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Sambaquis or shellmounds are archeological sites constructed by hunter-fisher-gatherers that inhabited the Brazilian coast about 10,000–2000 yrs BP. Jabuticabeira II (JABII: 2890 ± 55 to 1805 ± 65 BP) is one of dozens of contemporaneous sambaquis of the Santa Catarina state, South Brazil, and contains hundreds of neatly organized burials, indicating great population density. In order to gather information about the health, diet and way of life of people in JABII, a paleoparasitological, paleogenetic, and micro-human remains investigation was carried out. Pelvic region and environmental control samples from six individuals exhumed from JABII were submitted to microscopic and ancient DNA (aDNA) investigation. Paleoparasitological analyses based on light microscopy were negative. However, a variety of informative microremains were found. Diatoms, fish scales, and algae characterize the marine and estuarine environment. Ipomoea batatas and Zea mays starch grains suggested cultivated items as part of their diet in agreement with the literature. The finding of Podocarpus sp. pollen grain, characteristic of highlander vegetation, suggests human mobility of JABII individuals which were settlement in the coast. Paleogenetic analyses showed Ascaris sp. helminth infection based on nad1 gene fragment detected from an individual excavated at L3 FS7 burial (1826 ± 40 BP). This aDNA result places the antiquity of Ascaris sp. infection, and haplotypes that are circulating in humans and other animals nowadays, in Pre-Columbian South American times.

Experimental paleoparasitological approaches have been used in order to optimize the methodology previously to the application in archeological samples. In this study we evaluated the action of dehydration and local soil (Central Argentina) on the loss of parasite eggs in experimental coprolites, using two parasitological techniques: spontaneous sedimentation and sucrose-flotation. Experimental coprolites comprised fresh human feces, positive for Hymenolepis nana, Ascaris sp., and Enterobius vermicularis, submitted to controlled artificial dehydration. Experimental coprolites with soil addition were prepared by mixing archeological sediment with equal mass of fresh feces. Helminth eggs were counted and eggs per gram were estimated in each subsample. Statistical analyses were applied to compare subsamples before and after desiccation and with and without addition of soil sediment. The performance of parasitological methods statistically differed, the sucrose flotation technique being the less effective when fresh feces and experimental coprolites were analyzed. Partial deformation of eggs was observed via both techniques only in subsamples containing H. nana eggs. However, this was not seen in Ascaris sp. subsamples, possibly due to eggshell composition. We found that sample desiccation significantly decreased the number of eggs in the experimental coprolites. Mixing archeological sediment with the fecal material also resulted in significantly fewer eggs surviving, independent of desiccation. This shows that climate and soil in which archeological fecal samples are found can strongly influence the survival of parasite eggs from past populations. The small amount of parasite evidence often found in paleoparasitological analyses, including Central Argentina, could be attributed to the action of taphonomic processes rather than to the real absence of infection in these ancient populations. Importantly, the study highlights the role of local soil, confirmed for the first time by empirical data. The research provides valuable insights into the understanding of the paleoparasitological results of the region and of general paleoparasitology.

Capillariidae is a group of nematode parasites of vertebrates with a complex taxonomy. The structure of the eggshell, which was indicated as the most important characteristic for identification of genus or species through eggs, is very diverse among genera. The visualization and characterization of eggshell by light microscopy (LM) are a challenging task since different planes of the egg surface are needed. Nevertheless, categories of eggshell ornamentation were proposed by LM: smooth, punctuated, reticulated type I, and reticulated type II. The present study aimed to characterize the eggshell structure of Capillariidae species, parasites of mammals and avians, deposited in a helminthological collection using scanning electron microscopy (SEM). Institutional Biological Collections are taxonomic repositories of specimens described and strictly identified at the species level by systematics specialists. SEM eggshell images were obtained from 12 species belonging to 5 genera (Aonchotheca, Baruscapillaria, Capillaria, Echinocoleus, Eucoleus) and compared to their respective LM images. Eggshell patterns observed using SEM were associated categories of eggshell ornamentation previously proposed by LM images. The SEM data indicate that eggshell categories are not in agreement with capillariid genera or sites of infection. However, the study provides previously unknown SEM eggshell information from curated species, which contributes with a specific and supplementary taxonomic feature at the species level of Capillariidae.

The present study aimed to identify endoparasites from faecal samples of Kerodon rupestris, an endemic rodent of the Brazilian semiarid region. Samples collected from 10 locations at the Parque Nacional Serra da Capivara, Piaui State, Brazil were analyzed. Each faecal samples was processed, rehydrated, homogenized, and processed by spontaneous sedimentation. Morphological and morphometric characteristics of gastrointestinal parasites were observed using light microscopy. A total of three taxa from helminth and protozoan parasites were found. The results demonstrated parasite remains in 6/10 localities, and a high occurrence of Trichuris cf. gracilis, in 5/10 localities. Egg morphometric data was compared with Thichuris spp. egg Brazilian rodents. Two new parasite records were identified in K. rupestris , a nematode, Helminthoxys sp. (Oxyuridae) and a possible coccidian. The findings are discussed with taxonomic and ecological literature of intestinal parasites from rodents. The role of K. rupestris as source of ecological information in the region is suggested.

The identification of Blood Meal Source (BMS) in hematophagous vectors contributes to a better understanding of the ecology of hemoparasite transmission. Those insects can endure long periods without feeding, waiting for a favorable setting. Although this represents an important behavior observed in those groups, such as triatomines, little is known about how time can affect BMS detection, especially considering extended periods. To comprehend to which extent this behavioral phenomenon can impact molecular detection, we submitted two groups of Rhodnius robustus to increasing periods of starvation under experimental conditions. It was possible to recover the BMS until the 12th week of the starvation process. Nymphs were more resistant to prolonged periods of starvation (up to more than 189 days) than adults (maximum of 137 days), with no significant difference between their weights after being fed. The study brought new insights to the understanding of Trypanosoma cruzi transmission by R. robustus in the nature, with a temporal perspective.

La Paleoparasitología en la Argentina ha producido un gran número de trabajos, especialmente en Patagonia. Sin embargo, hay ausencia de estos estudios en las poblaciones de la región central del país. En este sentido, realizamos la primera investigación sobre parasitosis en restos humanos de sitios arqueológicos de la provincia de Córdoba. Se estudiaron 13 individuos de 4 sitios arqueológicos, datados entre 2562 ± 47 y 533 ± 57 14C años AP. Los análisis paleoparasitológicos se aplicaron sobre 20 muestras de sedimentos provenientes de la cavidad pélvica, empleando las técnicas de sedimentación espontánea, disgregación en ácido clorhídrico y flotación en sacarosa. Se hallaron estructuras sugestivas de huevos de helmintos de las superfamilias Ascaridoidea y Heterakoidea, y otros de nemátodos no identificados. Debido a que los animales son los huéspedes definitivos de estos parásitos, los resultados podrían sugerir la presencia de falso parasitismo en los individuos de la región. La relación cercana de los humanos con los animales, utilizados como parte de la dieta, se sustenta en estudios isotópicos y zooarqueológicos previos. La escasez de hallazgos parasitarios en este estudio posiblemente se debe a factores tafonómicos y a las características ambientales de los sitios., Asociación de Antropología Biológica de la República Argentina

Background There are more than 300 species of capillariids that parasitize various vertebrate groups worldwide. Species identification is hindered because of the few taxonomically informative structures available, making the task laborious and genus definition controversial. Thus, its taxonomy is one of the most complex among Nematoda. Eggs are the parasitic structures most viewed in coprological analysis in both modern and ancient samples; consequently, their presence is indicative of positive diagnosis for infection. The structure of the egg could play a role in genera or species discrimination. Institutional biological collections are taxonomic repositories of specimens described and strictly identified by systematics specialists. Methods The present work aims to characterize eggs of capillariid species deposited in institutional helminth collections and to process the morphological, morphometric and ecological data using machine learning (ML) as a new approach for taxonomic identification. Specimens of 28 species and 8 genera deposited at Coleção Helmintológica do Instituto Oswaldo Cruz (CHIOC, IOC/FIOCRUZ/Brazil) and Collection de Nématodes Zooparasites du Muséum National d’Histoire Naturelle de Paris (MNHN/France) were examined under light microscopy. In the morphological and morphometric analyses (MM), the total length and width of eggs as well as plugs and shell thickness were considered. In addition, eggshell ornamentations and ecological parameters of the geographical location (GL) and host (H) were included. Results The performance of the logistic model tree (LMT) algorithm showed the highest values in all metrics compared with the other algorithms. Algorithm J48 produced the most reliable decision tree for species identification alongside REPTree. The Majority Voting algorithm showed high metric values, but the combined classifiers did not attenuate the errors revealed in each algorithm alone. The statistical evaluation of the dataset indicated a significant difference between trees, with GL + H + MM and MM only with the best scores. Conclusions The present research proposed a novel procedure for taxonomic species identification, integrating data from centenary biological collections and the logic of artificial intelligence techniques. This study will support future research on taxonomic identification and diagnosis of both modern and archaeological capillariids. Graphical abstract Supplementary Information The online version contains supplementary material available at 10.1186/s13071-021-04721-6.

Capillariidae is a group of nematode parasites of vertebrates with a complex taxonomy. The structure of the eggshell, which was indicated as the most important characteristic for identification of genus or species through eggs, is very diverse among genera. The visualization and characterization of eggshell by light microscopy (LM) are a challenging task since different planes of the egg surface are needed. Nevertheless, categories of eggshell ornamentation were proposed by LM: smooth, punctuated, reticulated type I, and reticulated type II. The present study aimed to characterize the eggshell structure of Capillariidae species, parasites of mammals and avians, deposited in a helminthological collection using scanning electron microscopy (SEM). Institutional Biological Collections are taxonomic repositories of specimens described and strictly identified at the species level by systematics specialists. SEM eggshell images were obtained from 12 species belonging to 5 genera (Aonchotheca, Baruscapillaria, Capillaria, Echinocoleus, Eucoleus) and compared to their respective LM images. Eggshell patterns observed using SEM were associated categories of eggshell ornamentation previously proposed by LM images. The SEM data indicate that eggshell categories are not in agreement with capillariid genera or sites of infection. However, the study provides previously unknown SEM eggshell information from curated species, which contributes with a specific and supplementary taxonomic feature at the species level of Capillariidae.

Los nematodos del género Helminthoxys Freitas, Lent & Almeida, 1937 son parásitos intestinales de roedores caviomorfos con una amplia distribución neotropical. Este estudio detalló la morfología de Helminthoxys freitasi Quentin, 1969 por microscopía óptica y microscopía electrónica de barrido (MEB), e infirió una filogenia para la tribu Protozoophagini con las secuencias del gene parcial Citocromo c Oxidasa subunidade 1 (MT-CO1). Los roedores Mesomys hispidus (Desmarest, 1817) fueron colectados en tres áreas distintas en el estado de Acre, Brasil. Los helmintos encuentrados observaron alas laterales que alcanzan el nivel del ano, la región posterior del cuerpo en el hombre con tres pares de papilas sésiles y un par de papilas pedunculadas. La secuencia COI de H. freitasi revelan una estrecha relación con el género Wellcomia Sambon, 1907, corroborando con la filogenia morfológica anterior. Además, reportamos una nueva especie huésped, M. hispidus, con una nueva localidad en la selva amazónica.

For archaeological studies it is always necessary to consider taphonomic factors that could have influenced in ancient material preservation. Parasite eggs are usually highly degraded in ancient sites dated from all periods of time and taphonomic factors are mentioned to explain absence and low quantity of eggs found. In this study, we compare parasite egg recovery of three Great House latrines: two from Aztec Ruins (Rooms 219 and 225) and one from Salmon Ruins. We compared through statistical regression the recovery of eggs with the abundance of two classes of decomposers: mites and nematodes. These microorganisms have relation with nematode larvae and parasites remains degradation, respectively, mostly in moist environments. Pinworm was the only parasite found in the sites studied. Prevalences were 32.8% at Salmon Ruins, 72.7% at Aztec Ruins Room 225 and 14.3% at Room 219. Egg preservation was considered good at Salmon Ruins and Aztec Ruins Room 225. At Room 219, the eggs were badly preserved and there was a significant statistical correlation with mites (multiple r (18) = 0.64/ P = 0.002 ). This correlation could indicate that mites could be involved in preservation, and consequently that pinworm prevalence in this latrine is underestimated. Abiotic factors such as humidity could have possibly favored the biological relation. Considering the extreme egg degradation at Room 219, it is not possible to strictly compare parasite data with other Ancestral Pueblo sites, even with Room 225, located at the same site. Room 225 prevalence is the highest found in Ancestral Pueblo sites until now. Taphonomic analysis shows that decomposers operated at different levels of severity, despite the latrines' close location.

Paleoparasitological studies have demonstrated that changes in environment or culture are reflected in the patterns of parasitic infection diseases in populations worldwide. The advent of agriculture and animal domestication, with its accompanying reduction in human mobility and expanding population involves changes in or emergence of, parasites, the so-called first epidemiological transition. Cultural processes related to territory occupation contribute to both loss and acquisition of parasites. The archaeological site Lluta 57 in the Lluta Valley, Chile, provides a chronology of the transition from the pre-Inca or Late Intermediate Period (LIP), through the Late or Inca Period (LP), to the Hispanic Contact Period (HCP), providing the possibility of evaluating this epidemiological transition. The aim of this study was to conduct a paleoparasitological investigation of to gain insight into the dynamics of parasitism in Lluta people throughout the Inca expansion. Fourteen human coprolites from the three periods were rehydrated, submitted to spontaneous sedimentation, and examined by light microscopy for the presence of intestinal parasite eggs, pollen grains, and micro-remains. Eggs of four parasites: Enterobius vermicularis, Trichostrongylus sp., Trichuris sp., and Eimeria macusaniensis were recovered. Frequency, diversity, and number of parasite eggs per sample increased over the studied time period. Trichostrongylus sp. and E. macusaniensis were recorded in the region for the first time. Enterobius vermicularis eggs, absent in the LIP, were present as a hyper-infection in LP. The presence of E. macusaniensis is likely related to exploitation of llamas, which were used for food and transport and as sacrificial offerings. The paleobotanical analysis revealed ten families of pollen grains, as well as phytoliths and floral remains. In contrast to parasitological results, a diachronic pattern was not detected. Evolution of the settlements, with the advent of larger, more densely populated, villages, could have influenced the emergence and intensification of transmission of parasites in the region. The study showed that the Inca expansion influenced host-parasite-environment relationships in the Lluta Valley.

Studies of nematofauna of teleost fish from the Brazilian coast are relatively scarce and limited to identification based on morphology. The objective of the present study was to determine the diversity and prevalence of nematode parasites in teleost fish from the southeast Atlantic coast of Rio de Janeiro, through morphological, molecular, and ecological approaches. Parasites were collected from sixty specimens each of Genypterus brasiliensis, Micropogonias furnieri, and Mullus argentinae obtained in winters and summers of 2012–2014. Morphological and genetic characterization was conducted using light microscopy and the molecular targets 18S rDNA, ITS1, and mtDNA cox2. Nematodes identified in M. furnieri were Cucculanus genypteri (n = 1575, P = 98.3%) and Hysterothylacium deardorffoverstreetorum (s.l.) (n = 2, P = 3.3%); in G. brasiliensis were Dichelyne (Cucullanellus) sciaenidicola (n = 99, P = 33.3%), Cucculanus pulcherrimus (n = 45, P = 18.3%), Hysterothylacium deardorffoverstreetorum (s.l.) (n = 3, P = 5%), and Anisakis typica (n = 1, P = 1.7%); and, in M. argentinae, were H. deardorffoverstreetorum (s.l.) (n = 146, P = 48.3%), and Procamallanus (Spirocamallanus) halitrophus (n = 4, P = 6.7%). DNA sequence data of C. genypteri, C. pulcherrimus, D. (C.) sciaenidicola, and P. (S.) halitrophus were reported for the first time. New host records are M. argentinae for P. (S.) halitrophus, M. furnieri for A. typica, while H. deardorffoverstreetorum (s.l.) was found in all three fish species. Intestine showed significantly higher intensity than other sites, and no significant seasonal variation in parasitological indices was observed. Hysterothylacium specimens (n = 6) were found in fish muscle, potentially a public health concern.

