In our study we present an overview of the use of Oxford Nanopore Technologies (ONT) sequencing technology on the background of Enteric fever. Unlike traditional methods (e.g., qPCR, serological tests), the nanopore sequencing technology enables virtually real-time data generation and highly accurate pathogen identification and characterization. Blood cultures were obtained from a 48-year-old female patient suffering from a high fever, headache and diarrhea. Nevertheless, both the initial serological tests and stool culture appeared to be negative. Therefore, the bacterial isolate from blood culture was used for nanopore sequencing (ONT). This technique in combination with subsequent bioinformatic analyses allowed for prompt identification of the disease-causative agent as Salmonella enterica subsp. enterica serovar Paratyphi A. The National Reference Laboratory for Salmonella (NIPH) independently reported this isolate also as serovar Paratyphi A on the basis of results of biochemical and agglutination tests. Therefore, our results are in concordance with certified standards. Furthermore, the data enabled us to assess some basic questions concerning the comparative genomics, i.e., to describe whether the isolated strain differs from the formerly published ones or not. Quite surprisingly, these results indicate that we have detected a novel and so far, unknown variety of this bacteria.

• Klíčová slova
• Enteric fever, Nanopore sequencing, Paratyphoid fever, Pathogen identification, Salmonella,
• MeSH
• břišní tyfus * MeSH
• lidé středního věku MeSH
• lidé MeSH
• nanoporové sekvenování * MeSH
• Salmonella paratyphi A genetika   MeSH
• Salmonella MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• přehledy MeSH

INTRODUCTION: Ubiquitination is an important protein modification that regulates various essential cellular processes, including the functions of innate immune cells. Deubiquitinases are enzymes responsible for removing ubiquitin modification from substrates, and the regulation of deubiquitinases in macrophages during infection with Salmonella Typhimurium and Yersinia enterocolitica remains unknown. METHODS: To identify deubiquitinases regulated in human macrophages during bacterial infection, an activity-based proteomics screen was conducted. The effects of pharmacological inhibition of the identified deubiquitinase, USP8, were examined, including its impact on bacterial survival within macrophages and its role in autophagy regulation during Salmonella infection. RESULTS: Several deubiquiitnases were differentially regulated in infected macrophages. One of the deubiquitinases identified was USP8, which was downregulated upon Salmonella infection. Inhibition of USP8 was associated with a decrease in bacterial survival within macrophages, and it was found to play a distinct role in regulating autophagy during Salmonella infection. The inhibition of USP8 led to the downregulation of the p62 autophagy adaptor. DISCUSSION: The findings of this study suggest a novel role of USP8 in regulating autophagy flux, which restricts intracellular bacteria, particularly during Salmonella infection.

• Klíčová slova
• Salmonella, USP8, autophagy, chemical proteomics, deubiquitinases, deubiquitinating enzymes,
• MeSH
• autofagie MeSH
• deubikvitinasy metabolismus   MeSH
• endopeptidasy genetika   MeSH
• endozomální třídící komplexy pro transport genetika   MeSH
• lidé MeSH
• Salmonella typhimurium metabolismus   MeSH
• salmonelóza * MeSH
• thiolesterasa ubikvitinu genetika   metabolismus   MeSH
• ubikvitinace MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Názvy látek
• deubikvitinasy MeSH
• endopeptidasy MeSH
• endozomální třídící komplexy pro transport MeSH
• thiolesterasa ubikvitinu MeSH
• USP8 protein, human MeSH Prohlížeč

In one flock, one-eye blindness was observed in the chronical course of disease after infection with the germs of Salmonella arizonae. Changes such as granulomatous iridocyclochorioretinitis with the finding of the bacterial causative agent directly in the pathologically affected tissue were diagnosed on the basis of detailed histological examination of the affected eyeballs. The changes affected almost all parts of the eyeball (iris, ciliary body, lens, retina and chorioid) and were characterized by mononuclear infiltrate with differently intensive admixture of heterophils. The lens was liable to degeneration and fragmentation and giant-cell granulomes of different sizes formed in the posterior chamber.

