Seroprevalence of Toxoplasma Gondii in Sheep, Cattle and Horses in Urmia North-West of Iran

Background: Toxoplasma gondii is a zoonotic protozoan parasite found worldwide and responsi­ble for major economic losses in most classes of livestock. This study was aimed to deter­mine the prevalence of T. gondii infection in sheep, cattle and horses in Urmia, north-west of Iran, using MAT.Methods: Blood samples of 276 livestock and 26 horses were collected from July 2009 till April 2010. The data were analyzed by the Chi-square, Fisher's Exact and Cochran's and Mantel-Haen­s­zel Tests. The level of significance was set at P < 0.05.Results: Thirty-three (21.1%) sheep, 2 (1.6%) cattle and 3 (11.5%) horses were seropositive to T. gondii. Analysis showed that sheep were 15 times more likely to be seropositive comparing to cattle also 2 times more likely to be seropositive than horses.Conclusion: This study showed seroprevalence of equine T. gondii infection with a considerable rate in sheep in Urmia, northwest of Iran. More comprehensive studies on livestock toxoplasmo­sis are required for further analysis of the parasite reservoir for human infection

Using Human Umbilical Cord Matrix Cells (HUCM) for Culturing Toxoplasma gondii for Serological and Molecular Assays

Introduction & Objective: Toxoplasma gondii is an intracellular parasite and one of the most common parasites between human and animals. Nowadays the molecular methods are being used for the diagnosis. Many molecular and drug assays have been performed on this parasite. Many serological and molecular methods are available for detection of this parasite. The aim of this study was to use human umbilical cord matrix cells for propagation of Toxoplasma gondii for serological and molecular assays. Materials & Methods: In this experimental study, performed in Kerman Medical University, RH strain of tachyzoite of Toxoplasma gondii was isolated from mice and cultured in HUMC, A549 and Hep2 media. The growth and generation conditions of cultured parasite in different times were studied. DMEM medium was used to compare the viability of parasite. The obtained data were analyzed by variance analysis using SPSS software. Results: Parasite entry to cell line was observed in the fist 48 hours of culturing in all media. In day 3-6, propagation of parasites in HUMC cell line was better than other cultures. In the late 6th day volume of cultivated parasites in virulence was acceptable. Conclusion: Use of human umbilical cord matrix cells is a suitable and inexpensive method for proliferation of Toxoplasma gondii. Using these cell lines are useful in providing live parasite for research purposes, drug assays or making laboratory kits

Genetic diversity and distribution of Fasciola hepatica haplotypes in Iran: Molecular and phylogenetic studies

Fasciolosis is a zoonotic disease caused by Fasciola hepatica and Fasciola gigantica. Over the last decade, diagnostic tools to detect and differentiate Fasciola species have improved, but our knowledge of the distribution of haplotypes and gene flow of this parasite is not comprehensive yet. The purpose of this study was to investigate this gap in the epidemiology of F. hepatica in different provinces of Iran between 2015 and 2017. Isolated Fasciola were collected from abattoirs in 9 provinces. The partial sequence of mitochondrial NADH dehydrogenase subunit 1 (ND1) gene was used for the identification and molecular analysis of F. hepatica isolates. The amplified PCR products were purified and subjected to direct sequencing for subsequent construction of phylogenetic tree and network analysis. In the 130 subjects analyzed, 37 ND1 haplotypes were detected. This is the first study in Iran which investigates F. hepatica population and its genetic structure, based on mitochondrial ND1 marker in different geographical regions of Iran. © 2019 Elsevier B.V

Genetic diversity analysis of Blastocystis subtypes and their distribution among the domestic animals and pigeons in northwest of Iran

Blastocystis is a unicellular, anaerobic, eukaryotic protist, a common parasite found in the intestinal tracts of animals and humans. During the last few years, the host fecal DNA analysis by nucleic acid-based method has led to significant advances in Blastocystis diagnostics and enabled subtypes (STs). The zoonotic transmission of Blastocystis to humans is not well understood, therefore the present study was conducted to identify Blastocystis subtypes in Iran from different animal hosts from northwest of Iran. A total of 427 fresh fecal specimens were collected from cattle, sheep, poultry and pigeon (40,150,132,105 respectively). To detect the Blastocystis sp., each fecal specimen was examined microscopically. Total DNA from the samples that were positive for Blastocystis sp. was isolated, and the barcoding region of the small subunit of ribosomal rRNA (18S rRNA) was amplified and sequenced. Subsequently, sequence analyses, genetic diversity indices and evolutionary relationships of Blastocystis subtype populations were carried out. In total, 14.98 of the analyzed samples were positive for Blastocystis sp. and the subtypes detected were ST3,7,10 and 14. Among these, the ST10 was the main subtype that was found only in the cattle, sheep and poultry and the zoonotic subtype ST3 was present only from cattle. Our study shows the presence of Blastocystis subtypes in the sheep in north west of Iran and also demonstrated that the genetic approaches are crucial to understand the host specify of subtypes and the mode of infection. The study suggests that the genetic approaches will help us to understand the host specificity of subtypes and their role in infection if they are obtained from human and animals from the same geographical locations. Therefore, it is important to study the zoonotic aspects of this parasite with large number of samples from different groups of animals and from different geographical locations. © 202

