From molecules to ecosystems: Insights into a network of interactions for a Chagas disease outbreak using Triatoma brasiliensis as natural samplers.

Exploring the dynamics of disease transmission involves an understanding of complex interactions within the eco-epidemiologic framework. In the context of Chagas disease (CD), elements are mainly represented by the interactions among the pathogen, insect vector, host, humans and the environment. We performed quantitative and qualitative analyses on a dataset derived from 98 Triatoma brasiliensis infected by trypanosomatids, which were linked to a CD outbreak in the semi-arid region of northeastern Brazil. We extracted invertebrate-derived DNA (iDNA) from these insects, comprising 18 populations around the outbreak area, each indicative of various strata of anthropogenic influence. Food source (FS) diversity, representing potential parasite reservoirs, was determined through mitochondrial gene (cyt b) sequencing of vertebrates, and parasite genotyping was accessed using fluorescent amplified fragment barcodes (FFLB) of trypanosomatids. We also assessed the residents' awareness of breeding sites for CD vectors in the inspected houses. The quantification of Trypanosoma cruzi was estimated via real-time PCR and is denominated here as the average parasite load (PL) per insect (T. cruzi/intestinal unit). We aimed to address vector-parasite-host-environment interactions that were discussed based on their significance among the components. Notably, among the significant interactions, we observed that the PL in the insects was significantly influenced by FS. Infected insects that fed on the classic reservoir, Didelphis albiventris, and Galea spixii exhibited higher PLs, compared to those that fed on Kerodon rupestris (p < 0.04)-a primary host. While D. albiventris is already recognized as a synanthropic species, we propose that G. spixii may also be undergoing a synanthropic process. Conversely, domestic cats are frequently identified as FS in infected insects from the sylvatic environment, suggesting a possible change in their behavior towards a wild state. Therefore, we propose that neglected anthropogenic actions have facilitated the reciprocal (sylvatic-peridomestic) circulation of T. cruzi-especially noted for TcI because it was predominant in insects found in peridomestic environments. Residents are often unaware of the existence of insect breeding grounds near their homes, particularly when it involves the storage of materials without planning for use, such as piles of tiles, bricks and wood. Although indirect inferences about the interaction among vector-parasite-host-environment are still incipient, we highlight the potential use of vectors as natural samplers of biological and ecological components in transmitting the disease.

"Mi Casa, Tu Casa": the coati nest as a hub of Trypanosoma cruzi transmission in the southern Pantanal biome revealed by molecular blood meal source identification in triatomines.

