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This study aimed to determine the chemical compositions of crude aquatic extracts of M. pulegium L. and R. idaeus L., and their anti‐Toxoplasma activity. Crude aquatic extraction of aerial parts of R. idaeus L. and M. pulegium L. was performed. GC‐MS and HTPLC analyses were carried out. MTT assay was performed on Vero cells treated by different concentrations (Log −10 from 10−1 to 10−6) of the extracts. The anti‐Toxoplasma activity of the concentrations was investigated using vital staining. Menthol (99.23%) and limonene (0.227%) were the major compounds of the aquatic extract of M. pulegium L. Phytochemical compositions of R. idaeus L. were terpenoids, esterols, and flavonoids. The cell toxicity of M. pulegium L. was lower than R. idaeus L. (CC50 > 10−2 versus. ≥ 10−4). Aquatic extract of M. pulegium L. showed higher anti‐Toxoplasma activity (LC50 ≥ 10−6) than R. idaeus L. (LC50 ≥ 10−5). Statistically significant cell toxicity and anti‐Toxoplasma activity (p

Blastocystis is a zoonotic protozoan parasite frequently identified in the intestinal tract of humans and a vast variety of animals, worldwide. Here, we assessed the prevalence of Blastocystis and its subtypes in stool samples of raccoons. Stool samples from 30 raccoons were collected. Total DNA was extracted, and the barcoding region of the small subunit ribosomal rRNA (SSU rRNA) gene was amplified and sequenced. Specific fragment for Blastocystis was successfully amplified in five samples (16.66%). Sequencing analysis revealed ST1, ST2, and ST3 among 1, 2, and 2 Blastocystis-positive samples. Our results documented the presence of Blastocystis subtypes 1-3 in raccoons. Subtype 1 showed higher similarity to the human isolates of Blastocystis. However, it seems that raccoons may emerge as reservoirs for Blastocystis and may be linked to zoonotic transmission of the protist.

Microsporidia are known opportunistic microorganisms and usually transmitted via the fecal-oral route. However, there is no information about human-infecting microsporidia in wildlife in Iran. This study aimed to investigate and analyze human-infecting microsporidia isolated from raccoons in north of Iran. Totally, 30 fecal samples were collected; then, DNA extraction was performed and specific fragments of the SSU rRNA gene of Enterocytozoon bieneusi and Encephalitozoon species were amplified. After amplification and sequencing the ITS, the results were compared to the GenBank database. Phylogenetic trees and network analysis were employed to explore probable relationships. E. bieneusi was the only detected microsporidia among samples. Genotyping showed the genotypes D, E, and RA in 15/18 (83.33%), 1/18 (5.55%), and 2/18 (11.11%) of samples, respectively. Novel genotypes RA1 and RA2 grouped together and apart from other genotypes. E. bieneusi genotypes D and E clustered with the genotypes previously reported from animals, humans, and environmental samples. Network analysis revealed six distinct sequence types among raccoon's isolates. This study demonstrated that E. bieneusi genotype D was the most prevalent microsporidia among raccoons. It seems that wildlife may play a role in dispersion of microsporidia spores.

Summary 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.

Background: The raccoon, Procyon lotor Linn. (Procyonidae) is native to North and Central America but has been introduced in several European and Asian countries including Japan, Germany and Iran. Objective of this study was to determine frequency of gastrointestinal and tissue helminthes from feral raccoons in Iran. Methods: During 2015-2017, 30 feral raccoons including 12 males and 18 females were collected from Guilan Province, northern Iran (the only region in Iran where raccoons are found). The gastrointestinal tracts and tissues such as lung, liver and muscles were examined for presence of helminthes. Results: Twenty raccoons (66.7%) were found infected with five intestinal helminth species. The prevalence of infection with Strongyloides procyonis Little, 1966 (Nematoda) was 63.3%, Plagiorchis koreanus Ogata, 1938 (Trematoda) (13.3%), Centrorhynchus sp. Lühe, 1911 (Acanthocephala) (10.0%), Camerostrongylus didelphis Wolfgang, 1951 (Nematoda) (3.3%), and Spirocerca lupi Rudolphi, 1809 (Nematoda) (3.3%). No larvae or adult worms were found in other tissues of the examined raccoons. Conclusion: Most of the raccoons were infected with S. procyonis. The public health importance of zoonotic parasites transmittable through raccoons, the rapid control and decrease of raccoon populations and their distribution in Iran are also discussed.

