Porcine cytomegalovirus (PCMV) causes lifelong latent infections in swine. The pathogen is occasionally associated with inclusion body rhinitis and pneumonia in piglets, reproductive disorders in pregnant sows and respiratory disease complex in older pigs. Immunosuppressive potential of PCMV infection is discussed. Macrophages were recognised as one of target cell types where propagation of virus occurs. The aim of present study was to set up model PCMV infection of monocyte derived macrophages (MDMs) in vitro for PCMV immunobiology research. Obtained results showed that PCMV is able to infect and propagate in MDMs. Possible immunosuppressive effect of PCMV on infected macrophages was evaluated by measurement of immune relevant gene expression in MDMs. Infection decreased expression of IL-8 and TNF-α (pro-inflammatory cytokines) and increased expression of IL-10 (anti-inflammatory cytokine) on mRNA transcription level. Obtained data support hypothesis that higher sensitivity of animals to coinfection with other swine pathogens and its more severe clinical manifestations could potentially be the consequence of PCMV infection.

• MeSH
• cytokiny imunologie   MeSH
• cytomegalovirové infekce imunologie   veterinární   MeSH
• Cytomegalovirus fyziologie   MeSH
• exprese genu imunologie   MeSH
• interleukin-10 imunologie   MeSH
• interleukin-8 imunologie   MeSH
• makrofágy imunologie   virologie   MeSH
• nemoci prasat imunologie   virologie   MeSH
• prasata MeSH
• přirozená imunita MeSH
• TNF-alfa imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• 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.

• 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

Porcine reproductive and respiratory syndrome virus (PRRSV) can predispose pigs to secondary respiratory infection with bacteria such as Haemophilus parasuis. Animals infected with both pathogens develop more severe clinical disease. The immune response of porcine alveolar macrophages (PAMs) to simultaneous infection with PRRSV and H. parasuis was analysed in vitro, describing cytokine production, expression of cell surface molecules, and production of reactive oxygen species (ROS). Concurrent infection with PRRSV and H. parasuis increased gene expression of pro-inflammatory cytokines (TNF-α, IL-1β, IL-8) in PAMs in comparison with PAMs infected with PRRSV or H. parasuis alone. An additive effect of dual infection on IL-1β production was confirmed at the protein level. PAMs infected with PRRSV showed increased production of ROS compared to controls. Conversely, simultaneous infection of PAMs with PRRSV and H. parasuis decreased production of ROS, indicating the presence of an H. parasuis defence mechanism against respiratory burst. Concurrent infection of PAMs with PRRSV and H. parasuis was shown to elicit a pro-inflammatory immune response represented by significant IL-1β production. Severe multifactorial respiratory disease in natural conditions caused by both pathogens could be the consequence of pro-inflammatory mediated immunopathology.

• MeSH
• alveolární makrofágy imunologie   mikrobiologie   virologie   MeSH
• biologické markery analýza   MeSH
• cytokiny genetika   imunologie   MeSH
• Haemophilus parasuis imunologie   MeSH
• hemofilové infekce komplikace   imunologie   veterinární   virologie   MeSH
• koinfekce veterinární   MeSH
• kultivované buňky MeSH
• messenger RNA metabolismus   MeSH
• nemoci prasat imunologie   mikrobiologie   virologie   MeSH
• prasata MeSH
• regulace exprese virových genů MeSH
• regulace genové exprese u bakterií MeSH
• reprodukční a respirační syndrom prasat imunologie   virologie   MeSH
• virus reprodukčního a respiračního syndromu prasat imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Haemophilus parasuis (H. parasuis) is associated with meningitis, polyserositis, polyarthritis and bacterial pneumonia. At present, its prevention and control is difficult because of the lack of suitable subunit vaccines. Nowadays, high-throughput methods, immunoproteomics, are available to screen for more vaccine candidates. A protein extraction method for H. parasuis and two-dimensional electrophoresis (2-DE) were optimized to provide high-resolution profiles covering pH 3 to 10. Twenty immunoreactive spots were excised from gels after strict comparison between 2-DE Western blot membranes and the relevant gels. Matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) and MALDI-TOF-TOF-MS successfully identified 16 different proteins. Fifteen of them were reported as immunoreactive proteins in H. parasuis for the first time. In addition, recombinant HP5-7 (ABC transporter, periplasmic-binding protein) showed immunoreactivity both with hyperimmune rabbit serum and convalescent swine serum. Four recombinants of the 14 successfully expressed genes showed immunoreactivity with hyperimmune rabbit serum.

