Trichuris sp. from 1,040 +/- 50-year-old Cervidae coprolites from the archaeological site Furna do Estrago, Pernambuco, Brazil

We present results of the paleoparasitological analysis of Cervidae coprolites that were recovered from the archaeological site Furna do Estrago, Pernambuco, Brazil. Trichuris sp. eggs were recovered from the coprolite samples dated 1,040 ± 50 years before present. This is the first record of Trichuris sp. in semiarid Cervidae, unexpectedly recorded in archaeological material.     

Paleoparasitological Surveys for Detection of Helminth Eggs in Archaeological Sites of Jeolla-do and Jeju-do

A paleoparasitological survey to detect helminth eggs was performed in archaeological sites of Jeolla-do and Jeju-do, the Republic of Korea. Total 593 soil samples were collected in 12 sites of Jeolla-do and 5 sites of Jeju-do from April to November 2011, and examined by the methods of Pike and coworkers. A total of 4 helminth eggs, 2 eggs each for Trichuris trichiura and Ascaris sp., were found in soil samples from 1 site, in Hyangyang-ri, Jangheung-eup, Jangheung-gun, Jeollanam-do. The egg-recovery layer was presumed to represent a 19th century farm, which fact suggested the use of human manures. This is the third archaeological discovery of parasite eggs in Jeolla-do. Additionally, no helminth eggs in archaeological sites of Jeju-do is an interesting problem to be solved in the further investigations.     

Toxocara canis (Werner, 1782) eggs in the pleistocene site of Menez-Dregan,France (300,000-500,000 years before present)

On the archaeological site of Menez-Dregan in Brittany, France, dated 300,000-500,000 years-old, paleoparasitological analysis of cave deposits led to the detection of well-preserved helminth eggs, which morphology and morphometry pointed to the diagnosis of Toxocara canis eggs, a parasite of carnivore mammals. Paleolithic remains suggested a parasitism of the hyena Crocuta spelaea or other canids that inhabited the region.     

First study of fossil rodent middens as source of paleoparasitological evidences (northwestern Patagonia,Argentina)

The present paper reports the first paleoparasitological results obtained from coprolites of fossil rodent middens and demonstrates the potential of rodent middens as a source of paleoparasitological evidences in South America. Ten fossil rodent middens from northwestern Patagonia, Argentina, were studied. Five coprolites of each midden were fully processed, rehydrated, homogenized, subjected to spontaneous sedimentation, and examined through light microscopy. Eight of the 10 examined rodent middens contained parasite eggs. The eggs of parasites were assigned to Heteroxynema (Cavioxyura) viscaciae Sutton & Hugot, 1989 and Helminthoxys sp. (Nematoda: Oxyuridae), Trichuris sp. (Nematoda: Trichuridae) and one unidentified nematode. Fossil rodent middens were assigned to Lagidium viscacia (Caviomorph: Chinchillidae). The excellent preservation of parasite remains in coprolites from fossil rodent middens provided an opportunity to perform paleoparasitological inferences. The results of this papers demonstrates that fossil rodent middens offer an excellent opportunity for the recovery of parasite remains for future paleoparasitological studies in the southwest of South America.     

Paleoparasitological report on the stool from a Medieval child mummy in Yangju, Korea

Previous studies have successfully shown evidence for parasitic infections in human remains from various archaeological sites. However, in the case of Korea, since there have been very few paleoparasitological reports published, pre-20th century parasitic infection patterns remain obscure. Therefore, in order to partly fill this gap, we are reporting on a case of paleoparasitic infection from the feces of a 15th century child mummy from Yangju, Korea. In the course of the present study, we found the eggs of Clonorchis sinensis, Ascaris lumbricoides, and Trichuris trichiura in the feces of the mummy. Trichuris trichiura eggs were found in far greater numbers than other parasite eggs; in fact, intact bipolar plugs were clearly observed and even the larvae were still visible in some eggs. The eggs of C. sinensis and A. lumbricoides were also well preserved, though not in as great a number. Since we could find a number of well-preserved larvae-containing eggs, we are encouraged that successful extraction, amplification, and sequence determination of ancient DNA from the paleoparasite eggs might be possible in future studies. With additional paleoparasitological investigation using feces from Korean mummies, we hope that a history of parasite infection in Korea will be reconstructed.     

