Salmonella Enteritidis is the main serovar of poultry origin in humans, but its complex interaction with certain avian cells is still not fully understood. Previously we identified several genes significantly induced in chicken embryo fibroblasts (CEFs) by the wild-type strain S. Enteritidis 11 (SE 11). In the present study, we raised the question whether virulence-attenuated mutants of this strain would induce altered expression of the newly identified fibroblast genes associated with immune and non-immune functions of CEFs. Gene expression was evaluated by real-time PCR following challenge by the parental strain SE 11 and its virulence attenuated mutants lacking flagellin gene fliD only or fliD and the serovar-specific virulence plasmid pSEV. As a result, deletion mutants induced a lower expression of all immune genes, but an increased expression of the non-immune genes G0S2 and ENO2 relative to the parental strain. Our data indicate the importance of flagella and pSEV in modulation of virulence and host response in this model. We demonstrated, for the first time ever, an increased induction of survival genes G0S2 and ENO2 by virulence-attenuated mutants of S. Enteritidis.

• MeSH
• bakteriální proteiny genetika   MeSH
• fibroblasty mikrobiologie   MeSH
• flagella genetika   MeSH
• interakce hostitele a patogenu MeSH
• kuřecí embryo MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• nemoci drůbeže imunologie   mikrobiologie   MeSH
• plazmidy genetika   MeSH
• Salmonella enteritidis genetika   patogenita   MeSH
• salmonelová infekce u zvířat imunologie   mikrobiologie   MeSH
• virulence genetika   MeSH
• zvířata MeSH
• Check Tag
• kuřecí embryo MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

A European multi-country outbreak of Salmonella Enteritidis phage type (PT) 14b occurred from March to November 2014 associated with the consumption of eggs. The outbreak involved more than 400 human cases from France, Luxembourg, Austria and the United Kingdom. In 2016-2017, it has been re-evaluated combining recent epidemiological results with latest molecular data. The outbreak was traced back to one large Bavarian egg producer with four distinct premises, three located in Bavaria, one in the Czech Republic. The outbreak isolates of S. Enteritidis PT 14b were grouped into three closely related clades by whole genome sequencing. Two of these clades could be referred to two Bavarian premises of the egg producer on the basis of epidemiological and molecular data, while epidemiological data presumably linked the third clade to another premises of the egg producer. Interestingly and in contrast to the situation in other European countries where several outbreaks were documented, all notified 91 laboratory-confirmed cases of S. Enteritidis PT 14b from Bavaria were sporadic, singular cases not belonging to any epidemiological outbreaks. In conclusion, as demonstrated here, the resolution of food-related outbreaks with such a high discriminatory power is rare in outbreak investigation.

• MeSH
• epidemický výskyt choroby * MeSH
• fagotypizace metody   MeSH
• jednonukleotidový polymorfismus MeSH
• lidé MeSH
• otrava salmonelou epidemiologie   MeSH
• Salmonella enteritidis klasifikace   genetika   izolace a purifikace   MeSH
• salmonelóza epidemiologie   MeSH
• sekvenování celého genomu MeSH
• vejce mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• Francie epidemiologie   MeSH
• Lucembursko MeSH
• Rakousko MeSH
• Spojené království MeSH

• Klíčová slova
• Salmonella enteritidis fagotyp 8 MLVA profil 2-9-7-3-2,
• MeSH
• Evropská unie MeSH
• Salmonella enteritidis * genetika   MeSH
• Geografické názvy
• Česká republika MeSH

