Search

The current investigation was carried out during the period from July 2022 to March 2023, aiming to investigate the prevalence of gastrointestinal helminths in domestic birds collected from traditional markets in Guilan province. One hundred forty-eight domestic birds, including chickens (Gallus gallus domesticus), domestic ducks (Anas platyrhynchos domesticus), greylag geese (Anser anser), and domestic turkeys (Meleagris gallopavo domesticus) were examined. Totally, 42.56% of the investigated birds were positive for helminthic parasites. Morphological analysis revealed varying infection rates among birds: Echinostoma revolutum (5.40%), Hypoderaeum conoideum (2.02%), Cloacotaenia megalops (0.67%), Hymenolepididae family (4.05%), Ascaridia galli (16.89%), and Heterakis gallinarum (4.72%). The investigation involved molecular analysis of the 18S and ITS1 + 5.8S + ITS2 rRNA gene regions. The findings indicated that the 18S region of nematode isolates exhibited a similarity of 92 to 100% with sequences in the GenBank, whereas trematode and cestode isolates showed a gene similarity ranging from 88 to 99%. The ITS regions of nematode, trematode, and cestode isolates exhibited genetic similarities ranging from 87 to 100%, 73-99%, and 75-99%, respectively. Furthermore, phylogenetic analysis confirmed the categorization of the identified species within the Ascaridiidae, Heterakidae, Hymenolepididae, and Echinostomatidae families, indicating their close affinity with previously documented species. Implementing precise control measures such as consistent monitoring, adequate sanitation protocols, and administering anthelmintic treatments is crucial for effectively managing parasitic infections in free-range and backyard poultry farms. Additionally, conducting further surveys is advisable to assess the impact of these parasites on the health and productivity of poultry in the investigated area., Competing Interests: Declaration of competing interest The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Objective: Ganoderma extracts have the potential to be used as anti-cancer, anti-inflammatory, immunomodulator, and antimicrobial agents, as evaluated in numerous studies. This study was aimed to determine the lethal and inhibitory effects of aqueous, hydroalcoholic, and alcoholic extracts of Ganoderma lucidum on Toxoplasma gondii RH strain tachyzoites, in vitro., Results: All three types of extracts showed toxoplasmacidal effects. The highest percentage of mortality was related to hydroalcoholic extract. The EC50 of Ganoderma extracts for tachyzoites were 76.32, 3.274, and 40.18 for aqueous, hydroalcoholic and alcoholic extracts, respectively. The selectivity index obtained for hydroalcoholic extract was 71.22, showing the highest activity compared to other extracts. According to our findings, the hydroalcoholic part was the most effective substance among the extracts. This basic study showed obvious anti-toxoplasma effect of Ganoderma lucidum extracts. These extracts can be used as candidates for further in-depth and comprehensive studies especially In vivo experiments to prevent toxoplasmosis., (© 2023. The Author(s).)

Background: The liver flukes of the Fasciola species and Dicrocoelium spp. are recognised as parasites of domestic and wild herbivores. Both species of F. hepatica and F. gigantica as well as D. dendriticum are distributed in Iran. The present study aimed to identify Fasciola spp. and Dicrocoelium spp. using mitochondrial Cox1 (cytochrome c oxidase I) gene by HRM method., Methods: Totally, thirty infected liver specimens were collected from the sheep (n:23) and cattle (n:7) at the abattoirs of Qazvin Province, northwest Iran in 2022. DNA extraction and PCR amplification of Cox1 gene were conducted by HRM technique. DnaSP v.5.0 was used for compression of diversity indices of ribosomal 28S rDNA and mitochondrial Cox1 markers of Dicrocoelium spp. The taxonomic status of Dicrocoelium spp. was performed by sequencing and p hylogenetic analysis., Results: Overall, 26 and 4 isolates were identified as F. hepatica and F. gigantica , respectively. D. dendriticum was the sole infecting species of Dicrocoelium revealed by HRM analysis. Genomic analysis showed a moderate ( 28S rDNA genes: 0.600±0.215) to high ( Cox1 : 0.733±0.155) haplotype diversity for D. dendriticum ., Conclusion: The parasite-dependent mitochondrial gene ( Cox1 ) could identify a higher genetic diversity of D. dendriticum compared to nuclear 28S rDNA gene. HRM technique in the present study found to be a reliable technique for identification and genetic diversity of liver flukes but more comprehensive and in-depth studies in different parts of the country are needed., Competing Interests: Conflicts of interest The authors declare no conflicts of interest., (Copyright © 2023 Hajialilo et al. Published by Tehran University of Medical Sciences.)

Objective: The objective of the present study was to identify Trichostrongylus species by molecular analysis and also phylogenetic relationships of Trichostrongylus species by mitochondrial Cytochrome c oxidase subunit 1 ( Cox1 ) gene in Guilan province, northern Iran., Methods: Abomasum and duodenum contents of 144 livestock were collected from sheep, goats, and cattle in Guilan province. Morphological survey was performed for initial screening. Total DNA was extracted, and the partial region of Cox1 gene was amplified and sequenced. Genetic diversity was calculated and phylogenetic analysis of the data on nucleotide sequence was conducted by MEGA7 software., Results: Three species of Trichostrongylus including T. colubriformis , T. vitrinus , and T. axei were identified by morphological characteristics. The genetic divergence within the species in the present study was observed for T. axei (0-2.5%), T. colubriformis (0.77%), and T. vitrinus (0%). The mean inter-species difference between the three species of Trichostrongylus obtained in this study was 14.4-15.4%., Conclusion: The Cox1 sequences of the members of Trichostrongylus spp. were highly variable and this could be used as a valuable measure to achieve a proper assessment on biodiversity. Sequence data generation from other species of Trichostrongylus will be needed to reconstruct the phylogenetic relationships of this genus of nematodes.