• MeSH
• krocani * MeSH
• nemoci drůbeže patologie   MeSH
• oči patologie   MeSH
• Salmonella arizonae MeSH
• salmonelová infekce u zvířat patologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH

Salmonella infections are still considered a persistent problem in veterinary medicine. Vaccination is one of the tools for decreasing the burden of many pathogens on animals. However, the efficiency of available commercial or experimental vaccines against non-typhoid Salmonella strains is not yet sufficient. We followed the path of an inactivated vaccine that is safe and well accepted, but whose presented antigen spectrum is limited. We improved this issue by using diverse cultivation conditions mimicking bacterial protein expression during the natural infection process. The cultivation process was set up to simulate the host environment to enhance the expression of SPI-1 (Salmonella pathogenicity island) proteins, SPI-2 proteins, siderophore-related proteins, and flagellar proteins. Three different cultivation media were used and subsequent cultures were mixed together, inactivated, and used for the immunization of post-weaned piglets. A mixture of recombinant Salmonella proteins was also used as a recombinant vaccine for comparison. The clinical symptoms during the subsequent experimental infection, antibody response, and organ bacterial loads were examined. One day after the infection, we observed an increased rectal temperature in the group of unvaccinated animals and the animals vaccinated with the recombinant vaccine. The increase in the temperature of the pigs vaccinated with the inactivated Salmonella mixture was significantly lower. In the same group, we also found lower bacterial loads in the ileum content and the colon wall. The IgG response to several Salmonella antigens was enhanced in this group, but it did not reach the titers of the group vaccinated with the recombinant vaccine. To summarize, the pigs vaccinated with an inactivated mixture of Salmonella cultures mimicking protein expression changes during the natural infection exhibited less serious clinical symptoms and lower bacterial load in the body after the experimental infection compared to the unvaccinated pigs and the pigs vaccinated with a mixture of recombinant Salmonella proteins.

• Klíčová slova
• Post-weaned piglets, Salmonella Typhimurium, Vaccine, Virulence proteins,
• MeSH
• antigeny bakteriální genetika   MeSH
• atenuované vakcíny MeSH
• inaktivované vakcíny MeSH
• nemoci prasat * mikrobiologie   MeSH
• prasata MeSH
• protilátky bakteriální MeSH
• Salmonella typhimurium MeSH
• salmonelová infekce u zvířat * mikrobiologie   MeSH
• salmonelové vakcíny * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigeny bakteriální MeSH
• atenuované vakcíny MeSH
• inaktivované vakcíny MeSH
• protilátky bakteriální MeSH
• salmonelové vakcíny * MeSH

OBJECTIVE: Salmonella Paratyphi B dT+ (or Salmonella Java) is an emerging public health problem. The study objective was to characterize phenotypically and genotypically 13 human isolates from sporadic cases of infection. MATERIAL AND METHODS: Phenotypic characteristics of 13 human isolates were determined by phage typing, disk diffusion method for testing antimicrobial susceptibility to 11 antibiotics and screening for selected virulence markers, i.e bacterial adherence to xylene, motility and biofilm formation. Genotypic characteristics of the study isolates were determined by PCR with primers for the detection of class 1 integrons, plasmid profile analysis and PFGE for the study of genetic diversity. RESULTS: The study isolates were classified into different phage types, with 3aI or 3aI variant being the most common (61.5%). All study isolates were resistant to streptomycin and sulfisoxazole, two of them were multiresistant (to streptomycin, sulfisoxazole, ampicillin and nalidixic acid). The study isolates showed low hydrophobicity, except for one isolate (2/08), and 5 isolates exhibited motility of > or = 50 mm. High biofilm formation was detected in 5 isolates. Class 1 integrons were not detected in any isolate and plasmid profile analysis revealed the presence of a 90-kb plasmid in 8 isolates. XbaI PFGE analysis differentiated the isolates into 6 pulsotypes (X1, X2, X2a, X3, X4,X5). CONCLUSION: Although the study set of S. Paratyphi B dT+ (S. Java) was small, the increasing emergence of this serovar in the human population in Slovakia is noteworthy. The results of gene analysis in PFGE suggest clonal diversity as well as a clonal link between strains of the predominant phage type. In view of possible increase in antimicrobial resistance and the spread of certain clones, continuous surveillance of strains of S. Paratyphi B dT+ is needed.