Introduction The role of parasites in the pathogenesis of appendicitis has been debated for a long time. To date, several gastrointestinal parasites have been reported as the causes of appendicitis in humans. Taenia infestation of the appendix is uncommon and few cases have been reported in the literature. Presentation of case We reported a case of acute eosinophilic appendicitis (AEA) in a 42-year-old woman caused by T. saginata in northern Iran. The patient was admitted to the emergency department with a 2-day history of acute abdominal pain in her lower right quadrant. Abdominal ultrasonography showed intra-abdominal bleeding and endometrium cysts. Routine hematological tests showed increases in white blood cell (WBC) count of 19.8 × 103 per mcL with 3% eosinophilia. During abdominal laparotomy, peritoneal fluid was bulked with abdominal bleeding due to rupture of the uterine cyst. After investigation of inflammation in the appendix region, patient underwent appendectomy. Histopathological findings showed acute inflammation with eosinophils and a large number of round eggs with flattened segments of the genus Taenia. It is impossible to distinguish between T. saginata and T. solium based solely on egg morphology in the specimens. Therefore, based on history of the patient, which included no consumption of pork, the species was identified as T. saginata. At the three months follow-up, the patient was in good health. Conclusion In the current study, a case of AEA by T. saginata was reported. However, this was not the first case of acute appendicitis by T. saginata. Further studies are necessary to show roles of parasites in pathogenesis of AEA., Highlights • T. saginata is distributed globally, and more frequent in developing countries. • Taenia infestation of the appendix is uncommon and few cases have been reported in the literature. • In the current study, a case of acute eosinophilic appendicitis caused by T. saginata was reported.

Identification of different genotypes of echinococcal cyst in various domestic herbivores and humans within the target area was the principal aim of the present study, performed using sequence data of cox1 and nad1 mitochondrial genes. A total of 57 cystic echinococcosis (CE) cysts were isolated from indigenous livestock including 45 cattle, 9 sheep and 3 goats from several slaughterhouses in Guilan Province. Moreover, 12 formalin-fixed paraffin-embedded (FFPE) CE cyst tissues from humans were also included, obtained from the archives of several hospitals in Rasht, the capital of Guilan. Genetic sequencing was conducted using mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes. Our results found that E. granulosus sensu stricto (s.s.) and E. ortleppi were present in 92.7% and 7.2% isolates, respectively. E. granulosus s.s. (genotypes G1 and G3) and E. ortleppi were isolated from various livestock whereas all CE cysts isolated from humans were E. granulosus s.s. G1 genotype. We found that E. granulosus s.s. G1 was the predominant genotype within the study region. This is the first study to report E. ortleppi in cattle in Iran.

We recognize four species in the Neoechinorhynchus agilis complex. We studied specimens of Neoechinorhynchus (Hebesoma) personatus Tkach, SarabeevShvetsova, 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, SarabeevShvetsova, 2014 from M. cephalus and Planiliza haematocheila in the Pacific, especially the Sea of Japan. Neoechinorhynchus dimorphospinus AminSey, 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.Sur le complexe Neoechinorhynchus agilis (Acanthocephala, Neoechinorhynchidae), avec la description de Neoechinorhynchus ponticus n. sp. de Chelon auratus en mer Noire.Nous reconnaissons quatre espèces dans le complexe Neoechinorhynchus agilis. Nous avons étudié des spécimens de Neoechinorhynchus (Hebesoma) personatus Tkach, SarabeevShvetsova, 2014 de Mugil cephalus en mer Méditerranée au large de la Tunisie et en Mer Noire et également des spécimens de Neoechinorhynchus ponticus n. sp. de Chelon auratus Risso en Mer Noire. Les spécimens chez M. cephalus dans les deux endroits étaient similaires. Toutes les structures de N. ponticus n. sp. étaient considérablement plus petites que celles de N. personatus. Deux autres espèces du complexe N. agilis sont reconnues: Neoechinorhynchus agilis (Rudolphi, 1819) sensu stricto provenant de divers hôtes de l’Atlantique, y compris la Méditerranée, et Neoechinorhynchus yamagutii Tkach, SarabeevShvetsova, 2014 de M. cephalus et Planiliza haematocheila dans le Pacifique, en particulier la Mer du Japon. Neoechinorhynchus dimorphospinus AminSey, 1996 provenant de poissons marins du golfe Persique et de l’océan Pacifique au large du Vietnam pourrait être candidat à l’appartenance au complexe N. agilis. Les radiographies des crochets coupés au gallium et des crochets intacts de N. personatus et N. ponticus ont montré des différences dans la teneur en minéraux des crochets avec des niveaux de soufre plus élevés dans les hameçons plus petits et dans les hameçons des spécimens de la Mer Noire par rapport aux spécimens de la Méditerranée. Les différences génétiques relativement élevées entre N. ponticus n. sp. et d’autres espèces de Neoechinorhynchus utilisant un ensemble de données d’ADNr 18S partiel soutiennent son statut indépendant. Neoechinorhynchus ponticus n. sp. et N. personatus ont un ancêtre commun avec des espèces de Neoechinorhynchus prélevées sur des poissons d’eau salée.