We aimed to present an alternate method instead of PCR-RFLP and also develop an optimized method for rapid, time-saving and affordable molecular-based approach to discriminate species of liver fluke, Fasciola hepatica and F. gigantica. Seventy-six samples of F. hepatica and 28 F. gigantica were collected from the slaughterhouses of endemic regions in Iran. Following a comprehensive analysis of the mitochondrial complete sequences of both F. hepatica and F. gigantica, the extracted DNAs from all samples were used as templates in multiplex PCR reactions containing two sets of primers specific for cytochrome c oxidase I (cox I) gene of both species. In a parallel experiment, PCR-RFLP was performed for each sample using internal transcribed spacer (ITS1) sequence. Furthermore, following a PCR amplification for cox I gene, the amplicons were purified for sequencing. To assess the validity of the multiplex PCR approach, the obtained data from the multiplex PCR and PCR-RFLP experiments were compared with each other. By sequence analysis of 104 samples, 76 and 28 samples were identified as F. hepatica and F. gigantica, respectively. Results revealed 100% and 92% of accuracy as for multiplex PCR and PCR-RFLP. The designed multiplex PCR strategy offers a valid alternative approach to the conventional methods with distinctive features including convenience, cost-effectiveness, time-saving (3 hours from sampling to obtain final results) and high efficacy.

The role of various parasitic infections in the occurrence of appendicitis is illustrated through cases recorded all over the world. The purpose of the current study was to estimate the global prevalence of parasite infestation (other than E. vermicularis ) in appendectomy specimens.In the setting of the PRISMA guidelines, multiple databases (Science Direct, Scopus, Web of Science, PubMed, and Google Scholar) were explored in articles published until 28 September 2020. Totally, 62 studies (106 datasets) with 77, 619 participants were included in the analysis.The pooled prevalence of parasites in appendectomy samples was as follows; 0.012% (95% CI; 0.004-0.025) for Ascaris lumbricoides , 0.004% (95% CI; 0.001-0.009) for Trichuris trichiura , 0.025% (95% CI; 0.007-0.052) for Schistosoma mansoni , 0.002% (95% CI; 0.001-0.005) for Taenia spp., 0.061% (95% CI; 0.020-0.122) for Entamoeba histolytica and 0.034% (95% CI; 0.018-0.056) for Giardia lamblia .Our results demonstrated that the risk of appendicitis may increase in the presence of helminth and protozoan infections. As such, the most cases of parasites in appendectomy specimens were reported in developing countries. Regular screening plans for diagnosis, treatment and prevention are needed for prevention of parasitic infection as well as parasitic associated appendicitis, especially in endemic regions of the world.

Background: We aimed to detect the genetic diversity of samples identified morphologically as Fasciola spp. from sheep, cattle and goat from Lorestan Province, western Iran using PCR-RFLP method. Besides, we evaluated the genetic diversity indices, sequencing and phylogenetic analysis using mitochondrial gene ( ND1 and CO1 )., Methods: PCR-RFLP analysis of ribosomal ITS1 fragment by RsaI restriction enzyme to investigate the genetic characteristics of Fasciola species obtained from different hosts (18 sheep, 21 cattle, and 17goats) was conducted. The samples were sequenced. Sequences were evaluated using BLAST software and the parasite species were identified with similarity percentage and overlap with the species registered in the gene bank. Then similarity and diversity of intra-species and intra-species diversity of Fasciola species were calculated., Results: In Lorestan, based on RFLP pattern, 93% (52) of the Fasciola spp. isolates had a RFLP pattern related to F. hepatica and 7% (4) were F. gigantica . No hybrid forms were detected. The CO1 gene could clarify 19 haplotypes against ND1 gene that found 22 haplotypes among livestock. Sequencing results of the mtDNA showed intra-species identity 98. 5%-100% and Intra-species-diversity: 0-1.5% compared to the GenBank sequences., Conclusion: Using PCR-RFLP method, two species of F. hepatica and F. gigantica , were present in Lorestan Province, but F. hepatica was more prevalent. Mitochondrial genes could better test variability indices in different hosts than ribosomal genes, consequently among mitochondrial genes, the ND1 gene could better examine differences and similarities than CO1., Competing Interests: Conflict of Interest The authors declare there is no conflict of interest., (Copyright © 2022 Heydarian et al. Published by Tehran University of Medical Sciences.)

The aim of the present study was to detect free-living amoeba (FLA) in the water resources of Arak, Iran using molecular tools. A total of 154 samples were collected from different water supplies. Molecular analyses, sequencing, and phylogenetic study were conducted to confirm the species and genotypes of FLA. Fisher exact test was used to determine the significance. Of 154 water samples, 19 (12.3%) samples were tested positive for FLA. Three genotypes of Acanthamoeba including T4, subtype D, and T5 were identified among the isolates. The pathogenicity assay showed that the isolate of Acanthamoeba in drinking water was highly pathogenic. Three species of Naegleria, including N. australiensis, N. pagei, and N. gruberi were found among the samples. Six isolates of Vermamoeba were identified as V. vermiformis. Meanwhile, three other species including Vannella sp., Vahlkampfia avara, and Stenamoeba polymorpha were also recovered from the water samples. Statistical analysis showed a significant difference between the various water resources contaminated with FLA. This is the first study to reveal the presence of S. polymorpha in water sources in Iran. According to the findings of the present study, health officials should be beware of potential public health impacts of FLA in water resources.

