Microbiome and virome on indoor surfaces of an Antarctic research ship.

BACKGROUND: Few studies have focused on microbial diversity in indoor environments of ships, as well as the role of the microbiome and its ecological interconnections. In this study, we investigated the microbiome and virome present on the internal surfaces of a polar ship in different stages (beginning, during, and at the end) of the Brazilian Antarctic expedition in order to evaluate abundance of microorganisms in different periods. OBJECTIVES AND METHODS: We used shotgun metagenomic analysis on pooled samples from sampling surfaces in the ship's interior to track the microbial diversity. FINDINGS: Considering the total fraction of the microbiome, the relative abundance of bacteria, eukaryotes, viruses, and archaea was 83.7%, 16.2%, 0.04%, and 0.002%, respectively. Proteobacteria was the most abundant bacterial phyla, followed by Firmicutes, Actinobacteria, and Bacteroidetes. Concerning the virome, the greatest richness of viral species was identified during the middle of the trip, including ten viral families after de novo assembly: Autographiviridae, Chrysoviridae, Genomoviridae, Herelleviridae, Myoviridae, Partitiviridae, Podoviridae, Potyviridae, Siphoviridae, and Virgaviridae. MAIN CONCLUSIONS: This study contributed to the knowledge of microbial diversity in naval transportation facilities, and variations in the abundance of microorganisms probably occurred due to factors such as the number of passengers and activities on the ship.

Genomic epidemiology unveils the dynamics and spatial corridor behind the Yellow Fever virus outbreak in Southern Brazil.

Despite the considerable morbidity and mortality of yellow fever virus (YFV) infections in Brazil, our understanding of disease outbreaks is hampered by limited viral genomic data. Here, through a combination of phylogenetic and epidemiological models, we reconstructed the recent transmission history of YFV within different epidemic seasons in Brazil. A suitability index based on the highly domesticated Aedes aegypti was able to capture the seasonality of reported human infections. Spatial modeling revealed spatial hotspots with both past reporting and low vaccination coverage, which coincided with many of the largest urban centers in the Southeast. Phylodynamic analysis unraveled the circulation of three distinct lineages and provided proof of the directionality of a known spatial corridor that connects the endemic North with the extra-Amazonian basin. This study illustrates that genomics linked with eco-epidemiology can provide new insights into the landscape of YFV transmission, augmenting traditional approaches to infectious disease surveillance and control.

INTESTINAL PARASITES IN PECARI TAJACU AND SUS SCROFA DOMESTICUS IN THE CAATINGA FROM SOUTHEASTERN PIAUÍ, BRAZIL.

This study identifies gastrointestinal parasites in the feces of Pecari tajacu (caititu) and Sus scrofa domesticus (domestic pig) in southeastern Piauí, Brazil. The region covers 2 protected areas, Serra da Capivara National Park and Serra das Confusões National Park, and surrounding communities. Fecal samples from 64 animals, 42 from domestic swine and 22 from caititu, collected between 1985 and 2013, were analyzed by optical microscopy. Helminths and/or protozoa were found in 64% of the domestic pig samples and 27% of the caititu samples, totaling 18 morphospecies: Nematoda, Spirurida (2 morphospecies), Trichostrongyloidea, Eimeriidae, Aspidodera sp., Bertiella sp., Metastrongylus sp., Trichostrongylus sp., Moniezia sp., Gongylonema sp., Trichuris suis, Spirocerca lupi, Macracanthorhyncus hirudinaceus, Globocephalus urosubulatus, Strongyloides cf ransomi, Balantioides coli, and Eimeria cf scabra. The highest parasite diversity was obtained in the pig samples, totaling 15 morphospecies, compared to only 6 in caititus, with S. cf ransomi, G. urosubulatus, and S. lupi present in both hosts. We discuss the presence of parasites associated with domestic animals around the Protected Areas and potentially zoonotic parasites close to human communities, which raise concerns about the conservation of wildlife, human health, and livestock in the region.

