Search

Additional file 1: Table S1. Primers designed for qRT-PCR validation of candidate miRNAs, mRNAs, circRNAs and lncRNAs in HCT-8 cells at 3 h and 12 h post infection with Cryptosporum parvum.

Additional file 1: Table S1. Primers used in RT-qPCR and sequences used for construct generating. Table S2. RNA oligonucleotides for miRNA and siRNA.

The functional mitochondrial protein alternative oxidase (AOX) gene was used as a marker to analyse the phylogenetic relationship betweenCryptosporidiumisolates. This gene was characterized, and the phylogentic tree was established fromCryptosporidiumisolates and compared to those generated from 18S rRNA and heat-shock protein 70 (HSP70) gene sequences. The present trial aimed at finding out whether the AOX gene is suitable for phylogenetic analysis of theCryptosporidiumgenus. The results revealed that the genusCryptosporidiumcontained the phylogenetically distinct speciesC. parvum,C. hominis,C. suisandC. baileyi, which were consistent with the biological characterization and host specificity reported earlier.Cryptosporidiumspecies formed two clades: one includedC. hominis,C. suis,C. parvumcattle genotypes andC. parvummouse genotype; and the other comprisedC. meleagridisandC. baileyiisolates. WithinC. parvum, both the mouse genotype and the pig genotype I (also known asC. suis) isolates differed from cattle and human (also known asC. hominis) genotypes, based on the aligned nucleotide sequences. The sequence identity of the AOX gene was higher betweenC. meleagridisandC. baileyithan betweenC. meleagridisandC. parvum. The phylogenetic trees showed thatC. meleagridiswas closer toC. baileyithan toC. parvum. This result was inconsistent with the phylogenetic analysis deduced from 18S rRNA and HSP70 gene sequences, respectively. The present results suggest that the AOX gene is not only equally suitable for the phylogenetic analysis ofCryptosporidium, but also provides an outstanding and new approach in determiningCryptosporidiumheredity.

Tick-borne diseases are one of the most important classes of disease in animal husbandry and cause severe economic losses. In this study, 660 adult female ticks were collected from sheep in nine localities in Henan Province, China. All were identified as Haemaphysalis longicornis, confirmed with light microscopy and PCR amplification. The pathogens identified in these ticks included bacteria of the genus Anaplasma (Anaplasma ovis, A. bovis, A. phagocytophilum) and piroplasmal protozoans (Theileria luwenshuni, Babesia motasi). Our results show the high prevalence of A. bovis (20.4%, 135/660) and T. luwenshuni (14.4%, 95/660) in the ticks from sheep, whereas A. ovis, A. phagocytophilum, and B. motasi occurred in 2.3% (15/660), 1.5% (10/660), and 0.75% (5/660) of ticks, respectively. This is the first report of B. motasi in H. longicornis in China. In contrast, no T. uilenbergi infection was found in this study. These results confirm that H. longicornis is the most common tick species in sheep in Henan Province and can transmit A. phagocytophilum, a well-known zoonotic pathogen of public-health and veterinary significance. Importantly, these results suggest that A. bovis, transmitted by H. longicornis, is a predominant pathogen of sheep in this region.

usc Cryptosporidium is one of the most common zoonotic waterborne parasitic diseases worldwide and represents a major public health concern of water utilities in developed nations. As animals in catchments can shed human-infectious Cryptosporidium oocysts, determining the potential role of animals in dissemination of zoonotic Cryptosporidium to drinking water sources is crucial. In the present study, a total of 952 animal faecal samples from four dominant species (kangaroos, rabbits, cattle and sheep) inhabiting Sydney's drinking water catchments were screened for the presence of Cryptosporidium using a quantitative PCR (qPCR) and positives sequenced at multiple loci. Cryptosporidium species were detected in 3.6% (21/576) of kangaroos, 7.0% (10/142) of cattle, 2.3% (3/128) of sheep and 13.2% (14/106) of rabbit samples screened. Sequence analysis of a region of the 18S rRNA locus identified C. macropodum and C. hominis in 4 and 17 isolates from kangaroos respectively, C. hominis and C. parvum in 6 and 4 isolates respectively each from cattle, C. ubiquitum in 3 isolates from sheep and C. cuniculus in 14 isolates from rabbits. All the Cryptosporidium species identified were zoonotic species with the exception of C. macropodum. Subtyping using the 5' half of gp60 identified C. hominis IbA10G2 (n = 12) and IdA15G1 (n = 2) in kangaroo faecal samples; C. hominis IbA10G2 (n = 4) and C. parvum IIaA18G3R1 (n = 4) in cattle faecal samples, C. ubiquitum subtype XIIa (n = 1) in sheep and C. cuniculus VbA23 (n = 9) in rabbits. Additional analysis of a subset of samples using primers targeting conserved regions of the MIC1 gene and the 3' end of gp60 suggests that the C. hominis detected in these animals represent substantial variants that failed to amplify as expected. The significance of this finding requires further investigation but might be reflective of the ability of this C. hominis variant to infect animals. The finding of zoonotic Cryptosporidium species in these animals may have important implications for the management of drinking water catchments to minimize risk to public health. © 2016 Zahedi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Refereed/Peer-reviewed