Genomic palaeoparasitology traced the occurrence of Taenia asiatica in ancient Iran (Sassanid Empire, 2th cent. CE-6th cent. CE).

Palaeoparasitology investigates parasitological infections in animals and humans of past distance by examining biological remains. Palaeofaeces (or coprolites) are biological remains that provide valuable information on the disease, diet, and population movements in ancient times. Today, advances in detecting ancient DNA have cast light on dark corners that microscopy could never reach. The archaeological site of the Chehrabad salt mine of Achaemenid (550-330 BC) and Sassanid (third-seventh century AD) provides remains of various biotic and abiotic samples, including animal coprolites, for multidisciplinary studies. In the present work, we investigated coprolites for helminth eggs and larvae by microscopy and traced their biological agents' DNA by Next Generation Sequencing. Our results revealed various helminths, including Taenia asiatica, the species introduced in the 1990s. Implementing advanced modern molecular techniques like NGS gives a paramount view of pathogenic agents in space and time.

Gastrointestinal Helminthic Parasites of Stray Cats (Felis catus) in Northwest Iran.

BACKGROUND: Stray cats are considered an important source of various human and animal diseases, particularly diseases of parasitic helminths. We aimed to investigate the distribution of zoonotic species of gastrointestinal helminths in stray cats in Meshkin-Shahr district in Ardabil Province in the northwest of Iran. METHODS: The gastrointestinal tract of 104 stray cats from villages of Meshkin-Shahr district were provided during 2014-2015. Each gastrointestinal tract was cut into distinct sections, including esophagus, stomach, small intestine, and large intestine, and each section was examined separately for detection of helminths. Helminths were collected and then identified at the species level after clearing and staining. RESULTS: Overall, 88 out of 104 cats (84.6%) were found to be infected with at least one gastrointestinal helminth. The rate of infection for each species was as follows: Toxocara mystax (syn. cati) (49%), Taenia taeniaeformis (44.2%), Joyexiella pasqualei (32.7%), Dipylidium caninum (23.1%), Rictularia cahirensis (4.8%), and Physaloptera praeputialis (4.8%). Among these parasites, only Ph. praeputialis was collected from the stomach, all other helminths were collected from the small intestine. CONCLUSION: The results demonstrate a high infection rate of stray cats with zoonotic helminths. The presence of zoonotic species in stray cats, particularly T. mystax, has public health importance.

First Report of Passerilepis parina (Cestoda: Hymenolepididae) from Parus major in Iran: A Probable Fatal Case Due to Hyperinfection.

Cestodes are important parasites that can affect the health of humans and wildlife. Among these, the genus Passerilepis is an important endoparasite of Passeriform birds while poorly studied in Iran. During a parasitological field survey in central parts of Iran in 2018, thirty-two cestodes, as an obstructive intertwined mass, recovered from the intestine of a recently dead Parus major (great tit). Morphological characteristics of recovered cestodes were drawn carefully by a camera lucida equipped microscope and identification was carried out using standard keys. All of the collected cestodes were identified as P. parina. In the current study, we recorded P. parina from great tit for the first time in Iran.

Echinococcoses in Iran, Turkey, and Pakistan: Old Diseases in the New Millennium.

Echinococcosis is considered a cosmopolitan zoonosis caused by different species of small taeniid tapeworms of the genus Echinococcus and is regarded as a neglected zoonosis. Cystic and alveolar echinococcoses are endemic diseases of Tibetan, Pamir, and Iranian plateaus. All of the countries within the Iranian plateau are affected by echinococcosis. Pakistan, Turkey, and Iran are the three most populous countries of the region, in which echinococcosis is highly endemic. The three neighboring countries share strong cultural and socioeconomic ties. The present study aimed to provide a broad review of the status of cystic and alveolar echinococcosis, summarizing the current knowledge about geographical distribution, molecular epidemiology, and transmission dynamics of Echinococcus granulosus sensu lato and Echinococcus multilocularis in this region. Additionally, we aimed to understand disease burden and risk factors as basic requirements for establishing a surveillance system and planning prevention and control programs. A considerable body of information is available on different aspects of echinococcosis in this region; however, several information and research gaps need to be filled before planning control programs. None of the countries in the region have an elaborate echinococcosis control program. Effective control programs require multi/intersectoral coordination within a One Health approach with a long-term political and administrative commitment and enhanced international collaboration among the three countries.

First report of Spirocerca lupi larva in dung beetles (Scarabaeus armeniacus) in the central region of Iran: A morphological and molecular identification.

Dung beetles as detritivores insects, naturally use feces of vertebrates as foods and reproduction beds. This leads to frequent contacts between dung beetles and parasitic helminths. The current study was carried out to assess infections of dung beetles with larval stages of helminths in rural areas of Taleqan County, Alborz Province, Iran. A total number of 200 dung beetles were collected randomly in June 2017 from the highlands of Taleqan County. Beetles were dissected in normal saline and carefully studied using a stereomicroscope. Morphological characteristics of the recovered larvae were drawn using a camera lucida equipped microscope at 400× magnification. Then, genomic DNAs of the recovered larvae extracted and PCR amplifications of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene were carried out and the amplicons were sequenced. All collected dung beetles identified as Scarabaeus armeniacus from the Scarabaeidae family (55.5 % were male and 44.5 % female). Three females of the beetles have been found to be naturally infected with the third stage larvae of Spirocerca lupi. The average length and width of the larvae were 2.95 (2.81-3.15; CI 95 %) and 0.12 (0.1-0.15; CI 95 %) mm, respectively. The phylogenetic analysis showed that S. lupi belonged to a clade within the Spirocercidae family. In the current study, S. armeniacus introduced as a potential biological vector for transmission of S. lupi to vertebrates in the region. To the best of the authors' knowledge, this is the first report on the larval stages of S. lupi in S. armeniacus.

Phylogenetic analysis of Trichostrongylus vitrinus isolates from southwest Iran.

BACKGROUND: Trichostrongylus is one of the most important zoonotic trichostrongylid nematodes, infecting mostly livestock. Data on its genetic characteristics are lacking in Iran. METHODS: We determined the phylogenetic relationships of Trichostrongylus species in three counties of Kohgiloyeh and Boyerahmad (K-B) province, southwest Iran. Small intestine and abomasum of 70 sheep and goats were investigated. RESULTS: A total of 35 isolates of Trichostrongylus worms were detected and all were genetically identified as Trichostrongylus vitrinus. Analysis of 321 bp of the internal transcribed spacer 2 (ITS2) of ribosomal DNA revealed 16 genotypes. All genotypes were single nucleotide polymorphisms, including some hypervariable points. All sequences were trimmed to 170 bp, compared with sequences on GenBank including short sequences from other endemic foci of Iran and other countries and all isolates were used to generate a maximum likelihood phylogenetic tree, which consisted of two clades A and B. Clade A included isolates from Iran, Russia, New Zealand, Australia and the UK; clade B only contained South African isolates. Most clade A isolates (north, southwest and west Iran, Russia, New Zealand, Australia and UK) were in a similar phylogenetic position. One subclade was detected in clade A (isolates from Southwest Iran, New Zealand and UK). CONCLUSIONS: We hypothesize that drug resistant T. vitrinus may account for its exclusive detection in our samples. The high similarity of genotypes from Iran, New Zealand and UK may be due to their close political relationships during the colonial era. More research is needed to understand better the phylogeny of T. vitrinus and its relationship with drug resistance and human transmission.

Intestinal helminthic parasites of rodents in the central region of Iran: first report of a capillariid nematode from Dryomys nitedula.

OBJECTIVES: Rodents play an important role to spread zoonotic diseases through society. The current study was carried out to collect informative data on the intestinal helminthic infections of wild rodents in Taleqan County, Alborz Province, the center of Iran, emphasizing their zoonotic aspects. RESULTS: Sixty-two killed rodents by local farmers belonging to five species were collected, among which 24 were identified as Mus musculus, 15 as Meriones persicus, 12 as Meriones libycus, 10 as Apodemus witherbyi, and 1 as Dryomys nitedula. Of them, 30 (48.4%) were infected with at least one helminth species. Rodents were infected with Hymenolepis diminuta (42%), Syphacia obvelata (21%), Hymenolepis nana (17.7%), Heligmosomoides polygyrus (9.6%), Trichuris muris (8%), and as well as a capillariid nematode that was isolated for the first time from D. nitedula in Iran. The findings of the present study revealed a significant intestinal helminthic infection of rodents in Taleqan County. Improving hygiene practices, and making a preventive attitude can be helpful to reduce the hazards of rodent-borne diseases in the area where humans, livestock, and synanthropic rodents are living close to each other.

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.

Herbivores Coprolites from Chehrabad Salt Mine of Zanjan, Iran (Sassanid Era, 224-651 AD) Reveals Eggs of Strongylidae and Anoplocephalidae Helminths.

BACKGROUND: The ancient Chehrabad Salt mine, a well-known archaeological site in Iran, has recently received increasing interest from Iranian and international archeologists. Also, the biological remains from this site have provided valuable sources for studying the pathogenic agents of ancient times. This study aimed to identify the parasitic helminth eggs preserved in the herbivores coprolites. METHODS: From 2011 to 2015, we received three coprolites belonging to herbivorous animals recovered during excavations in Chehrabad Salt mine of Zanjan, Iran. The coprolites were dated back to the Sassanid era (224-651 AD) by using radiocarbon accelerator mass spectrometry (AMS) and archeological stratigraphy methods. Following rehydration of the specimens in a 0.5% trisodium phosphate solution, the suspensions were mounted in glycerin jelly on glass slides and examined by a light microscope with 100x and 400x magnifications. RESULTS: Two coprolites belonged to donkeys and one to an unknown herbivore species. The recovered eggs belonged to members of two helminths families, Strongylidae, and Anoplocephalidae. Also, within the two coprolites, some mites, presumably of the order Oribatida, were observed. CONCLUSION: The presence of two different nematodes in the equids coprolites provide clues of the burden of helminths infection on working animal at the Sassanid time and demonstrates the appropriate preservation condition of biological remains in the ancient salt mine of Chehrabad as well.

Wild Rodents and Their Ectoparasites in an Enzootic Plague Focus, Western Iran.

Introduction: Entomological surveys of ectoparasites and their hosts are an essential tool for assessing the risks of rodent-borne diseases transmitted to humans by arthropod vectors. Objectives: This study was carried out to update the epidemiological data of plague with respect to species compositions of the rodents and their ectoparasites at enzootic foci located in Kurdistan Province, Iran. Methods: The rodents' habitats were selected based on past records of plague and subclimates in each study district with especial attention to the vegetation type. The trapped rodents were anesthetized using a chloroform chamber, and the animals were then examined for ectoparasites by brushing their hair over a pan containing water. The ectoparasites were collected with a fine brush and preserved in 70% ethanol in screw cap tubes. Results: A total of 208 rodents were trapped from three districts. Taxonomic ranking of the rodents indicated that the specimens belonged to 2 suborders of Myomorpha and Sciuromorpha, 4 families (Muridae, Muscardinidae, Cricetidae, and Sciuridae), 7 genera, including Meriones, Apodemus, Mus, Sciurus, Microtus, and Dryomys, and 15 species. Out of 208 rodents, only 56 (26.9%) were infested with 22 species of ectoparasites. Totally, 312 ectoparasites were isolated from 56 rodents, including 12 flea species (54.5%), 6 mite species (27.3%), 3 tick species (13.6%), and one louse species (4.6%). Five species of fleas were recorded for the first time in Kurdistan Province, including Ctenophthalmus iranus persicus, Paraceras melis melis, Nosopsyllus iranus iranus, Paraceras sp., and Ctenophyllus spp. Conclusion: The finding revealed new records for rodents and ectoparasites in Kurdistan Province, as well as changes in dominant rodent species and their ectoparasites compared to previous studies. This phenomenon can influence the changes in the incidence of plague and its epidemiology.

Echinococcus multilocularis and Echinococcus granulosus in canines in North-Khorasan Province, northeastern Iran, identified using morphology and genetic characterization of mitochondrial DNA.

BACKGROUND: Canids are definitive hosts of Echinococcus multilocularis and Echinococcus granulosus. This study aimed to survey these two Echinococcus species in canids of North-Khorasan Province, northeastern Iran, using morphological criteria and genetic characterization of mitochondrial DNA. METHODS: The 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 for Echinococcus species. 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 cytochrome c oxidase 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. RESULTS: Overall, 9.4% of the canids (eight jackals and two foxes) were found infected with E. multilocularis by molecular methods, of which seven cases were also confirmed using morphological description of the adult worms. Echinococcus granulosus was found in 6.6% of the canines (four dogs, two jackals and one wolf) as determined by both molecular methods and adult cestode morphology. All E. granulosus isolates were identified as the G1 genotype. Comparative sequence analysis indicated 0-0.7% and 0% intraspecific divergence within E. granulosus isolates and 0% and 0-0.2% within E. multilocularis isolates for cox1 and nad1, respectively. CONCLUSIONS: This study revealed the presence of E. multilocularis and E. granulosus in canids of North-Khorasan Province of Iran. Jackals were found infected with both E. multilocularis and E. granulosus, but infection with the former species was higher.

Esophageal-Crop Capillariasis and Proventriculus Ventriculus Hystrichisiasis in a Migratory Duck (Anas crecca).

BACKGROUND: The aim of the present study was to study pathological and parasitological characteristics of simultaneous capillariasis and hystrichiasis in a diseased Anas crecca captured in Mazandaran Province of Iran on Oct 2016. METHODS: Parasitology and histopathology techniques were used to detect the parasites of the carcass of the captive common teal. RESULTS: Macroscopically, severe chronic inflammatory reactions and nodular or granuloma formation and irregular thickening of the affected wall of the esophagus, crop, proventrculus and ventriculus were observed. Microscopically, apart from the chronic inflammatory reactions and granuloma formation, in the paraffin sections stained with Harris Hematoxyline and Eosin technique, the characteristics of the mature female and male Capillaria spp. and their barrel-shaped operculated embryonated eggs were found within the tunnels burrowed by the nematode in the hyperplastic stratified squamous epithelium of the inflamed crop and distal portion of the esophagus. The mature female Hystrichis spp. containing oval-shaped, embryonated non-operculated eggs and male parasite were found within the labyrinthus spaces formed in the submucosa granulomas of the ventriculus and proventriculus. CONCLUSION: Based on the parasitological and pathological studies, the species of the nematode parasites were identified as Capillaria (Thominx) contorta and Hystrichis. tricolor.

A case of fatal disseminated strongyloidiasis accompanied with intestinal obstruction.

Strongyloides stercoralis is an endemic parasite in some regions including the tropical and subtropical areas with high humidity. Most infections are asymptomatic with nonspecific signs and symptoms, making the final diagnosis complicated. Here, we report a patient referred to our hospital with signs consistent with sepsis, intestine obstruction, which finally died with the diagnosis of strongyloidiasis. The patient was from northern parts of Iran which are considered as endemic areas for S. stercoralis. In conclusion, there is an important message in this history, i.e. physicians should be aware of specific and non-specific signs of strongyloidiasis especially in people living in endemic areas to make an accurate final diagnosis by proper clinical and paraclinical examinations.

Infection of the Jackal (Canis aureus) by Haplorchis taichui (Trematoda: Heterophyidae) in Southwestern Iran: A Clue for Potential Human Infection.

BACKGROUND: We detected eight trematodes in the small intestine of a road-killed jackal (Canis aureus) from Hamidiyeh District near the city of Ahvaz, Khuzestan Province in 2010. METHODS: Three worms were stained with carmine acid, mounted in Canada balsam on glass slides and examined under a light microscope at 1000X magnification. PCR and sequencing of a partial ITS2 sequence were used to approve the diagnosis. RESULTS: The flukes measured ≈1 mm in length with an elongated ovoid shape resembling the members of heterophyid, and only one testis, characteristics of the genus Haplorchis. Sequencing of a 481-bp fragment of the ITS2 locus from the worms revealed 97%-98% identity with the similar sequences of the H. taichui flukes previously identified in the fish, cat, and humans from Thailand, China, and Vietnam. CONCLUSION: Further studies with the application of reliable molecular tools to diagnose trematode infections in wildlife and humans can bring more insight into the epidemiology of fish-borne flukes including H. taichui in this area.

A survey on endoparasites in wild rodents of the Jaz Murian depression and adjacent areas, southeast of Iran.

In this survey, rodents and their endoparasites were investigated in the Jaz Murian depression and adjacent areas, southeast Iran. In total, 146 specimens of rodents belong to 13 species were trapped. In general, 10 different genera of endoparasites including 11 species were collected. The endoparasites were identified as follows: (1) Nematodes: Trichuris muris, Syphacia obvelata, Labiostomum sp., Labiostomum naimi, Mastrophorus muris, Aspicularis tetraptera and Heligmosomoides skrjabini, Physaloptera sp. (2) Cestodes: Choanotaenia sp., Raillietina sp., and Hymenolepis diminuta. Of 146 captured rodents, Tatera indica was found with high parasitic infestation (with 93% infested) comparing to Acomys dimidiatus (66%), Rattus rattus (50%), Meriones libycus (15%), Jaculus blanfordi (14%) and Mus musculus (8%) whereas, seven rodent species, Nesokia indica, Gerbillus nanus, Golunda ellioti, Calomyscus hotsoni, Apodemus witherbyi, Cricetulus migratorius and Microtus mystacinus were free from any parasitic infestation. Those six infested rodent species were collected from the center of the Jaz Murian depression, whereas seven non-infested rodents' species except N. indica and G. nanus live in the marginal ranges of the Jaz Murian depression, therefore, these species inhabiting the central parts were supposed to be more important from the health aspect. The species, Labiostomum naimi collected from A. dimidiatus is the first report of this species in rodents from Iran.

Parasitic Helminths in Wild Boars (Sus scrofa) in Mazandaran Province, Northern Iran.

BACKGROUND: Wild boars (Sus scrofa) are distributed worldwide and found in many parts of Iran. Although S. scrofa is reservoirs for many parasites, there is little data on helminthic prevalence in them. We aimed to survey the status of helminthic infections in S. scrofa in the Mazandaran Province of northern Iran. METHODS: Twenty-one wild boars were captured and examined for helminth infection during Dec 2012-Mar 2014. Adult worms such as Macracanthorhynchus hirudinaceus were identified by helminth size and shape, and the arrangement of the proboscis hooks. The sedimentation and flotation techniques were used to detect parasite eggs and larvae in faecal samples. Muscle samples were also surveyed for Trichinella larvae by artificial digestion method. RESULTS: Of the 21 samples, 13 (61.9%) were infected with one or more helminth species. Seven helminth types were identified in the alimentary track, comprising 5 nematodes, 1 trematode, and 1 acanthocephalan, with prevalence rates of Macracanthorhynchus hirudinaceus (57.14%), Globocephalus spp. (33.33%), Trichuris suis (19.04), Gongylonema pulchrum (14.28%), Fasciola hepatica (14.28%), Dioctophyma renale (4.76%), and Ascaris suum (4.76%). CONCLUSION: Wild boars might be involved in transmitting zoonotic parasites to humans. The abundance of these animals near human habitation creates favorable conditions for infection. So the risk of parasitic helminth diseases increases in other animals and humans.

Frequency of Intestinal Parasites among Zoo Animal by Morphometric Criteria and First Report of the Bivitellobilharzia nairi from Elephant (Elephasmaximus maximus) in Iran.

BACKGROUND: Intestinal parasitic infections are major causative agents of wildlife health complications among different parts of the world. This study aimed to investigate the gastro-intestinal parasites in feces of the zoo animals based on parasitological and morphometric criteria. METHODS: One hundred fresh fecal samples were collected from 35 species of animal lived in Eram park zoo, Tehran, Central Iran during Oct 2015 to Jun 2015. All collected samples were examined by microscopic observation following direct wet mount preparation (normal saline and Lugol's iodine), formalin-ether concentration, and permanent staining. The morphometric aspects of the recovered eggs were surveyed with the aid of Camera Lucida (×400). RESULTS: 65.7% (23/35) of zoo animal species were infected with intestinal parasites. The superfamily Trichostrongyloidea (6/16) and Strongylus sp. (16/4) were the most prevalent helminthic infections, while Blastocystis sp. (6/14), Entamoeba cyst (3/14) and Eimeria sp. (3/14) were the common protozoan parasites. For the first time, Bivitellobilharzia nairi egg was identified an elephant at Iran. Intestinal parasitic infections were apparently circulating among animals of the Eram park zoo. CONCLUSION: Identified parasitic infections can consider as a threatening source to visitors and workers' health that have contact with animals or their feces. Therefore, the effectual preventive strategies should be addressed to determine the risk factors, mechanisms of cross-transmission of parasite, the importance of applying the hygienic practices and well adjusting deworming programs for the animals, zoo workers and visitors.

Prevalence of Corynosoma caspicum infection in Gasterosteus aculeatus fish in Caspian Sea, Northern Iran.

AIM: There is little information about the prevalence of Corynosoma caspicum in fish particularly Gasterosteus aculeatus in Iran and the world. The aim of the present study was to find out the prevalence of acanthocephalan infection in Babolsar district, southern coastal of Caspian Sea, Northern Iran. MATERIALS AND METHODS: Between September 2012 and August 2014, a total of 360 G. aculeatus fishes were randomly collected by drift nets from coastal regions in Babolsar and then examined the intestine and body cavity for worm infections. RESULTS: A total of 360 G. aculeatus fishes, 109 (30.3%) were found infected with at least one Corynosoma capsicum, and there was no significant association between genders and the prevalence infection of acanthocephalan. Moreover, there was a significant difference in infected rate between summer (79%, 86/109) and spring (21%, 23/109) (p<0.05). CONCLUSION: The high occurrence of Corynosoma infection in G. aculeatus indicates the enzootic constancy status of the infection in the southern coastal of Caspian Sea, Northern Iran.