Background: Free-living amoebae (FLA) such as Acanthamoeba spp., are considered as opportunistic and pathogenic protozoans. Acanthamoeba granulomatous encephalitis (AGE) is a serious threat for immunodeficient patients and Acanthamoeba keratitis (AK) for contact lens users. We aimed to identify the presence of free living amoebae in nasal swabs of patients and contact lens users in Qazvin, Iran., Methods: During 2019, 251 nasal and oral swabs (including the pharynx and mouth) were collected from patients with diabetes, AIDS and those under periodic dialysis in Qazvin, Iran. In addition, 27 soft contact lenses were collected from the participants. Following DNA extraction, PCR and sequencing were conducted to identify the genotypes of the amoeba. Phylogenetic analysis of the identified sequences was performed using MEGA 7 software., Results: A strain of Acanthamoeba belonging to the T3 genotype was isolated from hemodialysis patients. Two specimens of Acanthamoeba with T3 genotype were isolated from keratitis patients., Conclusion: The clinicians should pay attention to the possible complication of this organism because this amoeba is potentially pathogenic for immunocompromised patients. Since the amoeba is present in environmental resources, the use of contact lenses should be accompanied by considering proper hygiene., Competing Interests: Conflict of interest The authors declare that there is no conflict of interest., (Copyright © 2022 Jalilehvand et al. Published by Tehran University of Medical Sciences.)

Background: Strongyloides stercoralis, a soil-transmitted helminth, occurs in humans, non-human primates, dogs, cats and wild canids. The zoonotic potential between these hosts is not well understood with data available on prevalence primarily focused on humans. To increase knowledge on prevalence, this review and meta-analysis was performed to estimate the global status of S. stercoralis infections in dogs., Methods: Following the PRISMA guidelines, online literature published prior to November 2020 was obtained from multiple databases (Science Direct, Web of Science, PubMed, Scopus and Google Scholar). Prevalence was calculated on a global and country level, by country income and climate, and in stray/animal shelter dogs versus owned dogs. Statistical analyses were conducted using R-software (version 3.6.1)., Results: From 9428 articles, 61 met the inclusion criteria. The estimated pooled global prevalence of S. stercoralis in dogs was 6% (95% CI 3-9%). Infection was found to be the most prevalent in low-income countries with pooled prevalence of 22% (95% CI 10-36%). The highest pooled prevalence of S. stercoralis in dogs was related to regions with average temperature of 10-20 °C (6%; 95% CI 3-11%), an annual rainfall of 1001-1500 mm (9%; 95% CI 4-15%) and humidity of 40-75% (8%; 95% CI 4-13%). Prevalence was higher in stray and shelter dogs (11%; 95% CI 1-26%) than in owned dogs (3%; 95% CI 1-7%)., Conclusions: As with S. stercoralis in humans, higher prevalence in dogs is found in subtropical and tropical regions and lower-income countries, locations which also can have high dog populations. While this study presents the first estimated global prevalence of S. stercoralis in dogs, it is potentially an underestimation with 15 of 61 studies relying on diagnostic methods of lower sensitivity and a paucity of data from most locations. Standardized protocols (e.g. quantity of feces and number of samples for a Baermann) in future studies could improve reliability of results. More prevalence studies and raising veterinary awareness of S. stercoralis are needed for a One Health approach to protect humans and dogs from the impact of the infection., (© 2022. The Author(s).)

Trichuriasis is one of the most common soil-transmitted helminth (STH) infections, affecting populations globally. The condition is particularly prevalent in tropical and subtropical areas with low levels of sanitation and poor living conditions. The objective of this systematic review and meta-analysis was to evaluate the prevalence of Trichuris trichiura infection in Asia at the country and region level. Multiple databases/academic search engines (Web of Science, PubMed, ProQuest, Scopus, and Google Scholar) were searched for literature on T. trichiura prevalence in Asia published through January 2021. Pooled prevalence was determined using the meta-package in R (version 3.6.1). Out of 13,836 articles, 226 studies (5,439,500 individuals) from 26 countries met the inclusion criteria. Of the 226 studies, 151 were community-based studies that included individuals across the age spectrum, while 75 studies focused on school children (typically in the 5-16 years age range). The overall T. trichiura pooled prevalence was 15.3% (95% CI: 12.4-19.1%), with a pooled prevalence of 13.3% (95% CI: 10.0-17.1%) for the community studies and 20.9% (95% CI: 14.7-27.9%) for the studies only including school children. For studies including all age groups, individuals in the 1-15 years age group had the highest pooled prevalence at 23.4% (95% CI: 1.7-49.4%). There was a significant difference found in overall pooled prevalence by sex (p < 0.001) and community type (rural versus urban) (p < 0.001). Although prevalence appears to be decreasing, study findings suggest that T. trichiura infection continues to be a public health problem in Asia. Therefore, control programs focused on at-risk individuals in endemic areas are needed., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Trichomonas vaginalis is an anaerobic protozoan parasite that causes trichomonosis in human. It is one of the most common non-viral sexually transmitted infections. It has been found to be most prevalent in patients referred to sexually transmitted disease clinics. In recent years, molecular methods have been used to identify genotypes of this parasite in different parts of the world and so far 6 types of T. vaginalis have identified. The aim of this study was to investigate the prevalence and genotype identification of T. vaginalis from married women in northern Iran. A total of 450 vaginal specimens were taken from married women, referring to health centers in northern Iran. Demographic information of women was collected through a questionnaire. The samples were first examined microscopically and then monitored in Dorsch culture medium for up to 10 days. Actin genes of positive samples were amplified by PCR. Finally, PCR products were used to determine the sequence and genotype of the parasite. Overall, 0.7% (3/450) samples were positive for T. vaginalis. All of the three infected women were housewives. After sequencing, the genotype of these parasites were type H (66.7%) (Accession no; MW414672-MW414673) and type E (33.3%) (Accession no: MW414671). Low prevalence of T. vaginalis in north of Iran indicate high level of hygiene in sexual intercourse and avoiding from high risk sexual behaviors, and also it seems that genotype H is dominant type of the parasite in the study area.

