Molecular epidemiology and subtyping of Blastocystis sp. and its subtypes in celiac patients; a case control study.

Blastocystis sp. is a common intestinal protist, reported from symptomatic and asymptomatic subjects. Blastocystis sp. has been reported from a broad spectrum of gastrointestinal disorders. Celiac disease (CD) is an autoimmune disorder of the small intestine, which leads to the lack of tolerance against gluten. Long-term following of gluten-free diet in CD patients decreases the gut microbiota restoration and probably decreases the chance of Blastocystis sp. colonization. The current study aimed to investigate the prevalence of Blastocystis sp. and its subtypes in CD patients in comparison to healthy subjects. Stool samples were collected from 238 participants including 92 confirmed CD patients and 146 healthy subjects. Upon DNA extraction, the presence of Blastocystis sp. was evaluated using amplification of discriminative regions of the small ribosomal RNA (ssu rRNA) gene. To characterize subtypes and alleles, amplified fragments were sequenced. Phylogenetic trees were constructed to visualize subtype correlation. Our findings showed that 21% (50) of samples including 16.3% (15/92) and 23.97% (35/146) were positive for Blastocystis sp. in CD patients and healthy controls, respectively. Except family relationship, other variables were not statistical correlated with the presence of Blastocystis sp.. Totally, 25 samples were successfully sequenced. Accordingly, ST1, ST2, and ST3 were characterized in 8 (32%), 9 (36%), and 8 (32%) of samples, respectively. Allele discrimination showed that all ST1 were allele 4; alleles 11, 9, and 12 were retrieved from ST2, and alleles 34, 36, and 38 were observed in ST3. The relationship between colonization of Blastocystis sp. and alteration in the gut microbiota composition is indeterminate, however, this hypothesis that following gluten-free diet in CD patients may affect the colonization of Blastocystis sp. via alteration in the gut microbiota composition could be interesting for further investigations.

Molecular analysis of internal transcribed spacer 2 of Dicrocoelium dendriticum isolated from cattle, sheep, and goat in Iran.

BACKGROUND: Dicrocoelium dendriticum is a broadly distributed zoonotic helminth, which is mainly reported from domesticated and wild ruminants. There is little data covering the molecular features of this trematode; therefore, current study aimed to molecularly analyze D. dendriticum in livestock. METHODS: Totally, 23 samples of D. dendriticum were collected from cattle, sheep, and goat from Ilam, Lorestan, and Khuzestan, three west and south-west provinces of Iran from February to August 2018. After genomic DNA extraction, the internal transcribed spacer (ITS) 2 fragment was amplified and sequenced in samples. To investigate genetic variations through the ITS 2 fragment of obtained D. dendriticum, phylogenetic tree and network analysis were employed. RESULTS: All 23 samples were successfully amplified and sequenced. Phylogenetic tree showed that our samples were clearly grouped in a clade together with reference sequences. There was no grouping based on either geographical regions or hosts. Network analysis confirmed the phylogenetic findings and showed the presence of nine distinct haplotypes, while our samples together most of sequences, which were previously submitted to the GenBank, were grouped in the Hap1. CONCLUSIONS: Our findings indicated that although ITS 2 fragment discriminate D. dendriticum, this fragment is not suitable to study intra-species genetic variations. Therefore, exploring and describing new genetic markers could be more appropriate to provide new data about the genetic distribution of this trematode.

Molecular characterization of Cryptosporidium skunk genotype in raccoons (Procyon lotor) in Iran: concern for zoonotic transmission.

Cryptosporidium spp. are significant zoonotic parasites in humans and animals worldwide. This study aimed to investigate the prevalence of Cryptosporidium infection among raccoon (Procyon lotor) in north of Iran. The fecal samples (n = 30) were collected from raccoons. After DNA extraction, all samples were examined by nested PCR amplification of the 18S ribosomal RNA (rRNA) gene. From 30 raccoon samples, 4 (13.3%) were positive, and the isolates were identified as Cryptosporidium skunk genotype based on sequence analysis. The large distribution of raccoons in northern provinces of Iran and their potency for carrying some human-infecting parasites like Cryptosporidium spp. propose this mammalian as a source for zoonotic parasites.