Recovering ancient parasites from Andean herbivores: test of the Mini-FLOTAC technique in archaeological samples.

The optimization of techniques for recovering parasitic remains is key in paleoparasitology. The Mini-FLOTAC technique (MF) is based on passive flotation and is used for diagnosis of parasites and was never tested on ancient samples. Our objective was to assess the effectiveness of MF in paleoparasitology, aiming at improving the techniques for the recovery of parasitic remains in order to upgrade the interpretative potential of the paleoparasitological evidence. Three techniques were tested: MF, spontaneous sedimentation (SS), and centrifugation-sucrose flotation (CF) testing camelid and goat coprolites. Statistical tests were performed with the R software. Our result displayed that, in the case of SAC samples, MF recovered less number of parasitic species than SS, but obtained a greater number of positive samples for protozoa. For goat samples, MF recovered a higher number of positive samples and parasitic species than SS, added that it was the technique that recovered a greater number of parasite structures. Therefore, results vary according to the zoological origin of the samples and the parasitic species recorded. We suggest starting using MF on ancient samples as a complementary method to those traditionally used in paleoparasitology. It is important to highlight that MF was a simple and faster way. The incorporation of reliable quantitative techniques opens the door to a new way of analyzing archaeological remains, deepening the study of the parasite-host relationships and its evolution through time with an epidemiological approach. Although further studies are needed, our results suggest the complementarity of these techniques in future paleoparasitological studies.

Protozoan parasites of birds from the Tremembé formation (Oligocene of the Taubaté Basin), São Paulo, Brazil.

OBJECTIVE: To analyze the presence of protozoan parasites in bird coprolites from the Tremembé Formation (Oligocene of the Taubaté Basin). MATERIALS: Twenty avian coprolites embedded in pyrobituminous shale matrices. METHODS: Samples were rehydrated and subjected to spontaneous sedimentation. RESULTS: Paleoparasitological analyses revealed oocysts compatible with the Eimeriidae family (Apicomplexa) and one single Archamoebae (Amoebozoa) cyst. CONCLUSIONS: The present work increases the amount of information about the spread of infections throughout the Cenozoic Era and reveals that the Brazilian paleoavifauna played an important role in the Apicomplexa and Amoebozoa life cycles. SIGNIFICANCE: This is the first record of protozoans in avian coprolites from the Oligocene of Brazil. These findings can help in the interpretation of phylogenies of coccidian parasites of modern birds, as certain taxonomic characters observed in the Oligocene Protozoa characterize monophyletic groups in current molecular phylogenetic analyses. LIMITATIONS: None of the oocysts were sporulated; therefore, it is not possible to identify the morphotypes to genus or species. SUGGESTIONS FOR FURTHER RESEARCH: Our results create new perspectives related to biogeographic studies of the parasitic groups described and may improve the understanding of the temporal amplitude of parasitic evolutionary relationships between Protozoans and birds.

Retrieving ascarid and taeniid eggs from the biological remains of a Neolithic dog from the late 9th millennium BC in Western Iran

BACKGROUND Paleoparasitology reveals the status of parasitic infections in humans and animals in ancient times based on parasitic particles found in biological remains from archaeological excavations. This line of research emerged in Iran in 2013. OBJECTIVE The identification of parasites from Neolithic times is an attractive subject that shows the oldest origins of parasitic infections in a given geographical region. From an archaeological point of view, this archaeological site is well-known for animal domestication and agriculture in ancient Iran. METHODS In this study, soil deposited on the surface and in the pores of a dog pelvic bone was carefully collected and rehydrated using trisodium phosphate solution. FINDINGS The results showed ascarid and taeniid eggs retrieved from the biological remains of a dog excavated at the East Chia Sabz archaeological site, which dates back to the Neolithic period (8100 BC). MAIN CONCLUSION The current findings clearly illustrate the natural circulation of nematode and cestode parasites among dogs at that time. These ancient helminth eggs can also be used to track the oldest parasitic infections in the Iranian plateau and contribute to the paleoparasitological documentation of the Fertile Crescent.