Anisakidae are represented mainly by species of Anisakis Dujardin, 1845, Pseudoterranova Krabbe, 1878, and Contracaecum Railliet and Henry, 1913. Integrative taxonomy, based on morphological and genetic approaches, can provide a precise identification of species, increasing the knowledge of their systematics, biology, and evolution. In this study, integrative taxonomy was applied in order to identify, at generic and/or specific taxonomic level, 245 anisakids from 63 cetaceans of 12 different species, which stranded in the Brazilian coast. Parasites from nine hosts were preserved by freezing and, from 54 hosts, were fixed in 70% ethanol (n = 43) or alcohol-formalin-acetic (n = 11) for a period varying from 6 to 19 years. The morphological analysis allowed the identification of 171 specimens at genus level: 120 as Anisakis sp. clade I, 47 as Anisakis sp. clade II, and four as Pseudoterranova sp. From those 15, specimens were identified at species level, seven as Anisakis typica, seven as Anisakis paggiae and one as Anisakis brevispiculata. Moreover, 74 specimens were identified as belonging to Anisakidae family. The genetic analysis based on cox2 gene allowed the identification/confirmation of Anisakis typica (n = 33), Anisakis paggiae (n = 1), and for the first time in the Brazilian coast, Anisakis brevispiculata (n = 4) and Anisakis ziphidarum (n = 1). Additionally, a probably new Anisakis species, genetically close to A. paggiae, is reported infecting a dwarf sperm whale. The results clearly indicated that the preservation of nematodes influenced significantly on the level of taxonomic definition, in both morphological and, mainly, genetic analyses. Pseudoterranova sp. was identified for the first time in the spinner dolphin, Stenella longirostris. The detection of nine A. paggiae specimens, in three Kogia breviceps animals, substantiates the austral occurrence of this species. The identification of A. brevispiculata in Kogia sima and K. breviceps confirms Kogiidae whales as their most common hosts. Anisakis typica was corroborated as a generalist species, the most widely distributed in the Brazilian coast, infecting a broad variety of Delphinidae and Kogiidae, with two new geographical records in Stenella frontalis and Lagenodelphis hosei. The study showed a new scenery of parasite diversity of marine anisakids, updating the Anisakis species distribution.

Raillietina celebensis is a cestode that parasitizes the small intestine of rats and humans. Here, we detail the morphology and morphometry of R. celebensis based on specimens collected from Rattus norvegicus in the municipality of São Gonçalo, state of Rio de Janeiro, Brazil, by light and confocal scanning laser microscopies and also report the results of molecular phylogenetic analyses to determine its relationships within the family Davaineidae. Analysis of the number and size of testes, number and shape of rostellar hooks, cirrus sac length, capsules and eggs per capsule, and morphology of the mature proglottid allowed concluding that the present specimens constitute a new record of R. celebensis in South America. Our genetic and phylogenetic analyses, based on the partial small subunit 18S rRNA gene, revealed R. celebensis to be in the family Davaineidae within the genus Raillietina, in agreement with the morphological taxonomy. Phylogenetic trees obtained by neighbor-joining and maximum likelihood methods demonstrated R. celebensis as a unique taxonomic unit, and also demonstrated some taxonomic inconsistences. The incorporation of Brazilian R. celebensis sequences derived from mammals in the phylogeny of davaineids is consistent with the assertion that neither Raillietina nor Fuhrmannetta can be supported as distinct genera.

IntroductionPaleoparasitology, the study of parasites in the past, brings the knowledge of where and when they occurred in preterit populations. Some groups of parasites, as capillariids, have a complex and controversial systematic, hindering the paleoparasitological diagnosis. In this article, we synthesized the occurrence of capillariids in both the New and the Old World in ancient times, and discussed the difficulty of the diagnosis of species and the strategies for identification. The present review also shows the current status of the phylogeny in capillariids and indicates the necessity to try new approaches for a better understanding of capillariid paleodistribution.MethodsFor the systematic review, a predefined guideline defined by PRISMA was used. The articles collected were identified, screened, and included in the review following criteria for eligibility. The current status of the phylogeny of capillariids was accessed using MUSCLE, Bioedit v.7.0.5 and MEGA v. 7.0.21 programs.ResultsThe review discussed 38 articles that presented information about capillariids in past populations. Most of capillariid eggs found in the New and Old World were not identified. However, Calodium hepaticum eggs were the most identified, as some from Eucoleus genus. It was observed that sites from the New World had a better chance for capillariid egg identification, due to previous knowledge of its host, when compared to the Old World. In the 18S rDNA phylogenetic analyses, two datasets were constructed, one including sequences from 7 Moravec's genera, where 3 genus-specific clusters, with high bootstrap values, could be observed for Capillaria (ML = 99%, NJ = 96%), Eucoleus (ML / NJ = 100%) and Paratrichosoma (ML / NJ = 100%). A fourth cluster of 18S rDNA dataset I revealed lack of definition of Pearsonema and Aonchotheca genera. The 18S rDNA dataset II comprised 8 Moravec's genera and defined 3 clusters, 2 genus-specific for Eucoleus (ML = 99%, NJ = 100%) and Capillaria (ML / NJ = 98%). The third 18S rDNA dataset II cluster included 6 genera and exhibited, once again, Pearsonema and Aonchotheca poor discrimination. The cox1 gene data consist of 4 Moravec's genera, and in spite of grouping some species-specific clusters, did not show genera-specific definition.ConclusionsDespite the numerous archaeological findings, both in the New and the Old Worlds, the identification of capillariid species based on the morphology and morphometry of eggs remains imprecise, often resulting in a generic diagnosis of a group or morphotype of capillariid. Capillariid is one of the most diverse group of helminths recovered in archaeological sites. The phylogenetic trees produced in this study showed limited genetic information available, unresolved genera and incongruence with the classical taxonomy. The elucidation of the paleodistribution of capillariids can give insights of the ancient host-parasite associations but also in modern sceneries.

Pretos Novos cemetery (PNC), Rio de Janeiro, Brazil (1769-1830) was created exclusively to bury enslaved Africans who died upon arrival at the city or before being sold in the slave market. The PNC site may be unique in the Americas in allowing the study of African parasite infections acquired in Africa. We aimed to identify parasites infecting PNC individuals through paleoparasitological and paleogenetic analyses. The bodies had been dismembered, placed in mass graves, and burned, and most human remains collected from the site are highly fragmented and show extensive degradation. Sacrum and pelvic sediments were collected from five individuals along with seven samples of sediment from other areas of the body, as controls. Samples were submitted to three parasitological techniques and, in paleogenetic analysis, to four molecular targets. Larvae, mites, pollen grains, and structure suggestive of plants and fungus were observed, but we found no evidence of helminth infection. Ascaris sp. cytb sequence was recovered in one individual. We emphasize that, even with the extensive degradation of PNC human remains and the process of curation of samples, it was possible to recover helminth aDNA. The origin of PNC individuals confirms that these infections were brought to Brazil from western and central Africa during the colonial era.

Flounders are commercially and economically important fish. A total of 120 specimens of flounders (60 Paralichthys isosceles, 30 Paralichthys patagonicus and 30 Xystreurys rasile) were collected off the coast of the state of Rio de Janeiro, Brazil. The fish were measured, necropsied and filleted, and then had their organs investigated for acanthocephalans. Taxonomic identification of the parasites was based on morphological, morphometric and genetic characters. Paralichthys isosceles and P. patagonicus were parasitized by juveniles of Serrasentis sagittifer, Bolbosoma turbinella, Corynosoma australe and C. cetaceum; Xystreurys rasile was parasitized by C. australe. Genetic characterization confirmed the identification of specimens of Bolbosoma turbinella and Corynosoma australe, as demonstrated by phylogenetic analyses using both ITS and cox1 molecular targets. Parasite indices of prevalence, intensity, mean intensity, abundance, mean abundance, and range of infection, as well as infection site, were evaluated for each parasite species. This is the first report of S. sagittifer parasitizing P. isosceles and P. patagonicus, and B. turbinella parasitizing P. patagonicus. Resumo Os linguados são peixes comercial e economicamente importantes. Um total de 120 espécimes de linguados (60 Paralichthys isosceles, 30 P. patagonicus e 30 Xystreurys rasile) foram coletados no litoral do estado do Rio de Janeiro, Brasil. Os peixes foram medidos, necropsiados, filetados e tiveram seus órgãos investigados para a presença de acantocéfalos. A identificação taxonômica foi baseada em caracteres morfológicos, morfométricos e genéticos. Paralichthys isosceles e P. patagonicus estavam parasitados por acantocéfalos juvenis de Serrasentis sagittifer, Bolbosoma turbinella, Corynosoma australe e C. cetaceum; Xystreurys rasile estava parasitado com C. australe. A caracterização genética confirmou a identificação dos espécimes de Bolbosoma turbinella e Corynosoma australe, como demonstrado por análises filogenéticas usando ambos marcadores moleculares ITS e cox1. Foram analisados os índices parasitários: prevalência, intensidade, intensidade média, abundância, abundância média, amplitude de variação da infecção e sítio de infecção de cada espécie de parasito. Este é o primeiro registro de S. sagittifer parasitando P. isosceles e P. patagonicus, e de B. turbinella parasitando P. patagonicus.

The Guanches, ancient inhabitants of the Canary Islands, Spain, practiced mummification of their dead. A paleoparasitological and paleogenetic analysis was conducted on mummified bodies (n = 6) (AD 1200, Cal BP 750) belonging to the Guanche culture from Gran Canaria Island. Coprolite and sediment samples (n = 19) were removed from below the abdominal region or sacral foramina. The samples were rehydrated in 0.5% trisodium phosphate solution for 72 hr at 4 C, and the paleoparasitological investigation was conducted by spontaneous sedimentation method and microscopic examination. The results revealed the presence of well-preserved eggs of Ascaris sp., Trichuris trichiura , Enterobius vermicularis , and hookworms. Ancient DNA was extracted from sediment samples to elucidate the ancestry of the mummies and for molecular detection of Ascaris sp. infection. Results of paleogenetic analysis demonstrated Ascaris sp. infection using 2 molecular targets, cytb and nad1. The mtDNA haplotypes U6b, U6b1, and HV were identified, which confirmed records of Guanche ancestry. The excellent preservation of Guanche mummies facilitated the paleoparasitological and paleogenetic study, the results of which contribute to our knowledge of Guanche culture and their health status.

Background Trypanosoma cruzi is a protozoan parasite characterized by extensive genetic heterogeneity. There are currently six recognised, genetically distinct, monophyletic clades designated discrete typing units (DTUs). TcI has the broadest geographical range and most genetic diversity evidenced by a wide range of genetic markers applied to isolates spanning a vast geographical range across Latin America. However, little is known of the diversity of TcI that exists within sylvatic mammals across the geographical expanse of Brazil. Results Twenty-nine sylvatic TcI isolates spanning multiple ecologically diverse biomes across Brazil were analyzed by the application of multilocus sequence typing (MLST) using four nuclear housekeeping genes. Results revealed extensive genetic diversity and also incongruence among individual gene trees. There was no association of intralineage genotype with geography or with any particular biome, with the exception of isolates from Caatinga that formed a single cluster. However, haplotypic analyses of METIII and LYT1 constitutive markers provided evidence of recombination events in two isolates derived from Didelphis marsupialis and D. albiventris, respectively. For diversity studies all possible combinations of markers were assessed with the objective of selecting the combination of gene targets that are most resolutive using the minimum number of genes. A panel of just three gene fragments (DHFR-TS, LYT1 and METIII) discriminated 26 out of 35 genotypes. Conclusions These findings showed geographical association of genotypes clustering in Caatinga but more characteristically TcI genotypes widely distributed without specific association to geographical areas or biomes. Importantly, we detected the signature of recombination events at the nuclear level evidenced by haplotypic analysis and incongruence. Electronic supplementary material The online version of this article (10.1186/s13071-018-2696-9) contains supplementary material, which is available to authorized users.

Despite interest in the origins of syphilis, paleopathological analysis has not provided answers, and paleogenetic diagnosis remains a challenge. Even venereal syphilis has low infectivity which means there are few circulating bacteria for most of the individual’s life. Human remains recovered from the Nossa Senhora do Carmo Church (17th to 19th centuries) and the Praça XV Cemetery (18th to 19th centuries), Rio de Janeiro, Brazil, were subjected toTreponemapaleogenetic analysis. Historical data point to endemic treponemal infections in the city, including venereal syphilis. Based on the physiopathology ofTreponema palliduminfection, 25 samples, mostly from skull remains of young adults, with no visible paleopathological evidence of treponematoses, were analyzed. PCR with three molecular targets,tpp47,polA, andtpp15, were applied. Ancient DNAtpp15 sequences were recovered from two young adults from each archaeological site and revealed the polymorphism that characterizesT. p.subsp.pallidumin a female up to 18 years old, suggesting a probable case of syphilis infection. The results indicated that the epidemiological context and the physiopathology of the disease should be considered in syphilis paleogenetic detection. The findings ofTreponemasp. aDNA are consistent with historical documents that describe venereal syphilis and yaws as endemic diseases in Rio de Janeiro. Data on the epidemiological characteristics of the disease and its pathophysiology offer new perspectives in paleopathology.

Anisakid nematodes have been identified in a wide variety of fish and marine mammal species. In Brazil, Anisakis physeteris, A. insignis, A. typica, A. nascetti, and those of the A. simplex complex have been reported infecting fishes and cetaceans. In this study, specimens collected from a dwarf sperm whale Kogia sima (Owen, 1866) stranded on the northeastern coast of Brazil were identified through morphological and genetic analyses as A. paggiae. Anisakids were examined through differential interference contrast light and scanning electron microscopy (SEM). Morphological and morphometric analysis revealed that these specimens belonged to Anisakis sp. clade II and more specifically to A. paggiae, exhibiting a violin-shaped ventriculus and 3 denticulate caudal plates, which are taxonomic characters considered unique to this species. Genetic analysis based on the mtDNA cox2 gene confirmed our identification of A. paggiae. Phylogenetic trees using both maximum likelihood and neighbor-joining methods revealed a strongly supported monophyletic clade (bootstrap support = 100%) with all available A. paggiae sequences. Integrative taxonomic analysis allowed the identification of A. paggiae for the first time in Brazilian waters, providing new data about their geographical distribution. Moreover, here we present the first SEM images of this species.

Paleoparasitological studies of the Brazilian colonial period are scarce. A paleoparasitological analysis was performed on human remains from the archeological site Prac¸a XV Cemetery in Rio de Janeiro, dating from the early 18th to 19th Centuries. The samples were obtained from the Institute ofthe Brazilian Archaeology collection, and showed evidence of washing and brushing. Sediments were extracted from sacral foramina by scraping. Sediments from skulls were used as negative paleoparasitological controls. Spontaneous sedimentation method was performed prior to microscopic analysis. The results revealed that 8 of 10 individuals were infected with intestinal helminths and/or protozoa. Eggs ofthe nematodes Trichuris sp. and Ascaris sp. as well as a single taeniid egg were found. Protozoa cysts suggestive of Entamoeba sp. were also observed. Trichuris sp. was the most frequent and abundant parasite, found in 70% ofindividuals (26 eggs). The study showed the importance of analysis of sedimentfrom human remains preserved in museum or scientific collections, even those subjected to a curating procedure. The levels of infection revealed here should be considered underestimations. This is the first paleoparasitological study from Rio de Janeiro city for the Brazilian colonial period and the first report of human Taenia sp. in the NewWorld. Fil: Jaeger, Lauren Hubert. Fundacão Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia de Tripanosomatídeos. Laboratório de Genética Molecular de Microorganismos; Brasil Fil: Taglioretti, Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales; Argentina Fil: Fugassa, Martín Horacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales; Argentina Fil: Dias, Ondemar. Instituto de Arqueología Brasileira; Brasil Fil: Neto, Jandira. Instituto de Arqueología Brasileira; Brasil Fil: Mayo Iñiguez, Alena. Fundacão Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia de Tripanosomatídeos. Laboratório de Genética Molecular de Microorganismos; Brasil

Thirty-six Paralichthys patagonicus and 30 Xystreurys rasile were collected in the state of Rio de Janeiro, Brazil to investigate the presence of anisakid and raphidascaridid nematodes. Anisakis typica, Terranova sp., Contracaecum sp., Hysterothylacium deardorffoverstreetorum, and Raphidascaris sp. were identified using integrative taxonomy of morphological and genetic data. Morphological and morphometric analysis was conducted using bright field microscopy with scanning electron microscopy for topographic characterization of the cuticular surface. Phylogenetic analysis, using ITS and cox2 molecular targets, clearly demonstrated the species identification of A. typica and H. deardorffoverstreetorum and the high diversity of H. deardorffoverstreetorum. This is the first report of A. typica, H. deardorffoverstreetorum, and Raphidascaris sp. parasitizing P. patagonicus and X. rasile.

Paleogenetic analysis for tuberculosis (TB) was conducted on bone and sediment samples dating from the 17th to 19th centuries from the archeological site of Nossa Senhora do Carmo Church in Rio de Janeiro, Brazil. Forty samples were analyzed, corresponding to 32 individuals from 28 burials, 22 of primary type and 6 of secondary type. The samples were collected following strict paleogenetic investigation guidelines and submitted to ancient DNA (aDNA) extraction. In order to detect TB infection, aDNA hybridizations with the molecular targets of Mycobacterium tuberculosis complex (MTC) IS6110 and IS1081 were applied. Additionally, the ancestry of individuals was assessed by human mitochondrial DNA (mtDNA) analysis of hypervariable segment I (HVS-I) sequence polymorphisms. The results of aDNA hybridizations demonstrated varying levels of MTC intensity in 17/32 individuals (53.1%), using the IS6110 target. The IS1081 MTC target showed lower sensitivity, confirming TB positivity in 10/32 (31.2%) individuals. The mtDNA analysis allowed the recovery of HVS-I sequences in 23/32 individuals (71.8%). The majority of these individuals (21/23, 91.3%) were of European ancestry, especially in primary burials. Haplogroups U, J, V, T, K, N, H and R, were identified with haplogroup U being the most frequent at 6/23 (26.1%). African and Amerindian mtDNA haplogroups were observed in two individuals in secondary burials. In spite of the ecclesiastic and aristocratic bias of the population of the study, human ancestry analysis revealed the prominent contribution of Europeans in the introduction or spread of TB in the New World.