Background: The Islamic Republic of Iran is one of the most important endemic foci of cutaneous leishmaniasis in the world. Annually, a large number of Iranian Shia pilgrims travel to Iraq from this area in order to participate in one of their most important religious ceremonies. This trip has coincided with the seasonal activity of sand flies in recent years. So, cutaneous leishmaniasis could be a serious threat for pilgrims on these trips., Aims: To report cases of cutaneous leishmaniasis among Iranian Shia pilgrims attending a religious ceremony in Iraq during 2017., Methods: Sixteen patients were referred to our laboratory in the Department of Parasitology and Mycology at Qazvin University of Medical Sciences. Dermal scrapings and stained slides prepared of skin lesions were used to morphological diagnosis. DNA extraction and PCR amplification were optimized to identification of Leishmania species., Results: All of the patients were infected with cutaneous leishmaniasis in microscopic survey. L. major was detected by molecular approach. The number of lesions observed in patients were 1 (31%), 2 (25%), and ≥ 3 (44%)., Conclusions: Since a large number of Shia Muslims participate in the annual religious ceremonies, serious measures must be taken to prevent the disease., (Copyright © World Health Organization (WHO) 2021. Open Access. Some rights reserved. This work is available under the CC BY-NC-SA 3.0 IGO license (https://creativecommons.org/licenses/by-nc-sa/3.0/igo).)

Introduction: Hydatidiosis is a serious parasitic disease in humans and livestock, worldwide. Echinococcus granulosus shows notable genetic variation among intermediate hosts. Several genotypes of the worm have been reported from different parts of Iran, but no information on the parasite genotypes status in the study region is available. The current study investigated the presence of different genotypes of E. granulosus in the livestock of Qazvin, Iran, by sequencing the mitochondrial Cox1 genes., Methodology: One hundred twenty E. granulosus isolates, including 30 from goats, 40 from cattle and 50 from sheep, were collected from the slaughterhouses in Qazvin province. Mitochondrial Cox1 gene region was amplified by PCR and 30 isolates were sequenced. Phylogenetic analysis was performed by using the MEGA 7.0 software. Morphological analysis was performed on rostellar hook length of protoscoleces., Results: All isolates were identified as E. granulosus sensu stricto (G1-G3 complex) among 17% of the isolates clarified as G3 genotypes. G1 was the predominant genotype among the specimens. No significant difference between the rostellar hooks measurements of different genotypes was observed., Conclusion: Our findings confirmed the presence of E. granulosus sensu stricto in the region, although further studies are required to determine the haplotype diversity of E. granulosus using different mitochondrial and nuclear genes., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Introduction: Blastocystis is a common intestinal parasite of human and animal hosts. The parasite has 17 subtypes, and among those at least nine subtypes (ST1-ST9) are found in human hosts., Objective: The aim of the present study was to investigate the presence of different subtypes of Blastocystis spp. among the patients referred to Velayat hospital of Qazvin province, Iran., Methods: Overall, 864 stool samples were examined by using formalin-ethyl acetate concentration method and Trichrome staining. All specimens were cultured in clotted fetal bovine medium. Later, DNA extraction and PCR amplification of 18S ribosomal RNA gene region was conducted and phylogenetic tree constructed., Results: The results revealed 7.9% (68/864) of the study population were infected with Blastocystis . Intestinal symptoms were observed in 61% (36/59) of individuals positive for Blastocystis , with abdominal pain in 58% (21/36) of cases which was more frequent than other intestinal signs. No significant relationship was observed among the study variables. By molecular and phylogenetic analysis, three subtypes ST1 (45%), ST2 (30%) and ST3 (23%) of parasite were identified., Conclusion: This study showed ST1 subtype was the predominant subtype among the positive specimens, meanwhile the highest haplotype and nucleotide diversity were clarified in ST3 subtype., (© 2020 Delshad A et al.)

Background: Free-living amoeba (FLA) are widely distributed in different environmental sources. The most genera of the amoeba are Acanthamoeba , Naegleria and Vermamoeba. The most common consequences of the infections in immune-deficient and immuno-competent persons are amoebic encephalitis and keratitis. The aim of this study was to investigate the presence of Acanthamoeba spp. and Naegleria spp., isolated from the main agricultural water canal in Qazvin., Methods: Totally, 120 water specimens were collected and later the specimens were cultured and cloned to identify positive samples. PCR amplification and sequencing were carried out to identify the isolated species as well as the genotypes of amoeba., Results: According to morphological surveys, 41.7% (50/120) of water specimens were positive for FLA. Molecular analysis revealed that 68.6% and 31.4% of Acanthamoeba specimens were identified as T3 and T4 genotypes, respectively. Also, two species of Naegleria named as N. lovaniensis (57.1%) and Naegleria sp. (42.8%) were identified. The results of pathogenicity assays demonstrated that 38.5% of T3 and 61.5% of T4 genotypes of Acanthamoeba were highly pathogenic parasites., Conclusion: The water flowing in the agricultural canal of the area is contaminated with potential pathogenic FLA, therefore, it is recommended that more attention to be paid towards proper treatment of water sources to prevent possible risk of the disease., Competing Interests: Conflict of interest The authors declare that there is no conflict of interest., (Copyright© Iranian Society of Parasitology & Tehran University of Medical Sciences.)