Taenia sp. in human burial from Kan River, East Siberia

We present an arhaeoparasitological analysis of a unique burial from the Neftprovod II burial ground in East Siberia, which dated from the Bronze Age. Analysis of a sediment sample from the sacral region of the pelvis revealed the presence of Taenia sp. eggs. Because uncooked animal tissue is the primary source of Taenia, this indicated that the individual was likely consuming raw or undercooked meat of roe deer, red deer, or elk infected with Taenia. This finding represents the oldest case of a human infected with Taenia sp. from Eastern Siberia and Russia.

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.

PREHISTORICAL Pediculus humanus capitis INFESTATION: QUANTITATIVE DATA AND LOW VACUUM SCANNING MICROSCOPY / Infestação pré-histórica por Pediculus humanus capitis: análise quantitativa e por microscopia de varredura de baixo vácuo

A pre-Columbian Peruvian scalp was examined decades ago by a researcher from the Oswaldo Cruz Foundation. Professor Olympio da Fonseca Filho described nits and adult lice attached to hair shafts and commented about the origin of head lice infestations on mankind. This same scalp was sent to our laboratory and is the subject of the present paper. Analysis showed a massive infestation with nine eggs/cm2 and an impressive number of very well preserved adult lice. The infestation age was roughly estimated as nine months before death based on the distance of nits from the hair root and the medium rate of hair growth. A small traditional textile was associated with the scalp, possibly part of the funerary belongings. Other morphological aspects visualized by low-vacuum scanning electron microscopy are also presented here for adults and nits.

Há décadas um escalpo peruano, datado do período pré-colombiano, foi examinado por um pesquisador da Fundação Oswaldo Cruz. O Professor Olympio da Fonseca Filho descreveu lêndeas e adultos fixos a fios de cabelos e fez comentários sobre a origem da infecção por piolhos na espécie humana. Este mesmo escalpo foi enviado ao nosso laboratório e é objeto deste artigo. Sua análise mostrou maciça infestação, com nove lêndeas/cm2 em impressionante número de adultos muito bem preservados. O tempo de infestação foi estimado em cerca de nove meses antes da morte, baseado na maior distância entre lêndeas e o couro cabeludo, levando em consideração taxa média de crescimento capilar de 1 cm por mês. Um pequeno pedaço de tecido tradicional peruano foi encontrado associado ao escalpo, provavelmente pertencente ao conjunto de peças usado no ritual funerário. Aqui, apresentamos alguns aspectos morfológicos de adultos e lêndeas vizualizados por microscopia eletrônica de varredura de baixo vácuo.

Studies on protozoa in ancient remains - A Review

Paleoparasitological research has made important contributions to the understanding of parasite evolution and ecology. Although parasitic protozoa exhibit a worldwide distribution, recovering these organisms from an archaeological context is still exceptional and relies on the availability and distribution of evidence, the ecology of infectious diseases and adequate detection techniques. Here, we present a review of the findings related to protozoa in ancient remains, with an emphasis on their geographical distribution in the past and the methodologies used for their retrieval. The development of more sensitive detection methods has increased the number of identified parasitic species, promising interesting insights from research in the future.

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.

Paleoparasitology: the origin of human parasites / Paleoparasitologia: a origem dos parasitas humanos

Parasitism is composed by three subsystems: the parasite, the host, and the environment. There are no organisms that cannot be parasitized. The relationship between a parasite and its host species most of the time do not result in damage or disease to the host. However, in a parasitic disease the presence of a given parasite is always necessary, at least in a given moment of the infection. Some parasite species that infect humans were inherited from pre-hominids, and were shared with other phylogenetically close host species, but other parasite species were acquired from the environment as humans evolved. Human migration spread inherited parasites throughout the globe. To recover and trace the origin and evolution of infectious diseases, paleoparasitology was created. Paleoparasitology is the study of parasites in ancient material, which provided new information on the evolution, paleoepidemiology, ecology and phylogenetics of infectious diseases.