We describe morphological features not previously reported for this old acanthocephalan Nephridiacanthus major (Bremser, 1811 in Westrumb, 1821) Golvan, 1962 first described over 200 years ago. Our specimens were collected from long-eared hedgehog Hemiechinus auritus (Gmelin, 1770) (Erinaceidae) in Iran. We compare the morphometrics of our material with others previously reported from the same host in Iran, Russia, central Asia and Europe. Our specimens had markedly smaller proboscides, proboscis hooks and lemnisci than those reported from Russia and central Asia, but comparable measurements of other structures with specimens previously described from other collections. We document our new observations with scanning electron microscopy features not previously demonstrable by other observers and provide a chemical analysis of proboscis hooks using energy-dispersive X-ray analysis for the first time. The molecular profile of this acanthocephalan, based on 18S rDNA and cox1 genes, was generated for the first time. The phylogenetic analysis showed that N. major is placed in a clade of the family Oligacanthorhynchidae, well separated from the families Moniliformidae and Gigantorhynchidae.

This study is part of an effort to document the diversity of avian schistosomes in ducks and snails in Northern Iran, a major flyway (Black Sea/Mediterranean) for migratory birds and where cercarial dermatitis (CD) is prevalent in rice growing areas. CD is an allergic skin reaction from schistosome trematodes that emerge from aquatic snails. Most CD cases are reported from recreational swimmers or aquaculture farmers. Much of the work on the epidemiology of CD has focused in recreational waters in the Americas and Europe, with fewer studies in aquaculture, particularly in Iran. The artificial environment at aquaculture sites support dense populations of snails that are hosts to schistosomes, as well as domestic ducks. Thus, are domestic ducks reservoir hosts of species of Trichobilharzia, one of the main etiological agents of CD in Northern Iran? This study focused on a survey of domestic ducks for the presence of the nasal schistosome, T. regenti, that has been reported widely in Europe. Trichobilharzia regenti were found in domestic ducks in the Guilan Province of Iran based on morphological and molecular analyses. The presence of this species in Northern Iran indicates that the domestic duck can serve as a reservoir host for this species and that one of the local snail species is likely the intermediate host. The continued study and surveillance of this species is important because it is a neuropathic schistosome that can use a diversity of bird definitive hosts and Radix snails that are widespread across Eurasia.

