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.

Clinical conditions associated withintestinal strongyloidiasis in Rio de Janeiro, Brazil

INTRODUCTION: Strongyloides stercoralis is a soil-transmitted helminth that produces an infection that can persist for decades. The relationships between certain clinical conditions and strongyloidiasis remains controversial. This study aims to identify the clinical conditions associated with intestinal strongyloidiasis at a reference center for infectious diseases in Rio de Janeiro, Brazil. METHODS: The clinical conditions that were assessed included HIV/AIDS, HTLV infection, cardiovascular diseases, diabetes, obstructive respiratory diseases, viral hepatitis, tuberculosis, cancer, chronic renal disease, nutritional/metabolic disorders, psychiatric conditions, rheumatic diseases and dermatologic diseases. We compared 167 S. stercoralis-positive and 133 S. stercoralis-negative patients. RESULTS: After controlling for sex (male/female OR = 2.29; 95% (CI): (1.42 - 3.70), rheumatic diseases remained significantly associated with intestinal strongyloidiasis (OR: 4.96; 95% CI: 1.34-18.37) in a multiple logistic regression model. With respect to leukocyte counts, patients with strongyloidiasis presented with significantly higher relative eosinophil (10.32% ± 7.2 vs. 4.23% ± 2.92) and monocyte (8.49% ± 7.25 vs. 5.39% ± 4.31) counts and lower segmented neutrophil (52.85% ± 15.31 vs. 61.32% ± 11.4) and lymphocyte counts (28.11% ± 9.72 vs. 30.90% ± 9.51) than S. stercoralis-negative patients. CONCLUSIONS: Strongyloidiasis should be routinely investigated in hospitalized patients with complex conditions facilitate the treatment of patients who will undergo immunosuppressive therapy. Diagnoses should be determined through the use of appropriate parasitological methods, such as the Baermann-Moraes technique. .

Genetic and morphological characterisation of a new species of the genus Hysterothylacium (Nematoda) from Paralichthys isosceles Jordan, 1890 (Pisces: Teleostei) of the Neotropical Region, state of Rio de Janeiro, Brazil

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.

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

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

Pre-Columbian Chagas disease in Brazil: Trypanosoma cruzi I in the archaeological remains of a human in Peruaçu Valley, Minas Gerais, Brazil

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.

Gastric lesions associated with the presence of Anisakis spp. Dujardin, 1845 (Nematoda: Anisakidae) in Cetaceans stranded on the coast of Ceara, Brazil / Lesões gástricas associadas à presença de Anisakis spp. Dujardin, 1845 (Nematoda: Anisakidae) em Cetáceos encalhados no litoral do Ceará, Brasil

The gastric compartments of ten cetaceans stranded on the coast of Ceará State, Northeast Brazil were analyzed in this study. Gastric Anisakis spp. was diagnosed in all individuals involved in this study. Parasites and tissue samples were collected during necropsy. The presence of Anisakis parasites showed similar distribution across the three gastric compartments and the majority was free within the gastric lumen. Macroscopically, the lesions were predominantly characterized by the presence of ulcers (60 percent, 6/10) within the gastric mucosa, occasionally associated with edema and hemorrhage (30 percent, 3/10). Eight cetaceans (8/10 - 80 percent) presented gastric microscopic alterations and in 75 percent (6/8) of these animals, chronic lymphoplasmocytic gastritis was observed with varying degrees of distribution and severity. Additionally, eosinophilic and granulomatous inflammation with giant cells, hemosiderosis, fibrosis and areas of necrosis were associated with location of parasites within the gastric mucosa. In this study, it was shown that the majority of cetaceans with the presence of Anisakis parasites presented macro and microscopic gastric alterations. These nematodes are probably associated with the development of these alterations; however, more pathological approaches are still required.

Neste estudo, foram analisados os compartimentos gástricos de dez cetáceos encalhados na costa do Ceará, região nordeste do Brasil. Parasitos do gênero Anisakis foram diagnosticados em todos os indivíduos estudados. As amostras parasitárias e teciduais foram coletadas durante o procedimento de necropsia. A presença de Anisakis demonstrou distribuição similar nos três compartimentos gástricos, estando a maior parte dos parasitos livre na mucosa gástrica. Macroscopicamente, as lesões se caracterizaram principalmente pela presença de úlceras (60 por cento, 6/10) gástricas, associadas ocasionalmente a edema e hemorragia (30 por cento, 3/10). Oito cetáceos (8/10 - 80 por cento) apresentaram alterações gástricas microscópicas e em 75 por cento (6/8) destes animais, foi observada uma gastrite crônica linfoplasmocitária com vários graus de distribuição e severidade. Foram constatados ainda, focos de inflamação granulomatosa com a presença de eosinófilos, células gigantes e a formação de hemossiderose, fibrose e de necrose em torno dos parasitos. Neste estudo, a maioria dos cetáceos com a presença de parasitos do gênero Anisakis apresentaram alterações gástricas macro e microscópicas. Estes nematóides estão provavelmente associados ao desenvolvimento destas alterações, entretanto, mais estudos são ainda necessários.