O parasitismo é composto por três subsistemas: o parasito, o hospedeiro e meio ambiente. Não existe organismo que não possa ser parasitado. A relação entre um parasito e suas espécies de hospedeiro não resulta, na maior parte das vezes, em dano ou doença ao hospedeiro. Entretanto, numa doença parasitária, a presença de um determinado parasito é sempre necessária, pelo menos em um determinado momento da infecção. Algumas espécies de parasitos que infectam humanos foram herdados dos pré-hominídeos, e foram compartilhados com outras espécies de hospedeiro filogeneticamente próximas, mas outras espécies foram adquiridas pelo meio ambiente à medida que os humanos se desenvolveram. As migrações humanas espalharam os parasitos adquiridos por todo o mundo. Para recuperar e traçar a origem e evolução das doenças infecciosas foi criada a paleoparasitologia. Paleoparasitologia é o estudo dos parasitos em materiais antigos, que trouxe novas informações na evolução, paleoepidemiologia, ecologia e filogenética das doenças infecciosas.

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.

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.

Chagas disease in prehistory

The classical hypothesis proposes that Chagas disease has been originated in the Andean region among prehistoric people when they started domesticating animals, changing to sedentary habits, and adopting agriculture. These changes in their way of life happened nearly 6,000 years ago. However, paleoparasitological data based on molecular tools showed that Trypanosoma cruzi infection and Chagas disease were commonly found both in South and North American prehistoric populations long before that time, suggesting that Chagas disease may be as old as the human presence in the American continent. The study of the origin and dispersion of Trypanosoma cruzi infection among prehistoric human populations may help in the comprehension of the clinical and epidemiological questions on Chagas disease that still remain unanswered.

A hipótese clássica sobre a origem da doença de Chagas propõe que tenha surgido entre as populações pré-históricas dos Andes quando começaram a domesticar animais, mudaram para hábitos sedentários e adotaram a agricultura. Estas mudanças em seus hábitos de vida aconteceram há aproximadamente 6.000 anos. Entretanto, os dados da paleoparasitologia, baseados na biologia molecular, mostraram que a infecção por Trypanosoma cruzi e a doença de Chagas eram comuns tanto em populações pré-históricas da América do Sul e América do Norte muito antes deste período. De acordo com os dados paleoparasitológicos, a doença de Chagas pode ser tão antiga quanto a presença humana no continente americano. O estudo sobre a origem e dispersão da infecção por Trypanosoma cruzi entre populações humanas pré-históricas pode auxiliar na compreensão de questões clínicas e epidemiológicas sobre a doença de Chagas que ainda permanecem sem resposta.

Paleoparasitological results for rodent coprolites from Santa Cruz Province, Argentina

The aim of this study was to examine the parasite remains present in rodent coprolites collected from the archaeological site Alero Destacamento Guardaparque (ADG) located in the Perito Moreno National Park (Santa Cruz Province, 47º57'S 72º05'W). Forty-eight coprolites were obtained from the layers 7, 6 and 5 of ADG, dated at 6,700 ± 70, 4,900 ± 70 and 3,440 ± 70 years BP, respectively. The faecal samples were processed and examined using paleoparasitological procedures. A total of 582 eggs of parasites were found in 47 coprolites. Samples were positive for eggs of Trichuris sp. (Nematoda: Trichuridae), Calodium sp., Eucoleus sp., Echinocoleus sp. and an unidentified capillariid (Nematoda: Capillariidae) and for eggs of Monoecocestus (Cestoda: Anoplocephalidae). Quantitative differences among layer for both coprolites and parasites were recorded. In this study, the specific filiations of parasites, their zoonotic importance, the rodent identity, on the basis of previous zooarchaeological knowledge, and the environmental conditions during the Holocene in the area are discussed.

