairs were analyzed by routine bacteriological methods for Salmonella spp. and Listeria spp., as well as enumeration of CPS. In addition, two samples from each commercial brand were analyzed for water activity (aw). Samples of brand A showed aw values (0.938 and 0.942) above those set by the legislation, while brand B (0.849 and 0.860) and brand C (0.826 and 0.854) were compliant. Microbiological analyses showed that 67.7% were negative to all investigated bacteria. Salmonella Typhimurium was isolated from 4.4% (4/90) of salami samples, all from commercial brand A. ­Listeria monocytogenes was detected in 3.3% (3/90) of samples, from commercial brands B and C. Moreover, 7.7% (7/90) of samples contained CPS populations non-compliant with legislation. Although the great majority of salami sold at street fairs of Porto Alegre was compliant with standards, S. enterica, L. monocytogenes, and CPS ≥ 5 × 103 cfu.g-1 could be found in this RTE product. Therefore, control measures in the processing industry and consumer's education about foodborne illness prevention should be maintained.(AU)


Salame é um alimento pronto para o consumo frequentemente adquirido pela população em feiras livres de Porto Alegre. Salmonella enterica, Listeria monocytogenes e Staphylococcus coagulase positiva são importantes causas de doenças transmitidas por alimentos e podem ser veiculadas por alimentos prontos para o consumo. O objetivo desse estudo foi avaliar a presença desses patógenos em salames vendidos em feiras livres. Noventa amostras de salame pertencentes a três marcas comerciais foram analisados por métodos bacteriológicos de rotina quanto à presença de Salmonella spp. e Listeria spp., bem como enumeração de Staphylococcus coagulase positiva (SCP). Além disso, foi determinada a Atividade de Água (aw) de duas amostras de cada marca comercial. Amostras da marca A apresentaram valores de aw (0,938 e 0,942) acima do permitido na legislação, enquanto as amostras da marca B (0,849 e 0,860) e C (0,826 e 0,854) não violaram esse parâmetro. A análise microbiológica demonstrou que 67,7% das amostras foram negativas para todas as bactérias investigadas. Salmonella Typhimurium foi isolada de 4,4% (4/90) das amostras de salame, todas da marca comercial A. Listeria monocytogenes foi detectada em 3,3% (3/90) das amostras das marcas B e C. Além disso, 7,7% (7/90) das amostras apresentaram SCP acima do número permitido pela legislação. Apesar da grande maioria dos salames comercializados em feiras livres estarem de acordo com a legislação, S. enterica, L. monocytogenes e SCP ≥ 5 × 103 cfu.g-1 podem estar presentes nesse alimento pronto para o consumo. Dessa forma, o controle nas indústrias e a educação dos consumidores sobre a prevenção de doenças transmitidas por alimentos devem ser mantidos.(AU)


Assuntos
Salmonella/patogenicidade , Staphylococcus/patogenicidade , Suínos , Listeria/patogenicidade , Bactérias , /métodos , Indústria Alimentícia , Normas de Qualidade de Alimentos , Carne
17.
Cell ; 175(5): 1336-1351.e17, 2018 11 15.
Artigo em Inglês | MEDLINE | ID: mdl-30318148

RESUMO

As a critical step during innate response, the cytoplasmic ß subunit (IFN-γR2) of interferon-γ receptor (IFN-γR) is induced and translocates to plasma membrane to join α subunit to form functional IFN-γR to mediate IFN-γ signaling. However, the mechanism driving membrane translocation and its significance remain largely unknown. We found, unexpectedly, that mice deficient in E-selectin, an endothelial cell-specific adhesion molecule, displayed impaired innate activation of macrophages upon Listeria monocytogenes infection yet had increased circulating IFN-γ. Inflammatory macrophages from E-selectin-deficient mice had less surface IFN-γR2 and impaired IFN-γ signaling. BTK elicited by extrinsic E-selectin engagement phosphorylates cytoplasmic IFN-γR2, facilitating EFhd2 binding and promoting IFN-γR2 trafficking from Golgi to cell membrane. Our findings demonstrate that membrane translocation of cytoplasmic IFN-γR2 is required to activate macrophage innate response against intracellular bacterial infection, identifying the assembly of functional cytokine receptors on cell membrane as an important layer in innate activation and cytokine signaling.


Assuntos
Selectina E/metabolismo , Imunidade Inata , Receptores de Interferon/metabolismo , Animais , Proteínas de Ligação ao Cálcio/antagonistas & inibidores , Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/metabolismo , Membrana Celular/metabolismo , Selectina E/deficiência , Selectina E/genética , Complexo de Golgi/metabolismo , Interferon gama/sangue , Interferon gama/metabolismo , Listeria/patogenicidade , Ativação de Macrófagos , Macrófagos/citologia , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fosforilação , Transporte Proteico , Células RAW 264.7 , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Receptores de Interferon/deficiência , Receptores de Interferon/genética , Transdução de Sinais
19.
Microbiol Spectr ; 6(4)2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-30027884

RESUMO

For nearly a century the use of antibiotics to treat infectious diseases has benefited human and animal health. In recent years there has been an increase in the emergence of antibiotic-resistant bacteria, in part attributed to the overuse of compounds in clinical and farming settings. The genus Listeria currently comprises 17 recognized species found throughout the environment. Listeria monocytogenes is the etiological agent of listeriosis in humans and many vertebrate species, including birds, whereas Listeria ivanovii causes infections mainly in ruminants. L. monocytogenes is the third-most-common cause of death from food poisoning in humans, and infection occurs in at-risk groups, including pregnant women, newborns, the elderly, and immunocompromised individuals.


Assuntos
Anti-Infecciosos/farmacologia , Farmacorresistência Bacteriana/efeitos dos fármacos , Listeria/efeitos dos fármacos , Listeria/patogenicidade , Listeriose/microbiologia , Idoso , Animais , Animais Domésticos , Anti-Infecciosos/uso terapêutico , Farmacorresistência Bacteriana/genética , Fazendas , Feminino , Microbiologia de Alimentos , Doenças Transmitidas por Alimentos/microbiologia , Humanos , Recém-Nascido , Sequências Repetitivas Dispersas/genética , Listeria/classificação , Listeria/genética , Listeria monocytogenes/efeitos dos fármacos , Listeriose/tratamento farmacológico , Listeriose/veterinária , Gravidez
20.
Cell Host Microbe ; 24(1): 5-7, 2018 07 11.
Artigo em Inglês | MEDLINE | ID: mdl-30001525

RESUMO

STING is a key mediator of mammalian innate immunity in response to pathogens. In this issue of Cell Host & Microbe and the June 19th issue of Cell Reports, Liu et al. (2018) and Martin et al. (2018) reveal that Drosophila STING is required for antiviral and antibacterial defenses, respectively.


Assuntos
Interações Hospedeiro-Patógeno/imunologia , Imunidade Inata , Insetos/imunologia , Mamíferos/imunologia , Transdução de Sinais/imunologia , Animais , Antibacterianos , Antivirais , Autofagia , Infecções Bacterianas/imunologia , Drosophila , Interações Hospedeiro-Patógeno/genética , Humanos , Insetos/genética , Listeria/imunologia , Listeria/patogenicidade , Mamíferos/genética , Mycobacterium tuberculosis/imunologia , Mycobacterium tuberculosis/patogenicidade , Transdução de Sinais/genética , Viroses/imunologia , Zika virus/imunologia , Zika virus/patogenicidade
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