Prevalence and phylogenetic analysis of gastrointestinal helminths(Nematoda: Trichostrongylidae) in ruminant livestock of northwest Iran

Trichostrongylidae family is considered as a group of gastrointestinal nematodes of ruminants with widespread distribution in the world. The parasites are the major causes of productivity loss in many countries including Iran. This study was carried out to determine the distribution of abomasal Trichostrongylidae among livestock of Qazvin, Iran. Totally, 160 abomasum samples including 83 from sheep, 72 from cattle and 5 from goats were collected from slaughterhouses throughout the Qazvin Province. The ribosomal DNA internal transcribed spacer 2 (ITS2) region was amplified using PCR followed by sequencing 13 different isolates. Phylogenetic analysis of the identified sequences was performed using MEGA 7 software. The prevalence rate of the nematodes was 19.4% among the specimens. Also, two spices including Marshallagia marshalli and Teladorsagia circumcincta were identified among the isolates with M. marshalli at higher frequency. No Trichostrongylus spp. was detected in the region. Since livestock plays a major role in ecosystem balance and as some species of nematodes have a zoonotic nature, it is necessary to proceed with more comprehensive epidemiologic studies to clarify the infection rate among the human population living in the region

Comparison of six simple methods for extracting ribosomal and mitochondrial DNA from Toxocara and Toxascaris nematodes

Six simple methods for extraction of ribosomal and mitochondrial DNA from Toxocara canis, Toxocara cati and Toxascaris leonina were compared by evaluating the presence, appearance and intensity of PCR products visualized on agarose gels and amplified from DNA extracted by each of the methods. For each species, two isolates were obtained from the intestines of their respective hosts: T. canis and T. leonina from dogs, and T. cati from cats. For all isolates, total DNA was extracted using six different methods, including grinding, boiling, crushing, beating, freeze-thawing and the use of a commercial kit. To evaluate the efficacy of each method, the internal transcribed spacer (ITS) region and the cytochrome c oxidase subunit 1 (cox1) gene were chosen as representative markers for ribosomal and mitochondrial DNA, respectively. Among the six DNA extraction methods, the beating method was the most cost effective for all three species, followed by the commercial kit. Both methods produced high intensity bands on agarose gels and were characterized by no or minimal smear formation, depending on gene target; however, beating was less expensive. We therefore recommend the beating method for studies where costs need to be kept at low levels. © 2013 Elsevier Inc

Comparison of six simple methods for extracting ribosomal and mitochondrial DNA from Toxocara and Toxascaris nematodes

Six simple methods for extraction of ribosomal and mitochondrial DNA from Toxocara canis, Toxocara cati and Toxascaris leonina were compared by evaluating the presence, appearance and intensity of PCR products visualized on agarose gels and amplified from DNA extracted by each of the methods. For each species, two isolates were obtained from the intestines of their respective hosts: T. canis and T. leonina from dogs, and T. cati from cats. For all isolates, total DNA was extracted using six different methods, including grinding, boiling, crushing, beating, freeze-thawing and the use of a commercial kit. To evaluate the efficacy of each method, the internal transcribed spacer (ITS) region and the cytochrome c oxidase subunit 1 (cox1) gene were chosen as representative markers for ribosomal and mitochondrial DNA, respectively. Among the six DNA extraction methods, the beating method was the most cost effective for all three species, followed by the commercial kit. Both methods produced high intensity bands on agarose gels and were characterized by no or minimal smear formation, depending on gene target; however, beating was less expensive. We therefore recommend the beating method for studies where costs need to be kept at low levels. © 2013 Elsevier Inc

Molecular Characterization of Mitochondrial Cytochrome c oxidase subunit 1 (Cox1) gene from Trichostrongylus Species (Nematoda: Trichostrongylidae) in Northern Iran

Objective: The objective of the present study was to identify Trichostrongylus species by molecular analysis and also phylogenetic relationships of Trichostrongylus species by mitochondrial Cytochrome c oxidase subunit 1 (Cox1) gene in Guilan province, northern Iran. Methods: Abomasum and duodenum contents of 144 livestock were collected from sheep, goats, and cattle in Guilan province. Morphological survey was performed for initial screening. Total DNA was extracted, and the partial region of Cox1 gene was amplified and sequenced. Genetic diversity was calculated and phylogenetic analysis of the data on nucleotide sequence was conducted by MEGA7 software. Results: Three species of Trichostrongylus including T. colubriformis, T. vitrinus, and T. axei were identified by morphological characteristics. The genetic divergence within the species in the present study was observed for T. axei (0-2.5%), T. colubriformis (0.77%), and T. vitrinus (0%). The mean inter-species difference between the three species of Trichostrongylus obtained in this study was 14.4-15.4%. Conclusion: The Cox1 sequences of the members of Trichostrongylus spp. were highly variable and this could be used as a valuable measure to achieve a proper assessment on biodiversity. Sequence data generation from other species of Trichostrongylus will be needed to reconstruct the phylogenetic relationships of this genus of nematodes. Keywords: Trichostrongylus spp., ruminants, Cox1, phylogenetic analysis, Ira

On the Neoechinorhynchus agilis (Acanthocephala: Neoechinorhynchidae) complex, with a description of Neoechinorhynchus ponticus n. sp. from Chelon auratus in the Black Sea

We recognize four species in the Neoechinorhynchus agilis complex. We studied specimens of Neoechinorhynchus (Hebesoma) personatus Tkach, Sarabeev & Shvetsova, 2014 from Mugil cephalus in the Mediterranean Sea off Tunisia and in the Black Sea, and also specimens of Neoechinorhynchus ponticus n. sp. from Chelon auratus Risso in the Black Sea. Specimens from M. cephalus at both locations were similar. All structures of N. ponticus n. sp. were considerably smaller than those of N. personatus. Two other species of the N. agilis complex are recognized: Neoechinorhynchus agilis (Rudolphi, 1819) sensu stricto from various hosts in the Atlantic and the Mediterranean, and Neoechinorhynchus yamagutii Tkach, Sarabeev & Shvetsova, 2014 from M. cephalus and Planiliza haematocheila in the Pacific, especially the Sea of Japan. Neoechinorhynchus dimorphospinus Amin & Sey, 1996 from marine fish in the Persian Gulf and the Pacific Ocean off Vietnam may be a candidate for membership in the N. agilis complex. X-ray scans of gallium cut and intact hooks of N. personatus and N. ponticus showed differences in the mineral content of hooks with higher sulfur levels in smaller hooks and in hooks from specimens in the Black Sea compared to specimens from the Mediterranean. The relatively high genetic differences between N. ponticus n. sp. and other species of Neoechinorhynchus using a partial 18S rDNA dataset support its independent status. Neoechinorhynchus ponticus n. sp. and N. personatus have a common ancestor with species of Neoechinorhynchus collected from saltwater fish

Intestinal protozoan and helminthic infections among hemodialysis and cancer patients

Intestinal parasitic infections (IPIs) can be a severe threat to immunocompromised patients. This is particularly true for those undergoing chemotherapy and hemodialysis. The present research is aimed at identifying intestinal parasites that might be present in immunocompromised patients. In this cross-sectional study 1040 stool samples were collected (279 samples from hemodialysis patients, 362 samples from chemotherapy patients and 399 samples from the control group) from March to September 2017. The samples were tested by direct, formalin-ether methods for protozoa and ova of intestinal parasites and Ziehl-Neelsen staining methods for coccidian parasites such as Cryptosporidium species. The overall parasitic infection rate was higher (15%) in hemodialysis patients and 11.3% in chemotherapy patients whereas the lowest rate was observed (7.3%) in the control group. The infectivity rates were statistically significant (P = 0.008) when compared with the control group. The most prevalent parasites present were Blastocystis hominis (8.9% of the cases), Entamoeba coli (1.6%), Iodamoeba butschlii (0.8%), Endolimax nana (0.6%), Chilomastix mesnili (0.5%), Strongyloides stercoralis (0.5%) and Taenia species (0.15%), whereas Giardia lamblia was found present only in the control group. Statistical analyses revealed a significant correlation between gastrointestinal symptoms (such as some abdominal pain, diarrhea) and the ratio of chemotherapy frequency to scheduled frequency and parasitic infections (P = 0.001). There was not a correlation between prevalence of parasites with age or education levels of the infected individuals. Results of the present study suggest that a periodic stool examinations in special parasitological laboratories should be included as part of routine and general medical care

