Genetic diversity of Cryptosporidium spp. including novel identification of the Cryptosporidium muris and Cryptosporidium tyzzeri in horses in the Czech Republic and Poland

Abstract

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.

Occurrence and Genetic Characteristics of Cryptosporidium hominis and Cryptosporidium andersoni in Horses from Southwestern China

“…In the present study, six of the 333 (1.8%) fecal specimens were detected to be positive for Cryptosporidium by PCR amplification of the partial SSU rRNA gene, which is consistent with the previously reported low infection rate of Cryptosporidium in horses in Algeria (2.3%), Poland (3.4%), and New York state (5.1%) (Burton et al 2010;Laatamna et al 2015;Wagnerov a et al 2015). The age of the Cryptosporidium-positive horses was between 2 and 3 yr.…”

First description of Cryptosporidium hominis GP60 genotype IkA20G1 and Cryptosporidium parvum GP60 genotypes IIaA18G3R1 and IIaA15G2R1 in foals in Brazil

“…This genotype was also found in foals with diarrhea in New Zealand (Grinberg et al, 2008). The IIaA15G2R1 genotype was identified in the Czech Republic and in Poland in a 3 year old asymptomatic equine (Wagnerová et al, 2015) and also in fecal samples from the environment in Italy (Galuppi et al, 2015).…”

Multi-locus genotypes of Enterocytozoon bieneusi in captive Asiatic black bears in southwestern China: High genetic diversity, broad host range, and zoonotic potential

“…The internal transcribed spacer (ITS) of the rRNA gene, which possesses considerable genetic variation, has been widely used for genotyping E . bieneusi isolates in humans and animals [ 6 , 8 ]. To date, more than 240 genotypes of E .…”