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
Pseudorabies virus (PrV), a causative agent of Aujeszky's disease, is deadly to most mammals with the exception of higher primates and men. This disease causes serious economic loses among farm animals, especially pigs, yet many European countries are today claimed to be Aujeszky's disease free because of the discovery of an efficient vaccination for pigs. In reality, the virus is still present in wild boar. Current vaccines are neither suitable for dogs nor are there anti-PrV drugs approved for veterinary use. Therefore, the disease still represents a high threat, particularly for expensive hunting dogs that can come into close contact with infected boars. Here we report on the anti-PrV activities of a series of synthetic diaminopurine-based acyclic nucleoside phosphonate (DAP-ANP) analogues. Initially, all synthetic DAP-ANPs under investigation are shown to exhibit minimal cytotoxicity by MTT and XTT tests (1-100μM range). Thereafter in vitro infection models are established using PrV virus SuHV-1, optimized on PK-15 and RK-13 cell lines. Out of the six DAP-ANP analogues tested, analogue VI functionalized with a cyclopropyl group on the 6-amino position of the purine ring proves the most effective antiviral DAP-ANP analogue against PrV infection, aided by sufficient hydrophobic character to enhance bioavailability to its cellular target viral DNA-polymerase. Four other DAP-ANP analogues with functional groups introduced to the C2'position are shown ineffective against PrV infection, even with favourable hydrophobic properties. Cidofovir(®), a drug approved against various herpesvirus infections, is found to exert only low activity against PrV in these same in vitro models.
- MeSH
- 2-aminopurin analogy a deriváty chemie farmakologie MeSH
- antivirové látky chemie farmakologie MeSH
- buněčné linie MeSH
- buňky MDCK MeSH
- Herpesviridae účinky léků MeSH
- organofosfonáty chemie farmakologie MeSH
- pseudovzteklina farmakoterapie MeSH
- psi MeSH
- replikace DNA účinky léků MeSH
- techniky in vitro MeSH
- transmisní elektronová mikroskopie MeSH
- Vero buňky MeSH
- zvířata MeSH
- Check Tag
- psi MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Ingestion of raw or undercooked meat is a potential source of human toxoplasmosis. The aim of this study was to determine the viability of Toxoplasma gondii cysts in vacuum packed (VP) goat meat and in dry fermented sausages (DFS), and evaluate certain physical and chemical parameters, like water activity (aw), pH value, content of salt, dry matter and fat. A portion of muscle tissue from experimentally infected animals was used for production of VP meat with or without addition of 2.5% curing salt, and stored at 4 °C or at -20 °C. Results of bioassay showed that, samples of vacuum packed Toxoplasma positive meat without salt addition were alive after six weeks at 4 °C. Incubation at -20 °C supported the viability after 3 h, but not after 4 h. After 7 days in 2.5% of curing salt, samples of T. gondii VP goat meat were still viable, but not after 14 days at 4 °C. All the DFS samples were not positive for infective cysts which mean that, they do not pose a risk of T. gondii transmission. These data suggest that vacuum packaging increases the survival of T. gondii cysts.
- MeSH
- kosterní svaly parazitologie MeSH
- kozy MeSH
- lidé MeSH
- masné výrobky analýza parazitologie MeSH
- maso analýza parazitologie MeSH
- nemoci koz parazitologie MeSH
- obaly potravin MeSH
- spotřebitelská bezpečnost produktů MeSH
- Toxoplasma genetika růst a vývoj izolace a purifikace MeSH
- toxoplazmóza zvířat parazitologie MeSH
- vakuum MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
It has been suggested that passive shedding of Mycobacterium avium subsp. paratuberculosis (MAP) in faeces may occur, but reliable data are missing. Passive shedding assumes the ingestion of MAP in contaminated feed and passive passage through the gastrointestinal tract without causing infection. In this study the presence of MAP in faeces in a closed herd of Limousin cattle was monitored for 53 months using quantitative real time PCR (qPCR) and culture. The initial prevalence of MAP in the herd was determined to be 63.4% and 4.9% using qPCR and culture, respectively. After the removal of two culture- and qPCR-positive (>10(4) MAP cells/g) cows, the prevalence of MAP using qPCR decreased to 42.1% and later to 15.6% and 6.7%. The continuous removal of suspected animals from the herd during the monitoring period minimised the presence of MAP in faeces to sporadic, which may have resulted from a decrease in the environmental infectious pressure. The findings suggest that the presence of low numbers of MAP in bovine faeces may not necessarily be caused by real infection, but rather by passive passage of MAP. This phenomenon should therefore be considered when interpreting MAP qPCR data.