• MeSH
• 2D gelová elektroforéza MeSH
• bakteriální proteiny chemie   genetika   imunologie   MeSH
• Haemophilus parasuis chemie   klasifikace   genetika   imunologie   MeSH
• hemofilové infekce imunologie   mikrobiologie   veterinární   MeSH
• hmotnostní spektrometrie MeSH
• králíci MeSH
• molekulární sekvence - údaje MeSH
• nemoci prasat imunologie   mikrobiologie   MeSH
• prasata MeSH
• proteomika MeSH
• protilátky bakteriální analýza   imunologie   MeSH
• séroskupina MeSH
• western blotting MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Čína MeSH

Piglets from 4 to 8 weeks of age originated from a Cryptosporidium-free research breed were orally inoculated with 1 × 10(6) infectious oocysts of Cryptosporidium scrofarum. The number of shed oocysts per gram of faeces served to describe the infection intensity and prepatent period. In addition, faecal samples collected daily and tissue samples of the small and large intestine collected at 30 days post-inoculation were examined for the C. scrofarum small subunit ribosomal RNA gene using PCR. The piglets inoculated at 4-weeks of age remained uninfected, whereas 5-week-old and older animals were fully susceptible with a prepatent period ranging from 4 to 8 days. Susceptible pigs shed oocysts intermittently, and shedding intensity, reaching a mean maximum of 6000 oocysts per gram, did not differ significantly among infected animals. This study demonstrates that pigs become susceptible to C. scrofarum infection as late as 5-weeks of age. The mechanisms of age related susceptibility remain unknown.

• MeSH
• Cryptosporidium genetika   MeSH
• feces parazitologie   MeSH
• geny rRNA genetika   MeSH
• kryptosporidióza imunologie   patologie   veterinární   MeSH
• nemoci prasat imunologie   patologie   MeSH
• počet parazitárních vajíček MeSH
• prasata MeSH
• věkové faktory MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem 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.

• 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

Monocytes play an essential role in the defense against bacterial pathogens. Bone marrow (BM) and peripheral blood (PB) monocytes in pigs consist of the main "steady-state" subpopulations: CD14 hi/CD163-/SLA-DR- and CD14 low/CD163+/SLA-DR+. During inflammation, the subpopulation of "inflammatory" monocytes expressing very high levels of CD163, but lacking the SLA-DR molecule (being CD14 low/CD163+/SLA-DR-) appears in the BM and PB and replaces the CD14 low/CD163+/SLA-DR+ subpopulation. However, current knowledge of monocyte migration into inflamed tissues in pigs is limited. The aim of the present study was to evaluate the distribution of "inflammatory" CD14 low/CD163+/SLA-DR- monocytes during experimental inflammation induced by Actinobacillus pleuropneumoniae (APP) and a possible role for chemokines in attracting "inflammatory" CD14 low/CD163+/SLA-DR- monocytes into the tissues. Monocyte subpopulations were detected by flow cytometry. Chemokines and chemokine receptors were detected by RT-qPCR. The "steady-state" monocytes were found in the BM, PB, spleen and lungs of control pigs. After APP-infection, "inflammatory" monocytes replaced the "steady-state" subpopulation in BM, PB, spleen and moreover, they appeared in an unaffected area, demarcation zone and necrotic area of the lungs and in tracheobronchial lymph nodes. They did not appear in mesenteric lymph nodes. Levels of mRNA for various chemokines with their appropriate receptors were found to be elevated in BM (CCL3-CCR1/CCR5, CCL8-CCR2/CCR5, CCL19-CCR7), necrotic area of the lungs (CCL3-CCR1, CCL5-CCR1/CCR3, CCL11-CCR3, CCL22/CCR4) and tracheobronchial lymph nodes (CCL3-CCR1) and therefore they could play a role in attracting monocytes into inflamed tissues. In conclusion, "inflammatory" monocytes appear in different lymphoid tissues and the lungs after APP infection in pigs. Various chemokines could drive this process.

• MeSH
• Actinobacillus pleuropneumoniae fyziologie   MeSH
• antigeny diferenciační myelomonocytární metabolismus   MeSH
• CD antigeny metabolismus   MeSH
• chemokiny genetika   metabolismus   MeSH
• infekce bakteriemi rodu Actinobacillus imunologie   mikrobiologie   veterinární   MeSH
• lymfoidní tkáň metabolismus   MeSH
• messenger RNA genetika   MeSH
• monocyty cytologie   metabolismus   MeSH
• nemoci prasat imunologie   mikrobiologie   MeSH
• plíce metabolismus   MeSH
• prasata MeSH
• průtoková cytometrie veterinární   MeSH
• receptory buněčného povrchu metabolismus   MeSH
• receptory chemokinů genetika   metabolismus   MeSH
• zánět mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The aim of this work is to identify antigenic regions within the ORF1 protein of Torque teno sus virus 1 (TTSuV1) and Torque teno virus sus 2 (TTSuV2) that could be used as antigens to detect virus-specific antibodies following infection in pigs. Protein sequences of TTSuV ORF1 genes were analysed to predict linear antigenic epitopes. Synthesized peptides were analysed for serological reactivity with swine sera. Such an antigenic region was identified at the C terminus of the ORF1 protein of both viruses and showed serological reactivity with 78 % (TTSuV1) and 88 % (TTSuV2) of swine sera. An ELISA with an immunodominant peptide as antigen was used to examine the sera of piglets, aged 4-20 weeks, and adults. Results indicated that TTSuV1- and TTSuV2-specific antibodies were detectable at 4 weeks. Antibody titres increased from week 10 and peaked at week 20. A relatively high antibody titre persisted to adulthood.