BackgroundCanids are definitive hosts ofEchinococcus multilocularisandEchinococcus granulosus.This study aimed to survey these twoEchinococcusspecies in canids of North-Khorasan Province, northeastern Iran, using morphological criteria and genetic characterization of mitochondrial DNA.MethodsThe carcasses of 106 canids, namely 61 jackals (Canis aureus), 23 foxes (Vulpes vulpes), 19 dogs (Canis familiaris) and three wolves (Canis lupus) were collected from the study area in 2013–2014 and examined forEchinococcusspecies. Morphological features were assessed by microscopy of adult worms. For molecular characterization, DNA was extracted, mostly from the adult worms but also from eggs. DNA fragments of the cytochromecoxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) mitochondrial genes were amplified and sequenced. Sequences were aligned and compared with reference sequences. Intraspecific and interspecific diversity were calculated and phylogenetic analysis was performed.ResultsOverall, 9.4% of the canids (eight jackals and two foxes) were found infected withE. multilocularisby molecular methods, of which seven cases were also confirmed using morphological description of the adult worms.Echinococcus granulosuswas found in 6.6% of the canines (four dogs, two jackals and one wolf) as determined by both molecular methods and adult cestode morphology. AllE. granulosusisolates were identified as the G1 genotype. Comparative sequence analysis indicated 0–0.7% and 0% intraspecific divergence withinE. granulosusisolates and 0% and 0–0.2% withinE. multilocularisisolates forcox1 andnad1, respectively.ConclusionsThis study revealed the presence ofE. multilocularisandE. granulosusin canids of North-Khorasan Province of Iran. Jackals were found infected with bothE. multilocularisandE. granulosus, but infection with the former species was higher.

Trichostrongylusspp. are primarily parasites of ruminants, but humans can become infected as accidental hosts. Information about the clinical aspects of human trichostrongyliasis is limited. This study investigated the clinical and haematological characteristics of a large number of trichostrongyliasis patients. In the Fouman district of Guilan Province in northern Iran, during 2015–2016, 60 patients were identified as positive forTrichostrongylusspp., using stool examination methods. The clinical manifestations and demographic data of all patients were recorded and further analysed. Twenty-three patients (38.3%) were male and 37 (61.7%) were female. Among the individuals infected only withTrichostrongylus, only nine patients (16.4%) were asymptomatic. Forty-six patients (83.6%) presented with gastrointestinal (76.3%), pulmonary (30.9%) and cutaneous (12.7%) symptoms. No statistically significant relationship was found between clinical manifestations and sex or age groups. Ten patients (18.1%) revealed eosinophilia and five (9.1%) presented with hypochromic microcytic anaemia. The relationship between eosinophilia and age group, sex and clinical manifestations showed no statistical significance. Our study indicated that trichostrongyliasis may be a major parasitic aetiology for gastrointestinal symptoms and eosinophilia in rural residents of endemic areas.

The present study was performed to analyze molecularly the phylogenetic positions of human-infecting Trichostrongylus species in Mazandaran Province, Iran, which is an endemic area for trichostrongyliasis. DNA from 7 Trichostrongylus infected stool samples were extracted by using in-house (IH) method. PCR amplification of ITS2-rDNA region was performed, and products were sequenced. Phylogenetic analysis of the nucleotide sequence data was performed using MEGA 5.0 software. Six out of 7 isolates had high similarity with Trichostrongylus colubriformis, while the other one showed high homology with Trichostrongylus axei registered in GenBank reference sequences. Intra-specific variations within isolates of T. colubriformis and T. axei amounted to 0–1.8% and 0–0.6%, respectively. Trichostrongylus species obtained in the present study were in a cluster with the relevant reference sequences from previous studies. BLAST analysis indicated that there was 100% homology among all 6 ITS2 sequences of T. colubriformis in the present study and most previously registered sequences of T. colubriformis from human, sheep, and goat isolates from Iran and also human isolates from Laos, Thailand, and France. The ITS2 sequence of T. axei exhibited 99.4% homology with the human isolate of T. axei from Thailand, sheep isolates from New Zealand and Iran, and cattle isolate from USA.

Treated wastewater samples were collected, filtered using sterile 47-mm cellulose nitrate membrane and DNA extracted from the filtered materials. The presence of Blastocystis sp. was confirmed via polymerase chain reaction (PCR) targeting the SSU rRNA gene of Blastocystis sp. in 5/12 of samples. Based on the subtype analysis after sequencing, 2, 2 and 1 of ST2, ST6 and ST8 were detected among the isolates, respectively. Furthermore, both ST6s were allele 139, alleles 11 and 138 were identified in ST2 and the only ST8 was allele 95. The phylogenetic tree showed that one of ST2 was clustered together with those ST2 that were already reported from humans and animals. The presence of Blastocystis sp. in treated wastewater can indicate the potential role of this type of water for irrigation in the transmission of pathogenic microorganisms to downstream farmlands.

