An experimental group of one-day-old chicken from a commercial hatchery was given a defined mixture of 7 gut anaerobes. The next day the chicks were inoculated by an APEC strain O78:H4-ST117 resistant to ciprofloxacin, alongside with the control group and monitored for 4 wk after the inoculation for the presence of the colonizing strains and ciprofloxacin-resistant E. coli. Significant reduction of colonization rates in the first 2 wk was recorded in the experimental group for the numbers of ciprofloxacin-resistant E. coli. The results show that colonization of chicken by defined anaerobic mixtures may provide a decisive protection during the critical period of the chicken intestinal microflora development.
- MeSH
- Bacteroides MeSH
- ciprofloxacin farmakologie MeSH
- Escherichia coli MeSH
- infekce vyvolané Escherichia coli * prevence a kontrola veterinární MeSH
- kur domácí MeSH
- nemoci drůbeže * prevence a kontrola MeSH
- probiotika * farmakologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Drůbež a drůbeží produkty jsou považovány za potenciálně významný zdroj rezistentních kmenů E. coli a jejich determinant kódovaných mobilními elementy, jako jsou geny pro ESBL/AmpC betalaktamázy. Drůbež je také hostitelem kmenů s extraintestinální patogenitou (ExPEC). Podobnost drůbežích APEC s humánními kmeny vedla k zařazení ExPEC mezi bakterie se zoonotickým potenciálem. Populační diverzita různých patotypů E. coli, druhová specifita a cirkulace různých kmenů a genetických determinant mezi populacemi jsou však velmi složité a výše uvedená rizika nelze řešit odděleně jedno od druhého.
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The present pilot study aimed at evaluating air sampling as a novel method for monitoring Campylobacter in poultry farms. We compared the bacteriological isolation of Campylobacter from boot swabs and air filter samples using ISO 10272-1:2017. A secondary aim was to evaluate the use of molecular methods, i.e. real time PCR, on the same sample set. Samples from 44 flocks from five European countries were collected, and included air samples, in parallel with boot swabs. Campylobacter spp. was isolated from seven of 44 boot swabs from three of five partners using the enrichment method. Two of these positive boot swab samples had corresponding positive air samples. Using enrichment, one positive air sample was negative in the corresponding boot swabs, but Campylobacter spp. was isolated from direct plating of the boot swab sample. One partner isolated Campylobacter spp. from six of 10 boot swabs using direct plating. Overall, 33 air filter samples were screened directly with PCR, returning 14 positive results. In conclusion, there was a lack of correspondence between results from analysis of boot swabs and air filters using ISO 10272-1:2017. In contrast, the combination of air filters and direct real-time PCR might be a way forward. Despite the use of the detailed ISO protocols, there were still sections that could be interpreted differently among laboratories. Air sampling may turn into a multi-purpose and low-cost sampling method that may be integrated into self-monitoring programs.
- MeSH
- Campylobacter genetika izolace a purifikace MeSH
- drůbež mikrobiologie MeSH
- farmy statistika a číselné údaje MeSH
- feces mikrobiologie MeSH
- internacionalita MeSH
- kampylobakterové infekce veterinární MeSH
- kur domácí mikrobiologie MeSH
- mikrobiologie vzduchu normy MeSH
- nemoci drůbeže mikrobiologie prevence a kontrola přenos MeSH
- pilotní projekty MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- multicentrická studie MeSH
- Geografické názvy
- Evropa MeSH
Lymphocytes represent the key antigen-specific leukocyte subpopulation. Despite their importance in mounting an immune response, an unbiased description of proteins expressed by chicken lymphocytes has not been presented. In this study, we therefore intravenously infected chickens with Salmonella Enteritidis, sorted CD4, CD8 and γδ T-lymphocytes from the spleen by flow cytometry and determined the proteome of each population by LC-MS/MS. CD4 T-lymphocyte characteristic proteins included ubiquitin SUMO-like domain and BAR domain containing proteins. CD8 T-lymphocyte specific proteins were characterized by purine ribonucleoside triphosphate binding and were involved in cell differentiation, cell activation and regulation of programmed cell death. γδ T-lymphocyte specific proteins exhibited enrichment of small GTPase of Rab type and GTP binding. Following infection, inducible proteins in CD4 lymphocytes included ribosomal proteins and downregulated proteins localized to the lysosome. CD8 T-lymphocytes induced MCM complex proteins, proteins required for DNA replication and machinery for protein processing in the endoplasmic reticulum. Proteins inducible in γδ T-lymphocytes belonged to immune system response, oxidative phosphorylation and the spliceosome. In this study, we predicted the likely events in lymphocyte response to systemic bacterial infection and identified proteins which can be used as markers specific for each lymphocyte subpopulation.