Taking into account the difficulties of taxonomic identification of larval anisakid nematodes based on morphological characters, genetic analyses were performed, together with those usually applied, in order to identify anisakid larvae found in the flounder Paralichthys isosceles from the littoral of the state of Rio de Janeiro, Brazil. The analysis of 1,820 larvae revealed a new species, similar to Hysterothylacium MD, Hysterothylacium 2, Hysterothylacium KB and Hysterothylacium sp regarding the absence of the larval tooth, an excretory pore situated below the nerve ring level, and slender lateral alae. Moreover, the new species differs from Hysterothylacium fortalezae and Hysterothylacium reliquens with regard to the number and size of spines present on the tail end and from Hysterothylacium patagonicus by the absence of interlabia. The maximum parsimony and neighbour joining tree topologies based on the 18S ribosomal DNA gene, complete internal transcribed spacer region and cytochrome oxidase 2 (COII) gene demonstrated that the Brazilian larvae belong to Raphidascarididae and represent a unique genetic entity, confirmed as a new Hysterothylacium species. Furthermore, the new species presents COII genetic signatures and shares polymorphisms with Raphidascarididae members. This is the first description of a new anisakid species from Brazil through the integration of morphological and molecular taxonomy data.

Synthesium pontoporiae, exclusive parasite of the endangered dolphin, Pontoporia blainvillei, is endemic and restricted to the South Atlantic and belongs to the Brachycladiidae family Odhner, 1905. The study of this family has been limited by the difficulty of accessing the parasites from their marine mammal hosts and as a consequence there is a paucity of genetic information available. Herein we present a genetic analysis using 18S rDNA sequences of S. pontoporiae and S. tursionis and the ND3 mtDNA sequence of S. pontoporiae. The genetic analysis of 18S rDNA sequences of brachycladiids and acanthocolpids determined two major clusters associated with their definitive hosts, marine mammals and fishes, respectively. Considering the tree topology of brachycladiids ND3 mtDNA gene, two clusters were defined, one with the Synthesium species. This work contributes with fundamental genetic information on S. pontoporiae, and suggests a Brachycladiidae genetic evolution related to their hosts.

This study represents the first survey of the parasitic fauna of cetaceans off the northeastern coast of Brazil. Parasites were collected from 82 animals rescued from the states of Ceará to Bahia, including the archipelago of Fernando de Noronha. A total of 14 species of cetaceans were evaluated: Sotalia guianensis, Stenella sp., Stenella clymene, Stenella longirostris, Stenella coeruleoalba, Stenella frontalis, Megaptera novaeangliae, Peponocephala electra, Steno bredanensis, Kogia breviceps, Kogia sima, Globicephala macrorhynchus, Tursiops truncatus, Physeter macrocephalus and Lagenodelphis hosei. The parasites were fixed and preserved in 70% ethanol or alcohol–formalin–acetic acid solution (AFA), clarified in phenol and mounted on slides for morphological identification. In total, 11 species and 8 genera of endo- and ectoparasites were identified: Halocercus brasiliensis, Halocercus kleinenbergi, Stenurus globicephalae, Halocercus sp., Anisakis sp., Crassicauda sp. (Nematoda), Phyllobothrium delphini, Monorygma grimaldii, Scolex pleuronectis, Strobicephalus triangularis, Tetrabothrius forsteri, Tetrabothrius sp., Trigonocotyle sp., Diphyllobothrium sp. (Cestoda), Campula sp. (Trematoda), Bolbosoma sp. (Acanthocephala), Cyamus boopis, Syncyamus pseudorcae and Xenobalanus globicipitis (Crustacea). The identification of some species represented novel records for the country and increased the occurrence of some parasites to new hosts. The use of standardized methodologies for collecting and evaluating a larger number of animals is essential for a better understanding of host-parasite relationships in cetaceans and their use as biological indicators in the region.

Ascaris lumbricoides (giant roundworm) and Trichuris trichiura (whipworm) are the most common intestinal parasites found in humans worldwide today and they almost always co-occur. However, we find two distinct patterns in archaeological material. In historical North American and Old World contexts, the association of A. lumbricoides and T. trichiura is similar to the modern epidemiological picture. In contrast, the co-occurrence of A. lumbricoides and T. trichiura eggs in prehistoric South America is rare. For prehistoric contexts, T. trichiura is the most common parasite found in archaeological material. Recently molecular biology techniques pointed to a subdiagnosis of roundworm infection in pre-Columbian South American populations. This is contrary to the modern epidemiological picture in which A. lumbricoides infection is predominant. This is a paradox, especially when one considers the number of eggs laid by female daily, 200,000 and 20,000 thousand per day, for A. lumbricoides and T. trichiura respectively. By reviewing the records of these parasites, this paradox is presented and explanations for the paradox are explored. Taphonomy, prehistoric behavior patterns and medicinal plant use seem to be most relevant to the explanation of the paradox. Nematophagous fungi is a less likely factor creating the near absence of A. lumbricoides eggs in the prehistoric New World.