Microbiome and virome on indoor surfaces of an Antarctic research ship

BACKGROUND Few studies have focused on microbial diversity in indoor environments of ships, as well as the role of the microbiome and its ecological interconnections. In this study, we investigated the microbiome and virome present on the internal surfaces of a polar ship in different stages (beginning, during, and at the end) of the Brazilian Antarctic expedition in order to evaluate abundance of microorganisms in different periods. OBJECTIVES AND METHODS We used shotgun metagenomic analysis on pooled samples from sampling surfaces in the ship's interior to track the microbial diversity. FINDINGS Considering the total fraction of the microbiome, the relative abundance of bacteria, eukaryotes, viruses, and archaea was 83.7%, 16.2%, 0.04%, and 0.002%, respectively. Proteobacteria was the most abundant bacterial phyla, followed by Firmicutes, Actinobacteria, and Bacteroidetes. Concerning the virome, the greatest richness of viral species was identified during the middle of the trip, including ten viral families after de novo assembly: Autographiviridae, Chrysoviridae, Genomoviridae, Herelleviridae, Myoviridae, Partitiviridae, Podoviridae, Potyviridae, Siphoviridae, and Virgaviridae. MAIN CONCLUSIONS This study contributed to the knowledge of microbial diversity in naval transportation facilities, and variations in the abundance of microorganisms probably occurred due to factors such as the number of passengers and activities on the ship.

Molecular Detection of Histoplasma capsulatum in Antarctica.

We detected Histoplasma capsulatum in soil and penguin excreta in the Antarctic Peninsula by sequencing after performing species-specific PCR, confirming previous observations that this pathogen occurs more broadly than suspected. This finding highlights the need for surveillance of emerging agents of systemic mycoses and their transmission among regions, animals, and humans in Antarctica.

Virome analysis in lakes of the South Shetland Islands, Antarctica - 2020.

Polar freshwater ecosystems are characterized by a distinct microbiota. However, little is known about viral diversity and abundance, especially regarding the ecology of RNA viruses. We used shotgun metagenomic analysis on samples from Antarctic ecosystems, and report here the characterization of the virome fraction, from different lakes located in the South Shetland Islands (Penguin, Ardley, Deception and King George Island) in the Peninsula Antarctica, in the summer season 2020. DNA viruses (99.4 %) prevailed over RNA viruses (0.6 %) in the lake samples. Six viral orders were identified in the metagenomic libraries: Caudovirales (dsDNA), which was prevalent in most lakes; Picornavirales (ssRNA+); Sobelivirales (ssRNA+); Tolivirales (ssRNA+); Petitvirales (ssDNA) and Baphyvirales (ssDNA), including eight viral families (Herelleviridae, Siphoviridae, Myoviridae, Microviridae, Marnaviridae, Bacilladnaviridae, Barnaviridae and Tombusviridae) and several other, mainly non-classified ssRNA(+) viruses in the lakes of Ardley Island. Bacteriophages (dsDNA) (Herelleviridae family) infecting the phylum Firmicutes and Siphoviridae were predominant in most lakes evaluated. Functional analysis demonstrated a prevalence of unknown proteins (68 %) in the virome. Our prospective study provides virome analysis data from different lakes in the South Shetland Islands, Antarctica, opening exploratory lines for future research related to the biodiversity and viral ecology in this extreme ecosystem.

Influenza A(H11N2) Virus Detection in Fecal Samples from Adélie (Pygoscelis adeliae) and Chinstrap (Pygoscelis antarcticus) Penguins, Penguin Island, Antarctica.

Influenza A viruses infect a range of host species, including a large variety of mammals and more than a hundred species of birds. A total of 95 avian fecal samples were collected from penguin colonies in the South Shetland Islands, close to the Antarctic Peninsula, and tested by reverse transcription-PCR (RT-PCR) to detect avian influenza viruses (AIVs). Five out of seven samples collected from Penguin Island were positive for AIVs. Analysis of the genomes recovered from four samples revealed the detection of influenza A(H11N2) virus in fecal samples from Adélie penguins (Pygoscelis adeliae) and from a colony of chinstrap penguins (Pygoscelis antarcticus). Bayesian phylogeographic analysis revealed the clustering of all currently available H11N2 samples from Antarctica's avifauna in a single cluster that emerged at least in the early 2010s, suggesting its continued circulation on the continent. Our results reinforce the need for continuous surveillance of avian influenza on the Antarctic continent. IMPORTANCE Although wild birds play a role in the transmission and ecology of avian influenza viruses (AIVs) across the globe, there are significant gaps in our understanding of the worldwide distribution of these viruses in polar environments. In this study, using molecular analysis and full-genome sequencing, we describe the detection of distinct influenza A(H11N2) viruses in fecal samples of penguins in the Southern Shetland Islands, Antarctica. We emphasize the need for virus monitoring as AIVs may have implications for the health of endemic fauna and the potential risk of the introduction of highly pathogenic AIVs to the continent.