Molecular detection of anaplasma marginale and Anaplasma ovis (Rickettsiales: Anaplasmataceae) in ixodid tick species in Iran

The present study was conducted as the first molecular detection of Anaplasma species in tick samples based on the sequencing of major surface proteins 4 (msp4) gene fragments in different parts of Iran. A total of 130 tick specimens were collected from Hormozgan, Lorestan, and Guilan, Iran, within 2015 to 2017. Hyalomma asiaticum, Hyalomma dromedarii, Rhipicephalus sanguineus, and Rhipicephalus (Boophilus) species were identified in different geographical regions. An amplicon of 464-bp msp4 of Anaplasma was amplified using polymerase chain reaction in various tick species. Three sequences, including one Anaplasma marginale from R. (Boophilus) species and two Anaplasma ovis from Rhipicephalus sanguineus, were obtained after sequencing. It is concluded that bovine and ovine anaplasmosis agents are present in tick samples in Iran. The use of the gene families of six major surface proteins for the detection of various Anaplasma species is recommended. © 2020 by Razi Vaccine & Serum Research Institute

Infection of the Jackal (Canis aureus) by Haplor-chis taichui (Trematoda: Heterophyidae) in Southwestern Iran: A Clue for Potential Human Infection

Background: We detected eight trematodes in the small intestine of a road-killed jackal (Canis aureus) from Hamidiyeh District near the city of Ahvaz, Khuzestan Province in 2010. Methods: Three worms were stained with carmine acid, mounted in Canada balsam on glass slides and examined under a light microscope at 1000X magnification. PCR and sequencing of a partial ITS2 sequence were used to approve the diagnosis. Results: The flukes measured ≈1 mm in length with an elongated ovoid shape resembling the members of heterophyid, and only one testis, characteristics of the genus Haplorchis. Sequencing of a 481-bp fragment of the ITS2 locus from the worms revealed 97%-98% identity with the similar sequences of the H. taichui flukes previously identified in the fish, cat, and humans from Thailand, China, and Vietnam. Conclusion: Further studies with the application of reliable molecular tools to diagnose trematode infections in wildlife and humans can bring more insight into the epidemiology of fish-borne flukes including H. taichui in this area

Study of the effect of iron oxide nanoparticles on mouse testis development during the embryonic period in NMRI strain

Of different types of bio-compatible nano‌particles, iron oxide nanoparticle has attracted attention for its applications especially in medicine and magnetic resonance imaging (MRI). We decided to study histological changes in testis and spermatogenesis in mature male mice that are affected by ironoxide nanoparticles (Fe2O3) during fetal period. In this study, the mice were divided into three experimental and two control and sham groups. Iron oxide at do-ses of 10, 30, 50 mg/kg were injected into pregnant mice during 10th, 12th and 14th days of the critical period of fetus testis development. After the maturation of the mice which were born, slices with the thickness of 0.06 µm were prep-ared from adult male testis and epididymis, stained with hematoxylin and eosin. The cells were counted, then the obta-ined data was analyzed with one-way ANONA and Tukey testes. The results showed that the number of epididymis sp-erms at the doses of 10 and 30 mg/kg, testis weight at the dose of 30 mg/kg, primary spermatocytes at the dose 10mg/kg had increased, while the testis diameter in all the three experimental groups and the volume of the testis at the dose of 50 mg/kg had decreased. Disorganization and vacuolization were observed at high doses. Passing through the cell me-mbrane, and considering its oxidation and reduction potentials, it was observed that Iron oxide nanoparticle acts as anti-oxidant at low doses and shows toxicity at high doses. The fact obtained in the paper reflects the dual potential of the ir-on oxide nanoparticle