Identification and Subtyping of Cryptosporidium parvum and Cryptosporidium hominis in Cancer Patients, Isfahan Province, Central Iran.

Background: Cryptosporidium spp. are protozoan parasites that cause diarrhea in humans and animals. Subtyping data about Cryptosporidium spp. in Isfahan, Iran is limited; therefore, we aimed to study the prevalence rate of Cryptosporidium spp. in cancer patients, associated risk factors, and subtypes of Cryptosporidium spp. Methods: Fecal samples were collected from 187 cancer patients from the Oncology Department of Seyed-al-Shohada Hospital, Isfahan University of Medical Sciences during 2014-2020 and screened for Cryptosporidium spp. using microscopical techniques. Nested PCR amplifying 18S rRNA gene was used to detect Cryptosporidium spp. in samples, followed by subtyping using nested PCR amplifying gp60 sequences. Results: Overall, the rate of infection with Cryptosporidium spp. was 4.3% (n=8). Five samples out of eight samples were identified as Cryptosporidium spp. using a nested PCR for the 18S rRNA gene, two subtypes of C. parvum named IIaA18G3R1 (n = 2) and IIaA17G2R1 (n = 2), and one subtype of C. hominis named IbA6G3 were identified by sequencing of the gp60. The IbA6G3 subtype has rarely been detected in other investigations. Conclusion: This is the first survey on the subtyping of Cryptosporidium spp. in this region. The results of the present survey show both zoonotic and anthroponotic transmission routes in the region.

Effect of two hydatid cyst antigens on the growth of melanoma cancer in C57/black mice.

Hydatid cyst is the larval stage of Echinococcus granulosus. In previous studies inhibitory effect of this parasite on cancer cell growth in culture medium has been shown. In this study effect of hydatid cyst antigens on tumor growth in experimental animals has been investigated. Two antigens of hydatid cyst including protoscolices excretory secretory antigen and hydatid fluid absorbed on alum as adjuvant were injected to two groups of C57/black mice as case groups. Control groups were injected with only saline and alum. All mice then were injected with melanoma cells. Both antigens reduced the tumor size in mice in case groups. The difference of tumor size in mice in case groups and control group was statistically significant. In conclusion, anti-tumor effect of hydatid cyst antigens may be related to antigenic similarities which exist between hydatid cyst and cancer cells.

First report of Tasmanian sheep strain (G2) genotype isolated from Iranian goat using the high resolution melting (HRM) analysis.

AIM: The present study was aimed to evaluate E. granulosus genotypes isolated from goats using HRM analysis in Isfahan province. BACKGROUND: Cystic echincoccosis, so-called hydatidosis, is widespread infection caused by the larval stage of Echinococcus granulosus. This is an important zoonotic disease worldwide, especially in the developing countries such as Iran. To date, molecular studies mainly based on the mitochondrial DNA sequences have identified distinct genotypes termed G1-G10 which can differ in some characteristics such as the growth and infectivity to different intermediate hosts or the survival rate in the definitive hosts that are important for the development of control strategies. METHODS: From August to December 2014, 1341 goats were investigated and hydatid cysts were collected from the liver and lungs of 43 infected goats in Isfahan province abattoirs, Isfahan, Iran. Total genomic DNA was extracted from each sample, amplified for the presence of polymorphism of mitochondrial gene coding for cytochrome c oxidase subunit 1 (CO1), using high resolution melting curve (HRM) method. RESULTS: the results of HRM analysis using the sequence of CO1 gene for 43 Echinococcus granulosus isolates from goats showed 31, 2 and 10 isolates were identified as G1, G2, and G3 genotypes, respectively. CONCLUSION: G1 is the predominant genotype in the isolated goat samples in Isfahan province, and the presence of G2 strain was reported for the first time in goat in Iran.

Genetic diversity of Echinococcus granulosus in center of Iran.

Hydatid cyst caused by Echinococcus granulosus is one of the most important parasitic diseases around the world and many countries in Asia, including Iran, are involved with this infection. This disease can cause high mortality in humans as well as economic losses in livestock. To date, several molecular methods have been used to determine the genetic diversity of E. granulosus. So far, identification of E. granulosus using real-time PCR fluorescence-based quantitative assays has not been studied worldwide, also in Iran. Therefore, the aim of this study was to investigate the genetic diversity of E. granulosus from center of Iran using real-time PCR method. A total of 71 hydatid cysts were collected from infected sheep, goat, and cattle slaughtered in Isfahan, Iran during 2013. DNA was extracted from protoscolices and/or germinal layers from each individual cyst and used as template to amplify the mitochondrial cytochrome c oxidase subunit 1 gene (cox1) (420 bp). Five cattle isolates out of 71 isolates were sterile and excluded from further investigation. Overall, of 66 isolates, partial sequences of the cox1 gene of E. granulosus indicated the presence of genotypes G1 in 49 isolates (74.2%), G3 in 15 isolates (22.7%), and G6 in 2 isolates (3.0%) in infected intermediate hosts. Sixteen sequences of G1 genotype had microgenetic variants, and they were compared to the original sequence of cox1. However, isolates identified as G3 and G6 genotypes were completely consistent with original sequences. G1 genotype in livestock was the dominant genotype in Isfahan region, Iran.

Molecular identification of Giardia lamblia; is there any correlation between diarrhea and genotyping in Iranian population?

AIM: The aim of this study is to investigate the molecular identification of Giardia lamblia in patients with diarrhea. BACKGROUND: Giardiasis caused by Giardia lamblia is a common intestinal disease. Although this parasitic infection found in mammals including human, pets and livestock, but few species within the genus Giardia can infects humans. G. lamblia have seven complex genotypes termed (A-H). Genotype A and B the main causes of human infections. PATIENTS AND METHODS: Sixty seven microscopically positive G. Lamblia samples were collected from clinical laboratories in Isfahan province between June 2013 and February 2014. Extraction of genomic DNA was performed for 65 concentrated cysts and 2 cultured trophozoites. Partial sequences of tpi including 148-bp and 81-bp were amplified for detection the genotypes A and B using RFLP- PCR protocol respectively. RESULTS: PCR results showed that out of 67 patients with giardiasis infection, genotype A (148 bp) was detected in 40 isolates (59.70%) compared to genotype B (81 bp) isolated was detected in 25 isolates (37.31%). Also two isolates (2.98%) had mix infection infected with genotype A and B. By comparing the frequency of genotype A (81.8%) and genotype B (13.6%), we found that genotype A is six times higher prevalence than genotype B in patients with diarrhea. CONCLUSION: We suggest that using sensitive techniques and larger sample for detection of G. lamblia genotypes and their subtypes would be necessary for investigation the immune system respond and correlation with diarrhea in the future studies in Iran.