The effects of Fasciola hepatica recombinant proteins (peroxiredoxin and cathepsin L1) on Crohn's disease experimental model.

The immunomodulatory potential of the excretory-secretory (E/S) proteins of the helminths has been shown in previous investigations. This study evaluated the effects of the recombinants and excretory-secretory proteins of the Fasciola hepatica on induced colitis in Balb/c mice. The F. hepatica Recombinant proteins, Cathepsin L1 and Peroxiredoxin, and E/S proteins were intraperitoneally injected into the three mice groups as the case groups, while the control groups received PBS. Colitis was induced in mice by intraluminal administration of the 2, 4, 6-Trinitrobenzenesulfonic acid solution (TNBS). After 8 h, the case groups received the second dosage of the treatments, and it was repeated 24 h later. The immunological, pathological, and macroscopic changes were evaluated 3 days after colitis induction. The macroscopic evaluation revealed significantly lower inflammatory scores in the mice treated with recombinant Peroxiredoxin (rPRX) and recombinant Cathepsin L1 (rCL1). Despite the macroscopic observation, the pathological finding was insignificant between the groups. IFN-γ secretion was significantly lower in splenocytes of the groups that received rPRX, rCL1, and E/S than the controls. IL-10 showed significantly higher levels in groups treated with rPRX and rCL1 than controls, whereas the level of IL-4 was not statistically significant. Excretory-secretory proteins of the F. hepatica showed immunomodulatory potency and the main effects observed in this study were through the reduction of inflammatory cytokine and inflammation manifestation as well as induction of anti-inflammatory cytokines.

Eimeria leuckarti in equid coprolites from the Sassanid Era (2nd-6th century CE) excavated in Chehrabad Salt Mine archaeological site, Iran.

OBJECTIVE: This study reports coccidian oocysts in an equid coprolite dated to the Sassanid Empire (2nd-6th century CE) recovered in Chehrabad Salt Mine archaeological site, Iran. METHODS: Between 2015 and 2017, an archaeoparasitological investigation led to the discovery of an equid coprolite in the Chehrabad Salt Mine archeological site, (Douzlakh), western Iran. Samples were rehydrated using trisodium phosphate solution and were examined by light microscopy. RESULTS: Seven oocysts of Eimeria leuckarti (Flesch, 1883) were identified; they were in various stages of sporulation. CONCLUSION: This is the first report of ancient coccidian oocysts from equids. The importance of this observation is discussed, and current knowledge of eimeriid oocysts at archaeological sites is reviewed. SIGNIFICANCE: The observations of E. leuckarti increases current knowledge of parasite biodiversity in ancient Iran when it rested along the Silk Road, a network of trade routes connecting the East and West that was central to economic, cultural, political, and religious interactions between these regions, and to livestock movement that could contribute to the transmission of the parasites from/to other regions. LIMITATIONS: The contextual information about animal species present in and around the Salt Mine during its working periods, including Achaemenid dynasty (6th to 4th century BCE) and Sassanid era (2nd to 6th century CE), is very limited and does not allow secure conclusions regarding the host origin of the coprolites. SUGGESTIONS FOR FURTHER RESEARCH: Application of molecular biology tools to identify the correct host origin of the coprolites and to detect more parasite species is advocated.

Fasciola Infection Unexpectedly Found During Cholecystectomy: Review on How to Avoid Increasing Surgery Interventions in Non-human Endemic Areas.

PURPOSE: Fascioliasis is caused by Fasciola hepatica of almost worldwide distribution and F. gigantica in wide regions of Asia and Africa. Their adult stage develops in the biliary canals and gallbladder. Infection follows an initial, 3-4 month long invasive, migratory or acute phase, and a several year-long biliary, chronic or obstructive phase. METHODS: The unexpected finding of a fasciolid inside the gallbladder during a cholecystectomy for obstructive lithiasis suspicion in a patient is reported from an area of Iran where human infection had been never reported before and studies on fascioliasis in livestock are absent. RESULTS: The fluke obtained was phenotypically classified as F. hepatica by morphometry and genotypically as F. gigantica by mtDNA cox1 fragment sequencing, although with F. hepatica scattered mutations in species-differing nucleotide positions. The clinical, radiological, and biological signs observed at the acute and chronic phases often lead to some misdiagnosis. Serological methods may be useful in cases of negative coprology. Diagnostic techniques with insufficient resolution leading to unnecessary invasive interventions are analyzed. The way to avoid unnecessary surgery is described, including analyses to be made, diagnostic tools to be used, and aspects to be considered. CONCLUSION: Reaching a correct diagnosis in the confusing presentations avoids procedure delays and unnecessary surgery. A correct drug treatment may be sufficient. Except in extreme pathological presentations, lesions decrease in number and size and finally disappear or calcify after a successful treatment. Finally, the need to increase awareness of physicians about fascioliasis is highlighted, mainly in non-human endemic areas.

Detection of Enterobius vermicularis in archived formalin-fixed paraffin-embedded (FFPE) appendectomy blocks: It's potential to compare genetic variations based on mitochondrial DNA (cox1) gene.

Acute appendicitis represents one of the most common causes of emergency abdominal surgery worldwide. Meanwhile, Enterobius vermicularis has been suggested as one of the probable causes of appendicitis. In this study, the morphological characteristics of the remnant pinworms and pathologic changes were explored in old-archived FFPE tissues of appendectomies. Moreover, we provide the first molecular identification, genetic, and haplotype variation of this nematode from the old-archived FFPE tissue section of appendectomy using the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. Seventeen FFPE appendectomies with E. vermicularis infection, stored over 12-22 years, were collected from two different geographical areas of Iran. In the histopathological examination, tissue changes were observed in thirteen cases (76.4%) and inflammation in four blocks (23.5%). After DNA extraction, the cox1 gene was amplified in twelve (70.6%) cases using the nested polymerase chain reaction (PCR). Phylogenetic analysis and a median-joining network of 78 available cox1 sequences of E. vermicularis revealed 59 haplotypes. We identified five haplotypes that fell into type B. All Haplotypes are novel except for two haplotypes, Hap32 and Hap37, identical to E. vermicularis sequences from Iran, Greece, and Germany. The ranges of diversity distance and haplotype diversity within the isolates were 0-1.9% and HD:0.643-0.667, subsequently. Overall, the absence of inflammation or even tissue changes in some sections can suggest the possible non-inflammatory role of E. vermicularis in appendicitis. Although FFPE material suffers from PCR inhibition, we could successfully use nested PCR to characterize E. vermicularis in old-archived appendectomy blocks and suggest this method as a complementary diagnosis technique in pathology. While the predominant type was B in the Middle East and Europe, further studies on a larger sample size from different geographical regions could probably confirm the results obtained in the present study.

Tracking the existence of Dioctophyma renale in Parthian Empire of Iran (247 BC-224 AD).

Paleoparasitological investigation in the archeological site of the Caspian Sea littoral revealed the presence of human Dioctophyma renale infection, a ubiquitous kidney-residing helminth of mammals, during the Parthian Empire of Iran (247 BC-224 AD). This study reports the oldest human infection with giant kidney worm in the Middle East, mainly in an area where the infection is reported nowadays. The paucity of records throughout the literature suggests that the same species of parasite occurred with low frequency in the past, as well. The cemetery in the archeological site of Kiasar represented a unique opportunity of studying human and animal parasites of the Parthian Empire in Iran.

Dermatoparasitoses in Referral Patients to the Laboratory.

Background: Dermatoparasitic infestations due to the mites Demodex spp. and Sarcoptes scabie are prevalent dermatological disorders worldwide. Methods: Referral patients from the Departments of Dermatology, Infectious Diseases, and from the psychologists, in some cases, to the laboratory of Medical Helminthology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran were examined and documented for demodicosis and scabies from March 2009 to December 2020. All patients' data were collected and then analyzed statistically by SDATA version 14, using the Chi-square test. Results: Out of 494-suspected patients suffering from dermal disorders, 99 patients (20.04%) and 20 cases (4.04%) were found infested with demodicosis and scabies, respectively. Most demodicosis cases belonged to the 46-60 year age group while the infestation rate of scabies was higher in the age group under 5 years (P=<0.0001). Demodicosis was seen more prevalent in women than men, and scabies were higher in men (P =0.15). The cases of demodicosis in fall and scabies in winter and spring were more frequent. Demodicosis picked up in 2015 and 2017 (P=0.03), while the prevalent year for scabies was in 2016 (P=0.77). Both current ectoparasites declined dramatically by Covid-19 pandemic. Conclusion: Demodicosis and scabies have been found correlated with age, and no statistical association was seen between the gender and seasonal factors. Besides, the obvious decline of demodicosis and scabies infestation rates during the Covid-19 outbreak can mention that social distance and hygiene standards have negative effects on dermatoparasites transmission.

First evidence of Brachylaima sp. (Diplostomida; Brachylaimidae) infesting the Etruscan shrew (Suncus etruscus) in Iran.

Of the eleven species of shrews described in Iran, the Etruscan shrew (Suncus etruscus), the smallest known extant mammal by mass, is recorded in northern and southern provinces. During rodent control programs, a female S. etruscus was trapped and found naturally infected with a trematode which was morphologically identified as Brachylaima sp. This finding represents the first report of trematodiasis in this host species.

Assessment of the Endoparasite Fauna amongst the Rodents in Kurdistan Province, West of Iran.

Background: We aimed to evaluate the rate of infection by endoparasites amongst rodents in the western regions of Iran to enhance the level of knowledge amongst health authorities in this entity. Methods: This study was conducted in the west and southwest of Kurdistan Province, including the cities of Sanandaj, Marivan, and Sarvabad. The field mission of this work was performed in three seasons' spring, summer, and autumn. The rodents were captured alive and their gastrointestinal tracts were evaluated for the worm endoparasites. Results: Herein, 208 rodents from 15 types of seven species, including Apodemus, Meriones, Mus, Sciurus, Cricetulus, Microtus, and Dryomys, were captured. In addition, 67 (32.21%) rodents were infected with endoparasites and 10 types of worms endoparasites, including Syphacia muris, Streptophagus spp., Mastophorus muris, Skrjabinema spp., Trichostrongylus spp., Trichuris muris, Hymenolepis nana, Hymenolepis dimimuta, Heligmosomoide spp., and other oxiuros were isolated from their gastrointestinal tracts. Most of these parasites (60%) were isolated from their small intestine whereas they were least (10%) found in their stomach and cecum. Conclusion: Having compared the results of this study with other studies in different regions of Iran, there is a higher variety of rodents and worm parasites in these regions of Iran.

First paleoparasitological evidence of Muellerius sp. in sheep or goat dated to the Sasanian Empire (500 CE) in ancient Iran.

A fecal pellet was recovered in an ancient salt mine in Chehrabad located in western Iran (36.55° N, 47.51° E). Based on prior publications showing the success of the salt mine's environment to preserve various life forms, it was decided to try and ascertain whether this faex contained any parasites of paleoparasitologic interest. The rehydration involved placing the pellet in an aqueous solution of 0.5% trisodium phosphate for a week, followed by the examination of aliquots of the entire rehydrated sample on a total of 153 microscope slides. The examination of the rehydrated material revealed the presence of two larval nematodes; there were no eggs or oocysts recovered. The larvae were photographed and measured, and the decision was made not to submit the two larvae to grinding and DNA extraction for molecular diagnostics due to their excellent state of preservation. The larvae were identified as first-stage larvae that appear to represent the genus Muellerius. Herein are reported the finding of two first-stage larvae of Muellerius nematodes (Metastrongyloidea, Protostrongylidae) from the rehydrated fecal pellet collected in a Chehrabad salt mine from the era of the Sasanian Empire.

Dual-Source Dual-Energy CT-Scan Confirms the Diagnosis of Ancient Hydatid Cysts Recovered from a Late Roman Burial in Amiens, France.

Background: Two calcified objects recovered from an adolescent in a burial site in Amiens, France, have been previously identified as hydatid cysts using thin-section petrography. The importance of ancient hydatidosis besides the value of these unique archeological excavated materials encouraged the authors to look at this attractive subject more interdisciplinary by implementing medical radiology. Methods: In the current experiment, which has been carried out in the Radiology Department, Tehran Heart Center (THC), Tehran, Iran, the conventional and dual-energy dual-source Tomography, X-Ray Computed-scan was used in studying the remaining structures of the two calcified masses. The imaging procedure was carried out based on X-Ray attenuation by two different tube voltages. Results: A high concentration of calcium sediment in the cyst walls was revealed in Hounsfield units, the measuring of the elements in CT. Taking advantage of implementing this imaging technique the oxalate calcium was also shown as the dominant component of the samples. The results were all in favor of diagnosing hydatid cysts. Conclusion: The achieved pictorial results in the present paper have highlighted the important role of CT scan as a noninvasive confirming technique in paleopathological investigations. Using Dual-source dual-energy CT-scan in reconfirming these previously identified hydatid cysts, is an encouraging message towards the necessity of sequential studies on invaluable biological excavated pieces.

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.

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.

Serological Detection of Trichinellosis among Suspected Wild Boar Meat Consumers in North and Northeast of Iran.