Lutz's spontaneous sedimentation technique and the paleoparasitological analysis of sambaqui (shell mound) sediments

Parasite findings in sambaquis (shell mounds) are scarce. Although the 121 shell mound samples were previously analysed in our laboratory, we only recently obtained the first positive results. In the sambaqui of Guapi, Rio de Janeiro, Brazil, paleoparasitological analysis was performed on sediment samples collected from various archaeological layers, including the superficial layer as a control. Eggs of Acanthocephala, Ascaridoidea and Heterakoidea were found in the archaeological layers. We applied various techniques and concluded that Lutz''s spontaneous sedimentation technique is effective for concentrating parasite eggs in sambaqui soil for microscopic analysis.     

Molecular Paleoparasitological Hybridization Approach as Effective Tool for Diagnosing Human Intestinal Parasites from Scarce Archaeological Remains

Paleoparasitology is the science that uses parasitological techniques for diagnosing parasitic diseases in the past. Advances in molecular biology brought new insights into this field allowing the study of archaeological material. However, due to technical limitations a proper diagnosis and confirmation of the presence of parasites is not always possible, especially in scarce and degraded archaeological remains. In this study, we developed a Molecular Paleoparasitological Hybridization (MPH) approach using ancient DNA (aDNA) hybridization to confirm and complement paleoparasitological diagnosis. Eight molecular targets from four helminth parasites were included: Ascaris sp., Trichuris trichiura, Enterobius vermicularis, and Strongyloides stercoralis. The MPH analysis using 18^th century human remains from Praça XV cemetery (CPXV), Rio de Janeiro, Brazil, revealed for the first time the presence E. vermicularis aDNA (50%) in archaeological sites of Brazil. Besides, the results confirmed T. trichiura and Ascaris sp. infections. The prevalence of infection by Ascaris sp. and E. vermicularis increased considerably when MPH was applied. However, a lower aDNA detection of T. trichiura (40%) was observed when compared to the diagnosis by paleoparasitological analysis (70%). Therefore, based on these data, we suggest a combination of Paleoparasitological and MPH approaches to verify the real panorama of intestinal parasite infection in human archeological samples.     

Syphacia sp. (Nematoda: Oxyuridae) in coprolites of Kerodon rupestris Wied, 1820 (Rodentia: Caviidae) from 5,300 years BP in northeastern Brazil

We present the results of paleoparasitological analyses in coprolites of Kerodon rupestris, rodent endemic to rocky areas of Brazil's semiarid region. The coprolites were collected from excavations at the archaeological site of Toca dos Coqueiros, in the National Park of Serra da Capivara, southeastern of state of Piauí. Syphacia sp. (Nematoda: Oxyuridae) eggs were identified in coprolites dated at 5,300 ± 50 years before present. This is the first record of the genus Syphacia in rodent coprolites in the Americas.     

SL1 RNA gene recovery from Enterobius vermicularis ancient DNA in pre-Columbian human coprolites

Enterobius vermicularis, pinworm, is one of the most common helminths worldwide, infecting nearly a billion people at all socio-economic levels. In prehistoric populations the paleoparasitological findings show a pinworm homogeneous distribution among hunter-gatherers in North America, intensified with the advent of agriculture. This same increase also occurred in the transition from nomad hunter-gatherers to sedentary farmers in South America, although E. vermicularis infection encompasses only the ancient Andean peoples, with no record among the pre-Colombian populations in the South American lowlands. However, the outline of pinworm paleoepidemiology has been supported by microscopic finding of eggs recovered from coprolites. Since molecular techniques are precise and sensitive in detecting pathogen ancient DNA (aDNA), and also could provide insights into the parasite evolutionary history, in this work we have performed a molecular paleoparasitological study of E. vermicularis. aDNA was recovered and pinworm 5S rRNA spacer sequences were determined from pre-Columbian coprolites (4110 BC-AD 900) from four different North and South American archaeological sites. The sequence analysis confirmed E. vermicularis identity and revealed a similarity among ancient and modern sequences. Moreover, polymorphisms were identified at the relative positions 160, 173 and 180, in independent coprolite samples from Tulán, San Pedro de Atacama, Chile (1080-950 BC). We also verified the presence of peculiarities (Splicing leader (SL1) RNA sequence, spliced donor site, the Sm antigen biding site, and RNA secondary structure) which characterise the SL1 RNA gene. The analysis shows that the SL1 RNA gene of contemporary pinworms was present in pre-Columbian E. vermicularis by 6110 years ago. We were successful in detecting E. vermicularis aDNA even in coprolites without direct microscopic evidence of the eggs, improving the diagnosis of helminth infections in the past and further pinworm paleoepidemiological studies.     

A First Case of Human Trichuriasis from a Roman Lead Coffin in France

A paleoparasitological study was carried out on 2 lead coffins recovered from the Roman site of Jaunay-Clan (near Poitiers, France). For the first time, this particular type of burial gave positive parasitological results, and eggs of the whipworm Trichuris trichiura were identified in 1 individual. In the present case, thanatomorphose associated with funerary practices may explain the scarcity of the recovered eggs. However, human whipworm has now been observed in 9 individuals dated to the Roman period. The very high frequency of Trichuris sp. eggs in Roman archaeological sites (up to 80%) suggests that fecal peril, hygiene, and waste management were problematic during this period. Finally, due to the fact that very few analyses have been conducted on human bodies dated to the Roman period, more analyses must be performed in the future to provide further information about diseases in the Roman world.     

Quantitative paleoparasitology applied to archaeological sediments

Three techniques to extract parasite remains from archaeological sediments were tested. The aim was to improve the sensibility of recommended paleoparasitological techniques applied in archaeological remains. Sediment collected from the pelvic girdle of a human body found in Cabo Vírgenes, Santa Cruz, Argentina, associated to a Spanish settlement founded in 1584 known as Nombre de Jesús, was used to search for parasites. Sediment close to the skull was used as control. The techniques recommended by Jones, Reinhard, and Dittmar and Teejen were used and compared with the modified technique presented here, developed to improve the sensibility to detect parasite remains. Positive results were obtained only with the modified technique, resulting in the finding of Trichuris trichiura eggs in the sediment.     

Diphyllobothrium: Neolithic parasite?

During paleoparasitological analyses on several Neolithic sites in Switzerland (Arbon-Bleiche 3) and southwestern Germany (Hornstaad-H?rnle I, Torwiesen II, and Seekirch-Stockwiesen), numerous eggs of Diphyllobothrium sp. were recovered. This is one of the earliest occurrences of this parasite during the prehistoric period in the Old World. The prevalence of this helminth in the samples studied raises the question as to how important parasitic diseases were during the Neolithic period and what their actual consequences were.     

Detection Trend of Helminth Eggs in the Strata Soil Samples from Ancient Historic Places of Korea

For several years, we have conducted a series of studies on the patterns of ancient parasitism prevailing in the soil of rural and urban areas of past Kingdom of Korea. Actually, during our survey of paleoparasitology in archaeological sites of Korean peninsula, numerous ancient parasite eggs were discovered in the samples from the city districts of Hansung (Joseon) and Buyeo (Baikje), the palace moat at Gyeongju (Silla), shell-midden site at Bonghwang-dong (Silla to Joseon), and the reservoir found in Hwawangsansung fortress (Silla). By the paleoparasitological studies, with respect to parasitism in the high-density populations of ancient towns and cities, we have managed to catch glimpses of the patterns prevalent therein: a serious parasitic contamination of the soil in ancient urban areas, but not in rural areas of the past. Our historical research also proposed the plausible mechanism of parasite infection very serious indeed among urban populations in Korean history. Although city dwelling doubtless has accrued significant benefits for people and populations with agriculture, it can be equally supposed that living in such highly populated areas might have facilitated the spread of parasite infection.     

