Trichostrongyloid nematodes in ruminants of northern Iran: prevalence and molecular analysis.

BACKGROUND: This study was carried out to investigate the prevalence and analyze the molecular characteristics based on the internal transcribed spacer (ITS) 2 region of the ribosomal RNA (RNA) gene of trichostrongylid nematodes in different ruminants from Guilan province, northern of Iran. METHODS: The gastrointestinal tracts of 144 ruminants including 72 cattle, 59 sheep, and 13 goats were collected from an abattoir in Guilan province during July to September 2018. After isolation the helminths, male specimens were identified based on morphological parameters. PCR and partial sequencing of the ITS2 fragment were conducted. After phylogenetic analysis, the intraspecific and interspecific differences were calculated. RESULTS: The prevalence of total infections with the nematodes was 38.9, 74.6 and 84.6% among cattle, sheep and goats, respectively. Eleven species of trichostrongylid nematodes including Haemonchus contortus, Marshallagia marshalli, Trichostrongylus axei, T. colubriformis, T. vitrinus, Ostertagia trifurcata, Teladorsagia circumcincta, Marshallagia occidentalis, O. lyrata, O. ostertagi, and Cooperia punctate were recovered from the ruminants. The most prevalent trichostrongyloid nematodes in cattle, sheep and goats were O. ostertagi (26.4%), M. marshalli (64.4%) and T. circumcincta (69.2%), respectively. Phylogenetic tree was discriminative for Trichostrongylidae family, while phylogenetic analysis of the ITS2 gene represented low variations and no species identification of Haemonchidae and Cooperiidae families. CONCLUSIONS: This study suggests the high prevalence and species diversity of trichostrongyloid nematodes in different ruminants, indicating the importance of implement antiparasitic strategies in north regions of Iran. As well, this study showed that the ITS2 fragment is not a discriminative marker for Haemonchidae and Cooperiidae families, and investigation of other genetic markers such as mitochondrial genes would be more valuable for better understanding of their phylogenetic relationships.

Zoonotic transmission of Teladorsagia circumcincta and Trichostrongylus species in Guilan province, northern Iran: molecular and morphological characterizations.

BACKGROUND: Parasitic trichostrongyloid nematodes have a worldwide distribution in ruminants and frequently have been reported from humans in Middle and Far East, particularly in rural communities with poor personal hygiene and close cohabitation with herbivorous animals. Different species of the genus Trichostrongylus are the most common trichostrongyloids in humans in endemic areas. Also, Ostertagia species are gastrointestinal nematodes that mainly infect cattle, sheep and goats and in rare occasion humans. The aim of the present study was to identify the trichostrongyloid nematodes obtained from a familial infection in Guilan province, northern Iran, using morphological and molecular criteria. METHODS: After anthelmintic treatment, all fecal materials of the patients were collected up to 48 h and male adult worms were isolated. Morphological identification of the adult worms was performed using valid nematode keys. Genomic DNA was extracted from one male worm of each species. PCR amplification of ITS2-rDNA region was carried out, and products were sequenced. Phylogenetic analysis of the nucleotide sequence data was performed using MEGA 6.0 software. RESULTS: Adult worms expelled from the patients were identified as T. colubriformis, T. vitrinus and Teladorsagia circumcincta based on morphological characteristics of the males. Phylogenetic analysis illustrated that each species obtained in current study was placed together with reference sequences submitted to GenBank database. CONCLUSIONS: The finding of current study confirms the zoonotic aspect of Trichostrongylus species and T. circumcincta in inhabitants of Guilan province. The occurrence of natural human infection by T. circumcincta is reported for the first time in Iran and the second time in the world.

Genetic diversity of an avian nasal schistosome causing cercarial dermatitis in the Black Sea-Mediterranean migratory route.

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.

Occurrence and molecular characterization of free-living amoeba species (Acanthamoeba, Hartmannella, and Saccamoeba limax) in various surface water resources of Iran.