- MeSH
- ELISA veterinární MeSH
- feces mikrobiologie MeSH
- kvantitativní polymerázová řetězová reakce veterinární MeSH
- Mycobacterium avium subsp. paratuberculosis genetika izolace a purifikace MeSH
- nemoci skotu epidemiologie mikrobiologie MeSH
- paratuberkulóza epidemiologie mikrobiologie MeSH
- prevalence MeSH
- roční období MeSH
- skot MeSH
- vylučování bakterií z těla * MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Francie MeSH
The breeding of domestic rabbit (Oryctolagus cuniculus) for human consumption has a long tradition mainly in European and Asian countries. Infections that can affect the production of meat or even be transmitted from animals to humans are important to monitor, especially for public health reasons as well as for their impact on animals health. This study aimed to collect sera from rabbits bred in different conditions and test the presence of Toxoplasma gondii and Encephalitozoon cuniculi antibodies. Whether infections were active or latent was assessed by determining the occurrence of IgM or IgM together with IgG antibodies which indicated active infection whereas latent infection was characterized by finding IgG antibodies only. An ELISA test was performed with 1883 sera samples collected throughout the Czech and Slovak Republics. The seroprevalence of T. gondii in 902 samples from 6 commercial farms (CF) was very low with only 4 rabbits (0.4%) being positive. In total 99 (10.1%) individuals out of 981 samples from 29 household farms (HF) were positive for T. gondii antibodies. Only 2 (50%) of the T. gondii positive CF rabbits had active infections while the rest were latently infected. The serological results showed that 35 (35.4%) rabbits from the T. gondii positive HF group suffered from active infection. Out of CF samples 185 (20.5%) were positive for E. cuniculi. Antibodies of E. cuniculi were detected in 497 (50.7%) HF rabbits. Active E. cuniculi infections were determined in 85.9% of CF and 56.3% of HF rabbits; respectively. Interestingly, the E. cuniculi positive rabbits were significantly more often positive for anti-T. gondii antibodies in comparison to E. cuniculi negative individuals. Prevalence of T. gondii in CF rabbits was negligible. According to our results meat of HF rabbits still poses a risk of T. gondii infection. Nevertheless, the risk is on its lowest level in 20 years which is apparently caused due to changes in feeding practices. The occurrence of E. cuniculi antibodies was significantly lower in rabbits from commercial farms, apparently because of better hygiene conditions.
- MeSH
- ELISA veterinární MeSH
- Encephalitozoon cuniculi imunologie izolace a purifikace MeSH
- encephalitozoonóza veterinární MeSH
- králíci imunologie parazitologie MeSH
- prevalence MeSH
- protilátky protozoální krev MeSH
- séroepidemiologické studie MeSH
- Toxoplasma imunologie izolace a purifikace MeSH
- toxoplazmóza zvířat epidemiologie MeSH
- veřejné zdravotnictví MeSH
- zvířata MeSH
- Check Tag
- králíci imunologie parazitologie MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
- Slovenská republika MeSH
Undercooked meat containing tissue cysts is one of the most common sources of Toxoplasma gondii infection in humans. Goats are very susceptible to clinical toxoplasmosis, and especially kids are common food animals, thereby representing a risk for human infection. A sequence-specific magnetic capture method was used for isolation of T. gondii DNA from tissue samples from experimentally infected goat-kids and real-time PCR for the 529 bp repeat element allowed quantification of T. gondii DNA. The contamination level in different types of tissue and in two groups of goats euthanized 30 and 90 dpi was compared. The highest concentration of T. gondii DNA in both groups of goats was found in lung tissue, but only the higher parasite count in lung tissue compared to other organs in group A (euthanized 30 dpi) was statistically significant. T. gondii concentrations were higher in liver and dorsal muscle samples from goats euthanized 90 dpi than in goats euthanized at 30 dpi, while the T. gondii concentration in hearts decreased. This study describes for the first time distribution of T. gondii parasites in post-weaned goat kids. New information about T. gondii predilection sites in goats and about the progression of infection between 30 and 90 dpi was achieved.
- MeSH
- játra parazitologie MeSH
- kosterní svaly parazitologie MeSH
- kozy MeSH
- kvantitativní polymerázová řetězová reakce metody veterinární MeSH
- magnetismus MeSH
- mozek parazitologie MeSH
- nemoci koz diagnóza parazitologie MeSH
- plíce parazitologie MeSH
- slezina parazitologie MeSH
- srdce parazitologie MeSH
- Toxoplasma izolace a purifikace MeSH
- toxoplazmóza zvířat diagnóza MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
We designed and synthesised a series of new cationic lipids based on spermine linked to various hydrophobic anchors. These lipids could be potentially useful for the preparation of stable cationic liposomes intended for the construction of drug targeting systems applicable in the field of anticancer/antiviral therapy, vaccine carriers, and vectors for the gene therapy. Low in vitro toxicity was found for these compounds, especially for LD1, in several cell lines. The delivery of both a fluorescence marker (calcein) and antiviral drugs into cells has been achieved owing to a large extent of internalization of cationic liposomes (labelled by Lyssamine-Rhodamine PE or fluorescein-PE) as demonstrated by fluorescent microscopy and quantified by flow cytometry. The bovine herpes virus type 1 (BHV-1) virus infection in vitro model using MDBK cells was employed to study the effect of the established antiviral drug HPMPC (Cidofovir®) developed by Prof. A. Holý. Inhibition of BHV-1 virus replication was studied by quantitative RT-PCR and confirmed by both Hoffman modulation contrast microscopy and transmission electron microscopy. We found that in vitro antiviral activity of HPMPC was significantly improved by formulation in cationic liposomes, which decreased the viral replication by about 2 orders of magnitude.