- MeSH
- CD4-pozitivní T-lymfocyty imunologie metabolismus mikrobiologie MeSH
- CD8-pozitivní T-lymfocyty imunologie metabolismus mikrobiologie MeSH
- intraepiteliální lymfocyty imunologie metabolismus mikrobiologie MeSH
- kur domácí imunologie metabolismus MeSH
- nemoci drůbeže imunologie metabolismus mikrobiologie prevence a kontrola MeSH
- Salmonella enteritidis imunologie metabolismus MeSH
- salmonelové vakcíny imunologie MeSH
- salmonelóza imunologie metabolismus mikrobiologie prevence a kontrola MeSH
- tandemová hmotnostní spektrometrie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Clostridium perfringens-induced necrotic enteritis is generally controlled by antibiotics. However, because of increasing antibiotic resistance, other antibacterial agents are required, preferably ones that do not affect the beneficial intestinal microbiota of the host. This study evaluated the in vitro selective growth-inhibitory effect of 8-hydroxyquinoline (8HQ) on C. perfringens vs. bifidobacteria in a medium containing chicken ileal digesta. Prior to the experiments, the minimum inhibitory concentrations of 8HQ and penicillin G were determined by broth microdilution assay. The minimum inhibitory concentration values of 8HQ for C. perfringens were 16-32 times lower than the values for bifidobacteria. Treatment of autoclaved and non-autoclaved chicken ileal digesta with 8HQ showed a selective anticlostridial effect. After incubation of C. perfringens with autoclaved ileal digesta for 3 h, all 8HQ concentrations tested (32-2048 μg/mL) significantly reduced C. perfringens bacterial count. In contrast, the same treatment had no or only a slight effect on bifidobacteria counts. Unlike 8HQ, penicillin G did not exhibit any selectivity. Similar results were obtained after incubation for 24 h. In non-autoclaved ileal digesta, all 8HQ concentrations tested significantly reduced C. perfringens bacterial counts after incubation for 30 min and 3 h, while no effect was observed on bifidobacteria. These results suggest that 8HQ may serve as a prospective veterinary compound for use against necrotic enteritis in poultry.
- MeSH
- antibakteriální látky farmakologie MeSH
- Bifidobacterium účinky léků MeSH
- Clostridium perfringens účinky léků MeSH
- ileum mikrobiologie MeSH
- klostridiové infekce prevence a kontrola veterinární virologie MeSH
- kur domácí mikrobiologie MeSH
- nemoci drůbeže prevence a kontrola virologie MeSH
- oxychinolin farmakologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The colonization of poultry with different Salmonella enterica serovars poses an issue throughout the world. In this study we therefore tested the efficacy of a vaccine consisting of attenuated strains of Salmonella enterica serovars Enteritidis, Typhimurium and Infantis against challenge with the same serovars and with S. Agona, Dublin and Hadar. We tested oral and aerosol administration of the vaccine, with or without co-administration of cecal microbiota from adult hens. The protective effect was determined by bacterial counts of the challenge strains up to week 18 of life and by characterizing the immune response using real-time PCR specific for 16 different genes. We have shown that a vaccine consisting of attenuated S. Enteritidis, S. Typhimurium and S. Infantis protected chickens against challenge with the wild type strains of the same serovars and partially protected chickens also against challenge with isolates belonging to serovars Dublin or Hadar. Aerosol vaccination was more effective at inducing systemic immunity whilst oral vaccination stimulated a local immune response in the gut. Co-administration of cecal microbiota increased the protectiveness in the intestinal tract but slightly decreased the systemic immune response. Adjusting the vaccine composition and changing the administration route therefore affects vaccine efficacy.