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

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

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

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

Enterobius vermicularis: ancient DNA from north and south American human coprolites

A molecular paleoparasitological diagnostic approach was developed for Enterobius vermicularis. Ancient DNA was extracted from 27 coprolites from archaeological sites in Chile and USA. Enzymatic amplification of human mtDNA sequences confirmed the human origin. We designed primers specific to the E. vermicularis 5S ribosomal RNA spacer region and they allowed reproducible polymerase chain reaction identification of ancient material. We suggested that the paleoparasitological microscopic identification could accompany molecular diagnosis, which also opens the possibility of sequence analysis to understand parasite-host evolution

Diagnóstico molecular de Enterobius vermicularis (Lannaeus, 1758) em populações pré-históricas: análise da região intergênica do gene ribossomal 5S RNA e do gene SL 1 RNA / Molecular diagnosis of Enterobius vermicularis (Lannaeus, 1758) in prehistoric populations: analysis of the intergenic region of the ribossomal gene 5S RNA and gene SL 1 RNA

As análises de DNA antigo (aDNA) recuperado de espécimes arqueológicos têm revolucionado áreas das Ciências especialmente na evolução humana e na origem das doenças humanas. Estudos paleoparasitológicos têm mostrado a presença do parasito Enterobius vermicularis em coprólitos de populações antigas das Américas. Até agora o diagnóstico de E. vermiculares em restos arqueológicos depende do exame microscópico. O propósito deste trabalho foi desenvolver uma abordagem metodológica para a recuperação de aDNA e o diagnóstico molecular de E. vermicularis a partir de coprólitos humanos. Propomos o diagnóstico molecular usando como alvo a região intergênica do gene ribossomal 5S RNA de E. vermicularis. Esta estratégia foi analisada quanto a sua especificidade e sensibilidade em parasitos recuperados de amostras fecais e em coprólitos experimentais...Cinqüenta e uma amostras fecais coletadas de pacientes de distintas regiões geográficas do Brasil, e positivas microscopicamente para diversos enteroparasitos incluindo 24 para E. vermicularis fizeram parte deste estudo. O diagnóstico molecular, considerando 420pb da região intergênica do gene 5S rRNA de E. vermicularis identificou 20 das 24 amostras positivas microscopicamente para o parasito, e quando a região de 198pb foi considerada, 100 por cento das amostras analisadas positivas pela análise microscópica, inclusive amostras co-infetadas, foram positivas. A PCR da de 198pb foi sensível para diagnosticar E. vermicularis em amostras fecais com apenas um único ovo do parasito. Os métodos de polimerização reconstrutora e de Nested PCR previamente aplicados em coprólitos experimentais mostraram se úteis na recuperação de aDNA de E. vermicularis em 9 amostras de coprólitos do total de 27 amos5S rRNA de 9 amostras fecais de distintas regiões do Brasil e 7 de coprólitos da América do Norte e da América do Sul, revelou um alto grau de conservação nesta região e a presença do gene SL1 RNA. Nós sugerimos a estrutura secundária do gene SL1 de E. vermicularis, a qual é similar à estrutura modelo de três hastes-alças, e com as mesmas peculiaridades que, em outros organismos são essenciais para a reação de trans-splincing. Pela primeira vez é demonstrada a recuperação de seqüências de aDNA de E. vermicularis provenientes de restos arqueológicos. O diagnóstico molecular específico e sensível para o parasito e a identificação da seqüência e estrutura secundária do gene SL1 RNA de E. vermicularis, também são resultados inéditos.

Paleoparasitology: perspectives with new techniques

Paleoparasitologia e o estudo de parasitos encontrados em material arqueologico. O desenvolvimento deste campo da pesquisa teve inicio com a identificacao de ovos de helmintos em tecidos mumificados, analise de coprolitos e, recentemente, atraves da biologia molecular. Neste artigo faz-se uma breve revisao da historia da paleoparasitologia com referencia especial aos estudos de ADN antigo (ancient DNA) em material arqueologico