Abstract Background: Due to the similarity of Strongyloides stercoralis with free-living nematodes of Rhabditis species they might be miss-diagnosed with each other in microscopical examination of stool samples. The aim of this study was molecular characterization and differentiation of human derived isolates of S. stercoralis and Rhabditis species based on the mitochondrial gene of cytochrome c oxidase subunit 1 ( cox1 ) amplification. Methods: Using parasitological methods, ten isolates of S. stercoralis and three isolates of Rhabditis spp. were obtained from fresh stool samples of patients and the genomic DNA of the samples were extracted. PCR amplification of cox1 gene was carried out for all the isolates and the products were sequenced. Results: The phylogenetic analysis illustrated that S. stercoralis and Rhabditis spp. isolates were placed in two distinguishable separate clades. Inter-species genetic variation between isolates of S. stercoralis and Rhabditis spp . were ranged from 13.5 to 14.5%. Conclusions: Cox1 gene was a suitable marker for discrimination of S. stercoralis from Rhabditis spp. retrieved from human in the current study. The availability of gene sequence information will be helpful in the future development and validation of discriminatory PCR-based assays of these nematodes.

OBJECTIVES: The objective of this study was to evaluate the prevalence of Strongyloides stercoralis and other intestinal parasites in patients receiving immunosuppressive drugs in northern Iran and to investigate related risk factors.METHODS: This cross-sectional study was conducted among 494 patients receiving immunosuppressive drugs, including cancer patients undergoing chemotherapy (n=188) and those treated with prolonged corticosteroid administration (n=306). All fresh fecal samples were examined using the direct wet-mount, formalin ethyl acetate concentration, and agar plate culture techniques.RESULTS: In total, 16.8% of patients were positive for at least 1 intestinal parasite; the helminthic and protozoan infection rates were 5.1% and 12.3%, respectively. The infection rate was significantly higher in corticosteroid-treated individuals (19.6%) than cancer patients (12.2%) (pS. stercoralis among patients receiving chemotherapy and those treated with corticosteroids were 4.3% and 5.2%, respectively. The prevalence rate of S. stercoralis infection was significantly higher in older patients (p

One of the major migration routes for birds going between Europe and Asia is the Black Sea-Mediterranean route that converges on the Volga Delta, continuing into the area of the Caspian Sea. Cercarial dermatitis is a disorder in humans caused by schistosome trematodes that use aquatic birds and snails as hosts and is prevalent in areas of aquaculture in Northern Iran. Before the disorder can be addressed, it is necessary to determine the etiological agents and their host species. This study aimed to document whether domestic mallards are reservoir hosts and if so, to characterize the species of schistosomes. Previous work has shown that domestic mallards are reservoir hosts for a nasal schistosome.In 32 of 45 domestic mallards (Anas platyrhynchos domesticus) (71.1%), the schistosome Trichobilharzia franki, previously reported only from Europe, was found in visceral veins. Morphological and molecular phylogenetic analysis confirmed the species designation. These findings extend the range of T. franki from Europe to Eurasia.The occurrence of cercarial dermatitis in Iran is high in areas of aquaculture. Previous studies in the area have shown that domestic mallards are reservoir hosts of T. regenti, a nasal schistosome and T. franki, as shown in this study. The genetic results support the conclusion that populations of T. franki from Iran are not differentiated from populations in Europe. Therefore, the schistosomes are distributed with their migratory duck hosts, maintaining the gene flow across populations with compatible snail hosts in Iran.Routes migratoires, oiseaux domestiques et dermatite cercarienne : répartition de Trichobilharzia franki dans le nord de l’Iran.Contexte : L’une des principales voies de migration des oiseaux à destination et en provenance de l’Europe et de l’Asie est la route mer Noire-Méditerranée qui converge vers le delta de la Volga et se poursuit dans la région de la mer Caspienne. La dermatite cercarienne est une affection causée chez l’homme par des trématodes Schistosomatidae utilisant des oiseaux aquatiques et des mollusques comme hôtes, qui est répandue dans les zones d’aquaculture du nord de l’Iran. Avant de pouvoir lutter contre cette parasitose, il est nécessaire de connaître les agents étiologiques et leurs espèces hôtes. Ce travail vise à documenter si les canards domestiques sont des hôtes réservoirs et si oui, à caractériser les espèces de schistosomes. Des travaux antérieurs ont montré que les canards domestiques sont des hôtes réservoirs pour un schistosome nasal. Résultats : Chez 32 de 45 canards domestiques (Anas platyrhynchos domesticus) (71,1 %), le schistosome Trichobilharzia franki, précédemment signalé uniquement en Europe, a été trouvé dans les veines viscérales. L’identification de l’espèce a été vérifiée par une analyse morphologique et phylogénétique moléculaire. Cela étend l’aire de répartition de l’Europe à l’Eurasie. Conclusion : La dermatite cercarienne est répandue dans les zones d’aquaculture en Iran. Des travaux antérieurs dans la région ont montré que les canards domestiques sont des hôtes réservoirs de T. regenti, un schistosome nasal et de T. franki, comme le montre cette étude. Les résultats génétiques soutiennent que les populations de T. franki d’Iran ne sont pas différenciées des populations d’Europe. Par conséquent, les schistosomes se dispersent avec leur hôte canard lors de la migration, maintenant un flux génétique entre les populations avec des mollusques hôtes compatibles en Iran.