This study was conducted to determine the presence and molecular identity of Acanthamoeba species in the surface water resources of four provinces in Iran, namely Guilan, Mazandaran (North of Iran), Alborz, and Tehran (capital city), using culture- and molecular-based methods. During March to November 2014, 49 surface water samples were collected from environmental water sources-the distinct surface waters of Guilan, Mazandaran, Alborz, and Tehran provinces, in Iran. For the isolation of Acanthamoeba species, approximately 500 ml of the water samples were filtered through a cellulose nitrate membrane with a pore size of 0.45 µ. The filter was transferred onto non-nutrient agar plates seeded with Gram-negative bacteria (Escherichia coli) as a food source. The presence of Acanthamoeba was confirmed by the genus-specific primer pair JDP1 and 2, and/or NA primers were used to identify Acanthamoeba and certain other free-living amoebae. In total, 38 out of 49 samples were positive by culture and/or PCR for Acanthamoeba and other free-living amoebae from all three provinces. By sequencing the positive isolates, the strains were shown to belong to Acanthamoeba (16 isolates belonged to T4 and 2 isolates belonged to T5), Hartmannella vermiformis (3/24), and Saccamoeba limax (2/24). The T4 and T5 genotypes were detected in Guilan and Mazandaran provinces. Two isolates from Guilan and Tehran provinces belonged to S. limax, and H. vermiformis was detected in Guilan province. The results of this study highlight the need to pay more attention to free-living amoebae, as human activity was observed in all of the localities from which these samples were taken. These surface waters can be potential sources for the distribution and transmission of pathogenic Acanthamoeba in the study areas, and free-living amoebas (FLA) (particularly the Acanthamoeba species) can serve as hosts for and vehicles of various microorganisms.

Current status of intestinal parasitic infections and associated risk factors in rural population of Guilan province, northern Iran: trichostrongyliasis is the most prevalent helminthic infection.

Aim: This study aimed to determine the distribution of enteric parasitic infections and related risk factors among rural communities of Guilan province, Northern Iran, and to compare the results with the situation in the past. Background: Intestinal parasitic infections are still considered as a major public health concern, particularly in human communities with poor economy and sanitation. Methods: This cross-sectional study was performed in rural areas of Masal and Shanderman district from February to December 2020. A total of 917 stool samples were collected and examined for presence of intestinal helminthes and protozoa using direct, formalin-ether and Kato-Katz techniques. Results: A total of 156 (17%) out of 917 examined individuals were infected with intestinal parasites. The overall prevalence of protozoa, helminths and mixed infections were 11.8% (108/917), 4.5% (41/917) and 0.8% (7/917), respectively. Blastocystis was the most prevalent intestinal protozoa (9.6%) followed by Giardia lamblia (1.9%), Endolimax nana (1.1%), E. coli (0.8%) and Entamoeba hartmani (0.1%). The highest prevalence of intestinal helminths belonged to Trichostongylus spp. (3.5%) and Strongyloides stercoralis (1.3%). Statistical analysis showed significant association between giardiasis and sex (P<0.03). On the other hand, prevalence of enteric helminths was influenced by close contact with livestock, keeping herbivorous animals at home, job, education, and consumption of uncooked vegetables (P<0.05). Conclusion: The findings indicate a decreasing trend in the prevalence of intestinal parasitic infections in Guilan province in comparison to the past few decades. Hookworm infections, which was very prevalent in the area, are now rare, while trichostrongylosis showed a high prevalence in rural residents of the study area.

Hymenolepis diminuta Infection in a Rural Child from North of Iran.

We report a case of Hymenolepis diminuta infection in a two years old boy living in Guilan Province, northern Iran diagnosed in 2019. The patient was complained of anorexia, weight loss, weakness and disturbed sleep. Stool examination revealed numerous eggs of H. diminuta. After treatment with a single dose of oral praziquantel, the patient recovered without evidence of the egg shedding in follow-up stool samples. Moreover, we performed detailed phylogenetic analysis of the H. diminuta comparing with other isolates deposited in GenBank database based on Cox1 gene. Based on BLAST analysis of Cox1 gene our sequence showed 97.4-99.2% similarity with those of H. diminuta available in GenBank. The present study recommends the importance of reporting the infection cases, in order to improve knowledge on epidemiology and control of the neglected disease.

Comparison of Molecular and Parasitological Methods for Diagnosis of Human Trichostrongylosis.

