Early detection of biohazardous bacteria that can be misused as biological weapons is one of the most important measures to prevent the spread and outbreak of biological warfare. For this reason, many instrument platforms need to be introduced into operation in the field of biological warfare detection. Therefore the purpose of this study is to establish a new detection panel for biothreat bacteria (Bacillus anthracis, Yersinia pestis, Francisella tularensis, and Brucella spp.) and confirm it by collaborative validation by using a multiplex oligonucleotide ligation followed by polymerase chain reaction and hybridization to microspheres by MagPix detection platform (MOL-PCR). Appropriate specific sequences in bacterial DNA were selected and tested to assemble the detection panel, and MOLigo probes (short specific oligonucleotides) were designed to show no cross-reactivity when tested between bacteria and to decrease the background signal measurement on the MagPix platform. During testing, sensitivity was assessed for all target bacteria using serially diluted DNA and was determined to be at least 0.5 ng/µL. For use as a diagnostic kit and easier handling, the storage stability of ligation premixes (MOLigo probe mixes) was tested. This highly multiplex method can be used for rapid screening to prevent outbreaks arising from the use of bacterial strains for bioterrorism, because time of analysis take under 4 h.
- Publikační typ
- časopisecké články MeSH
Problems with parasitic infections are common in zoological gardens and circuses. In some animals it can lead to several disorders such as systemic disease, reproductive disorders (abortions and neonatal mortality), and even to death if severe illness is untreated. Thus, the aim of this study was to evaluate the prevalence of three common parasites in 74 animals from three zoos, and four circuses in Southern Italy. Antibodies to Toxoplasma gondii, Neospora caninum, and Encephalitozoon cuniculi were detected in 51%, 12%, and 20% of animals, respectively. Co-infections of T. gondii and N. caninum were reported in seven animals (9%) and co-infection of T. gondii and E. cuniculi in one animal. T. gondii, N. caninum and E. cuniculi seroprevalence differed in type of diet (P ≤ 0.0001; P ≤ 0.037 and P ≤ 0.004, respectively). T. gondii and E. cuniculi seroprevalence also differed in animal families (P ≤ 0.0001) and according to type of housing (P ≤ 0.003), respectively. Statistical differences were not found in other characteristics (gender, age, country of birth, origin, and contact with cats or dogs). This is the first serological study focusing on protozoan and microsporidian parasites in zoo and circus animals from Southern Italy and the first detection of antibodies to E. cuniculi in camels in Europe.
- MeSH
- Encephalitozoon cuniculi izolace a purifikace MeSH
- encephalitozoonóza epidemiologie parazitologie veterinární MeSH
- kokcidióza epidemiologie parazitologie veterinární MeSH
- Neospora izolace a purifikace MeSH
- prevalence MeSH
- savci * MeSH
- séroepidemiologické studie MeSH
- Toxoplasma izolace a purifikace MeSH
- toxoplazmóza zvířat epidemiologie parazitologie MeSH
- zvířata v ZOO MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Itálie MeSH
The aim of this study was to determine the seroprevalence of antibodies to B. burgdorferi s.l. in wild small mammals in the Czech Republic and compare sensitivity of PCR and cultivaton. Wild small mammals (n = 691) were trapped in years 2010-2014 in three localities of the Czech Republic. Heart rinses (n = 340) and sera (n = 351) were examined by modified indirect ELISA. Seventy animals were randomly selected for comparison of results of cultivation and PCR. Mean annual antiborelian positivity was 12% with statistical difference (p < 0.05) between Bank Vole (Clethrionomys glareolus) and other six animal species, while there was no statistical difference (p > 0.05) between rodentia and insectivora, gender and localities. The cultivation revealed one positive sample (1.4%), negative in both PCR and ELISA. Method PCR revealed seven positive samples (10%); two of them were simultaneously dubious in ELISA. Eleven animals, negative in cultivation and PCR, had antibodies in ELISA. Method of PCR compared to cultivation seems to be more sensitive for detection of Borrelia.
- MeSH
- Arvicolinae imunologie mikrobiologie MeSH
- Borrelia burgdorferi genetika růst a vývoj imunologie MeSH
- divoká zvířata imunologie mikrobiologie MeSH
- ELISA MeSH
- hlodavci imunologie mikrobiologie MeSH
- imunoglobulin G krev MeSH
- imunoglobulin M krev MeSH
- lymeská nemoc epidemiologie imunologie veterinární MeSH
- polymerázová řetězová reakce MeSH
- protilátky bakteriální krev MeSH
- savci imunologie mikrobiologie MeSH
- senzitivita a specificita MeSH
- séroepidemiologické studie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
Toxoplasma gondii, Neospora caninum, and Encephalitozoon cuniculi are important infectious agents, with T. gondii and E. cuniculi having zoonotic potential. There are two main clonal lineages (types I and II) of T. gondii in Europe, but little is known about genotypes of T. gondii in wild animals. The aim of our study was molecular detection of these three pathogens in tissues of wild red foxes ( Vulpes vulpes) from the Czech Republic. Using PCR (B1 gene), we detected T. gondii in 10% of the animals that we tested ( n=100); N. caninum and E. cuniculi were not detected. The T. gondii samples were genotyped by single multiplex PCR assay with 15 microsatellite markers. Five samples were successfully genotyped as genotype II, a unique finding for T. gondii isolated from red foxes from the Czech Republic.
- MeSH
- divoká zvířata MeSH
- Encephalitozoon cuniculi genetika izolace a purifikace MeSH
- encephalitozoonóza epidemiologie mikrobiologie veterinární MeSH
- genotyp MeSH
- kokcidióza epidemiologie parazitologie veterinární MeSH
- lišky parazitologie MeSH
- Neospora genetika izolace a purifikace MeSH
- Toxoplasma genetika izolace a purifikace MeSH
- toxoplazmóza zvířat epidemiologie parazitologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
INTRODUCTION AND OBJECTIVES: Encephalitozoon cuniculi is an obligate intracellular parasite infecting especially domestic rabbits; however, spontaneous infections have been documented in other mammalian species such as dogs, cats, rabbits, horses, cows and sheep all over the world. Encephalitozoonosis is a chronic and latent disease leading to renal failure, encephalitis, disorders of brain and urinary tract, and may lead to death. There are limited reports on encephalitozoonosis in wildlife, which is why the aim of this study was to detect the prevalence of antibodies to E. cuniculi in European hares. MATERIALS AND METHODS: Samples of blood sera from 701 wild hares from the Czech Republic (n = 245), the Slovak Republic (n = 211) and Austria (n = 245) were examined by indirect immunofluorescence antibody test (IFAT); samples with titer ≥ 40 were marked as positive. RESULTS: The total seroprevalence of E. cuniculi antibodies was 1.42% with titres in the range 40-640. Antibodies to E. cuniculi were detected in 2.9% (7/245), 0.8% (2/245) and 0.47% (1/211) hares from the Czech Republic, Austria and the Slovak Republic, respectively. CONCLUSIONS: This is the first detection of antibodies to E. cuniculi in hares from Europe showing that hares could be exposed to E. cuniculi infection, however with a low rate.
- MeSH
- Encephalitozoon cuniculi izolace a purifikace MeSH
- encephalitozoonóza epidemiologie mikrobiologie veterinární MeSH
- fluorescenční protilátková technika nepřímá veterinární MeSH
- prevalence MeSH
- protilátky fungální analýza MeSH
- séroepidemiologické studie MeSH
- zajíci * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
- Rakousko MeSH
- Slovenská republika MeSH
