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.     

Molecular paleoparasitological diagnosis of Ascaris sp. from coprolites: new scenery of ascariasis in pre-Colombian South America times

Paleoparasitological studies using microscopy showed that Ascarisand Trichuris trichiura are the human intestinal parasites most found in archaeological sites. However, in pre-Columbian South American archaeological sites, Ascaris is rare. In this work we standardized a molecular methodology for Ascaris diagnosis directly from ancient DNA retrieved from coprolites. Using cythochrome b gene (142 bp) target, ancient DNA sequences were retrieved from South American samples, negative by microscopy. Moreover, the methodology applied was sensitive enough to detect ancient DNA extracted from 30 Ascaris eggs from an European coprolite. These results revealed a new scenery for the paleodistribution of Ascaris in South America.     

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.     

Paleomicrobiology: Revealing Fecal Microbiomes of Ancient Indigenous Cultures

Coprolites are fossilized feces that can be used to provide information on the composition of the intestinal microbiota and, as we show, possibly on diet. We analyzed human coprolites from the Huecoid and Saladoid cultures from a settlement on Vieques Island, Puerto Rico. While more is known about the Saladoid culture, it is believed that both societies co-existed on this island approximately from 5 to 1170 AD. By extracting DNA from the coprolites, followed by metagenomic characterization, we show that both cultures can be distinguished from each other on the basis of their bacterial and fungal gut microbiomes. In addition, we show that parasite loads were heavy and also culturally distinct. Huecoid coprolites were characterized by maize and Basidiomycetes sequences, suggesting that these were important components of their diet. Saladoid coprolite samples harbored sequences associated with fish parasites, suggesting that raw fish was a substantial component of their diet. The present study shows that ancient DNA is not entirely degraded in humid, tropical environments, and that dietary and/or host genetic differences in ancient populations may be reflected in the composition of their gut microbiome. This further supports the hypothesis that the two ancient cultures studied were distinct, and that they retained distinct technological/cultural differences during an extended period of close proximity and peaceful co-existence. The two populations seemed to form the later-day Taínos, the Amerindians present at the point of Columbian contact. Importantly, our data suggest that paleomicrobiomics can be a powerful tool to assess cultural differences between ancient populations.     

Paleoparasitological finding of eggs of nematodes in rodent coprolites dated at the early Holocene from the archaeological site Cerro Casa de Piedra 7, Santa Cruz, Argentina

The aim of the present study was to examine the parasite remains present in rodent coprolites collected from the archaeological site Cerro Casa de Piedra 7 (CCP7), located in the Perito Moreno National Park (47°57'S, 72°05'W), Santa Cruz Province, Argentina. Eight coprolites obtained from the layer 17, dated at 10,620 ± 40 to 9,390 ± 40 yr B.P., were examined for parasites. Feces were processed whole, rehydrated, homogenized, subjected to spontaneous sedimentation, and examined via light microscopy. Eggs of parasites were measured and photographed. Seven of 8 coprolites possessed 199 eggs of 2, probably new, species of nematodes, including 43 eggs of Heteroxynema sp. Hall, 1916 (Cavioxyura sp. Quentin, 1975) (Oxyurida, Heteroxynematidae), and 156 eggs of Trichuris sp. Roederer, 1761 (Trichinellida, Trichuridae). Heteroxynema sp. is cited for the first time from ancient material worldwide. The finding of Trichuris spp. in both rodents and other host samples from the area under study is indicative of the stability of the biological and environmental conditions for this nematode genus to establish in the Patagonian Early Holocene. The rodent host was assigned to an unknown species of Caviomorpha (Hystricognathi) that lived during the Pleistocenic transition in Patagonia.     

Paleoparasitological analysis of rodent coprolites in holocenic samples from Patagonia, Argentina

The aim of the present study was to examine the parasite fauna present in rodent coprolites collected from Cerro Casa de Piedra (CCP7), located in Perito Moreno National Park (P.N.PM., 47 degrees 57'S and 72 degrees 05'W), Santa Cruz Province, Argentina. Eight coprolites obtained from the layer XIII of CCP7, with an antiquity considered as 7,920 +/- 130 yr B.P, were examined for parasites. Each coprolite was whole processed, rehydrated, homogenized, spontaneously sedimented, and examined using light microscopy. Eggs of parasites were measured and photographed. All the samples were parasitized by nematodes, with 267 eggs of Trichuris sp., 24 eggs of an aspidoderid, and 3 capillariid eggs. The rodent host was tentatively identified as a species of Ctenomys, the hypogeic rodents endemic to South America. The finding of Paraspidodera in Patagonian samples represents new evidence that strengthens the co-phylogenies between nematodes of this genus and Ctenomys and reinforces the value of parasites as tags in paleoparasitology.     