- MeSH
- atenuované vakcíny terapeutické užití MeSH
- kombinované vakcíny terapeutické užití MeSH
- kur domácí imunologie mikrobiologie MeSH
- nemoci drůbeže imunologie prevence a kontrola MeSH
- Salmonella enteritidis imunologie MeSH
- Salmonella typhimurium imunologie MeSH
- Salmonella imunologie MeSH
- salmonelová infekce u zvířat imunologie mikrobiologie prevence a kontrola MeSH
- salmonelové vakcíny imunologie terapeutické užití 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
In order to design a new Salmonella enterica vaccine, one needs to understand how naive and immune chickens interact differently when exposed to S. enterica. In this study we therefore determined the immune response of vaccinated and non-vaccinated chickens after intravenous infection with Salmonella enterica serovar Enteritidis (S. Enteritidis). Using flow cytometry we showed that 4 days post infection (DPI), counts of CD4 and B-lymphocytes did not change, CD8 and γδ T-lymphocytes decreased and macrophages and heterophils increased in the spleen. When vaccinated and non-vaccinated chickens were compared, only macrophages and heterophils were found in significantly higher counts in the spleens of the non-vaccinated chickens. The non-vaccinated chickens also expressed higher anti-LPS antibodies than the vaccinated chickens. The expression of interleukin (IL)1β, IL6, IL8, IL18, LITAF, IFNγ and iNOS did not exhibit any clear pattern in the cells sorted from the spleens of vaccinated or non-vaccinated chickens. Only IL17 and IL22 showed a differential expression in the CD4 T-lymphocytes of the vaccinated and non-vaccinated chickens at 4 DPI, both being expressed at a higher level in the non-vaccinated chickens. Due to a similar IFNγ expression in the CD4 T-lymphocytes in both the vaccinated and non-vaccinated chickens, and a variable IL17 expression oscillating around IFNγ expression levels, the IL17∶IFNγ ratio in CD4 T-lymphocytes was found to be central for the outcome of the immune response. When IL17 was expressed at higher levels than IFNγ in the non-vaccinated chickens, the Th17 immune response with a higher macrophage and heterophil infiltration in the spleen dominated. However, when the expression of IL17 was lower than that of IFNγ as in the vaccinated chickens, the Th1 response with a higher resistance to S. Enteritidis infection dominated.
- MeSH
- CD4-pozitivní T-lymfocyty cytologie MeSH
- cytokiny metabolismus MeSH
- ELISA metody MeSH
- interferon gama metabolismus MeSH
- interleukin-17 metabolismus MeSH
- kur domácí MeSH
- leukocyty cytologie MeSH
- lipopolysacharidy metabolismus MeSH
- makrofágy metabolismus MeSH
- nemoci drůbeže imunologie prevence a kontrola MeSH
- polymerázová řetězová reakce metody MeSH
- průtoková cytometrie metody MeSH
- Salmonella enteritidis metabolismus MeSH
- salmonelové vakcíny metabolismus MeSH
- salmonelóza imunologie prevence a kontrola MeSH
- slezina cytologie MeSH
- Th1 buňky cytologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
In this study we were interested in the vaccine potential of two attenuated mutants of Salmonella enterica serovar Enteritidis for poultry. The first mutant was attenuated by the removal of the whole Salmonella Pathogenicity Island 1 (SPI1) and the second mutant was devoid of the whole SPI2. These 2 mutants were used for oral vaccination of 2 chicken lines; Lohmann Brown and ISA Brown. Chickens were vaccinated orally on day 1 of life, revaccinated on day 21 and challenged on day 42. The challenge was performed either orally or intravenously. Despite a slightly different response between the two chicken lines, both the mutants gave protection to poultry against S. Enteritidis challenge as documented by findings such as the bacterial counts in tissues, spleen weight, antibody production and cytokine response (namely IL-17 and IL-22). When the 2 mutants were compared, vaccination with the SPI1 mutant proved to be more effective in the protection of poultry against S. Enteritidis challenge than the vaccination with the SPI2 mutant. On the other hand, vaccination with the SPI2 mutant stimulated a slightly higher antibody production and such a mutant might therefore be a better choice if Salmonella is used as a vector for the delivery of heterologous antigens with a desired stimulation of the humoral part of the immune system.
- MeSH
- aplikace orální MeSH
- atenuované vakcíny aplikace a dávkování genetika imunologie MeSH
- bakteriální nálož MeSH
- cytokiny sekrece MeSH
- genomové ostrovy MeSH
- kur domácí MeSH
- leukocyty mononukleární imunologie MeSH
- nemoci drůbeže imunologie prevence a kontrola MeSH
- protilátky bakteriální krev MeSH
- Salmonella enteritidis imunologie patogenita MeSH
- salmonelová infekce u zvířat imunologie mikrobiologie patologie prevence a kontrola MeSH
- salmonelové vakcíny aplikace a dávkování genetika imunologie MeSH
- sekundární imunizace metody MeSH
- slezina patologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH