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.     

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.     

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.     

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.     

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.     

Paleoparasitological analysis of coprolites from K2, an Iron Age archaeological site in South Africa: the first finding of Dicrocoelium sp. eggs

Until now, Dicrocoelium sp. eggs have only been recorded from European and 1 North American archaeological sites. We present evidence for the first record of Dicrocoelium sp. from an African archaeological site. A paleoparasitological study was conducted on 7 coprolite samples from K2, a Late Iron Age site on the farm Greefswald, in the Northern Province of South Africa. Standard parasitological analysis revealed the presence of Dicrocoelium sp. and Trichuris sp. eggs. Today, the parasite does not occur in this region. Trichurid eggs are a relatively common find in paleoparasitological analysis. The presence of Dicrocoelium sp. provides new clues about the antiquity of this parasite, as well as aspects of ancient environment, climate, and interactions among humans, animals, and parasites.     

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.     

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.     

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.     

New record of anoplocephalid eggs (Cestoda: Anoplocephalidae) collected from rodent coprolites from archaeological and paleontological sites of Patagonia,Argentina

Results of paleoparasitological examination of rodent coprolites collected from archaeological and paleontological sites from Patagonia, Argentina, are present. Each coprolite was processed, rehydrated, homogenized, spontaneously sedimented and examined using light microscope. Coprolites and eggs were described, measured and photographed, and were compared with current faeces of Lagidium viscacia. Eggs with morphological features, attributed to an anoplocephalid cestode were found in samples collected from Cueva Huenul 1 (36°56′45″S, 69°47′32″W, Neuquén Province, Holocene) and Los Altares Profile (43º53′35″S, 68º23′21″W, Chubut Province, Late Holocene). These are the first findings of this anoplocephalid from faecal material from patagonic rodents.     

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.     

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.     

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.     

First paleoparasitological study of micromammal coprolites from the holocene of the Somuncurá Plateau Protected Natural Area (Patagonia Argentina)

The Somuncurá Plateau is a Protected Natural Area located in the middle of the northern extra-Andean arid Patagonia. Inhabited by at least 20 small mammal species, is the place with the uppermost species richness in Patagonia. The aim of this study was to examine the parasite remains from micromammal coprolites collected in association with a bone sequence recovered at the east of the Somuncurá Plateau (site “Alero Las Lechuzas”). Coprolites came from the four temporal units previously defined: unit I (4790?±?100?yrs. ¹⁴C B.P.), unit II, unit III (7840?±?120?yrs. ¹⁴C B.P.) and unit IV. Each coprolite was processed, rehydrated, homogenized, processed by spontaneous sedimentation and examined using a light microscope. Coprolites and eggs were described, measured and photographed. Samples were positive for two nematode species: Helminthoxys caudatus Freitas, Lent & Almeida, 1937 (Oxyurida, Oxyuridae) and Trichuris spp. (Trichinellida: Trichuridae). This is the first paleoparasitological study developed for the Somuncurá Plateau Protected Area. Moreover, this is the first time that the genus Helminthoxys is reported from ancient times worldwide. Coprolites were attributed to the mountain cavy Microcavia australis (Rodentia, Caviidae).The presence of H. caudatus for the Middle Holocene of northern Patagonia contributes to the study of the history of the histricomorphs and pinworms relationships.     

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.     

Reappraisal of the Genus Tapirus (Perissodactyla, Tapiridae): Systematics and Phylogenetic Affinities of the South American Tapirs

The record of the genus Tapirus in South America is associated with the faunistic events of the Great American Biotic Interchange (GABI). The taxon is considered an immigrant of Holarctic origin. Although remains are scarce and incomplete during the Pleistocene, an analysis of these materials allowed us to consider valid seven fossil species : Tapirus tarijensis, T. cristatellus, T. greslebini, T. rioplatensis, T. oliverasi, T. mesopotamicus, and T. rondoniensis. A phylogenetic analysis was carried out in order to elucidate the relationships of the American fossil and extant species. Our result is consistent with a paraphyletic hypothesis for South American tapirs and suggests that a second dispersal event would have occurred from South America to North America, of a form closely related to T. cristatellus, resulting in the derived forms of North America.     

A South American snake lineage from the Eocene Greenhouse of North America and a reappraisal of the fossil record of “anilioid” snakes

“Anilioidea” is a likely paraphyletic assemblage of pipe snakes that includes extant Aniliidae from equatorial South America, Uropeltoidea from South and Southeast Asia, and a fossil record that consists primarily of isolated precloacal vertebrae ranging from the earliest Late Cretaceous and includes geographic distributions in North America, South America, Europe, and Africa. Articulated precloacal vertebrae from the middle Eocene Bridger Formation of Wyoming, attributed to Borealilysia nov. gen., represent an unambiguous North American aniliid record and prompts a reconsideration of described pipe snakes and their resultant biogeographic histories. On the basis of vertebral apomorphies, the vast majority of reported fossils cannot be assigned to “Anilioidea”. Instead, most records represent stem taxa and macrostomatans erroneously assigned to anilioids on the basis of generalized features associated with fossoriality. A revised fossil record demonstrates that the only extralimital distributions of fossil “anilioids” consist of the North American aniliid record, and there is no unambiguous fossil record of Old World taxa. The occurrence of aniliids in the mid-high latitudes of the late early Eocene of North America is consistent with histories of northward shifts in equatorial ecosystems during the early Paleogene Greenhouse.     

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.     

Paleo‐metagenomics of North American fossil packrat middens: Past biodiversity revealed by ancient DNA

Fossil rodent middens are powerful tools in paleoecology. In arid parts of western North America, packrat (Neotoma spp.) middens preserve plant and animal remains for tens of thousands of years. Midden contents are so well preserved that fragments of endogenous ancient DNA (aDNA) can be extracted and analyzed across millennia. Here, we explore the use of shotgun metagenomics to study the aDNA obtained from packrat middens up to 32,000 C¹⁴ years old. Eleven Illumina HiSeq 2500 libraries were successfully sequenced, and between 0.11% and 6.7% of reads were classified using Centrifuge against the NCBI “nt” database. Eukaryotic taxa identified belonged primarily to vascular plants with smaller proportions mapping to ascomycete fungi, arthropods, chordates, and nematodes. Plant taxonomic diversity in the middens is shown to change through time and tracks changes in assemblages determined by morphological examination of the plant remains. Amplicon sequencing of ITS2 and rbcL provided minimal data for some middens, but failed at amplifying the highly fragmented DNA present in others. With repeated sampling and deep sequencing, analysis of packrat midden aDNA from well‐preserved midden material can provide highly detailed characterizations of past communities of plants, animals, bacteria, and fungi present as trace DNA fossils. The prospects for gaining more paleoecological insights from aDNA for rodent middens will continue to improve with optimization of laboratory methods, decreasing sequencing costs, and increasing computational power.