Systematic Review and Meta-analysis of Role of Felids as Intermediate Hosts in the Life Cycle of Neospora caninum Based on Serological Data

Purpose Neosporosis is an important widespread parasitic infection caused by N. caninum. It infects a wide range of warmblooded animals as intermediate hosts and dogs as the definitive host. Nevertheless, there are a number of questions regarding the life cycle and epidemiological aspects of N. caninum. Also, the role of felids ( domestic and non-domestic) in the life cycle of N. caninum has been little described. Therefore, this study was conducted to evaluate the global prevalence of N. caninum in domestic and wild felids. Methods PubMed, Scopus, Google Scholar, Web of Science, and ScienceDirect databases were searched for articles published on the prevalence of N. caninum in felids until Aprill 2, 2022 and the reference lists of retrieved articles were screened. A random-effects meta-analysis model was used to estimate the pooled prevalence and 95 confidence interval. Heterogeneity among studies was evaluated using Cochran's Q and the I-2 statistic. Results After exclusion of irrelevant articles and duplication removal, 30 studies were eligible for quantitative analysis including 20 studies on domestic cats and 10 studies on wild felids. The overall prevalence of neosporosis infection in cats was 15 (95 CI 10-21) that was significantly higher in wild felids (26, 95 CI 13-38) than in domestic cats (11, 95 CI 6-16) (P = 0.03). There was no significant difference in pooled prevalence between male and female domestic cats (P = 0.75). Regarding continent, the lowest prevalence of neosporosis infection was in Asia (9, 95 CI 1-20) and the highest was in North America (43.6, 95 CI 33.9-53.2) and Africa (18, 95 CI 9-46). Higher prevalence was obtained when using the NAT with 22 (95 CI 7-37), compared to the IFAT (17, 95 CI 9-24) and ELISA (6, 95 CI 2-9) (P = 0.01). Conclusion The findings highlighted the importance of felids as potential intermediate hosts of neosporosis despite the fact that the source of the parasite for these animals was unknown. Further studies should be performed to investigate the role of this top predator (felids) in maintaining both domestic and sylvatic cycles of Neospora caninum

Assessment of genetic markers for multilocus sequence typing (MLST) of Fasciola isolates from Iran

Background Several markers have been described to characterise the population structure and genetic diversity of Fasciola species (Fasciola hepatica (F. hepatica) and Fasciola gigantica (F. gigantica). However, sequence analysis of a single genomic locus cannot provide sufficient resolution for the genetic diversity of the Fasciola parasite whose genomes are similar to 1.3 GB in size. Objectives To gain a better understanding of the gene diversity of Fasciola isolates from western Iran and to identify the most informative markers as candidates for epidemiological studies, five housekeeping genes were evaluated using a multilocus sequence typing (MLST) approach. Methods MLST analysis was developed based on five genes (ND1, Pepck, Pold, Cyt b and HSP70) after genomic DNA extraction, amplification and sequencing. Nucleotide diversity and phylogeny analysis were conducted on both concatenated MLST loci and each individual locus. A median joining haplotype network was created to examine the haplotypes relationship among Fasciola isolates. Results Thirty-three Fasciola isolates (19 F. hepatica and 14 F. gigantica) were included in the study. A total of 2971 bp was analysed for each isolate and 31 sequence types (STs) were identified among the 33 isolates (19 for F. hepatica and 14 for F. gigantica isolates). The STs produced 44 and 42 polymorphic sites and 17 and 14 haplotypes for F. hepatica and F. gigantica, respectively. Haplotype diversity was 0.982 +/- 0.026 and 1.000 +/- 0.027 and nucleotide diversity was 0.00200 and 0.00353 +/- 0.00088 for F. hepatica and F. gigantica, respectively. There was a high degree of genetic diversity with a Simpson's index of diversity of 0.98 and 1 for F. hepatica and F. gigantica, respectively. While HSP70 and Pold haplotypes from Fasciola species were separated by one to three mutational steps, the haplotype networks of ND1 and Cyt b were more complex and numerous mutational steps were found, likely due to recombination. Conclusions Although HSP70 and Pold genes from F. gigantica were invariant over the entire region of sequence coverage, MLST was useful for investigating the phylogenetic relationship of Fasciola species. The present study also provided insight into markers more suitable for phylogenetic studies and the genetic structure of Fasciola parasites