• MeSH
• infekce DNA virem imunologie   veterinární   virologie   MeSH
• mapování epitopu MeSH
• molekulární sekvence - údaje MeSH
• nemoci prasat imunologie   virologie   MeSH
• prasata MeSH
• protilátky virové imunologie   MeSH
• sekvence aminokyselin MeSH
• Torque teno virus chemie   genetika   imunologie   MeSH
• virové plášťové proteiny chemie   genetika   imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The virulence factors EAST1 and AIDA are often detected in ETEC/VTEC strains isolated from pigs and their role in diarrhoeal infections is discussed. In order to elucidate the pathogenesis of AIDA, the colonisation patterns of F4 positive and AIDA positive strains were investigated. Two wild-type Escherichia coli strains AIDA/EAST1 and F4/EAST1 isolated from diarrhoeal piglets were used for animal experiment to evaluate the ability of the EAST1 toxin to be involved in induction of diarrhoea. Gnotobiotic piglets were supplemented with normal porcine serum and orally inoculated with the strains. Faecal bacterial shedding of the challenge strains was observed during the experiment. Light microscopy and scanning electron microscopy were used to detect the colonisation pattern of both challenge strains. Although bacterial isolation demonstrated shedding of the challenge strains until the end of the experiment, diarrhoea did not develop in any piglet. Based on histological examination, piglets were more heavily colonised in the case of infection with E. coli O149/F4/EAST1 strain. Scanning electron microscopy showed bacterial cells of F4/EAST1 E. coli adhering to enterocytes, in contrast to AIDA/EAST1 which were poorly present on the intestinal surface. The EAST1 toxin alone was not able to induce diarrhoea in animals. Therefore our results demonstrate that the function/role of EAST1 and AIDA in colibacillosis of pigs remains to be elucidated.

• MeSH
• bakteriální adheziny imunologie   MeSH
• bakteriální toxiny imunologie   MeSH
• enterotoxiny imunologie   MeSH
• Escherichia coli imunologie   MeSH
• feces mikrobiologie   MeSH
• gnotobiologické modely imunologie   MeSH
• imunohistochemie veterinární   MeSH
• infekce vyvolané Escherichia coli imunologie   mikrobiologie   veterinární   MeSH
• mikroskopie elektronová rastrovací veterinární   MeSH
• náhodné rozdělení MeSH
• nemoci prasat imunologie   mikrobiologie   MeSH
• nemoci střev imunologie   mikrobiologie   veterinární   MeSH
• neparametrická statistika MeSH
• prasata MeSH
• proteiny z Escherichia coli imunologie   MeSH
• průjem imunologie   mikrobiologie   veterinární   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The significance of maternal immunity against non-typhoid Salmonella spp. acquired by piglets via colostrum and milk was evaluated in a Salmonella enterica serovar Typhimurium challenge experiment. Piglets from sows vaccinated with an experimental inactivated vaccine exhibited high levels of serum immunoglobulins G and A against S. Typhimurium 4 days after birth, just prior to experimental oral challenge. The S. Typhimurium load in the ileal and caecal wall of piglets 3 days after experimental inoculation was lower by a 2-log magnitude compared to unvaccinated controls. Such a vaccine, delivering colostral/lactogenic immunity to piglets thus has the potential to reduce the prevalence non-typhoid Salmonella spp. infection.

• MeSH
• bakteriální nálož veterinární   MeSH
• bakteriální vakcíny imunologie   MeSH
• cékum mikrobiologie   MeSH
• ileum mikrobiologie   MeSH
• imunita získaná od matky * MeSH
• imunoglobulin A krev   MeSH
• imunoglobulin G krev   MeSH
• inaktivované vakcíny imunologie   MeSH
• kolostrum imunologie   MeSH
• mléko imunologie   MeSH
• nemoci prasat imunologie   mikrobiologie   MeSH
• novorozená zvířata MeSH
• prasata MeSH
• Salmonella typhimurium imunologie   MeSH
• salmonelová infekce u zvířat imunologie   mikrobiologie   MeSH
• slizniční imunita MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH