The prevalence of Cryptosporidium and microsporidia in feral horses, which have minimal contact with livestock and humans, is not currently known. We report the findings of a study on Cryptosporidium and microsporidia in 34 Mustangs and 50 Chincoteague ponies in the USA. Fecal samples were screened for presence of Cryptosporidium spp. by analysis of the small-subunit rRNA (SSU) and 60-kDa glycoprotein (gp60) genes, and Enterocytozoon bieneusi and Encephalitozoon spp. by analysis of the ribosomal internal transcribed spacer region (ITS). Cryptosporidium spp. and E. bieneusi were detected in 28/84 (33.3%) and 7/84 (8.3%) samples, respectively. Sequence analysis of SSU and ITS revealed the presence of Cryptosporidium parvum (n = 20) and E. bieneusi genotype horse 1 (n = 7), respectively. Subtyping of C. parvum isolates at the gp60 locus showed the presence of subtype IIaA17G2R1 in Mustangs and subtypes IIaA13G2R1 and IIaA15G2R1 in Chincoteague ponies. Enterocytozoon bieneusi genotype horse 1 was detected in Mustangs (n = 2) and Chincoteague ponies (n = 5). No Cryptosporidium or E. bieneusi positive animals had diarrhea. The finding that Mustangs and Chincoteague ponies are host to the zoonotic pathogen C. parvum suggests that their infrequent contact with humans and livestock is sufficient to maintain transmission; however, we should also consider the possibility that C. parvum is an established parasite of Mustangs and Chincoteague ponies that persists in these animals independently of contact with humans or livestock.
- MeSH
- Cryptosporidium parvum klasifikace genetika izolace a purifikace MeSH
- divoká zvířata MeSH
- Enterocytozoon klasifikace genetika izolace a purifikace MeSH
- feces parazitologie MeSH
- fylogeneze MeSH
- genotyp MeSH
- glykoproteiny genetika MeSH
- koně MeSH
- kryptosporidióza epidemiologie parazitologie přenos MeSH
- lidé MeSH
- mezerníky ribozomální DNA chemie MeSH
- mikrosporidióza epidemiologie parazitologie veterinární MeSH
- nemoci koní epidemiologie parazitologie MeSH
- prevalence MeSH
- RNA ribozomální genetika MeSH
- zoonózy parazitologie přenos MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Geografické názvy
- Spojené státy americké MeSH
Faecal samples were collected from 352 horses on 23 farms operating under six different management systems in the Czech Republic and Poland during 2011 and 2012. Farms were selected without previous knowledge of parasitological status. All faecal samples were screened for Cryptosporidium spp. presence using microscopy, following aniline-carbol-methyl violet staining and PCR analysis of the small-subunit (SSU) rRNA and the 60-kDa glycoprotein (gp60) genes. Cryptosporidium muris-positive samples were additionally genotyped at four minisatellite markers: MS1 (encoding a hypothetical protein), MS2 (encoding a 90-kDa heat shock protein), MS3 (encoding a hypothetical protein) and MS16 (encoding a leucine-rich repeat family protein). Cryptosporidium spp. was detected by PCR in 12/352 (3.4%) samples from 4 out of 13 farms. None of the samples tested by microscopy was positive. There was no relationship between Cryptosporidium prevalence and age, sex, diarrhoea or management system; however, Cryptosporidium was found only on farms where horses were kept on pasture during the day and in a stable overnight. Sequence analyses of SSU and gp60 genes revealed the presence of C. muris RN66 (n = 9), Cryptosporidium parvum IIaA15G2R1 (n = 1), Cryptosporidium tyzzeri IXbA22R9 (n = 1), and Cryptosporidium horse genotype VIaA15G4 (n = 1). The C. muris subtypes were identified as MS1-M1, MS2-M4, novel MS2-M7 and MS16-M1 by multilocus sequence of three minisatellite loci. The MS3 locus was not amplified from any isolate. This is the first report of C. tyzzeri and C. muris subtypes from horses.
- MeSH
- Cryptosporidium klasifikace genetika izolace a purifikace MeSH
- feces parazitologie MeSH
- genetická variace * MeSH
- genotyp MeSH
- koně MeSH
- kryptosporidióza epidemiologie MeSH
- nemoci koní parazitologie MeSH
- prevalence MeSH
- průjem parazitologie veterinární MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
- Polsko MeSH
A total of 219 and 124 individual fecal samples of horses and donkeys, respectively, were screened for the presence of Cryptosporidium spp., Encephalitozoon spp., and Enterocytozoon bieneusi DNA by genus-specific nested PCR. Isolates were genotyped by sequence analysis of SSU rRNA, GP60, TRAP-C1, COWP, and HSP70 loci in Cryptosporidium, and the ITS region in microsporidia. Cryptosporidium spp. was detected on 3/18 horse farms and 1/15 farms where donkeys were kept. Overall, five (2.3%) horse and two (1.6%) donkey specimens were PCR positive for Cryptosporidium. Genotyping at SSU and GP60 loci revealed that three isolates from horses and donkeys were C. parvum subtype family IIaA16G1R1, one isolate from a horse was, C. muris RN66, and one isolate from a donkey was C. muris TS03. An isolate from a horse shared 99.4% and 99.3% similarity with Cryptosporidium hominis and C. cuniculus, respectively, at the SSU locus. This isolate shared 100% identity with C. hominis at the TRAP-C1, COWP, and HSP70 loci, and it was from the novel gp60 subtype family IkA15G1. Microsporidia were found on 6/18 horse and 2/15 donkey farms. E. bieneusi was identified in 6.8% (15/219) and 1.6% (2/124), and Encephalitozoon cuniculi was identified in 1.8% (4/219) and 1.6% (2/124), of horses and donkeys, respectively. Three genotypes of E. cuniculi (I, II and III) were detected in horses, and E. cuniculi genotype II was detected in donkeys. Four genotypes of E. bieneusi (horse1, horse 2, CZ3, D) were described in horses. An additional five horses and two donkeys were positive for E. bieneusi, but the isolated were not genotyped. Neither Cryptosporidium nor microsporidia prevalence were affected by sex, age, type of breeding, or whether the host was a horse or a donkey.
- MeSH
- Cryptosporidium klasifikace genetika izolace a purifikace MeSH
- Equidae * mikrobiologie parazitologie MeSH
- feces mikrobiologie parazitologie MeSH
- fylogeneze MeSH
- genotyp MeSH
- koně MeSH
- kryptosporidióza epidemiologie parazitologie MeSH
- Microsporidia klasifikace genetika izolace a purifikace MeSH
- mikrosporidióza epidemiologie mikrobiologie veterinární MeSH
- nemoci koní epidemiologie mikrobiologie parazitologie MeSH
- prevalence MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Alžírsko 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
Nine ponies, aged 12 months at the time of infection, were randomly allocated to one of three groups. Each pony was infected with 30,000 infective cyathostome larvae stored 4 weeks under the natural environmental conditions of the Czech Republic. Horses of Group A were infected with larvae conditioned from August 16 to September 12 (the first phase of the experiment). Horses of Group B were inoculated with larvae conditioned from October 10 to November 4 (the second phase of the experiment). In the third phase, horses from Group C received larvae stored under natural conditions (from 8.11 to 18.11.2011). The larvae were then kept under laboratory conditions (5 °C, the light reduction according to the nature condition). In the first and second phase of the experiment, 98.57% and 95.48% of adult nematodes were found, respectively. However, only 29.07% of adults were found in the third phase of experiment, and the proportion of arrested larvae (EL3) was 67.73%. The share of these larvae in the cecum during last phase of the experiment comprised 92.59% of all cyathostomes.
- MeSH
- cékum parazitologie MeSH
- koně MeSH
- larva MeSH
- náhodné rozdělení MeSH
- nemoci koní parazitologie MeSH
- Strongyloidea růst a vývoj MeSH
- strongyloidóza koní parazitologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
We describe the morphological, biological, and molecular characteristics of Cryptosporidium pig genotype II and propose the species name Cryptosporidium scrofarum n. sp. to reflect its prevalence in adult pigs worldwide. Oocysts of C. scrofarum are morphologically indistinguishable from C. parvum, measuring 4.81-5.96 μm (mean=5.16)×4.23-5.29 μm (mean=4.83) with a length to width ratio of 1.07±0.06 (n=400). Oocysts of C. scrofarum obtained from a naturally infected pig were infectious for 8-week-old pigs but not 4-week-old pigs. The prepatent period in 8-week-old Cryptosporidium-naive pigs was 4-6 days and the patent period was longer than 30 days. The infection intensity of C. scrofarum in pigs was generally low, in the range 250-4000 oocysts per gram of feces. Infected pigs showed no clinical signs of cryptosporidiosis and no pathology was detected. Cryptosporidium scrofarum was not infectious for adult SCID mice, adult BALB/c mice, Mongolian gerbils (Meriones unguiculatus), southern multimammate mice (Mastomys coucha), yellow-necked mice (Apodemus flavicollis), or guinea pigs (Cavia porcellus). Phylogenetic analyses based on small subunit rRNA, actin, and heat shock protein 70 gene sequences revealed that C. scrofarum is genetically distinct from all known Cryptosporidium species.
- MeSH
- Cryptosporidium klasifikace cytologie genetika MeSH
- druhová specificita MeSH
- feces parazitologie MeSH
- fylogeneze * MeSH
- genotyp MeSH
- Gerbillinae MeSH
- kryptosporidióza patologie veterinární MeSH
- morčata MeSH
- myši inbrední BALB C MeSH
- myši SCID MeSH
- myši MeSH
- nemoci prasat parazitologie patologie MeSH
- prasata MeSH
- protozoální geny genetika MeSH
- zvířata MeSH
- Check Tag
- morčata MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
A total of 9 (8 stallions and 1 mare) 1 year old ponies were used for the experimental infection caused by Encephalitozoon cuniculi genotype II (10(7) spores per animal). Subsequently, individual horses were slaughtered 7, 14, 21, 28, 35, 42, 49, 56, and 63 days post infection. Immediately after slaughter, tissues samples of stomach, duodenum, jejunum, ileum, caecum, colon, spleen, liver, kidney, bladder, heart, lungs, and brain were sampled. In addition, urine, feces and blood specimens were collected. Enzyme-linked immunosorbent assay was used for determination of humoral immune response and nested PCR targeting 16S rDNA, whole ITS, and 5.8S rDNA was used for detection of E. cuniculi in collected organs, blood, feces and urine. No clinical signs of microsporidiosis including diarrhea or colic, neurological signs and fever were observed in any horses during whole experiment. Acute microsporidiosis in ponies was characterized by the dissemination of microsporidia into almost all organs and significant increase of concentration of specific antibodies in blood was observed from 28 to 42 DPI. After this acute stage microsporidia disappeared from most organs with the exception of the kidney, which was positive up to 63 DPI when the experiment was terminated. No pathological changes were observed in any organs with exception of one mare's brain, where E. cuniculi-positive cavity measuring 5 cm × 3 cm in diameter formed in the lobus piriformis.
- MeSH
- Encephalitozoon cuniculi imunologie MeSH
- encephalitozoonóza imunologie veterinární MeSH
- humorální imunita fyziologie MeSH
- koně MeSH
- nemoci koní imunologie mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Faecal samples were collected from 377 horses on 23 farms with varying management systems in the Czech Republic. Microsporidia were found on 16 farms and the overall prevalence of Enterocytozoon bieneusi and Encephalitozoon cuniculi was 17.3% (66/377) and 6.9% (26/377), respectively. The prevalence of E. cuniculi in horses over 3 years of age was significantly higher (10.0%) compared to younger horses (4.0%). No significant differences in prevalence were observed among stallions, geldings, and mares for both microsporidia. Significantly higher infection rates of E. bieneusi and E. cuniculi were recorded in horses kept in stables than those on pasture. Two genotypes of E. cuniculi (I and II) and 15 genotypes of E. bieneusi including six previously described and nine novel genotypes were detected. The most common genotype detected was E. bieneusi genotype D identified in 51.5% (34/66) of positive horses. The identification of E. bieneusi genotypes D, EpbA, G and WL15, which were previously reported in pigs, humans, raccoons and horses, indicates that horses could be a potential source of zoonotic infection in humans.
- MeSH
- chov zvířat * MeSH
- Encephalitozoon cuniculi izolace a purifikace MeSH
- encephalitozoonóza epidemiologie veterinární MeSH
- Enterocytozoon izolace a purifikace MeSH
- fylogeneze MeSH
- koně MeSH
- mikrosporidióza epidemiologie veterinární MeSH
- nemoci koní epidemiologie mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
