INTRODUCTION: Hammondia hammondi and Toxoplasma gondii are closely related protozoan parasites, but only T. gondii is zoonotic. Both species use felids as definitive hosts and cannot be differentiated by oocyst morphology. In T. gondii, a 529-base pair (bp) repetitive element (TgREP-529) is of utmost diagnostic importance for polymerase chain reaction (PCR) diagnostic tests. We identified a similar repetitive region in the H. hammondi genome (HhamREP-529). METHODS: Based on reported sequences, primers and probes were selected in silico and optimal primer probe combinations were explored, also by including previously published primers. The analytical sensitivity was tested using serial dilutions of oocyst DNA. For testing analytical specificity, DNA isolated from several related species was used as controls. The newly established TaqMan PCR (Hham-qPCR1) was applied to tissues collected from H. hammondi-infected gamma-interferon gene knockout (GKO) mice at varying time points post-infection. RESULTS: Ten forward and six reverse primers were tested in varying combinations. Four potentially suitable dual-labelled probes were selected. One set based on the primer pair (Hham275F, Hham81R) and the probe (Hham222P) yielded optimal results. In addition to excellent analytic specificity, the assay revealed an analytical sensitivity of genome equivalents of less than one oocyst. Investigation of the tissue distribution in GKO mice revealed the presence of parasite DNA in all examined organs, but to a varying extent, suggesting 100- to 10,000-fold differences in parasitic loads between tissues in the chronic state of infection, 42 days post-infection. DISCUSSION: The use of the 529-bp repeat of H. hammondi is suitable for establishing a quantitative real-time PCR assay, because this repeat probably exists about 200 times in the genome of a single organism, like its counterpart in T. gondii. Although there were enough sequence data available, only a few of the primers predicted in silico revealed sufficient amplification; the identification of a suitable probe was also difficult. This is in accord with our previous observations on considerable variability in the 529-bp repetitive element of H. hammondi. CONCLUSIONS: The H. hammondi real-time PCR represents an important novel diagnostic tool for epidemiological and cell biological studies on H. hammondi and related parasites.
- MeSH
- diferenciální diagnóza MeSH
- feces parazitologie MeSH
- kočky parazitologie MeSH
- kokcidióza veterinární MeSH
- kvantitativní polymerázová řetězová reakce metody MeSH
- molekulární patologie metody MeSH
- myši parazitologie MeSH
- oocysty genetika izolace a purifikace MeSH
- protozoální geny MeSH
- repetitivní sekvence nukleových kyselin MeSH
- Sarcocystidae * genetika izolace a purifikace MeSH
- Toxoplasma * genetika izolace a purifikace MeSH
- toxoplazmóza zvířat MeSH
- zvířata MeSH
- Check Tag
- kočky parazitologie MeSH
- myši parazitologie MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Parasite hybrid zones resulting from host secondary contact have never been described in nature although parasite hybridization is well known and secondary contact should affect them similarly to free-living organisms. When host populations are isolated, diverge and recontact, intimate parasites (host specific, direct life cycle) carried during isolation will also meet and so may form parasite hybrid zones. If so, we hypothesize these should be narrower than the host's hybrid zone as shorter parasite generation time allows potentially higher divergence. We investigate multilocus genetics of two parasites across the European house mouse hybrid zone. We find each host taxon harbours its own parasite taxa. These also hybridize: Parasite hybrid zones are significantly narrower than the host's. Here, we show a host hybrid zone is a suture zone for a subset of its parasite community and highlight the potential of such systems as windows on the evolutionary processes of host-parasite interactions and recombinant pathogen emergence.
- MeSH
- fylogeneze MeSH
- genetické markery MeSH
- genotyp MeSH
- hlístice genetika MeSH
- hybridizace genetická * MeSH
- mitochondriální DNA genetika MeSH
- myši genetika parazitologie MeSH
- paraziti genetika MeSH
- Pneumocystis genetika MeSH
- populační genetika * MeSH
- zvířata MeSH
- Check Tag
- myši genetika parazitologie MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
- Německo MeSH
We undertook a study on Cryptosporidium spp. in wild cricetid rodents. Fecal samples were collected from meadow voles (Microtus pennsylvanicus), southern red-backed voles (Myodes gapperi), woodland voles (Microtus pinetorum), muskrats (Ondatra zibethicus) and Peromyscus spp. mice in North America, and from bank voles (Myodes glareolus) and common voles (Microtus arvalis) in Europe. Isolates were characterized by sequence and phylogenetic analyses of the small subunit ribosomal RNA (SSU) and actin genes. Overall, 33·2% (362/1089) of cricetids tested positive for Cryptosporidium, with a greater prevalence in cricetids from North America (50·7%; 302/596) than Europe (12·1%; 60/493). Principal Coordinate analysis separated SSU sequences into three major groups (G1-G3), each represented by sequences from North American and European cricetids. A maximum likelihood tree of SSU sequences had low bootstrap support and showed G1 to be more heterogeneous than G2 or G3. Actin and concatenated actin-SSU trees, which were better resolved and had higher bootstrap support than the SSU phylogeny, showed that closely related cricetid hosts in Europe and North America are infected with closely related Cryptosporidium genotypes. Cricetids were not major reservoirs of human pathogenic Cryptosporidium spp.
- MeSH
- Arvicolinae parazitologie MeSH
- Cryptosporidium klasifikace izolace a purifikace patogenita fyziologie MeSH
- divoká zvířata parazitologie MeSH
- feces parazitologie MeSH
- fylogeneze MeSH
- fylogeografie MeSH
- genotyp MeSH
- hlodavci parazitologie MeSH
- kryptosporidióza epidemiologie parazitologie MeSH
- myši parazitologie MeSH
- RNA ribozomální genetika MeSH
- sekvenční analýza DNA MeSH
- zdroje nemoci parazitologie MeSH
- zvířata MeSH
- Check Tag
- myši parazitologie MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Evropa MeSH
- Severní Amerika MeSH
Toxoplasma gondii and Neospora caninum are closely related coccidian parasites infecting a wide range of wild and domestic animals as intermediate hosts, and rodents serve as important reservoir hosts during the life cycles of these parasites. The present study is aimed at identifying T. gondii and N. caninum infection in 360 wild house mice (Mus musculus) collected across the Czech-German border, where 2 genetically distinct mouse subspecies meet and hybridize. Toxoplasma gondii or N. caninum DNA was detected in the brains of individual mice by PCR, but mixed infections were never observed. No significant differences in gender or trapping localities were found in the positive mice. The survey reveals a low frequency of T. gondii (0.6%) and N. caninum (3.6%) occurrence in the house mice population of the monitored part of the hybrid zone.
- MeSH
- chiméra MeSH
- divoká zvířata MeSH
- kokcidióza epidemiologie parazitologie veterinární MeSH
- mozek parazitologie MeSH
- myši parazitologie MeSH
- nemoci hlodavců epidemiologie parazitologie MeSH
- Neospora izolace a purifikace MeSH
- polymerázová řetězová reakce veterinární MeSH
- prevalence MeSH
- protozoální DNA izolace a purifikace MeSH
- Toxoplasma izolace a purifikace MeSH
- toxoplazmóza zvířat epidemiologie parazitologie MeSH
- zdroje nemoci MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši parazitologie MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Československo MeSH
- Německo MeSH
- MeSH
- deratizace metody MeSH
- dezinsekce MeSH
- infestace ektoparazity * prevence a kontrola MeSH
- ivermektin aplikace a dávkování terapeutické užití MeSH
- kongresy jako téma MeSH
- kontaminace potravin prevence a kontrola MeSH
- lidé MeSH
- malárie prevence a kontrola MeSH
- myši parazitologie růst a vývoj MeSH
- ptáci parazitologie růst a vývoj MeSH
- štěnice účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši parazitologie růst a vývoj MeSH
- zvířata MeSH
Wormy mice in a hybrid zone have been interpreted as evidence of low hybrid fitness, such that parasites contribute to species separation. However, because of its natural heterogeneity, observations of parasite load must be numerous with good field area coverage. We sampled 689 mice from 107 localities across the Bavaria-Bohemia region of the European house mouse hybrid zone and calculated their hybrid indices using 1401 diagnostic single nucleotide polymorphisms (SNPs). We tested whether hybrids have greater or lesser diversity and load of parasite helminths than additive expectations, performing load analyses on the four most common taxa. We found hybrids have significantly reduced diversity and load of each of the commonest helminths; rarer helminths further support reduced load. Although within-locality comparisons have little power, randomization tests show the repeated pattern is unlikely to be due to local parasite heterogeneity, and simulations show a patch of low parasite diversity is unlikely to fall by chance just so in the field area, such that it produces the observed effects. Our data therefore contradict the idea that helminths reduce hybrid fitness through increased load. We discuss a vicariant Red Queen model that implies immune genes tracking parasites will escape Dobzhansky-Muller incompatibilities, generating hybrid variants untargeted by parasites.