Paleoparasitological remains revealed by seven historic contexts from "Place d'Armes", Namur, Belgium

Human occupation for several centuries was recorded in the archaeological layers of "Place d'Armes", Namur, Belgium. Preventive archaeological excavations were carried out between 1996/1997 and seven historical strata were observed, from Gallo-Roman period up to Modern Times. Soil samples from cesspools, latrines, and structures-like were studied and revealed intestinal parasite eggs in the different archaeological contexts. Ascaris lumbricoides, A. suum, Trichuris trichiura, T. suis. Taenia sp., Fasciola hepatica, Diphyllobothrium sp., Capillaria sp. and Oxyuris equi eggs were found. Paleoparasitology confirmed the use of structures as latrines or cesspit as firstly supposed by the archaeologists. Medieval latrines were not only used for rejection of human excrements. The finding of Ascaris sp. and Trichuris sp. eggs may point to human's or wild swine's feces. Gallo-Roman people used to eat wild boar. Therefore, both A. suum and T. suis, or A. lumbricoides and T. trichuris, may be present, considering a swine carcass recovered into a cesspit. Careful sediment analysis may reveal its origin, although parasites of domestic animals can be found together with those of human's. Taenia sp. eggs identified in latrine samples indicate ingestion of uncooked beef with cysticercoid larvae. F. hepatica eggs suggest the ingestion of raw contaminated vegetables and Diphyllobothrium sp. eggs indicate contaminated fresh-water fish consumption. Ascaris sp. and Trichuris sp. eggs indicate fecal-oral infection by human and/or animal excrements.     

Paleoparasitological analysis of a raptor pellet from southern Patagonia

Organic remains attributable to one regurgitated pellet were examined. The pellet, belonging to a bird of prey and collected from a cave of Southern Patagonia, was dated at 6,540 +/- 110 yr. With standard paleoparasitological procedures, eggs of Capillaria sp. and a mite, Demodex sp., were found. The parasites found in the pellet belong to a rodent ingested by the bird. The present report constitutes the first paleoparasitological study of a regurgitated pellet.     

A palaeoparasitological analysis of rodent coprolites from the Cueva Huenul 1 archaeological site in Patagonia (Argentina)

The aim of the present study was to examine the parasite fauna present in rodent coprolites collected from Cueva Huenul 1 (CH1), northern Neuquén (Patagonia, Argentina), an archaeological site that provides stratified sequences of archaeological and palaeontological remains dating from the Late Pleistocene/Early Holocene Transition to the Late Holocene period. Twenty rodent coprolites collected from different sedimentary units from the site, with ages ranging from 13.844 ± 75-1.416 ± 37 years BP, were examined for parasites. Each coprolite was processed as a whole: rehydrated, homogenised, spontaneously sedimented and examined using light microscopy. The coprolites and the eggs of any parasites present were described, measured and photographed. In all, 158 parasite eggs were found in 10 coprolites. The faeces were positive for Viscachataenia quadrata Denegri, Dopchiz, Elissondo & Beveridge and Monoecocestus sp. Beddard (Cestoda: Anoplocephalidae) and for Heteroxynema (Cavioxyura) viscaciae Sutton & Hugot (Nematoda: Oxyuridae). The coprolites examined were tentatively attributed to Lagidium viscacia Molina (Mammalia, Rodentia, Caviomorpha, Chinchillidae). The life cycles of these parasites are discussed.     