Poultry is the most frequent reservoir of non-typhoid Salmonella enterica for humans. Understanding the interactions between chickens and S. enterica is therefore important for vaccine design and subsequent decrease in the incidence of human salmonellosis. In this study we therefore characterized the interactions between chickens and phoP, aroA, SPI1 and SPI2 mutants of S. Enteritidis. First we tested the response of HD11 chicken macrophage-like cell line to S. Enteritidis infection monitoring the transcription of 36 genes related to immune response. All the mutants and the wild type strain induced inflammatory signaling in the HD11 cell line though the response to SPI1 mutant infection was different from the rest of the mutants. When newly hatched chickens were inoculated, the phoP as well as the SPI1 mutant did not induce an expression of any of the tested genes in the cecum. Despite this, such chickens were protected against challenge with wild-type S. Enteritidis. On the other hand, inoculation of chickens with the aroA or SPI2 mutant induced expression of 27 and 18 genes, respectively, including genes encoding immunoglobulins. Challenge of chickens inoculated with these two mutants resulted in repeated induction of 11 and 13 tested genes, respectively, including the genes encoding immunoglobulins. In conclusion, SPI1 and phoP mutants induced protective immunity without inducing an inflammatory response and antibody production. Inoculation of chickens with the SPI2 and aroA mutants also led to protective immunity but was associated with inflammation and antibody production. The differences in interaction between the mutants and chicken host can be used for a more detailed understanding of the chicken immune system.

• MeSH
• atenuované vakcíny imunologie   MeSH
• buněčné linie MeSH
• cékum imunologie   mikrobiologie   MeSH
• kur domácí MeSH
• makrofágy imunologie   MeSH
• mutace MeSH
• nemoci drůbeže imunologie   mikrobiologie   MeSH
• Salmonella enteritidis * genetika   imunologie   MeSH
• salmonelová infekce u zvířat imunologie   mikrobiologie   MeSH
• salmonelové vakcíny imunologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The prevalence of Salmonella enterica serovar Enteritidis is gradually decreasing in poultry flocks in the EU, which may result in the demand for a vaccine that allows for the differentiation of vaccinated flocks from those infected by wild-type S. Enteritidis. In this study, we therefore constructed a (Salmonella Pathogenicity Island 1) SPI1-lon mutant with or without fliC encoding for S. Enteritidis flagellin. The combination of SPI1-lon mutations resulted in attenuated but immunogenic mutant suitable for oral vaccination of poultry. In addition, the vaccination of chickens with the SPI1-lon-fliC mutant enabled the serological differentiation of vaccinated and infected chickens. The absence of fliC therefore did not affect the immunogenicity of the vaccine strain and allowed for serological differentiation of the vaccinated chickens. The SPI1-lon-fliC mutant is therefore a suitable marker vaccine strain for oral vaccination of poultry.

• MeSH
• bakteriální proteiny genetika   imunologie   MeSH
• flagelin genetika   imunologie   MeSH
• kur domácí MeSH
• mutace * MeSH
• nemoci drůbeže prevence a kontrola   MeSH
• proteasa La genetika   imunologie   MeSH
• protilátky bakteriální imunologie   MeSH
• Salmonella enteritidis genetika   růst a vývoj   imunologie   ultrastruktura   MeSH
• salmonelová infekce u zvířat prevence a kontrola   MeSH
• salmonelové vakcíny aplikace a dávkování   imunologie   MeSH
• vakcinace veterinární   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

In this study we were interested in the serovar cross-protection potential of Salmonella Pathogenicity Island 1 (SPI1) attenuated vaccine strains of Salmonella enterica serovars Enteritidis and Typhimurium and immune response of vaccinated and naive chickens to Salmonella infection. The immune response was characterized by real time PCR quantifying transcripts of interleukins IL1β, IL17, IL22, interferon gamma (IFNγ), inducible NO synthase (iNOS), immunoglobulins IgM, IgA, IgY and Ig light chain, and six genes of acute phase response including avidin, serum amyloid A, extracellular fatty acid-binding protein (Ex-FABP), immune responsive gene 1, chemokine AH221 and trappin-6. Vaccination with SPI1 mutants of both serovars protected chickens against Salmonella infection, independent of the serovar used for the challenge and the time post infection. However, expressions of all interleukins, iNOS and Ex-FABP showed that protection against homologous serovars was significantly higher than against heterologous serovars after intravenous challenge at 4 days post infection. The vaccination with a mixture of S. Enteritidis and S. Typhimurium SPI1 mutants induced an intermediate protection against challenge with both serovars, i.e. the mixed vaccine provided an additional protective effect when compared with the chickens vaccinated with a vaccine formed by only a single Salmonella serovar.