BACKGROUND: The study of the ecology of Trypanosoma cruzi is challenging due to its extreme adaptive plasticity, resulting in the parasitism of hundreds of mammal species and dozens of triatomine species. The genetic analysis of blood meal sources (BMS) from the triatomine vector is an accurate and practical approach for gathering information on which wild mammal species participate in a local transmission network. South American coatis, Nasua nasua, act as important reservoir host species of T. cruzi in the Pantanal biome because of their high rate of infection and elevated parasitemia, with the main discrete typing unit (DTU) lineages (TcI and TcII). Moreover, the carnivore coati is the only mammal species to build high arboreal nests for breeding and resting that can be shared by various vertebrate and invertebrate species. Herein, we applied the sensitive and specific methodology of DNA barcoding and molecular cloning to study triatomines found in a coati nest to access the diversity of mammal species that explore this structure, and therefore, may be involved in the parasite transmission network. METHODS: Twenty-three Triatoma sordida were collected in one coati's nest in the subregion of Nhecolândia, Pantanal. The DNA isolated from the gut of insects was subjected to BMS detection by PCR using universal primers that flank variable regions of the cytochrome b (cytb) and 12S rDNA mitochondrial genes from vertebrates. The Trypanosoma spp. diagnosis and DTU genotyping were based on an 18S rDNA molecular marker and also using new cytb gene primers designed in this study. Phylogenetic analyses and chord diagrams were constructed to visualize BMS haplotypes, DTU lineages detected on vectors, and their interconnections. RESULTS: Twenty of 23 triatomines analyzed were PCR-positive (86.95%) showing lineages T. cruzi DTU TcI (n = 2), TcII (n = 6), and a predominance of TcI/TcII (n = 12) mixed infection. Intra-DTU diversity was observed mainly from different TcI haplotypes. Genetic analyses revealed that the southern anteater, Tamandua tetradactyla, was the unique species detected as the BMS of triatomines collected from the coati's nest. At least three different individuals of T. tetradactyla served as BMS of 21/23 bugs studied, as indicated by the cytb and 12S rDNA haplotypes identified. CONCLUSIONS: The identification of multiple BMS, and importantly, different individuals of the same species, was achieved by the methodology applied. The study demonstrated that the southern anteaters can occupy the South American coati's nest, serving as the BMS of T. sordida specimens. Since anteaters have an individualist nonsocial behavior, the three individuals detected as BMS stayed at the coati's nest at different times, which added a temporal character to BMS detection. The TcI and TcII infection, and significantly, a predominance of TcI/TcII mixed infection profile with different TcI and TcII haplotypes was observed, due to the discriminatory capacity of the methodology applied. Tamandua tetradactyla, a host which has been little studied, may have an important role in the T. cruzi transmission in that Pantanal subregion. The data from the present study indicate the sharing of coatis' nests by other mammal species, expanding the possibilities for T. cruzi transmission in the canopy strata. We propose that coatis' nests can act as the true hubs of the T. cruzi transmission web in Pantanal, instead of the coatis themselves, as previously suggested.

Molecular detection of blood meal source up to three months since the last meal: Experimental starvation resistance in triatomines.

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.

Machine learning approach to support taxonomic species discrimination based on helminth collections data.

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.

New data on eggshell structure of capillariid species: a SEM perspective.

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.

African helminth infection out of Africa: Paleoparasitological and paleogenetic investigations in Pretos Novos cemetery, Rio de Janeiro, Brazil (1769-1830).

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.

Acanthocephalan parasites of the flounder species Paralichthys isosceles, Paralichthys patagonicus and Xystreurys rasile from Brazil / Acantocéfalos parasitos de espécies de linguados Paralichthys isosceles, Paralichthys patagonicus e Xystreurys rasile no Brasil

Abstract 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.

Acanthocephalan parasites of the flounder species Paralichthys isosceles, Paralichthys patagonicus and Xystreurys rasile from Brazil.

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.

Worldwide paleodistribution of capillariid parasites: Paleoparasitology, current status of phylogeny and taxonomic perspectives.

INTRODUCTION: Paleoparasitology, 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. METHODS: For 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. RESULTS: The 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. CONCLUSIONS: Despite 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.

Acanthocephalan parasites of the flounder species Paralichthys isosceles, Paralichthys patagonicus and Xystreurys rasile from Brazil / Acantocéfalos parasitos de espécies de linguados Paralichthys isosceles, Paralichthys patagonicus e Xystreurys rasile no Brasil

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.(AU)

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.(AU)

First Paleogenetic Evidence of Probable Syphilis and Treponematoses Cases in the Brazilian Colonial Period.

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 to Treponema paleogenetic analysis. Historical data point to endemic treponemal infections in the city, including venereal syphilis. Based on the physiopathology of Treponema pallidum infection, 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, and tpp15, were applied. Ancient DNA tpp15 sequences were recovered from two young adults from each archaeological site and revealed the polymorphism that characterizes T. p. subsp. pallidum in 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 of Treponema sp. 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.

Tuberculosis in post-contact Native Americans of Brazil: Paleopathological and paleogenetic evidence from the Tenetehara-Guajajara.