Paleoparasitologic, paleogenetic and paleobotanic analysis of XVIII century coprolites from the church La Concepción in Santa Cruz de Tenerife, Canary Islands, Spain

We present the results of a paleoparasitologic, paleogenetic and paleobotanic analysis of coprolites recovered during the excavation of the church La Concepción in Santa Cruz de Tenerife. Coprolites (n = 4) were rehydrated and a multidisciplinary analysis was conducted. The paleobotanic analysis showed numerous silicates, seeds and fruits of the family Moraceae. In the paleoparasitologic study, Ascaris sp. eggs (n = 344) were identified. The paleogenetic results confirmed the Ascaris sp. infection as well as the European origin of human remains. These findings contribute to our knowledge of ancient helminthes infections and are the first paleoparasitological record of Ascaris sp. infection in Spain.

Parasites in rodent coprolites from the historical archaeological site Alero Mazquiarán, Chubut Province, Argentina

The aim of this study was to examine the parasitic remains that were found in rodent coprolites collected from the archaeological site Alero Mazquiarán (Chubut Province, 45º44'15"S, 70°25'9"W), which is assigned to the interface of the Araucanian and Tehuelche cultures, dated at 212 ± 35 years B.P. The faecal material from two unidentified rodent species (X-10 and X-11) was collected from one human pelvic cavity found in a multiple burial. The faecal samples were processed and examined using paleoparasitological procedures. The X-10 coprolites were positive for eggs of Monoecocestus sp. (Cestoda: Anoplocephalidae) and the X-11 faeces were positive for Pterygodermatites sp. (Nematoda: Rictulariidae), Trichosomoides sp. (Nematoda: Trichosomoididae) and Monoecocestus sp. In this study, we discuss parasitic life cycles, the zoonotic importance of parasites and the behaviour of the aboriginal people.

Animal helminths in human archaeological remains: a review of zoonoses in the past: [revision] / Helmintos animais em vestígios arqueológicos humanos: revisão de zoonoses no passado: [revisão]

The authors present a review of records of intestinal parasitic helminths from animals in human archaeological remains, reported since the emergence of paleopathological studies. The objective was to relate paleoparasitological findings to geographic, biotic, and abiotic factors from the environment in which the prehistoric populations lived, and understand some aspects related to the process of human dispersion and biological and cultural evolution. Modification of eating habits and the incorporation of new cultural practices are analyzed from the perspective of zoonoses from prehistory to the present day, especially in Brazilian indigenous populations. Three tables identifying the helminths, their natural hosts, dates, and sites of archaeological findings complete this review. In conclusion, various zoonoses known today have occurred since antiquity, and these data, combined with studies on the emergence and reemergence of diseases, could make possible to compose scenarios for the future.

São revistos os registros de ocorrência de helmintos intestinais parasitos de animais em vestígios arqueológicos humanos, relatados desde o surgimento dos estudos paleopatológicos. Busca-se relacionar os achados em paleoparasitologia com fatores geográficos, bióticos e abióticos do ambiente em que as populações pré-históricas viviam, e com aspectos do processo de dispersão e evolução biológica e cultural humana. A modificação de hábitos alimentares e a incorporação de novas práticas culturais são analisadas sob o ponto de vista das zoonoses desde a pré-história até a atualidade, em especial em populações indígenas brasileiras. Três tabelas identificando os helmintos, seus hospedeiros naturais, datações e local dos achados arqueológicos complementam esta revisão. Conclui-se que várias zoonoses conhecidas hoje ocorrem desde a antiguidade e que estes dados, combinados a estudos de emergência e reemergência de doenças, podem auxiliar a compor cenários para o futuro.