The oncoprotein Tax was characterized genetically from a large cohort of human T-cell lymphotropic virus type 1 (HTLV-1) seropositive individuals from the most endemic region of HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and HTLV-1 infection in Argentina, the province of San Salvador de Jujuy. Sixteen HAM/TSP patients and 47 HTLV-1 healthy carriers were evaluated. Six Tax genetic polymorphisms were identified and observed in 70.8% of healthy carriers and 62.5% of HAM/TSP patients. Tax genetic polymorphisms were not associated with clinical status but A8344C polymorphism statistically provide a borderline protective effect of HAM/TSP outcome. Nucleotide diversity in healthy carriers was 0.00549, whereas HAM/TSP virus population revealed a low diversity of 0.00379, suggests a positive selection for Tax protein conservation in this group. It is concluded that tax genetic polymorphisms do not increase the risk of developing HAM/TSP in this endemic region. However, in spite of the low prevalence of HTLV-1aB genotype, statistical analysis revealed an important correlation of tax genetic signatures with HTLV-1aA trans-continental subgroup.

The province of San Salvador de Jujuy, located in the northwest of Argentina, is a highly endemic area for HTLV-1 infection and a foci of tropical spastic paraparesis/HTLV-1-associated myelopathy (HAM/TSP). Therefore, to better understand this, we carried out a genetic characterization of a large set of HTLV-1 strains (n = 65) of descendants of Amerindians from this region. The LTR and env regions were analyzed. The genetic analysis showed that all of these new HTLV-1 isolates from Argentina belong to the Transcontinental subgroup A of the HTLV-1a Cosmopolitan subtype, with the exception of three isolates that cluster within the Japanese subgroup B. Interestingly, the majority of the sequences from Jujuy province belonged to a distinct cluster within the Latin America Transcontinental subgroup, referred to here as the Jujuy subcluster, and were characterized by specific signatures in the LTR. Given that the samples analyzed in this study belong to the Amerindian population and the high prevalence of HTLV-1 in Jujuy in contrast to the low prevalence of this virus in the country, it could be that HTLV-1aA was spread in Argentina from the Amerindians to the cosmopolitan population. Moreover, this is the first report of an HTLV-1aB or Japanese subgroup in descendants of non-Japanese people in South America.

Molecular analysis of coprolites from Hinds Cave, Texas recovered chloroplast DNA sequences. The sequences were interpreted as evidence of diet. We analyzed 19 Hinds Cave coprolites to evaluate the potential sources of the chloroplast DNA (cpDNA) and compared our results to previous studies. This review shows that some cpDNA sequences could be from well-known prehistoric plants foods. Some other sequences could have come from ambient plant material in the guts of small animals eaten by humans in antiquity. Using pollen concentration analysis, we identify sources of ambient plant material which could have been inhaled or imbibed. It is even possible that cpDNA sequences are from proplastids within ambient pollen grains themselves. However, three sequence types cannot be explained as resulting from only dietary or ambient sources. We suggest instead that these might be from medicinal or hallucinogenic plants. We compared these three sequences to existing sequences in the GenBank. We found that these sequences are 100% matches for Rhamnus, Fouquieria, and Solanum. 2007 Published by Elsevier Ltd.

p12 pX ORF-I (p12I) of simian T cell lymphotropic virus 1 from Central and West Africa is a 91-amino acid (aa) protein, contrasting with the known p12 sequences from human T cell lymphotropic virus type 1, which are 99 aa in length. An in silico sequence analysis shows a premature termination codon in HTLV-1 p12I sequences from South American and Japanese strains. A translation analysis indicated the presence of 86-aa and 81-aa p12I proteins, shorter than the 99-aa protein found in the majority of HTLV-1 strains around the world. This report shows one more common feature between these two viruses. Additionally, the HTLV-1 p12 signature reported as a genetic marker of Brazilian sequences (p12I P63 and P91) is also highly prevalent in p12 Argentinean sequences. Because p12I may confer a proliferative advantage on HTLV-1-infected cells the existence of naturally truncated proteins might supply insights into its pathogenic mechanisms.

Anisakid nematodes are parasites with an indirect life cycle, involving many hosts, including cetaceans that act as definitive hosts. Identification at species level is crucial for a better understanding of the epidemiology and ecology of these parasites, but an accurate diagnosis based only on morphology is difficult. In Brazil, genetic characterization of anisakids is still rare, with Anisakis typica and A. physeteris being the only two species identified by genetic markers until now. The definitive hosts of A. nascettii are Mesoplodon grayi, M. bowdoini, M. layardii and M. mirus. Geographical locations of A. nascettii include the coasts of New Zealand, South Africa and Europe. In this study, a nematode was collected from a Gervais' beaked whale, Mesoplodon europaeus, stranded in Ceará State, north-east Brazil. Genetic analysis based on 18S rDNA, internal transcribed spacer (ITS) region and mtDNA cox2 gene revealed 100% identity with Anisakis sp. U94365, 99% with Anisakis sp. (MP-2005 EU718477 A. nascettii) and 99% with A. nascettii DQ116431, respectively. Phylogenetic analyses of ITS and cox2 sequences using both neighbour-joining and maximum-likelihood methods provided strong support for a clade with only A. nascettii specimens. This study demonstrated for the first time the identification of A. nascettii in the Brazilian Atlantic Coast, revealing a wider geographical distribution of this parasite worldwide and extending the range of its definitive hosts to a new Mesoplodon species, M. europaeus, the Gervais' beaked whale.

We evaluated the presence and distribution of Trypanosoma cruzi DNA in a mummy presenting with megacolon that was dated as approximately 560 +/- 40 years old. The mummy was from the Peruaçu Valley in the state of Minas Gerais, Brazil. All samples were positive for T. cruzi minicircle DNA, demonstrating the presence and broad dissemination of the parasite in this body. From one sample, a mini-exon gene fragment was recovered and characterized by sequencing and was found to belong to the T. cruzi I genotype. This finding suggests that T. cruzi I infected humans during the pre-Columbian times and that, in addition to T. cruzi infection, Chagas disease in Brazil most likely preceded European colonization.