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

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

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

The molecular profile of Paratrajectura longcementglandatus Amin, Heckmann et Ali, 2018 (Acanthocephala: Transvenidae) from percid fishes in the marine waters of Iran and Iraq

Paratrajectura longcementglandatus Amin, Heckmann et Ali, 2018 (Transvenidae) was recently described from two species of percid fishes collected from the marine territorial waters of Iraq and Iran in the Persian Gulf. The genus Paratrajectura Amin, Heckmann et Ali, 2018 is a close relative to transvenid genera TrajecturaPichelin et Crib, 2001 and TransvenaPichelin et Crib, 2001. Morphologically, Paratrajectura is characterised by having apical proboscis cone, long, tubular cement glands, short lemnisci, prominent roots on all proboscis hooks, subterminal female gonopore, and males with long pre-equatorial testes. Molecular studies of P. longcementglandatus using 18S rDNA and cox1 genes compared with available data of members of other families of Echinorhynchida showed that P. longcementglandatus is grouped with species of the genus Transvena forming a clade within the family Transvenidae

Identification of Acanthamoeba spp. from water and soil of public parks in north of Iran

Acanthamoeba, a free-living and opportunistic protozoan parasite, is a causative agent of severe human infections of the cornea and brain. The present study evaluated the distribution and genotyping of Acanthamoeba spp. in water and soil of recreational places in various areas in Guilan province in northern Iran. Eighty water and 20 soil samples were collected from the study area. Water samples were vacuum filtered through a 0.45 mu m pore-size membrane filter. Soil samples were washed with sterile distilled water, and washings were similarly filtered, as mentioned for water samples. The filtered material was cultured on non-nutrient agar plates seeded with heat-killed Escherichia coli. Molecular analysis was performed by PCR and sequencing using specific primers for Acanthamoeba. Finally, 26 isolates were successfully sequenced. According to culture and PCR methods, 54% of water and 100% of soil samples were contaminated with Acanthamoeba. Based on the sequencing data, genotypes T4 (47%), T5 (35.29%) and, T3 (11.76%), T11 (5.88%) were identified in water samples. GenotypesT4 (66.6%), T5 (22.2%) and T15 (11.1%) identified in water samples. Most isolates might present a potential health hazard for humans in this region. To the best of our knowledge, this is the first comprehensive survey of water and soil of recreational areas in northern Iran and the first report on identifying genotype T15 from soil sources

On the

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