Human trichostrongyliasis is a zoonotic disease that is prevalent among rural populations in some countries. This study was performed to evaluate various parasitological methods and polymerase chain reaction (PCR) for the diagnosis of human trichostrongyliasis. A total of 206 fresh stool samples were collected from residents of endemic villages of Northern Iran. All samples were examined using conventional parasitological methods, including wet mount, formalin ethyl acetate concentration (FEAC), agar plate culture (APC), Harada-Mori culture (HMC), and Willis, along with the PCR technique. Among the total of 206 individuals examined, 72 people (35%) were found infected with Trichostrongylus species using combined parasitological methods. By considering the combined results of parasitological methods as the diagnostic gold standard, the Willis technique had a sensitivity of 91.7% compared with 52.8% for the APC, 40.3% for the HMC, 37.5% for FEAC, and 5.6% for the wet mount technique. The diagnostic specificity of all the parasitological methods was 100%. Furthermore, the PCR method detected Trichostrongylus spp. DNA in 79 fecal samples (38.3%) with a sensitivity of 97.2% and a specificity of 93.3%. According to the current findings, the Willis method was more sensitive than are the other parasitological methods in the diagnosis of human trichostrongyliasis. However, the PCR assay was more sensitive and more reliable in the detection of human trichostrongyliasis in comparison with the parasitological methods.

Strongyloides stercoralis and other intestinal parasites in patients receiving immunosuppressive drugs in northern Iran: a closer look at risk factors.

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%) (p<0.05). The prevalence rate of S. 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<0.05). CONCLUSIONS: Strongyloidiasis is one of the most common parasites among patients receiving immunosuppressive drugs in northern Iran. Early diagnosis and proper treatment of these patients are necessary to minimize the complications of severe strongyloidiasis.

Helminthes in Feral Raccoon (Procyon lotor) as an Alien Species in Iran.

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.

Prevalence of Strongyloides stercoralis and Other Intestinal Parasites among Institutionalized Mentally Disabled Individuals in Rasht, Northern Iran.

BACKGROUND: We aimed to determine the status of strongyloidiasis in mentally disabled population in the institutional places in Rasht City, the capital of Guilan Province, northern Iran. METHODS: This cross-sectional study was conducted in 8 institutions for mentally retarded population in Rasht in 2013. Before collecting the samples, a questionnaire was filled out for each participant by an expert person. A single stool sample was obtained from each of the 173 subjects and examined using direct wet mount, formalin-ether concentration technique and agar plate culture method. RESULTS: A total of 173 mentally disabled individuals aged 2-57 (25.69±11.56) yr old were studied. Stool examination showed that 51 (29.5%) cases were infected with at least one parasite. Of 173 studied cases only 10 (5.8%) individuals were infected with pathogenic parasites, of which 2 (1.2%) cases were infected with Strongyloides stercoralis and 8 (4.6%) with Giardia lamblia. On the other hand, 42 (24.3%) of the studied population were infected with non-pathogenic intestinal protozoa such as Blastocystis hominis (n=29, 16.8%), Entamoeba coli (n=16, 9.2%) and Endolimax nana (n=4, 2.3%). Mixed protozoal infections were observed in 8 (4.6%) individuals. CONCLUSION: The prevalence rate of S. stercoralis in mentally disabled individuals in Rasht was somewhat higher than those of normal population of the province. The same picture was seen when the prevalence of G. lamblia and non-pathogenic protozoa in normal and mentally disabled populations were compared.

The Endemicity of Human Fascioliasis in Guilan Province, Northern Iran: the Baseline for Implementation of Control Strategies.

BACKGROUND: The largest global outbreaks of liver fluke disease (Fascioliasis) in humans, caused by species of the genus Fasciola, have occurred in Guilan Province of Iran, affecting more than 15000 people. Although, different aspects of fascioliasis have been the subject of various researches during last two decades, nevertheless no community-based study has been performed in endemic regions of Guilan. The aim of present study was to obtain the basic information needed to develop future control strategies. METHODS: Fecal and blood samples were collected from 1,984 volunteers in the Bandar-Anzali district, the region where previous epidemics occurred. Fecal samples were examined by Kato-Katz and formalin-ether methods for the presence of Fasciola eggs. Sera samples were analyzed by ELISA to detect anti-cathepsin L antibodies. RESULTS: Twenty-seven (1.36%) individuals were seropositive, 9 (0.45%) individuals were egg positive (mean egg count 50.7 (±30.36) eggs per gram of faeces) and 30 individuals (1.51%) were positive using both methods. No statistical association was observed between infection and age, gender, location, occupation, educational status and dietary habits. The prevalence of intestinal parasites is also included. CONCLUSION: Human fascioliasis is hypoendemic in this region and recommends a passive case-finding approach, effective primary prevention measures, health education through mass media and effective veterinary public health measures for control of human disease.