• MeSH
• atenuované vakcíny genetika   imunologie   MeSH
• chemokiny imunologie   MeSH
• genomové ostrovy MeSH
• kur domácí imunologie   MeSH
• nemoci drůbeže imunologie   prevence a kontrola   MeSH
• protilátky bakteriální krev   imunologie   MeSH
• Salmonella enterica genetika   imunologie   patogenita   MeSH
• Salmonella enteritidis genetika   imunologie   MeSH
• Salmonella typhimurium genetika   imunologie   MeSH
• salmonelová infekce u zvířat imunologie   prevence a kontrola   MeSH
• salmonelové vakcíny genetika   imunologie   MeSH
• vakcinace veterinární   MeSH
• zkřížená ochrana * MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Molecular typing and resistotyping coupled with gyrA single nucleotide polymorphism (SNP) of 60 Salmonella Enteritidis (SE) isolates originated from poultry, food, and humans in Serbia is described. Molecular fingerprinting was performed by randomly amplified polymorphic DNA (RAPD) using four primers, and the diversity index (D) was 0.688. In combination with resistotyping and gyrA SNP, D increased to 0.828. A total of 23 genetic groups were obtained. When four RAPD primers were combined, epidemic isolates from a fast-food restaurant outbreak were clustered in a distinctive genetic group. Among 60 SE strains, three had multiple resistances to three or more antibiotics. Nine strains were resistant to nalidixic acid (NAL; a non-fluorinated quinolone). The mutations in quinolone resistance-determining region (QRDR) found in NAL-resistant strains were attributed to Asp(87) → Asn in six strains, Asp(87) → Gly in one strain, and Ser(83) → Phe in one strain. One NAL-resistant strain had no mutations in QRDR, suggesting another mechanism of resistance.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální léková rezistence MeSH
• bakteriální proteiny genetika   MeSH
• DNA gyráza genetika   MeSH
• jednonukleotidový polymorfismus MeSH
• kur domácí MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• molekulární typizace MeSH
• nemoci drůbeže mikrobiologie   MeSH
• potravinářská mikrobiologie MeSH
• Salmonella enteritidis klasifikace   účinky léků   genetika   izolace a purifikace   MeSH
• salmonelová infekce u zvířat mikrobiologie   MeSH
• salmonelóza mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Srbsko MeSH

Genes localized at Salmonella pathogenicity island-1 (SPI-1) are involved in Salmonella enterica invasion of host non-professional phagocytes. Interestingly, in macrophages, SPI-1-encoded proteins, in addition to invasion, induce cell death via activation of caspase-1 which also cleaves proIL-1β and proIL-18, precursors of 2 proinflammatory cytokines. In this study we were therefore interested in whether SPI-1-encoded type III secretion system (T3SS) may influence proinflammatory response of macrophages. To test this hypothesis, we infected primary porcine alveolar macrophages with wild-type S. Typhimurium and S. Enteritidis and their isogenic SPI-1 deletion mutants. ΔSPI1 mutants of both serovars invaded approx. 5 times less efficiently than the wild-type strains and despite this, macrophages responded to the infection with ΔSPI1 mutants by increased expression of proinflammatory cytokines IL-1β, IL-8, TNFα, IL-23α and GM-CSF. Identical macrophage responses to that induced by the ΔSPI1 mutants were also observed to the infection with sipB but not the sipA mutant. The hilA mutant exhibited an intermediate phenotype between the ΔSPI1 mutant and the wild-type S. Enteritidis. Our results showed that the SPI-1-encoded T3SS is required not only for cell invasion but in macrophages also for the suppression of early proinflammatory cytokine expression.