Controlling cellular orientation, proliferation, and differentiation is valuable in designing organ replacements and directing tissue regeneration. In the present study, we developed a hybrid microfluidic system to produce a dynamic microenvironment by placing aligned PDMS microgrooves on surface of biodegradable polymers as physical guidance cues for controlling the neural differentiation of human induced pluripotent stem cells (hiPSCs). The neuronal differentiation capacity of cultured hiPSCs in the microfluidic system and other control groups was investigated using quantitative real time PCR (qPCR) and immunocytochemistry. The functionally of differentiated hiPSCs inside hybrid system's scaffolds was also evaluated on the rat hemisected spinal cord in acute phase. Implanted cell's fate was examined using tissue freeze section and the functional recovery was evaluated according to the Basso, Beattie, and Bresnahan (BBB) locomotor rating scale. Our results confirmed the differentiation of hiPSCs to neuronal cells on the microfluidic device where the expression of neuronal-specific genes was significantly higher compared to those cultured on the other systems such as plain tissue culture dishes and scaffolds without fluidic channels. Although survival and integration of implanted hiPSCs did not lead to a significant functional recovery, we believe that combination of fluidic channels with nanofiber scaffolds provides a great microenvironment for neural tissue engineering, and can be used as a powerful tool for in situ monitoring of differentiation potential of various kinds of stem cells. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1534-1543, 2016.

Intestinal microsporidiosis is known as an opportunistic infection in immunocompromised patients. The current study aimed to investigate intestinal microsporidia infection in human subjects with/without immunodeficiency. Totally, 600 stool samples were collected from immunocompromised (254) and immunocompetent (346) subjects. DNA extraction was performed and the SSU rRNA and the ITS genes were amplified to detect and characterize microsporidia and the relevant genotypes. Phylogenetic trees were drawn using MEGA7 software to illustrate the correlation between isolates. From 600 enrolled subjects, 283 and 317 were male and female, respectively. The average age ± SD of all tested subjects was 28.85 ± 26.92. The results of PCR demonstrated the presence of E. bieneusi and Encephalitozoon sp., among 10/600 (1.67%) and 26/600 (4.33%) of samples, respectively. Accordingly, E. bieneusi was seen among 4/346 (1.15%), 1/53 (1.88%), 3/124 (2.42%), and 2/63 (3.17%), and Encephalitozoon sp., was detected from 17/346 (4.91%), 3/53 (5.36%), 4/124 (3.22%) and 2/63 (3.17%) of healthy subjects, RA patients, cancer patients, and transplantation recipients, respectively. Statistical significant correlation was not seen between the presence of microsporidia and age, gender, stool appearance, and geographical region. Molecular analysis showed that all E. bieneusi were the genotype D. Phylogenetic tree demonstrated no classification according to the presence/absence of immunodeficiency, geographical locations and presence of diarrhea. The high prevalence of Encephalitozoon sp., in comparison to E. bieneusi in this study suggested the importance of this genus alongside with E. bieneusi in Iran. In addition, predominance of the genotype D highlighted the wide distribution of this genotype in Iran.