Zoonotic parasites associated with felines from the Patagonian Holocene

Feline coprolites were examined for parasites with the aim of studying ancient infections that occurred in the Patagonian region during the Holocene period. Eggs compatible to Trichuris sp., Calodium sp., Eucoleus sp., Nematodirus sp., Oesophagostomum sp. (Nematoda), Monoecocestus sp. (Cestoda) and Eimeria macusaniensis (Coccidia) were recovered from faecal samples. The results obtained from the analysis provide evidence of consumption by felids of the viscera of both rodents and camelids. This knowledge allows for improved explanations as to the distribution of parasitism and its significance to the health of humans and animals inhabiting the area under study during the Middle Holocene.

Paleoparasitology of Chagas disease: a review

One hundred years since the discovery of Chagas disease associated with Trypanosoma cruzi infection, growing attention has focused on understanding the evolution in parasite-human host interaction. This interest has featured studies and results from paleoparasitology, not only the description of lesions in mummified bodies, but also the recovery of genetic material from the parasite and the possibility of analyzing such material over time. The present study reviews the evidence of Chagas disease in organic remains excavated from archeological sites and discusses two findings in greater detail, both with lesions suggestive of chagasic megacolon and confirmed by molecular biology techniques. One of these sites is located in the United States, on the border between Texas and Mexico and the other in state of Minas Gerais, in the Brazilian cerrado (savannah). Dated prior to contact with Europeans, these results confirm that Chagas disease affected prehistoric human groups in other regions outside the Andean altiplanos and other transmission areas on the Pacific Coast, previously considered the origin of T. cruzi infection in the human host.

Paleoparasitology at "Place d'Armes", Namur, Belgium: a biostatistics analysis of trichurid eggs between the Old and New World / Paleoparasitologia na "Praça das Armas", Namur, Bélgica: uma análise bioestatística de ovos de tricurídeos entre o Velho e o Novo Mundo

Paleoparasitological findings about human occupation and their domestic animals, from Gallo-Roman period up to recent times, were described at the archaeological site of "Place d'Armes", Namur, Belgium, by preventive archaeological excavations. Organic sediment samples from cesspools, latrines and structures-like were analyzed and revealed intestinal parasite eggs (helminthes) in all of the different archaeological contexts. The parasitic association Ascaris sp. and Trichuris sp. was found although it is not easy to determinate the specific parasite species. Trichurids were described inboth New and Old Worlds since the prehistorical time. To elucidate the zoological origin of the organic remains, the parasites and the prior function of the sanitary structures Trichuris eggs were statistically analyzed for morphometric parameters, and suggest that it could be an important tool to help the whole paleoparasitological diagnosis.

Achados paleoparasitológicos sobre a ocupação humana e seus animais domésticos, desde o período Galo-Romano até períodos recentes, foram registrados no sitio arqueológico da "Praça das Armas", em Namur, Bélgica, através de excavações de salvamento arqueológico. Amostras de sedimentos orgânicos coletados de fossas, latrinas e estruturas semelhantes foram analisadas e revelaram ovos de helmintos intestinais, em todos os diferentes contextos arqueológicos. A associação parasitária Ascaris sp. e Trichuris sp. foi encontrada, sem identificação de espécie. Tricurídeos foram descritos em ambos, Novo e Velho Mundo, desde tempos pré-históricos. Para contribuir na elucidação da origem zoological dos vestígios orgânicos, dos parasitos e a função primárias das estruturas sanitárias, ovos de Trichuris foram analisados estatisticamente através dos seus parâmetros morfométricos, e sugerem que esta pode ser uma importante ferramenta para o diagnóstico paleoparasitológico.

New finding of Giardia intestinalis (Eukaryote, Metamonad) in Old World archaeological site using immunofluorescence and enzyme-linked immunosorbent assays

In this study, nine organic sediment samples from a medieval archaeological site at Pineuilh, France, were examined for Giardia intestinalis using two commercially available immunological kits [enzyme-linked immuno sorbent and immunofluorescence (IFA) assays]. Both techniques detected G. intestinalis in one sample, dated to 1,000 Anno Domini. This is the first time IFA was successfully used to detect protozoa in Old World archaeological samples. Such immunological techniques offer important perspectives concerning ancient protozoa detection and identification.