- MeSH
- biodiverzita MeSH
- cizopasní červi imunologie MeSH
- hybridizace genetická * MeSH
- jednonukleotidový polymorfismus MeSH
- myši genetika imunologie parazitologie MeSH
- parazitární zátěž * MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši genetika imunologie parazitologie MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Three and 8 week old pigs were inoculated with Cryptosporidium muris HZ206 (Mus musculus musculus isolate), Cryptosporidium tyzerri CR2090 (M. m. musculus isolate) or C. tyzzeri CR4293 (isolate from a hybrid between Mus musculus domesticus and M. m. musculus) at a dose of 1 × 10(7) oocysts per animal. Inoculated pigs showed no detectable infection and no clinical symptoms of cryptosporidiosis during 30 days post infection (DPI), and no macroscopic changes were detected in the digestive tract following necropsy. Developmental stages were not detected in gastrointestinal tract tissue by histology or PCR throughout the duration of the experiment. The infectivity of isolates was verified on SCID mice, in which oocysts shedding started from 4 to 8 DPI. Based on our findings, it can be concluded that pigs are not susceptible to C. muris or C. tyzzeri infection.
- MeSH
- Cryptosporidium klasifikace genetika fyziologie MeSH
- feces parazitologie MeSH
- genotyp MeSH
- hostitelská specificita MeSH
- kryptosporidióza parazitologie veterinární MeSH
- myši SCID MeSH
- myši parazitologie MeSH
- nemoci hlodavců parazitologie MeSH
- nemoci prasat parazitologie MeSH
- prasata MeSH
- protozoální DNA chemie izolace a purifikace MeSH
- RNA ribozomální genetika MeSH
- sekvenční seřazení veterinární MeSH
- zvířata MeSH
- Check Tag
- myši parazitologie MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
We studied morphological and molecular polymorphism of 53 Tetratrichomonas isolates obtained from amphibian, reptilian, mammalian hosts, and from a slug with the aid of protargol staining and analyses of ITS1-5.8S rRNA-ITS2, SSU rRNA, and alpha-tubulin gene sequences. The phylogenetic tree based on the concatenate of all sequences showed the monophyly of the genus Tetratrichomonas with respect to the genus Trichomonas. Our data suggest that two parabasalid genera, Pentatrichomonoides and Trichomonoides, may belong to the genus Tetratrichomonas. Tetratrichomonas isolates were divided into 16 robust host-specific and monophyletic groups that probably represent separate, mostly new, species. As only five Tetratrichomonas species were described from the examined host taxa so far, our study uncovered considerable species diversity within the genus. The wide host range, high level of species-specific host specificity, and newly revealed biodiversity make the genus Tetratrichomonas a valuable model for studying evolution of parasites.
- MeSH
- Bayesova věta MeSH
- financování organizované MeSH
- fylogeneze MeSH
- genetická variace imunologie MeSH
- interakce hostitele a parazita genetika MeSH
- mezerníky ribozomální DNA genetika MeSH
- molekulární evoluce MeSH
- molekulární sekvence - údaje MeSH
- myši parazitologie MeSH
- prasata parazitologie MeSH
- protozoální DNA genetika chemie MeSH
- RNA ribozomální 5.8S genetika MeSH
- sekvenční analýza DNA MeSH
- Trichomonadida genetika klasifikace MeSH
- zvířata MeSH
- Check Tag
- myši parazitologie MeSH
- zvířata MeSH
- MeSH
- analýza přežití MeSH
- antiprotozoální látky farmakologie terapeutické užití MeSH
- Encephalitozoon účinky léků MeSH
- finanční podpora výzkumu jako téma MeSH
- fluorochinolony farmakologie terapeutické užití MeSH
- mikrosporidióza farmakoterapie MeSH
- myši parazitologie MeSH
- techniky in vitro MeSH
- Vittaforma účinky léků MeSH
- zvířata MeSH
- Check Tag
- myši parazitologie MeSH
- zvířata MeSH