Free-living amoeba (FLA), including Acanthamoeba and Naegleria are facultative parasites in humans. The amoeba have widespread distribution in various water sources. The aim of this study was isolation and molecular identification of Acanthamoeba and Naegleria isolated from swimming pools and also hot and cold tub waters in Qazvin province. The samples (166 water samples) were cultured to isolate and identify positive specimens. PCR (polymerase chain reaction) amplification, sequencing and phylogenetic analysis were conducted to confirm the isolated species and genotypes of amoeba. According to morphological characterizations, 18.6% of specimens were identified as FLA, which in 71% were Acanthamoeba by PCR method. Molecular analysis revealed that 36.3%, 18.1% and 4.5% of Acanthamoeba specimens were identified as T3, T4 and T11 Acanthamoeba genotypes, respectively. Protacanthamoeba bohemica (27.2%) and Acanthamoeba sp. (4.5%) were found among the specimens. The results of osmo-tolerance and thermo-tolerance assays demonstrated that 50% of T3 and 25% of T4 genotypes of Acanthamoeba were highly pathogenic parasites. The molecular approach showed the presence of Naegleria lovaniensis (9%) in hot tub water of swimming pools. This study demonstrated that the swimming pools and hot tub water in Qazvin province were contaminated with Acanthamoeba and Naegleria species.

Blastocystis sp. is a polymorphic intestinal parasite in humans and animals. The parasite has a worldwide distribution, especially in developing countries with poor sanitation, exposure to animals, and improper disposal systems. The aim of this study was to identify the subtypes of Blastocystis sp. among children of Qazvin, northwest Iran. Totally, 864 stool samples were collected from the children referred to Qods hospital in Qazvin, Iran. Fecal specimens were investigated by formalin-ethyl acetate concentration method and trichrome staining as well as cultivation of all samples in clotted fetal bovine medium. DNA extraction of culture-positive specimens and PCR amplification of 18S ribosomal RNA gene region was performed. The sequences detected were compared with reference genes in the GenBank, and the sequences further deposited in the GenBank database. Data analysis was performed by Chi square test while a p value of < 0.05 was considered as significant. Of 864 isolates, 4.1% (36/864) were positive for Blastocystis sp. with infection rate insignificantly higher among the females than males. The highest infection rate was estimated at 6.8% in 6-9 years old age group with abdominal pain as the most common (33%) gastrointestinal sign. No statistically significant difference was found between the variables and Blastocystis infection. Molecular analysis clarified the presence of three subtypes of Blastocystis including ST1 (56%), ST2 (28%), and ST3 (16%) of among specimens with ST1 as the predominant subtype. A significant association between intestinal signs and the subtypes was not found. Considering ST1 as the predominant subtype, it seems that zoonotic transmission is a main route of human infections with Blastocystis sp. in the area studied., Competing Interests: Conflict of interestThe authors declare no conflict of interest., (© Indian Society for Parasitology 2019.)

Introduction: Fascioliasis and dicrocoeliasis are the most frequent zoonotic diseases with increasing human health problems in different parts of Iran. Two species, Fasciola hepatica (F. hepatica) and Fasciola gigantica (F. gigantica), are spread in the country. Molecular approaches have a decisive role in identifying both the species. The aim of this study was to detect Fasciola spp. and Dicrocoelium spp. by amplifying the ITS-2 and 28S rDNA gene sequence., Methods: Overall, 30 infected liver samples were collected from the livestock of Qazvin, Iran. The adult flukes were collected from different livestock. DNA extraction and PCR amplification of ribosomal RNA gene region (ITS2) and 28S rDNA gene fragment were conducted and a phylogenetic tree was constructed., Result: All the isolates obtained from the cattle (No: 7) and 82.6% (No: 19) of sheep isolates were infected with F. hepatica species, whereas 17.4% (No: 4) of sheep isolates were infected with F. gigantica. It was also shown that F. hepatica was the predominant species of Fasciola present in the region. All the specimens were infected with Dicrocoelium dendriticum (D. dendriticum)., Conclusion: Both the species of Fasciola were found in Qazvin. D. dendriticum was the sole infecting species of the Dicrocoelium genus in the livestock of the city of Qazvin. Further research studies are needed to determine the intermediate host of the parasites in the region., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Background: We aimed to investigate the genotypes of Giardia intestinalis among the food handlers in Qazvin, Iran., Methods: Overall, 1530 stool specimens were collected from the food handlers who visited Shahid Bolandian Health Center, Qazvin, Iran during 2016. Specimens were evaluated by microscopic and concentration methods. Twenty specimens with appropriate number of giardia cysts were selected followed by DNA extraction . Determination of giardia genotypes was achieved through PCR and sequencing the glutamate dehydrogenase gene. The phylogenetic tree was drawn using the MEGA7 software. Finally, the data were analyzed statistically with a P -value<0.05 was considered as significant., Results: Twenty stool samples (1.3%) were positive for Giardia cyst. All positive specimens were obtained from male participants with abdominal cramp being their most common symptoms. The mean age for infected individuals was 32 yr. Molecular characterization was successfully performed for 17 isolates and two genotypes A (AII, 65%) and B (BIII, 35%) were identified., Conclusion: The most prevalent giardia genotypes among the food handlers in Qazvin were A (AII) and B (BIII) genotypes with A (AII) genotype as the dominant one in the region. Considering the direct association between the food handlers and public health as well as the impact of geographical and host conditions on dispersion and pathogenicity of various genotypes and their zoonotic aspects, further investigations are necessary., Competing Interests: Conflict of interests The authors declare that there is no conflict of interests., (Copyright© Iranian Society of Parasitology & Tehran University of Medical Sciences.)

Free-living amoebae belong to the genus Acanthamoeba; can feed on microbial population by phagocytosis, and with the capability to act as a reservoir and a vehicle of microorganisms to susceptible host. Therefore, the role of endosymbiosis in the pathogenesis of Acanthamoeba is complex and not fully understood. The aim of the present study was to identify bacterial, fungal, and human adenovirus (HADV) endosymbionts as well as evaluating the endosymbionts role of such organisms in the pathogenesis of Acanthamoeba in keratitis patients living in Iran. Fifteen Acanthamoeba (T4 genotype) isolates were recovered from corneal scrapes and contact lenses of patients with keratitis. Cloning and purification was performed for all isolate. Gram staining was performed to identify bacterial endosymbionts. DNA extraction, PCR, and nested PCR was set up to identify endosymbiont of amoeba. Evaluation of pathogenicity was conducted by osmo-tolerance and thermo-tolerance assays and cell culture, and then CPE (cytopathic effect) was survey. Statistical analysis was used between Acanthamoeba associated endosymbionts and Acanthamoeba without endosymbiont at 24, 48, 72, and 96 h. A p value < 0.05 was considered as significant, statistically. A total of 9 (60%) Acanthamoeba (T4 genotypes) isolates were successfully cloned for detecting microorganism endosymbionts. The only isolate negative for the presence of endosymbiont was ICS9. ICS7 (Pseudomonas aeruginosa, Aspergillus sp., and human adenovirus endosymbionts) and ICS2 (Escherichia coli endosymbiont) isolates were considered as Acanthamoeba associated endosymbionts. ICS7 and ICS2 isolates were highly pathogen whereas ICS9 isolate showed low pathogenicity in pathogenicity evaluated. Positive CPE for ICS7 and ICS2 isolates and negative CPE for ICS9 isolate were observed in cell culture. The average number of cells, trophozoites, and cysts among ICS7, ICS2, and ICS9 isolates at 24, 48, 72, and 96 h was significant. This is the first survey on microbial endosymbionts of Acanthamoeba in keratitis patients of Iran, and also the first report of Aspergillus sp, Achromobacter sp., Microbacterium sp., Brevibacillus sp, Brevundimonas sp and Mastadenovirus sp in Acanthamoeba as endosymbionts. Our study demonstrated that microbial endosymbionts can affect the pathogenicity of Acanthamoeba; however, further research is required to clarify the exact pattern of symbiosis, in order to modify treatment protocol., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Objective: Food handlers play an important role in the production and distribution of parasitic contaminations in these societies. The aim of this study was to assess the prevalence of intestinal parasitic infections among the food handlers in Qazvin, Iran., Methods: Totally, 1530 stool samples were collected from food handlers in Qazvin, Iran. Direct feces examination and formalinethyl acetate concentration method followed by Trichrome staining were performed to detect the parasites., Results: The prevalence of intestinal parasites was 82 (5.4%) among food handlers. The species of protozoan parasites found were Blastocystis hominis, Giardia lamblia, Entamoeba coli , and Endolimax nana . The outbreaks of Blastocystis hominis (2.9%), and Endolimax nana (0.3%) were the most and the least protozoan infections observed among the specimens, respectively. Also, there was no helminthic infection detected among the samples. The prevalence of intestinal parasites among the food handlers, based on variables including sex, age, job, and education, did not differ, whereas the prevalence of intestinal symptoms was shown to be higher in infected food handlers (p<0.05)., Conclusions: Despite a decrease in the incidence of intestinal parasites, the protozoan infections are still among the health concerns in these areas. Therefore, continuous monitoring and training of food handlers by local health authorities should be performed on a regular basis.

Introduction: Hymenolepis nana is a zoonotic tapeworm with widespread distribution. The goal of the present study was to identify the parasite in the specimens collected from NorthWestern regions of Iran using PCR-sequencing method., Methods: A total of 1521 stool samples were collected from the study individuals. Initially, the identification of hymenolepis nana was confirmed by parasitological method including direct wet-mount and formalin-ethyl acetate concentration methods. Afterward, PCR-sequencing analysis of ribosomal ITS2 fragment was targeted to investigate the molecular identification of the parasite., Results: Overall, 0.65% (10/1521) of the isolates were contaminated with H. nana in formalin-ethyl acetate concentration. All ten isolates were succefully amplified by PCR and further sequenced. The determined sequences were deposited in GenBank under the accession numbers MH337810 -MH337819., Conclusion: Our results clarified the presence of H. nana among the patients in the study areas. In addition, the molecular technique could be accessible when the human eggs are the only sources available to identify and diagnose the parasite.

This study was aimed at determining the prevalence of parasitic infections among the food handlers of Saqqez County in 2016. Stool samples collected from 1530 food handlers. All samples were examined by both direct technique and sedimentation technique using formalin-ethyl acetate for detecting the intestinal parasites. Of total participants, 1462 (95.6%) were male and 62 (4.4%) female. Intestinal parasitic infections were found in 92 (6.01%) individuals. The highest infection rate was due to Giardia lamblia (4.1%) and the lowest associated with Endolimax nana , Iodamoeba butschlii , and Hymenolepis nana (0.1%). No significant correlation between the intestinal parasitic infections and age, gender, education level, gastrointestinal signs and symptoms, and the type of profession, was established. The results of the present study revealed that only a small number of food handlers in Saqqez County were infected with intestinal parasites with G. lamblia as the most common cause of infection. More strict health control for those working in food industries is of vital importance and continuous education to improve the level of general health in these people is undoubtedly considered as a key determinant towards lower number of parasitic infections., Competing Interests: Compliance with ethical standardsThe authors declare there is no conflict of interests.

Naegleria species are the ubiquitous free-living amoebas that are found worldwide in soil and water. Among Naegleria spp., N. fowleri can cause primary amebic meningoencephalitis (PAM). Ninety water samples were collected from the pond of parks. Also, the water quality parameters were measured at the sampling site (such as temperature, pH, total dissolved solids (TDS), electrical conductivity (EC) and Turbidity). After filtering, the samples were cultured on Bacto-agar enriched with Escherichia coli . A PCR assay was conducted on the culture-positive samples in the ITS1, 5.8SrDNA and ITS2 regions, and then the PCR products were sequenced. The pond water of parks was contaminated with some Naegleria spp. (except N. fowleri ) and a Vahlkampfia avara . There was no significant relationship between water quality parameters and the presence of Naegleria (p > 0.05). Our protocol investigates to detect Naegleria spp. from ponds water of parks in Mashhad city and the relations between the water quality parameters and its presence.

Background: Free-living amoeba (FLA) are wide-spread protozoa that are found in different environmental sources including water, soil, dust, hospital units and ventilation areas. These amoebas can act as opportunistic or non-opportunistic pathogens. Among FLAs, some genera such as Acanthamoeba are important because of their potential pathogenic ability in humans. The purpose of this study is to identify of Acanthamoeba isolated from pond water of parks in a tropical and subtropical region in the Middle East, and its relation with physicochemical parameters.From August to December 2015, 90 samples were collected from pond water of parks of 13 regions of Mashhad City. Physicochemical parameters were measured in situ. After filtering, the samples were cultured on Bacto-agar enriched with Escherichia coli. PCR analysis was conducted on the culture-positive samples, and then the PCR products were sequenced. Statistical analysis was performed by SPSS software and Fisher's Exact and Mann-Whitney test., Results: Among the samples that were examined, 19 samples (21.1%) were positive for Acanthamoeba. The sequencing revealed that Acanthamoeba isolates belonged to T4 genotype of Acanthamoeba. There was no significant relationship between physicochemical parameters and the presence of Acanthamoeba., Conclusion: The prevalence of Acanthamoeba in pond water of parks was relatively high, but there was no significant relationship between physicochemical parameters and the presence of Acanthamoeba.

Background: This study aimed to investigate the anti- Acanthamoeba effects of the most used marketed disinfecting solutions in Iran. Moreover, the efficacy of some nano-compounds was tested against pathogenic Acanthamoeba ., Methods: The present study was conducted in the School of Public Health, Tehran University of Medical sciences, Tehran, Iran during 2015-2016. Cysts of Acanthamoeba T4 genotype (7 × 10 4 /ml) mixed at the equal volume with contact lens solutions including Opti-free, Ginza, ReNu, Maxima, Light, and Cyclean for the recommended time by the manufacturers. Nano-silver and nano-gold compounds were also treated with the amoebae. Chlorhexidine 0.02% and normal saline were used as positive and negative controls, respectively. Dead and alive amoebae were determined using vital stain and suspension was cultured in non-nutrient agar. The entire process was repeated at least three times., Results: In none of the solutions in the manufacturer's brochure recommended time, full cytotoxic effect was observed on the cysts of Acanthamoeba . Opti free express solution destroyed the cysts after 6 days. Nanosilver and nano-gold compounds showed no cytotoxic effect on the cysts of Acanthamoeba ., Conclusion: None of the Nanoparticles compounds as well as contact lenses disinfecting solutions which studied was effective on Acanthamoeba cysts in the manufacturer's brochure recommended time. However, continuing study on Nano-silver and Nano-gold compounds to find effective ingredients against Acanthamoeba are highly recommended., Competing Interests: Conflict of interest The authors declare that there is no conflict of interest.

Background: Acanthamoeba spp. is potentially pathogenic free-living amoeba that can exist in various water sources. The presence of this amoeba in water sources could be a health hazard as Acanthamoeba could lead to severe diseases such as Acanthamoeba keratitis and encephalitis. This study aimed to determine the genotypes of isolated Acanthamoeba spp. in raw wastewater samples in Tehran, Iran., Methods: Overall, 90 raw wastewater samples were collected from water treatment facilities in west and south of Tehran, Iran during 2014-2016. Water samples were filtered and cultured on non-nutrient agar (NNA) medium enriched with Escherichia coli . Morphological and molecular analyses were done on positive strains. The pathogenic ability of the isolated strains was determined using physical assays., Results: Totally, 6 out of 90 (6.7%) samples were positive for Acanthamoeba , according to morphological characteristics of double-walled cysts. Genotyping and sequencing of the positive strains showed Acanthamoeba belonging to T4 (83%) and T11 (17%) genotypes. In vitro pathogenicity tests were revealed that five isolates were classified as non-pathogenic strains and one strain belonging to T4 genotype was classified as the highly pathogenic amoebae., Conclusion: The current research reflected a low contamination of wastewater sources to Acanthamoeba . More studies regarding the contamination of wastewaters before and after treatment are required in different places of the country., Competing Interests: Conflict of Interests The authors declare that there is no conflict of interest.

