Human ocular onchocerciasis caused by Onchocerca lupi (Spirurida, Onchocercidae) in Iran.

Cases of canine onchocerciasis caused by Onchocerca lupi are increasingly reported from Europe and the western United States of America. The zoonotic role of this parasite had already been suspected in Europe as the clinical signs and histopathology seen in two ocular cases from Albania and the Crimean region were very similar to those of canine ocular onchocerciasis. In the most recent reports of human onchocerciasis, O. lupi has been morphologically and molecularly identified as the causative agent of ocular infestation in two patients from Turkey, and one patient from Tunisia. Here, we report an additional case of nodular lesions involving two, and possibly more, immature worms in a patient from Iran. The parasite was found to belong to the genus Onchocerca based on morphological features and the species was confirmed as O. lupi from a partial sequence analysis of 12S ribosomal DNA.

Trichinella britovi in the jackal Canis aureus from south-west Iran.

Trichinellosis is an important helminthic food-borne zoonosis, which is caused by nematodes of the genus Trichinella. Although, Trichinella spp. has been detected frequently in Iranian wildlife, this parasitic infection is not considered a major public health problem. This is largely because Islamic codes forbid consumption of pork meat in this country. However, knowledge about this zoonotic pathogen is important because human trichinellosis has been documented in countries where most of the population is Muslim. The aims of the present work were to investigate whether Trichinella spp. was still circulating in wildlife of the Khuzestan Province (south-west Iran) about 30 years after the first investigation, to identify the aetiological agent at the species level by molecular analyses, and to review the literature on Trichinella spp. in animals of Iran. During the winter 2009-2010, muscle samples from 32 road-killed animals (14 dogs and 18 jackals, Canis aureus) were collected. Muscle samples were digested and Trichinella sp. larvae were isolated from two jackals. The Trichinella sp. larvae have been identified as Trichinella britovi by molecular analyses. These results confirm that T. britovi is the prevalent species circulating in wild animals of Iran.

Comparison of PCR-Based Diagnosis with Centrifuged-Based Enrichment Method for Detection of Borrelia persica in Animal Blood Samples.

BACKGROUND: The mainstay of diagnosis of relapsing fever (RF) is demonstration of the spirochetes in Giemsa-stained thick blood smears, but during non fever periods the bacteria are very scanty and rarely detected in blood smears by microscopy. This study is aimed to evaluate the sensitivity of different methods developed for detection of low-grade spirochetemia. METHODS: Animal blood samples with low degrees of spirochetemia were tested with two PCRs and a nested PCR targeting flaB, GlpQ, and rrs genes. Also, a centrifuged-based enrichment method and Giemsa staining were performed on blood samples with various degrees of spirochetemia. RESULTS: The flaB-PCR and nested rrs-PCR turned positive with various degrees of spirochetemia including the blood samples that turned negative with dark-field microscopy. The GlpQ-PCR was positive as far as at least one spirochete was seen in 5-10 microscopic fields. The sensitivity of GlpQ-PCR increased when DNA from Buffy Coat Layer (BCL) was used as template. The centrifuged-based enrichment method turned positive with as low concentration as 50 bacteria/ml blood, while Giemsa thick staining detected bacteria with concentrations ≥ 25000 bacteria/ml. CONCLUSION: Centrifuged-based enrichment method appeared as much as 500-fold more sensitive than thick smears, which makes it even superior to some PCR assays. Due to simplicity and minimal laboratory requirements, this method can be considered a valuable tool for diagnosis of RF in rural health centers.

Molecular typing and phylogenetic analysis of some species belonging to phlebotomus (larroussius) and phlebotomus (adlerius) subgenera (Diptera: psychodidae) from two locations in iran.

BACKGROUND: Haematophagous females of some phlebotomine sandflies are the only natural vectors of Leishmania species, the causative agents of leishmaniasis in many parts of the tropics and subtropics, including Iran. We report the presence of Phlebotomus (Larroussius) major and Phlebotomus (Adlerius) halepensis in Tonekabon (Mazanderan Province) and Phlebotomus (Larroussius) tobbi in Pakdasht (Tehran Province). It is the first report of these species, known as potential vectors of zoonotic visceral leishmaniasis in Iran, are identified in these areas. METHODS: In 2006-2007 individual wild-caught sandflies were characterized by both morphological features and sequence analysis of their mitochondrial genes (Cytochrome b). The analyses were based on a fragment of 494 bp at the 3' end of the Cyt b gene (Cyt b 3' fragment) and a fragment of 382 bp CB3 at the 5' end of the Cyt b gene (Cyt b 5' fragment). We also analysed the Cyt b Long fragment, which is located on the last 717 bp of the Cyt b gene, followed by 20 bp of intergenic spacer and the transfer RNA ser(TCN) gene. RESULTS: Twenty-seven P. halepensis and four P. major from Dohezar, Tonekabon, Mazanderan province and 8 P. tobbi from Packdasht, Tehran Province were identified by morphological and molecular characters. Cyt b 5' and Cyt b 3' fragment sequences were obtained from 15 and 9 flies, respectively. Cyt b long fragment sequences were obtained from 8 out of 27 P. halepensis. CONCLUSION: Parsimony analyses (using heuristic searches) of the DNA sequences of Cyt b always showed monophyletic clades of subgenera and each species did form a monophyletic group.

Molecular characterization of Anopheles fluviatilis species complex in the Islamic Republic of Iran.

A species-specific polymerase chain reaction (PCR) assay was used to identify the species composition of the Anopheles fluviatilis complex in the Islamic Republic of Iran. All the amplified DNA samples from specimens collected from different areas yielded a fragment of 450 bp size, a PCR product corresponding to that of the species denoted as Y. The sequence data from 21 ITS2 [second internal transcribed spacer] regions were compared with those publicly available in the GenBank database and confirmed that the specimens were 100% identical to species Y of India. Species Y is presumably the same as species T that has no role in transmission of malaria in India, whereas An. fluviatilis is known as a secondary vector of malaria in the Islamic Republic of Iran.

Molecular characterization of Anopheles fluviatilis species complex in the Islamic Republic of Iran

A species-specific polymerase chain reaction [PCR] assay was used to identify the species composition of the Anopheles fluviatilis complex in the Islamic Republic of Iran. All the amplified DNA samples from specimens collected from different areas yielded a fragment of 450 bp size, a PCR product corresponding to that of the species denoted as Y. The sequence data from 21 ITS2 [second internal transcribed spacer] regions were compared with those publicly available in the GenBank database and confirmed that the specimens were 100% identical to species Y of India. Species Y is presumably the same as species T that has no role in transmission of malaria in India, whereas An. fluviatilis is known as a secondary vector of malaria in the Islamic Republic of Iran