• MeSH
• fagotypizace MeSH
• gastroenteritida mikrobiologie   MeSH
• genotyp MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• plazmidy MeSH
• Salmonella paratyphi B klasifikace   genetika   izolace a purifikace   fyziologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH

BACKGROUND: Salmonella enterica serovar Typhimurium is one of the most common enteropathogenic bacteria found in pigs in Europe. In our previous work, we demonstrated the protective effects in suckling piglets when their dams had been vaccinated with an S. Typhimurium-based inactivated vaccine. This study is focused on a procedure leading to serological discrimination between vaccinated and infected pigs. As we supposed, distinct environment during natural infection and in bacterial cultures used for vaccine preparation led to a slightly different spectrum of expressed S. Typhimurium proteins. The examination of porcine antibodies produced after the experimental infection with S. Typhimurium or after vaccination with S. Typhimurium-based inactivated vaccine by affinity chromatography and mass spectrometry revealed differences in antibody response applicable for serological differentiation of infected from vaccinated animals. RESULTS: Antibodies against Salmonella SipB, SipD and SseB proteins were detected at much higher levels in post-infection sera in comparison with control and post-vaccination sera. On the other hand, proteins BamB, OppA and a fragment of FliC interacted with antibodies from post-vaccination sera with a much higher intensity than from control and post-infection sera. In addition, we constructed ELISA assays using post-infection antigen - SipB protein and post-vaccination antigen - FliC-fragment and evaluated them on a panel of individual porcine sera. CONCLUSIONS: The analysis of antibody response of infected and vaccinated pigs by proteomic tools enabled to identify S. Typhimurium antigens useful for distinguishing infected from vaccinated animals. This approach can be utilized in other challenges where DIVA vaccine and a subsequent serological assay are required, especially when genetic modification of a vaccine strain is not desirable.

• Klíčová slova
• DIVA vaccine, Mass spectrometry, Porcine, Recombinant protein, Salmonella Typhimurium,
• MeSH
• antigeny bakteriální metabolismus   MeSH
• ELISA veterinární   MeSH
• inaktivované vakcíny imunologie   MeSH
• nemoci prasat diagnóza   imunologie   MeSH
• prasata MeSH
• proteomika * MeSH
• protilátky bakteriální krev   MeSH
• Salmonella typhimurium genetika   MeSH
• salmonelová infekce u zvířat diagnóza   imunologie   MeSH
• salmonelové vakcíny imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigeny bakteriální MeSH
• inaktivované vakcíny MeSH
• protilátky bakteriální MeSH
• salmonelové vakcíny MeSH

Interaction between pigs and Salmonella enterica serovar Derby (Salmonella Derby) is much less understood in comparison with Salmonella enterica serovar Typhimurium (Salmonella Typhimurium). To study interactions of weaned piglets with Salmonella Derby, we compared the course of infections with Salmonella Derby De1 and Salmonella Typhimurium DT104 strains, both isolated from pig herds with a long history of asymptomatic infection. Salmonella Derby strain used was shed during the 28-day experiment period, while Salmonella Typhimurium strain was not found in faeces after day 17 post-infection. When the piglets were co-infected with both strains, Salmonella Derby was present in faeces until the end of the experiment, whilst Salmonella Typhimurium disappeared after day 21 post-infection. At the end of the experiment, Salmonella Derby was present in more tissues when compared with Salmonella Typhimurium. Piglets infected with Salmonella Typhimurium responded earlier with synthesis of anti-lipopolysaccharide IgM and IgG antibodies and with higher antibody levels compared to piglets infected with Salmonella Derby. Cellular immune response to both strains was very low and was detected later than was the onset of IgG antibody production.