Human infections with Trichostrongylus species have been reported in most parts of Iran. The aim of this study was the identification, molecular characterization and phylogenetic analysis of human Trichostrongylus species based on ITS2 region of ribosomal DNA from Guilan Province, northern Iran. Stool samples were collected from rural inhabitants and examined by formalin-ether concentration and agar plate culture techniques. After anthelmintic treatment, male adult worms were collected from five infected cases. Genomic DNA was extracted from one male worm of each species in every treated individual and one filariform larva isolated from each case. PCR amplification of ITS2-rDNA region was performed and the products were sequenced. Among 1508 individuals, 46 (3.05%) were found infected with Trichostrongylus species using parasitological methods. Male worms of T. colubriformis, T. vitrinus and T. longispicularis were expelled from five patients after treatment. Out of 41 filariform larvae, 40 were T. colubriformis, and the other one was T. axei. Phylogenetic analysis showed that each species was placed together with reference sequences submitted to GenBank database. Intra-species similarity for all species obtained in the current study was 100%. T. colubriformis was found to be probably the most common species in this region of Iran. For the first time, the authors of the present study report the occurrence of natural human infection by T. longispicularis in the world. Therefore, the number of Trichostrongylus species infecting human in Iran now increased to ten.

A case of Strongyloides stercoralis hyperinfection in a patient with dermatomyositis and diabetes mellitus is herein reported. The case was a 60-year-old female admitted due to watery diarrhea and unconsciousness. She had a 10-year history of chronic immunosuppressive therapy including methotrexate and prednisolone for dermatomyositis. Stool parasitological examination revealed numerous rhabditiform larvae of threadworm “S. stercoralis.” Larva in stool sample was characterized by sequencing of mitochondrial DNA. After treatment with ivermectin, the patient recovered without evidence of S. stercoralis in follow-up stool samples. In endemic areas, stool examination for detection of S. stercoralis should be performed on a regular basis for all patients receiving immunosuppressive therapy, as early detection and treatment are necessary to minimize complications of severe strongyloidiasis.

Toxoplasma gondii is an obligate intracellular protozoan parasite that infects humans and animals. T. gondii surface antigen 1 (SAG1) is an appropriate antigen with high specificity and sensitivity for the detection of T. gondii infection in humans and animal hosts. The aim of this study was to determine the seroprevalence of T. gondii infection using SAG1 antigen (P30) in ownership dogs in Meshkin-Shahr district in the northwestern Iran. The sera samples were collected from 171 domestic dogs and tested using indirect ELISA (SAG1 antigen). The data were analyzed using SPSS software version 13. From a total of 171 dogs, 82 (48 %) of them were sero-positive. No statistical significant difference was seen between T. gondii infection and gender (P = 0.995). The highest sero-prevalence of rate was observed in >5 years animals; but no statistical significant difference was seen between T. gondii infection and age (P = 0.589). Our findings indicate that Toxoplasma seropositivity rate is high in ownership dogs in northwest of Iran. This is probably due to high exposure to contaminated food, soil, or water sources with sporulated Toxoplasma oocysts.

Rodentolepis nana (syn. Hymenolepis nana) is a cyclophyllidean zoonotic enteric parasite with worldwide distribution. In humans, it is more prevalent in children, especially in temperate zones. A morphologically similar species of R. nana occurs in different rodents, including gerbils. In the present study molecular characterization of five isolates of R. nana from Rhombomys opimus in the Golestan Province (n= 2) and Razavi Khorasan Province (n= 3), both in north-eastern Iran, were analysed. After DNA extraction, the internal transcribed spacer 2 (ITS2) region of the R. nana isolates was amplified and sequenced, and genetic variation was analysed. Accordingly, two isolates from the Razavi Khorasan Province showed ITS2 signatures that differed from the isolates from the Golestan Province. The third isolate from Razavi Khorasan did not reveal these specific signatures, but exhibited sparse nucleotide polymorphisms in the ITS2 sequence. The ITS gene is conserved, and variation in this fragment could be an interesting subject for further inspection of existing variation in the genome of R. nana, among different domestic and wildlife host species and from different areas.