• MeSH
• alveolární makrofágy imunologie   metabolismus   MeSH
• cytokiny genetika   metabolismus   MeSH
• genomové ostrovy * MeSH
• nemoci prasat imunologie   mikrobiologie   MeSH
• prasata MeSH
• Salmonella enteritidis genetika   MeSH
• Salmonella typhimurium genetika   metabolismus   MeSH
• salmonelová infekce u zvířat imunologie   mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Natural variation in the presence or the absence of STM0517-0529 genes allowing allantoin utilisation has been described in field isolates of the multidrug resistant Salmonella enterica serovar Typhimurium belonging to the phage type DT104. Interestingly, S. enterica subspecies enterica serovar Typhimurium DT104 is quite frequent in pigs and cattle, but rarely present in egg-laying hens. Taking into account the different mode of allantoin metabolism in birds and mammals, we were interested in whether the absence of STM0517-0529 genes may disable this clone in poultry colonisation. We have therefore constructed the allB (also designated as STM0523) mutants in S. enterica subspecies enterica serovar Typhimurium and S. enterica subspecies enterica serovar Enteritidis, and with these, we infected mice, newly hatched chickens and adult egg-laying hens to show that the defect in allantoin utilisation does not influence S. enterica virulence for mice or adult hens, but slightly decreases virulence of S. enterica for chickens. The decrease in virulence of the allB mutant was relatively minor as it could be observed only after a mixed infection model, consistent with a lower prevalence, but not a total absence of such clones in poultry flocks.

• MeSH
• alantoin metabolismus   MeSH
• drůbež MeSH
• kyselina močová metabolismus   MeSH
• mnohočetná bakteriální léková rezistence MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• nemoci drůbeže mikrobiologie   patologie   MeSH
• Salmonella enteritidis genetika   metabolismus   patogenita   MeSH
• Salmonella typhimurium genetika   metabolismus   patogenita   MeSH
• salmonelová infekce u zvířat mikrobiologie   patologie   MeSH
• virulence MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

In this study, we were interested in the association of attenuated mutants of Salmonella enterica serovar Enteritidis with subpopulations of porcine white blood cells (WBC). The mutants included those with inactivated aroA, phoP, rfaL, rfaG, rfaC and fliC genes and a mutant with five major pathogenicity islands removed (ΔSPI1-5 mutant). Using flow cytometry, we did not observe any difference in the interactions of the wild-type S. Enteritidis, aroA and phoP mutants with WBC. ΔSPI1-5 and fliC mutants had a minor defect in their association with granulocytes and monocytes, but not with T- or B-lymphocytes. All three rfa mutants associated with granulocytes, monocytes and B-lymphocytes more than the wild-type S. Enteritidis did. Electron microscopy confirmed that the association correlated with the intracellular presence of S. Enteritidis and that the Salmonella-containing vacuole in the WBC infected with the rfa mutants, unlike all other strains, did not develop into a spacious phagosome. Intact lipopolysaccharide, but not the type III secretion system encoded by SPI-1, SPI-2 or the flagellar operon, is important for the initial interaction of S. Enteritidis with porcine leukocytes. This information can be used for the design of live Salmonella vaccines preferentially targeting particular cell types including cancer or tumor cells.

• MeSH
• bakteriální geny genetika   MeSH
• glykosyltransferasy genetika   MeSH
• leukocyty mikrobiologie   ultrastruktura   MeSH
• mutace genetika   MeSH
• nemoci prasat imunologie   mikrobiologie   patologie   MeSH
• prasata MeSH
• Salmonella enteritidis enzymologie   genetika   imunologie   MeSH
• salmonelová infekce u zvířat imunologie   mikrobiologie   patologie   MeSH
• salmonelové vakcíny imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH