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.

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.

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.

Fasciola hepatica eggs in paleofaeces of the Persian onager Equus hemionus onager, a donkey from Chehrabad archaeological site, dating back to the Sassanid Empire (224-651 AD), in ancient Iran.

Fascioliasis is a highly pathogenic zoonotic disease caused by the liver trematodes Fasciola hepatica and F. gigantica. Within the multidisciplinary initiative against this disease, there is the aim of understanding how this disease reached a worldwide distribution, with important veterinary and medical repercussions, by elucidating the spreading steps followed by the two fasciolids from their paleobiogeograhical origins. Fasciola eggs were detected in paleofaeces of a donkey, probably the present-day endangered Persian onager Equus hemionus onager, found in the Chehrabad salt mine archaeological site, Zanjan province, northwestern Iran. The biological remains dated back to the Sassanid period, 224-651 AD. Egg characteristics allowed for their specific ascription to F. hepatica. The interest of this finding relies on the fact of being the first archaeological finding of Fasciola in Asia and the Near East. Moreover, it allows to reach many conclusions about historical, epidemiological and spreading aspects of the disease. The finding in Chehrabad indicates that, at that time, this fasciolid had already spread through the Zagros mountains eastward from the Fertile Crescent. In that region and in ancient Egypt, livestock domestication played a crucial role in facilitating the disease spread during the postdomestication period. Donkeys appear at present to be usually infected by fasciolids in countries of the Fertile Crescent - Ancient Egypt region or neighbouring that region, with prevalences from low to very high. The high pathogenicity and mortality induced by Fasciola in these equines should be considered as an additional potential factor among the causes of the extinctions of E. h. hemippus in Syria, E. h. hydruntinus in the Anatolia-Balkans area, E. h. onager in the Caucasus and maybe also its decline in Iran. Indeed, Eurasiatic wild asses were present in the region and neighbourhood of the Fertile Crescent when the domestication of the livestock reservoirs of Fasciola began.

Genotyping and molecular analysis of Enterocytozoon bieneusi isolated from immunocompromised patients in Iran.

Microsporidia are known as opportunistic unicellular pathogens, particularly so in individuals with congenital or acquired immunodeficiency. Enterocytozoon bieneusi is one of the most common species infecting both immunocompromised and immunocompetent individuals. The aim of this study was to assess the distribution of E. bieneusi genotypes among immunocompromised patients in Iran. From 329 stool samples referred for parasitological analysis during 2011-2014, 14 samples from immunocompromised patients proving positive for E. bieneusi by SSU rDNA analysis were selected. Genotyping was carried out using specific primers targeting the Internal Transcribed Spacer (ITS) region. Subsequently, all samples were sequenced and results queried against the GenBank database. Moreover, sequences were subject to phylogenetic analysis. The expected amplification product was generated for all samples. Genotype D was identified in patients with HIV+/AIDS, transplant recipients, and cancer patients, while Genotype E was identified only in cancer and HIV+/AIDS patients. Phylogenetic analysis revealed that there was no relationship between genotypes and types of immunosuppression, whereas most genotype D isolates grouped with those described previously from cattle, horses, birds, and humans. E. bieneusi genotype D appears to be the most frequent genotype in immunocompromised patients, while Genotype E was observed only in HIV+/AIDS patients and cancer patients, not transplant recipients.