- MeSH
- antivirové látky aplikace a dávkování farmakologie MeSH
- bovinní herpesvirus 1 účinky léků fyziologie MeSH
- buněčné kultury MeSH
- buněčné linie MeSH
- cytopatogenní efekt virový MeSH
- cytosin aplikace a dávkování analogy a deriváty farmakologie MeSH
- fluorescenční mikroskopie MeSH
- kationty MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- ledviny cytologie virologie MeSH
- lipidy chemie MeSH
- liposomy MeSH
- nosiče léků chemie MeSH
- organofosfonáty aplikace a dávkování farmakologie MeSH
- replikace viru účinky léků MeSH
- skot MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Encephalitozoon cuniculi is an obligate intracellular pathogen that has a wide host distribution, but primarily affects rabbits. The aim of this study was to characterize both the cell-mediated and the antibody response in rabbits after experimental infection using 2 different infection routes: oral and ocular. SPF rabbits were infected with low (10³ spores) and high (10⁷ spores) infection doses. Monitored parameters included clinical signs, detection of spores in urine, antibody response detected with ELISA, and cell-mediated immunity detected by antigen-driven lymphocyte proliferation. At week 13 post-infection, half of the rabbits in each group were suppressed by intramuscular administration of dexamethasone. At week 18 post-infection, animals were euthanized. Clinical signs were mild with exacerbation after immunosuppression. Spores in urine and antigen-specific cell-mediated immunity were detected from weeks 5 and 4 post-infection, respectively. Specific IgM was detected 1 week after infection, and IgG antibodies followed 1 week later in rabbits infected with the high dose. Immunological responses were dose dependent. The authors can conclude that both oral and ocular experimental infection with E. cuniculi resulted in an immune response of the infected animals. Rabbits could be used as an experimental model for the study of ocular microsporidiosis.
- MeSH
- aktivace lymfocytů MeSH
- buněčná imunita MeSH
- Encephalitozoon cuniculi imunologie patogenita MeSH
- encephalitozoonóza imunologie parazitologie patologie MeSH
- imunoglobulin G krev MeSH
- imunoglobulin M krev MeSH
- králíci MeSH
- modely nemocí na zvířatech MeSH
- nemoci úst imunologie parazitologie patologie MeSH
- oční infekce imunologie parazitologie patologie MeSH
- outbrední kmeny zvířat MeSH
- protilátky protozoální krev MeSH
- tvorba protilátek MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Encephalitozoon cuniculi is an obligate intracellular pathogen that has wide host distribution, but primary affects rabbits. This study presents a seroepidemiological study of E. cuniculi infection in 500 pet rabbits from the Czech Republic using ELISA capable of measuring IgM and IgG antibodies. Specific IgM antibodies, reflecting acute, reactivated infection or reinfection, were detected in 32.4% of all rabbits. IgG antibodies indicating chronic infection, were presented in 68.0% of all rabbits. The highest detection rate of IgM (54.4%) and IgG (86.1%) antibodies was ascertained in rabbits with neurological symptoms (n=79, group I). In rabbits with renal disorders (n=47, group II) 36.2% animals were specific IgM and 80.9% IgG positive. Out of 9 rabbits with ocular disorders (group III), 44.4% were positive for anti-E. cuniculi IgM and 77.8% for IgG antibodies. In rabbits with multiple signs (neurological and renal or ocular, n=16, group IV), 43.8% animals were specific IgM and 68.8% IgG positive. Out of 287 rabbits with other disease (group V), 26.5% were positive for anti-E. cuniculi IgM and 64.1% for IgG antibodies. However, the high presence of IgM (24.2%) and IgG (51.6%) antibodies was detected in clinically healthy rabbits (n=62, group VI). Toxoplasma gondii infection should be considered as a differential diagnosis for neurological and ocular disorders in rabbits. Using ELISA, 19.2% from all rabbits were positive for specific anti-T. gondii IgG. The highest seropositivity was detected in group III (44.4%). Simultaneous testing of IgM and IgG specific antibodies give an indication of the infection status. Presence of IgM antibodies is indicative for active infection with requirement to institute proper antimicrosporidial therapy. As active infection was detected in considerably high numbers of rabbits with clinical signs that are not usually associated with E. cuniculi, and even in asymptomatic rabbits, detection of both isotypes of specific antibodies should be a routine part of a health check in rabbits.
- MeSH
- ELISA veterinární MeSH
- Encephalitozoon cuniculi izolace a purifikace MeSH
- encephalitozoonóza diagnóza imunologie mikrobiologie veterinární MeSH
- hospodářská zvířata imunologie mikrobiologie MeSH
- imunoglobulin G krev MeSH
- imunoglobulin M krev MeSH
- králíci imunologie mikrobiologie MeSH
- protilátky bakteriální krev MeSH
- rozdělení chí kvadrát MeSH
- séroepidemiologické studie MeSH
- zvířata MeSH
- Check Tag
- králíci imunologie mikrobiologie MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