Acanthocefalan eggs in animal coprolites from archaeological sites from Brazil

An important point in paleoparasitology is the correct diagnosis of the origin of coprolites found in archaeological sites. The identification of human and animal coprolites, through the study of the shape, size, characteristics after rehydration, alimentary contents, and the presence of parasites, has proved to be accurate for human coprolites. For non-human ones we compared coprolites with recent faeces of animals collected near the archaeological sites, following the methodology above mentioned. In this paper anteaters coprolites (Tamandua tetradactyla; Myrmecophaga tridactyla) with eggs of Gigantorhynchus echinodiscus (Archiancanthocephala; Gigantorynchidae) were identified.     

粪化石在古生态研究中的应用综述

粪化石(coprolite)是石化的动物粪便,而广义上粪化石(bromalite)是石化的动物消化物、排泄物和排出物的总称。作为一类重要遗迹化石,粪化石蕴含了大量实体化石难以提供的生物学信息,为古生态学研究提供以下依据:1)粪化石作为媒介帮助理解远古动物的行为习性、消化系统的结构和功能、食谱特征以及古生态系统食物链/食物网;2)粪化石中保存的古寄生虫线索能有效地解读古生物寄生关系和某些常见肠道寄生虫的起源问题;3)新生代以来古人类粪化石证据可以直接解开早期古人类的食谱、疾病特征及其迁移路径;4)中、新生代以来的粪化石中保存的植物残留信息(孢粉化石和植硅体)是重建古植被面貌、恢复古气候和探索早期动植物关系的重要依据。本文回顾了粪化石研究历史,并针对粪化石在上述古生态各研究方向的最新进展进行了系统总结和综述,认为粪化石可有效解读古生态。文章最后对粪化石的最新研究技术方法(如CT扫描技术)及未来发展方向进行了展望。     

The finding of Echinostoma (Trematoda: Digenea) and hookworm eggs in coprolites collected from a Brazilian mummified body dated 600-1,200 years before present

The identification of parasites from ancient cultures expands our list of parasites infective to extant humans. A partially mummified human body from the archeological site of Lapa do Boquete, Minas Gerais State, Brazil, was recently discovered. It was interred between 600 and 1,200 yr ago. Dietary analysis showed that the mummified body was from a society that had a mixed subsistence of agriculture and gathering of wild foods. Coprolites from the body contained numerous helminth eggs. The eggs were identified as those of Echinostoma sp. and hookworm. Hookworm infection in pre-Columbian populations is already established, but this is the first evidence of Echinostoma sp. eggs found in human coprolites. The diagnosis of a true infection, as opposed to false parasitism, is discussed. The possibility of Echinostoma ilocanum infection is discussed, as this is a common species found in humans in the Asiatic region, which could have been introduced in South America in the pre-Columbian period. Alternative possibilities are also considered, including indigenous Brazilian Echinostoma species.     

河南灵井许昌人遗址鬣狗粪化石的初步研究

河南灵井许昌人遗址是我国近年来发掘的最为重要的古人类遗址之一。该遗址中出土了大量的石器、骨器、动物化石、粪便化石等遗存以及人类头盖骨化石等。我们对该遗址出土的鬣狗粪化石进行了类型学及其包含的微体化石等方面研究,从分析统计结果来看,鬣狗粪化石中包含了丰富的古信息,如古寄生虫卵、动物毛发、孢粉、植硅体、真菌等。本文主要针对鬣狗粪化石中古寄生虫卵及动物毛发进行分析,探讨了作为寄主鬣狗罹患的寄生虫病,以及鬣狗的食物来源等情况,为深入理解更新世晚期人类适应环境与气候提供新的证据。     

Trichuris eggs in animal coprolites dated from 30,000 years ago

Trichuris eggs were found in Kerodon rupestris (Rodentia, Caviidae) coprolites collected in archaeological layers dated from 30,000 yr BP (before present) to 8,450 yr BP. Adult worms and eggs of this genus were not found in a search of living mammals of the region. Results indicate that K. rupestris was a host for an unknown Trichuris species not found in this rodent presently. Climate changes that occurred by 10,000 yr ago in the region could be the cause of its disappearance. The finding of parasites in archaeological material can show the antiquity of host-parasite relationships and parasite losses through time.     

A Megafauna’s Microfauna: Gastrointestinal Parasites of New Zealand’s Extinct Moa (Aves: Dinornithiformes)

We perform the first multidisciplinary study of parasites from an extinct megafaunal clade using coprolites from the New Zealand moa (Aves: Dinornithiformes). Ancient DNA and microscopic analyses of 84 coprolites deposited by four moa species (South Island giant moa, Dinornis robustus; little bush moa, Anomalopteryx didiformis; heavy-footed moa, Pachyornis elephantopus; and upland moa, Megalapteryx didinus) reveal an array of gastrointestinal parasites including coccidians (Cryptosporidium and members of the suborder Eimeriorina), nematodes (Heterakoidea, Trichostrongylidae, Trichinellidae) and a trematode (Echinostomida). Parasite eggs were most prevalent and diverse in coprolites from lowland sites, where multiple sympatric moa species occurred and host density was therefore probably higher. Morphological and phylogenetic evidence supports a possible vicariant Gondwanan origin for some of the moa parasites. The discovery of apparently host-specific parasite taxa suggests paleoparasitological studies of megafauna coprolites may provide useful case-studies of coextinction.     

Parasitism of prehistoric humans and companion animals from Antelope Cave, Mojave County, northwest Arizona

Previously, we reported a tick recovered from Antelope Cave in extreme northwest Arizona. Further analyses of coprolites from Antelope Cave revealed additional parasitological data from coprolites of both human and canid origin. A second tick was found. This site is the only archaeological locality where ticks have been recovered. We also discovered an acanthocephalan in association with Enterobius vermicularis eggs in the same coprolite. This association shows that the coprolite was deposited by a human. This discovery expands our knowledge of the range of prehistoric acanthocephalan infection. In addition, findings from canid coprolites of Trichuris vulpis are reported. This is the first published discovery of T. vulpis from a North American archaeological context. The close association of dogs with humans at Ancestral Puebloan (Anasazi) sites raises the potential that zoonotic parasites were transferred to the human population. The archaeological occupation is associated with the Ancestral Pueblo culture 1,100 yr ago.     

Zoonotic and human parasites of inhabitants of Cueva de los Muertos Chiquitos, Rio Zape Valley, Durango, Mexico

We present the first reconstruction of the parasitoses among the people of the Loma San Gabriel culture, as represented by 36 coprolites excavated from the Cueva de los Muertos Chiquitos in Durango, Mexico. The coprolites date to approximately 1,400-yr-ago. Species identified based on eggs recovered include the trematode Echinostoma sp., the tapeworms Hymenolepis sp. and Dipylidium caninum , and the nematodes Ancylostoma duodenale, Enterobius vermicularis, and Trichuris trichiura. After rehydration and screening, 2 methods were used to recover eggs from these samples including spontaneous sedimentation and flotation. Samples were analyzed by 3 different laboratories for independent verification and comparison of methods. Spontaneous sedimentation resulted in the discovery of hymenolepidid eggs that were not found with flotation. Sedimentation was a more-sensitive indicator of prevalence as well. The modified method of flotation permitted estimation of egg concentration and resulted in the detection of a few specimens not found by sedimentation. The results of both methods showed that 19 (of 36) coprolites contained helminth eggs. Our results detected the presence of pathogenic helminths including hookworms and whipworms. The cestodes found do not cause severe pathology in humans. The early dates of hookworm and whipworm, relative to other findings in the southwest United States, indicate that these parasites arrived relatively late in prehistory in Arizona and New Mexico, probably moving into the area with travelers from Mesoamerica.     

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.     