BACKGROUND: Trichinellosis is a foodborne zoonosis disease worldwide. Humans acquire infection by ingesting raw or uncooked animal flesh containing viable Trichinella larvae. The most common reservoirs of this helminth are pigs and wild boars. In northern Iran, hunting and consuming wild boars meat by some communities, including ethnic Armenians, may expose them to trichinellosis. Here, we investigated anti-Trichinella IgG antibodies in high-risk individuals in northeastern Iran. METHODS: From Mar to Aug 2020, we collected 189 blood samples from individuals with a history of wild boar meat consumption and examined the sera for anti-Trichinella IgG antibodies using a commercial ELISA kit (NovaTec Immunodiagnostica GmbH, Germany). Sera from 30 individuals with no history of eating wild boar meat was used to determine the range of actual negative values and possible cross-reactivity with other similar antigens. RESULTS: Of the 189 participants, 5 (2.6%) had anti-Trichinella IgG antibodies (OD, 1.176 ±0.154). None of the 30 negative controls became positive (OD, 0.198 ± 0.044). The age, gender, occupation, and education showed no significant association with Trichinella seropositivity rate (P>0.05). All five seropositive cases were among 112 individuals (4.46% seropositivity) that resided in the western part of the study area, stretching from Behshar to Gorgan. CONCLUSION: Eating wild boar meat might expose individuals to trichinellosis in the north and northeast of Iran. Further studies with more individuals from different parts of the country and confirmation of the ELISA by additional tests like Western blot will give a more in-depth insight into human trichinellosis epidemiology in Iran.

Occurrence of Dioctophyme renale (Goeze, 1782) in road-killed canids of Iran and its public health implication.

Dioctophyme renale, is the largest of parasitic nematodes, which infects different species of fish-eating carnivores worldwide. The northern provinces of Iran (Guilan and Mazandaran) located in south of the Caspian Sea are suitable for parasitic infections due to the mild and humid climatic conditions. From separate surveys of road-killed canids in various parts of the Caspian Sea littoral area in Iran, 70 carcasses were collected along the roads of Guilan and Mazandaran from 2015 to 2017. Dioctophyme renale detected by direct observation and molecular methods based on Cytochrome c oxidase subunit 1 (COX1 gene) sequencing analysis. Molecular investigation was also performed to validate prevalence and reduce false negative concerns. Dioctophyme renale was found in eight of 70 carnivores, mostly in the right kidneys, as well as two cases in the abdominal cavity of a dog and a golden jackal. More carcasses on the roads were seen with lacerated internal organs. Given the frequent number of giant kidney worms in canids in the region, the transmission of this zoonotic helminth to humans seems possible, since the area is a tourism hub in the country. The infection burden of this helminth should be investigated using DNA analysis of kidney tissue of road-killed carnivores in Iran.

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.

Identification of Fasciola Species Using Tegumental Spines in Tissue Sections.

BACKGROUND: Efforts to find a reliable non-molecular means of identification has been the main purpose of the current work that always is persuaded by researchers interested in the field of parasitology. METHODS: Adult fasciolids were obtained from the slaughterhouses in different parts of Iran in 2017, and investigated using the classical old fashion morphological appearances of the worms implementing a camera lucida equipped microscope. Histological procedure was subsequently performed for almost the entire collected adult worms followed by Hematoxylin and Eosin (H&E) staining technique. DNA extraction and RFLP-PCR technique were carried out for the entire fasciolid liver flukes. To attain more comparable morphological conclusions, Scanning Electron Micrographs were also implemented for two molecularly identified fasciolids. RESULTS: Based on spine morphology observed in worm's tissue sections two types of tegumental spines, "pointed" and "molar" shapes have been identified addressing to distinguish F. hepatica and F. gigantica species respectively. The present identification has been also supported by Molecular analysis using RFLP-PCR technique. CONCLUSION: There are some hidden morphological characters implemented in species identification for certain helminths. Meanwhile, the emergence of computer image analysis system (CIAS) on the scene of taxonomy, has revolutionized the accuracy of measurement in morphology by employing detailed parameters that have not been regarded before. The current study has illustrated the tegumental spines of two Fasciola species in tissue sections which has not been enough considered in helminthological publications so far.

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.

Confusing a Pollen Grain with a Parasite Egg: Infection or Traditional Medicine?

Camacho & Reinhard stated in the December 2019 issue of the KJP (57: 621-625) that we confused a pollen grain with an Enterobius egg found in the grave of a female adolescent residing in ancient Tehran 7,000 years ago. We want here to clarify and answer to the outlined points in their article.