Historical geographic overlap of human rickettsiosis with animal reservoirs in Juiz de Fora, Minas Gerais, Brazil / Sobreposição geográfica histórica da riquetsiose humana com reservatórios animais em Juiz de Fora, Minas Gerais, Brasil

Background and objectives: we investigated the existence of a historical geographic overlap between the location of spotted fever group rickettsiosis human cases, a disease caused by the gram-negative bacterium Rickettsia sp., and that of mammalian reservoirs, specifically domestic horses and capybaras, in the urban perimeter of the city of Juiz de Fora, Minas Gerais, Brazil. Methods: cases of human rickettsiosis that occurred during a period of 17 years (2003-2020) were geolocated and the distribution of cases in time and geographic space was assessed using 1st and 2nd order geospatial association indicators. We also analyzed the overlap between the locations of human rickettsiosis cases and the area of occurrence of domestic horses and capybaras. Results: men were diagnosed more often than women, but a large proportion of affected women died. The results indicate an aggregation of human rickettsiosis cases in time (cases tend to occur close to each other at each epidemic event) and in geographic space (cases are concentrated in a specific geographic region of the urban perimeter). Human cases seem to be more associated with city regions with: i) higher local frequency of domestic horses and not capybaras; ii) lower rates of family development. Conclusion: it is suggested that, in the local epidemiological scenario, domestic horses appear to be the main sources of the rickettsia infecting humans, not capybaras.(AU)

Justificativa e objetivos: foi investigada a existência de uma sobreposição geográfica histórica entre a localização dos casos de riquetsiose humana do grupo da febre maculosa, um grupo de doenças causadas pela bactéria Gram-negativa Rickettsia sp., e dos reservatórios mamíferos, especificamente cavalos domésticos e capivaras, no perímetro urbano do município de Juiz de Fora, Minas Gerais, Brasil. Métodos: foram geolocalizados os casos de riquetsiose humana ocorridos durante um período de 17 anos (2003-2020), sendo avaliada a distribuição dos casos no tempo e espaço geográfico através de indicadores de associação geoespacial de 1a e 2a ordem. Também analisamos a superposição dos locais dos casos de riquetsiose humana com a área de ocorrência de cavalos domésticos e capivaras. Resultados: homens foram diagnosticados mais frequentemente que as mulheres, mas grande proporção das mulheres acometidas faleceu. Os resultados indicam uma agregação dos casos de riquetsiose humana no tempo (os casos tendem a ocorrer próximos entre si a cada evento epidêmico) e no espaço geográfico (os casos se concentram em uma região geográfica específica do perímetro urbano). Os casos humanos aparentam ser mais associados às regiões da cidade com: i) maior frequência local de cavalos domésticos e não das capivaras; ii) menores índices de desenvolvimento familiar. Conclusão: sugere-se que, no cenário epidemiológico local, são os cavalos domésticos que aparentam ser as principais fontes da riquétsia infectando os humanos, não as capivaras.(AU)

Justificación y objetivos: investigamos la existencia de una superposición geográfica histórica entre la localización de casos de rickettsiosis exantemáticas humana, enfermedad causada por la bacteria gramnegativa Rickettsia sp., y la de mamíferos reservorios, específicamente caballos domésticos y capibaras, en el perímetro urbano de la ciudad de Juiz de Fora, Minas Gerais, Brasil. Métodos: se geolocalizaron los casos de rickettsiosis humana ocurridos durante un período de 17 años (2003-2020), y se evaluó la distribución de casos en el tiempo y espacio geográfico utilizando indicadores de asociación geoespacial de 1er y 2do orden. También analizamos la superposición entre las ubicaciones de los casos de rickettsiosis humana y el área de ocurrencia de los caballos domésticos y capibaras. Resultados: los hombres fueron diagnosticados con más frecuencia que las mujeres, pero una gran proporción de mujeres afectadas fallecieron. Los resultados indican una agregación de casos de rickettsiosis humana en el tiempo (los casos tienden a ocurrir cerca uno del otro en cada evento epidémico) y en el espacio geográfico (los casos se concentran en una región geográfica específica del perímetro urbano). Los casos humanos parecen estar más asociados con regiones urbanas con: i) mayor frecuencia local de caballos domésticos y no de capibaras; ii) menores tasas de desarrollo familiar. Conclusión: se sugiere que, en el escenario epidemiológico local, los caballos domésticos parecen ser las principales fuentes de la rickettsia que infecta a los humanos, no los capibaras.(AU)

Statistically enriched geospatial datasets of Brazilian municipalities for data-driven modeling.