The continuous increase in Acanthamoeba keratitis, a severe corneal infection, worldwide is mainly due to the increase in the number of soft contact lens users. In the present study, which involves a 5-year study, a total of 138 corneal scraps and contact lenses together with their paraphernalias were obtained from suspected amoebic keratitis patients. All samples were cultured using culture-enrichment method. Pathogenic assay, using thermotolerance and osmotolerance tests were also performed on the positive strains. Sequencing of the isolated strains was done by targeting the DF3 region of 18s rRNA gene. The results revealed that 18 (13 %) of patients were infected with Acanthamoeba spp. As expected, T4 genotype was the most common genotype among the clinical samples; however, in three cases, Acanthamoeba belonging to T11 and T9 were detected. Interestingly, T9 genotype, commonly classified as non-pathogenic amoebae, was identified as a causal agent of a patient with amoebic keratitis. From the pathogenic assay, four strains belonging to T4 genotypes were highly pathogenic. This is the first report of Acanthamoeba T9 genotypes isolated in Iran and the first report of T9 type occurring in amoebic keratitis patients worldwide. Due to the increasing trend of amoebic keratitis (AK) and the identification of new genotypes, such as T9 as the causative agent of AK, more researches in this field are necessary in the region and the world at large.

Background: Free-living amoeba (FLA)-related keratitis is a progressive infection of the cornea with poor prognosis. The present study aimed to investigates the contact lenses of patients with keratitis for pathogenic free-living amoebae., Methods: Overall, 62 contact lenses and their paraphernalia of patients with keratitis cultured and tested for the presence of free-living amoebae using morphological criteria. Unusual plates including plates containing mix amoebae and Vermamoeba were submitted to molecular analysis., Results: Out of 62 plates, 11 revealed the outgrowth of free living amoeba of which 9 were Acanthamoeba, one plates contained mix amoebae including Acanthamoeba and Vermamoeba and one showed the presence of Vermamoeba. These two latter plates belonged to patients suffered from unilateral keratitis due to the misused of soft contact lenses. One of the patients had mix infection of Acanthamoeba (T4) and V. vermiformis meanwhile the other patient was infected with the V. vermiformis., Conclusion: Amoebic keratitis continues to rise in Iran and worldwide. To date, various genera of free-living amoebae such as Vermamoeba could be the causative agent of keratitis. Soft contact lens wearers are the most affected patients in the country, thus awareness of high-risk people for preventing free-living amoebae related keratitis is of utmost importance.

During a long term of road survey for detection of the zoonotic helminthes in carnivores, a male Vulpes vulpes was naturally found infected by Dioctophyme renale in Caspian Sea littoral in Iran. Since the parasite transmission usually occurs through consumption of fish and other accidental food items, investigation of the worm amongst fish-eating animals in the areas with potential circulation of the helminth life cycle, has been regarded as a public health importance so far. Concerning the rout of parasite transmission to humans, different epidemiological aspects should be studied in the country. Present paper describes a destructive lesion of the kidney in the fox due to D. renale after almost a decade of monitoring for the parasite reservoirs in the region.

Objective: Echinococcus granulosus is a globally important cestode parasite causing remarkable medical and economical losses in the world. Ten genotypes (G1-G10) have been identified within this complex species. Protoscoleces rostellar hook characters e.g. total large and small hook lengths may be useful to differentiate genotypes. This study investigates the value of rostellar hook morphometry on genetically identified isolates of E. granulosus using mitochondrial cox1 and nad1 sequencing., Methods: In total, 24 hydatid cyst samples of livestock and human origin were collected. The isolates were then sequenced for the mitochondrial cox1 and nad1 genes and total large and small rostellar hook lengths of protoscoleces were measured., Results: Total large and small hook lengths could differentiate between G1 and G6 genotypes; however, G1 and G3 were not distinguishable by hook morphometry. Only large hook length was significantly different between the G3 and G6 isolates., Conclusion: The G6 genotype is readily distinguishable from G1 by using both small and large hook lengths; however, only total large hook length was significantly different between the G3 and G6 genotypes.

Nineteen hydatid cyst isolates collected from camels in central Iran were subjected to sequences analysis of mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes. A consensus sequence obtained containing 366 nucleotides for cox1 and 471 nucleotides for nad1 genes. Overall, the camel isolates indicated five different sequences in cox1 and nine in nad1 genes. The sequences analysis indicated that 26.3%, 42.1%, and 31.6% of isolates belonging to G1, G3, and G6 genotypes of Echinococcus granulosus, respectively. The isolates with G3 genotype indicated one cox1 sequence having 100% homology with reference G3 sequence (AN: M84663) and two different nad1 sequences, one having 100% homology with reference G3 sequence (AN: AJ237634) and the other with a silent mutation (G to A) in position 279. The presence of G3 genotype (buffalo strain) of E. granulosus as dominant genotype in camels is emphasized. As G3 genotype has formerly been reported in human, the epidemiological role of camels is warranted in future surveys.

In the present study, the prevalence of antibodies to Toxoplasma gondii in sport horses of Qazvin was examined using modified agglutination test (MAT). On 52 horse sera totally examined for anti-Toxoplasma antibodies, 37 horses (71.2%) were seropositive by MAT. Results of the present study showed a high rate of Toxoplasma infection in horses in Qazvin area. More comprehensive study on equine toxoplasmosis is recommended.