Morphological and Molecular Characteristic of Megaselia scalaris (Diptera: Phoridae) Larvae as the Cause of Urinary Myiasis.

We report a case of urinary myiasis occurring in a 60-yr-old Iranian male patient with urinary tract problems and a history of travel to Thailand who was referred to Shafagh Medical Laboratory in Tehran (Iran). Larvae excreted in the patient's urine were confirmed by morphological identification key and DNA barcoding to belong to the species Megaselia scalaris Loew, which is known as the scuttle fly. Based on the patient's history, he was infected with M. scalaris in Thailand. To the best of our knowledge, this is the first report of urinary myiasis caused by M. scalaris in Thailand.

The Anopheles stephensi odorant binding protein 1 (AsteObp1) gene: a new molecular marker for biological forms diagnosis.

Anopheles (Cellia) stephensi Liston 1901 is known as an Asian malaria vector. Three biological forms, namely "mysorensis", "intermediate", and "type" have been earlier reported in this species. Nevertheless, the present morphological and molecular information is insufficient to diagnose these forms. During this investigation, An. stephensi biological forms were morphologically identified and sequenced for odorant-binding protein 1 (Obp1) gene. Also, intron I sequences were used to construct phylogenetic trees. Despite nucleotide sequence variation in exon of AsteObp1, nearly 100% identity was observed at the amino acid level among the three biological forms. In order to overcome difficulties in using egg morphology characters, intron I sequences of An. stephensi Obp1 opens new molecular way to the identification of the main Asian malaria vector biological forms. However, multidisciplinary studies are needed to establish the taxonomic status of An. stephensi.

First record of a new member of Anopheles Hyrcanus Group from Iran: molecular identification, diagnosis, phylogeny, status of kdr resistance and Plasmodium infection.

The current study aimed to provide further evidence on the status of species composition, insecticide resistance, and vectorial capacity within the members of Anopheles (Anopheles) Hyrcanus Group in Ardebil, Giulan, and Khuzestan provinces of Iran. Sequencing the internal transcribed spacer 2 (ITS2) of ribosomal DNA gene led to identification of two members of Hyrcanus complex: Anopheles hyrcanus Pallas and a new species/form, hereafter called Anopheles hyrcanus sp(IR) as a world record. Furthermore, we identified and compared partial sequences of exons I and II and the whole intron I region of insecticide resistance-related voltage-gated sodium channel (vgsc) gene in populations of Hyrcanus Group and other main old world Anopheles species. The ITS2 and vgsc sequences in members of Hyrcanus Group and other Anopheles species were used for construction of phylogenetic tree, which demonstrated the evolutionary relatedness among Western and Eastern Palearctic taxa within the Hyrcanus Group. A nested polymerase chain reaction assay for detection of Plasmodium species revealed the infection of Plasmodium falciparum within An. hyrcanus collected from Fooman district in Guilan province. The data from this study led to the introduction of a new member/form within the Hyrcanus Group, identification and definition of the status of knockdown resistance related to pyrethroids and DDT in their vgsc gene, detection of Plasmodium infection, and further evidence on genetic relatedness within these taxa. The overall results may suggest reconsidering the role ofAn. hyrcanus in malaria transmission, which would be useful for implementation and evaluation of malaria control programs in Western Palearctic region.

Molecular characteristics of Anopheles maculipennis Meigen in Zanjan, north west of Iran, inferred from ITS2 sequence analysis.

The present study has been designed in order to verify the species composition within Anopheles maculipennis complex in North West of Iran. We determined ribosomal DNA sequences of the second internal transcribed spacer (ITS2) region from samples of Anopheles maculipennis Complex in Zanjan province. A total of 1536 specimens within the Complex were tested by Multiplex PCR, only An. maculipennis was found in this area. One clone out of four different individual mosquitoes of each field was generated with ITS2 PCR and half of them (192 samples) selected randomly for RFLPs. PCR-RFLP assay identified 2 haplotypes; haplotype I (99%) and haplotype II (1%). Twenty five sequences were generated comprising the 5.8S gene, the ITS2 and the 28S ribosomal gene. The alignment was 422 in length and percentage of GC content was 50.3% (26.07% A, 23.59% T, 26.78% C, 23.7% G). The ITS2 was 290 bp in length and two haplotypes were revealed varying by a single base (T<-->C) at site 378. An. maculipennis is the dominant species anopheline of the province. ITS2 analysis revealed evidence of a slightly interaspecific variation among populations. However, further investigations on the genetic polymorphism among An. maculipennis populations and in particular within those belonging to the continental haplotype are required to support any hypothesis on differences in behavior across the distribution range for this potential malaria vector.

Inter and intra-specific genetic variation of avian Eimeria isolated from Iran by random amplified polymorphic DNA--polymerase chain reaction.

Eimeria species from poultry breeder farms without previous exposure to anticoccidial vaccines in five distinct geographical regions of Iran were examined for genetic relatedness by the random amplified polymorphic DNA (RAPD) assay. Eight different oligonucleotide decamers with arbitrary DNA sequences were tested as primers to amplify DNA from five isolates of each E. acervulina, E. tenella, and E. maxima. Depending on the species/isolate-primer combination, between 1 and 14 DNA fragments ranging in size from 240 to 3000 bp were amplified. The two isolates originated from Northeast and North parts of Iran showed minor differences and two isolates originated from Northeast and Southwest of Iran showed major differences in their amplified DNA patterns. The intra-specific similarity coefficient within five isolates of each species of, E. acervulina, E. tenella, E. maxima was 74, 82 and 72%, respectively. The distance indices observed between species were greater than those found between isolates (80-90%) with all examined primers. The inferred phylogenetic tree on the fingerprinting of all species revealed that the RAPD-PCR can easily differentiate within and between species and could be a useful and valuable tool in future epidemiological studies, designing and developing of vaccines against avian coccidosis, here in Iran and neighboring countries.