• Klíčová slova
• Derby, Immune response, Pig, Salmonella, Typhimurium,
• MeSH
• feces mikrobiologie   MeSH
• koinfekce imunologie   MeSH
• nemoci prasat imunologie   MeSH
• prasata MeSH
• protilátky bakteriální krev   MeSH
• Salmonella enterica imunologie   izolace a purifikace   MeSH
• Salmonella typhimurium imunologie   izolace a purifikace   MeSH
• salmonelová infekce u zvířat imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• protilátky bakteriální MeSH

Salmonella spp. is a common zoonotic pathogen, causing gastrointestinal infections in people. Pigs and pig meat are a major source of infection. Although farm biosecurity is believed to be important for controlling Salmonella transmission, robust evidence is lacking on which measures are most effective. This study enrolled 250 pig farms across nine European countries. From each farm, 20 pooled faecal samples (or similar information) were collected and analysed for Salmonella presence. Based on the proportion of positive results, farms were categorised as at higher or lower Salmonella risk, and associations with variables from a comprehensive questionnaire investigated. Multivariable analysis indicated that farms were less likely to be in the higher-risk category if they had '<400 sows'; used rodent baits close to pig enclosures; isolated stay-behind (sick) pigs; did not answer that the hygiene lock/ anteroom was easy to clean; did not have a full perimeter fence; did apply downtime of at least 3 days between farrowing batches; and had fully slatted flooring in all fattener buildings. A principal components analysis assessed the sources of variation between farms, and correlation between variables. The study results suggest simple control measures that could be prioritised on European pig farms to control Salmonella.

• Klíčová slova
• Salmonella, biosecurity, effectiveness, pig, risk factor,
• MeSH
• biologická bezpečnost MeSH
• chov zvířat metody   MeSH
• farmy MeSH
• nemoci prasat * epidemiologie   prevence a kontrola   MeSH
• prasata MeSH
• Salmonella MeSH
• salmonelová infekce u zvířat * epidemiologie   prevence a kontrola   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Evropa epidemiologie   MeSH

Salmonellae are widely distributed but nomenclaturally controversial pathogens of both humans and animals. Despite elaborate studies, much still remain to be discovered about these organisms. Although Salmonella nomenclature has proved to be rather complex, in 2005, Salmonella enterica finally gained official approval as the type species of the genus Salmonella. In addition, one other species has been approved and recognised in the genus Salmonella, namely, Salmonella bongori. New serovars (serotypes) are continually being discovered each year and reported in the journal Research in Microbiology. Salmonella serovars and their antigenic formulae are listed in the White-Kauffmann-Le Minor scheme and updated by the World Health Organization's Collaborating Centre for Reference and Research on Salmonella at the Pasteur Institute, Paris, France.

• MeSH
• lidé MeSH
• Salmonella enterica klasifikace   genetika   izolace a purifikace   MeSH
• salmonelová infekce u zvířat mikrobiologie   MeSH
• salmonelóza mikrobiologie   MeSH
• směrnice jako téma MeSH
• terminologie jako téma * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

A set of 487 clinical isolates of Salmonella Enteritidis were phage typed and analyzed for virulence markers, i.e. bacterial cell surface hydrophobicity, motility, biofilm formation and the presence of a 60 kb serovar-specific virulence plasmid. The most frequent phage type was PT8 (48.3%), followed by PT13a (7.2%), PT15 (6.4%), and PT4 (4.5%). Thirty-one (6.4%) strains were non-typeable. As many as 128 (26.3%) strains showed hydrophobicity in the hydrocarbon xylene adherence assay, with the highest percentages of highly hydrophobic strains being found among the following phage types: PT9a (84.6%), PT25 (81.8%), PT15 (54.8%) and PT8 (23.4%). Motility > or = 50 mm was observed in 294 (60.4%) strains and visible biofilm was formed in the test tube assay in different degrees by 448 (91.9%) strains. The capacity of in vitro biofilm formation is indicative of a high virulence potential of the study strains. The 60 kb serovar-specific virulence plasmid was present in 467 (95.9%) strains. Clear correlation between phage types and particular virulence markers was not revealed in the study set of strains. The obtained in vitro results are suggestive of flexibility of Salmonella Enteritidis in infecting the host.

• MeSH
• fagotypizace * MeSH
• fágy salmonel MeSH
• faktory virulence analýza   MeSH
• gastroenteritida mikrobiologie   MeSH
• lidé MeSH
• Salmonella enteritidis klasifikace   izolace a purifikace   patogenita   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Názvy látek
• faktory virulence MeSH