Scanning electron microscope study of ancient parasite eggs recovered from Korean mummies of the Joseon Dynasty

We have previously shown that parasite eggs have been identified in the coprolites of Korean mummies. These eggs have shed light on parasitic infection patterns in Korean populations living several hundred years ago. We conducted a scanning electron microscopy (SEM) study on ancient Trichuris trichiura, Ascaris lumbricoides, Metagonimus yokogawai, Paragonimus westermani, and Gymnophalloides seoi eggs recovered from Korean mummies of the Joseon Dynasty. We anticipated that the taphonomic conditions of mummification would alter the eggs of certain species but not of others. Our SEM data show that each species of ancient egg exhibited different degrees of preservation. Thus, some of them, for example, M. yokogawai, exhibited a better preservation status than others, suggesting that they should be the first candidates considered when choosing subjects for future paleoparasitological studies.     

Nuclear gene sequences from a late pleistocene sloth coprolite

The determination of nuclear DNA sequences from ancient remains would open many novel opportunities such as the resolution of phylogenies, the sexing of hominid and animal remains, and the characterization of genes involved in phenotypic traits. However, to date, single-copy nuclear DNA sequences from fossils have been determined only from bones and teeth of woolly mammoths preserved in the permafrost. Since the best preserved ancient nucleic acids tend to stem from cold environments, this has led to the assumption that nuclear DNA would be retrievable only from frozen remains. We have previously shown that Pleistocene coprolites stemming from the extinct Shasta sloth (Nothrotheriops shastensis, Megatheriidae) contain mitochondrial (mt) DNA from the animal that produced them as well as chloroplast (cp) DNA from the ingested plants. Recent attempts to resolve the phylogeny of two families of extinct sloths by using strictly mitochondrial DNA has been inconclusive. We have prepared DNA extracts from a ground sloth coprolite from Gypsum Cave, Nevada, and quantitated the number of mtDNA copies for three different fragment lengths by using real-time PCR. We amplified one multicopy and three single-copy nuclear gene fragments and used the concatenated sequence to resolve the phylogeny. These results show that ancient single-copy nuclear DNA can be recovered from warm, arid climates. Thus, nuclear DNA preservation is not restricted to cold climates.     

Critical analysis of coprolite evidence of medicinal plant use, Piauí, Brazil

Human coprolites, dating from 8500 to 7000 years BP, were collected from the rock-shelter of Boqueirão da Pedra Furada, in Piauí, Brazil. These dates fall within the “Serra Talhada” cultural traditions I and II of the Tradição Nordeste, a paleohuman culture of northeastern Brazil. Archaeoparasitological analysis of the coprolites revealed whipworm eggs and hookworm eggs. Analysis of Tradição Nordeste skeletons and hair shows a variety of diseases including dental problems, osteoarthritis, and head louse infestation. Palynological analysis of the coprolites revealed 12 genera that were potentially medicinal. The pollen data were critically analyzed to assess the potentiality that the pollen represented medicinal use. Strong cases could be made that three of the genera, Anacardium, Borreria, and Terminalia, were actually used for medicinal purposes, probably to treat symptoms caused by the intestinal parasites.     

The state of the art of paleoparasitological research in the Old World

Paleoparasitology in the Old World has mainly concerned the study of latrine sediments and coprolites collected from mummified bodies or archaeological strata, mostly preserved by natural conditions. Human parasites recovered include cestodes, trematodes, and nematodes. The well preserved conditions of helminth eggs allowed paleoepidemiological approaches taking into account the number of eggs found by archaeological stratum. Tentatively, sanitation conditions were assessed for each archaeological period.     

Analysis of ancient DNA from coprolites: a perspective with random amplified polymorphic DNA-polymerase chain reaction approach

The aim of this work was to determine approaches that would improve the quality of ancient DNA (aDNA) present in coprolites to enhance the possibility of success in retrieving specific sequence targets. We worked with coprolites from South American archaeological sites in Brazil and Chile dating up to 7,000 years ago. Using established protocols for aDNA extraction we obtained samples showing high degradation as usually happens with this kind of material. The reconstructive polymerization pretreatment was essential to overcome the DNA degradation and the serial dilutions helped with to prevent polymerase chain reaction (PCR) inhibitors. Moreover, the random amplified polymorphic DNA-PCR has been shown to be a reliable technique for further experiments to recover specific aDNA sequences.