Insights about echinostomiasis by paleomolecular diagnosis

Echinostomiasis is a zoonosis caused by intestinal trematodes and transmitted by the ingestion of mollusks, crustaceans, fish, amphibians, and reptiles, either raw or poorly cooked. Today human infection is endemic in Southeast Asia and the Far East, but has been reported more recently in other regions of the world. Interestingly eggs identified as Echinostoma sp. were found in coprolites from a mummified body human in Brazil, dated 560 ± 40 BP (before present). However, the specific diagnosis based on morphology of the eggs has not been resolved at the species level. As a follow-up to the previous finding, the current study now aims to standardize the methodology for molecular diagnosis and apply it to the coprolite, using current Echinostoma paraensei-positive feces as the reference, and also the same fecal material dried in a stove as an experimental coprolite model. Isolated eggs of E. paraensei and adult worm were included to verify the sensibility and as positive control, respectively. An adult worm of E. luisreyi was used for comparison. PCR using primers in-house for ITS1 region (126 bp) and cox1 (123 bp) of Echinostoma spp. and subsequent nucleotide sequencing were performed. This is the first molecular paleoparasitological diagnosis for echinostomiasis. The methodology was able to amplify specific DNA fragments for the genus Echinostoma sp. in all samples: adult worm, feces, and a single egg of the parasite, in both the experimental coprolite and archaeological sample. Additionally we observed that ancient DNA can also be retrieved without rehydrating the material. The nucleotide sequences from E. paraensei and E. luisreyi are very similar in the fragment analyzed that difficult the differentiation these species, but DNA sequence analysis recovered in the parasite found in the mummy showed more similarity with the species E. paraensei.     

The finding of eggs of Diphyllobothrium in human coprolites (4,100-1,950 B.C.) from northern Chile

Twenty six coprolites from an archaeological site in the province of Iquique, northern Chile, were examined for parasites. Coprolites were found in two excavation units, I and II (Tiliviche site), dated respectively at 5,900 B.C. to 4,110 B.C. and 4,110 B.C. to 1,950 B.C., and identified as of human origin. Only at the unit II coprolites containing helminth eggs identified as Diphyllobothrium pacificum were found. The presence of this tapeworm, a parasite of the American Sea Lion, in human coprolites, points to a diet which included marine fishes and provides information on the antiquity of infection by Diphyllobothrium pacificum. It is interesting to note that Baer (1969) suggests the presence of this tapeworm in pre-Columbian populations when diagnosing the first human cases in today's population in Peru.     

V-shaped Pits in Regions of Ancient Baekje Kingdom Paleoparasitologically Confirmed as Likely Human-Waste Reservoirs

In a paleo-parasitological analysis of soil samples obtained from V-shaped pits dating to the ancient Baekje period in Korean history, we discovered Ascaris lumbricoides, Trichuris trichiura, and Clonorchis sinensis eggs. In light of the samples'' seriously contaminated state, the V-shaped pits might have served as toilets, cesspits, or dung heaps. For a long period of time, researchers scouring archaeological sites in Korea have had difficulties locating such structures. In this context then, the present report is unique because similar kind of the ancient ruins must become an ideal resource for successful sampling in our forthcoming paleoparasitological studies.     

Opisthorchiasis in infant remains from the medieval Zeleniy Yar burial ground of XII-XIII centuries AD

We present a paleoparasitological analysis of the medieval Zeleniy Yar burial ground of the XII-XII centuries AD located in the northern part of Western Siberia. Parasite eggs, identified as eggs of Opisthorchis felineus, were found in the samples from the pelvic area of a one year old infant buried at the site. Presence of these eggs in the soil samples from the infant’s abdomen suggests that he/she was infected with opisthorchiasis and imply consumption of undercooked fish. Ethnographic records collected among the population of the northern part of Western Siberia reveal numerous cases of feeding raw fish to their children. Zeleniy Yar case of opisthorchiasis suggests that this dietary custom has persisted from at least medieval times.