Tuberculosis (TB) has been described in Native American populations prior to the arrival of European explorers, and in Brazilian populations dating from the Colonial Period. There are no studies demonstrating TB infection in native Brazilians, and the history and epidemiological scenario of TB in Brazil is still unknown. The aim of this study was to verify the presence of TB infection among the native Tenetehara-Guajajara population from Maranhão State, Brazil, 210 ± 40 years ago. A Tenetehara-Guajajara skeleton collection was submitted to paleopathological analysis, and rib bone samples (n = 17) were used for paleogenetic analysis based on Mycobacterium tuberculosis complex (MTC) targets. Porotic hyperostosis and cribra orbitalia were found in 10 and 13 individuals, respectively. Maternal ancestry analysis revealed Native American mtDNA haplogroups A and C1 in three individuals. Three samples showed osteological evidence suggestive of TB. katG and mtp40 sequences were detected in three individuals, indicating probable TB infection by two MTC lineages. Tuberculosis infection in the Tenetehara-Guajajara population since the 18th century points to a panorama of the disease resulting, most probably, from European contact. However, the important contribution of African slaves in the population of Maranhão State, could be also considered as a source of the disease. This study provides new data on TB during the Brazilian Colonial Period. This is the first report integrating paleopathological and paleogenetic data for the study of TB in Brazil.

First record of the nematode Libyostrongylus dentatus Hoberg, Lloyd & Omar, 1995 (Trichostrongylidae) in ostriches (Struthio camelus Linnaeus, 1758) (Struthionidae) outside the Americas.

BACKGROUND: Libyostrongylus douglassii, Libyostrongylus dentatus and Libyostrongylus magnus are nematodes that infect ostriches. The first species has been identified in ostriches from Africa, Europe, Americas and Oceania. Although the natural range of ostriches is Africa, L. dentatus was first described in birds from the USA and later identified in Brazil, where co-infections with L. douglassii have been commonly reported. Libyostrongylus magnus is known from the original description only. There are a few reports on infections with L. douglassii in ostriches from Africa and all farmed birds examined are from the southern region of the continent. The aim of this report was to verify Libyostrongylus spp. infections in wild ostriches from Ethiopia. Fecal samples from ostriches, Struthio molybdophanes, were collected and submitted to coproculture. Infective larvae were identified to the species level based on general morphology and morphometry. In addition, phylogenetic analysis of the first and second internal transcribed spacer (ITS1 and ITS2) of the nuclear ribosomal DNA was performed. RESULTS: Infective larvae from Ethiopian ostriches had the morphological characteristics of L. dentatus. Confidence interval estimate for sheath tail length from Ethiopian Libyostrongylus sp. isolates overlapped one for Brazilian L. dentatus. Neighbor-joining and Maximum Likelihood phylogenetic trees based on sequences of the ITS1 and ITS2 regions revealed that the Ethiopian samples belong to the L. dentatus species clade. Monospecific infections with L. dentatus were confirmed in Ethiopian wild ostriches, opposed to the co-infections typically found in the Americas. CONCLUSIONS: To our knowledge, this is the first record of L. dentatus from African ostriches, the region from which this parasite originated.

Multilocus sequence typing: genetic diversity in Trypanosoma cruzi I (TcI) isolates from Brazilian didelphids.

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.

New paleoparasitological investigations from the pre-inca to hispanic contact period in northern Chile.

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.

Nematode parasites of commercially important fish from the southeast coast of Brazil: Morphological and genetic insight.

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 2012­2014. 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.

Integrative taxonomy of anisakid nematodes in stranded cetaceans from Brazilian waters: an update on parasite's hosts and geographical records.

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.

First Record of Raillietina celebensis (Cestoda: Cyclophyllidea) in South America: Redescription and Phylogeny.

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.

Integrative taxonomy of Anisakidae and Raphidascarididae (Nematoda) in Paralichthys patagonicus and Xystreurys rasile (Pisces: Teleostei) from Brazil.

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.

Evidence of Helminth Infection in Guanche Mummies: Integrating Paleoparasitological and Paleogenetic Investigations.

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.