The lack of georeferencing in geospatial datasets hinders the accomplishment of scientific studies that rely on accurate data. This is particularly concerning in the field of health sciences, where georeferenced data could lead to scientific results of great relevance to society. The Brazilian health systems, especially those for Notifiable Diseases, in practice do not register georeferenced data; instead, the records indicate merely the municipality in which the event occurred. Typically in data-driven modeling, accurate disease prediction models based on occurrence requires socioenvironmental characteristics of the exact location of each event, which is often unavailable. To enrich the expressiveness of data-driven models when the municipality of the event is the best available information, we produced datasets with statistical characterization of all 5,570 Brazilian municipalities in 642 layers of thematic data that represent the natural and artificial characteristics of the municipalities' landscapes over time. This resulted in a collection of datasets comprising a total of 11,556 descriptive statistics attributes for each municipality.

PARASITES OF THE BRAZILIAN ROCK CAVY, KERODON RUPESTRIS: REVEALING THEIR HISTORY IN THE BRAZILIAN SEMIARID REGION.

Using microscopy and/or immunodiagnosis, the authors analyzed 284 fecal samples from the Brazilian rock cavy, Kerodon rupestris, that were collected between 1984 and 2015 in Serra da Capivara National Park for the presence of helminths and protozoa. Fourteen morphospecies of helminth eggs of the following taxa were found: Trematoda, Nematoda, Strongylidae, Lagochilascaris sp., Strongylida, Trichuris (2 species), Oxyuridae (3 species), Ancylostomatidae (2 species), and Ascarididae (2 species), along with 3 protozoan taxa: Coccidia, Cryptosporidium sp., and Balantidium sp. During the last 30 yr, the population of K. rupestris has increased in the region as a consequence of the creation and management of the National Park, and data from this study show a concurrent increase in the diversity of intestinal parasites in this host, including new reports. Some of these species have zoonotic potential, which suggests that K. rupestris may be in contact with domestic farm animals and/or human feces. These results show the importance of integrating different diagnostic approaches for the identification of protozoa in the region and indicate that further methods need to be employed to increase recovery. This work highlights the usefulness of parasite studies in assessing the health of ecosystems, especially in protected areas, which should be considered by park managers and health agencies.

Chagas disease in the context of the 2030 agenda: global warming and vectors.

The 2030 Agenda for Sustainable Development is a plan of action for people, planet and prosperity. Thousands of years and centuries of colonisation have passed the precarious housing conditions, food insecurity, lack of sanitation, the limitation of surveillance, health care programs and climate change. Chagas disease continues to be a public health problem. The control programs have been successful in many countries in reducing transmission by T. cruzi; but the results have been variable. WHO makes recommendations for prevention and control with the aim of eliminating Chagas disease as a public health problem. Climate change, deforestation, migration, urbanisation, sylvatic vectors and oral transmission require integrating the economic, social, and environmental dimensions of sustainable development, as well as the links within and between objectives and sectors. While the environment scenarios change around the world, native vector species pose a significant public health threat. The man-made atmosphere change is related to the increase of triatomines' dispersal range, or an increase of the mobility of the vectors from their sylvatic environment to man-made constructions, or humans getting into sylvatic scenarios, leading to an increase of Chagas disease infection. Innovations with the communities and collaborations among municipalities, International cooperation agencies, local governmental agencies, academic partners, developmental agencies, or environmental institutions may present promising solutions, but sustained partnerships, long-term commitment, and strong regional leadership are required. A new world has just opened up for the renewal of surveillance practices, but the lessons learned in the past should be the basis for solutions in the future.

Callitrichine gammaherpesvirus 3 and Human alphaherpesvirus 1 in New World Primate negative for yellow fever virus in Rio de Janeiro, Brazil.

BACKGROUND: Herpesvirus transmission between humans and non-human primate (NHP) can occur through contact scratches with lesions, infected saliva, and mainly through contaminated food. Therefore, cross-infection can lead to severe illness or even death for both the animal and human. In 2017, during the yellow fever (YF) outbreak in Brazil, species of the New World Primates (NWP) from Rio de Janeiro state, tested negative for yellow fever virus (YFV) detection. OBJECTIVES: To evaluate herpesvirus in the population NWP in Rio de Janeiro. METHODS: To investigate, liver samples of 283 NWP, from several regions of the state of Rio de Janeiro, were tested for the herpesvirus family using a Pan-polymerase chain reaction (Pan-PCR) and sequencing. FINDINGS: 34.6% (98/283) tested positive for at least one herpesvirus; 29.3% (83/283) tested positive to Human alphaherpesvirus 1 (HSV-1), this virus from humans can be lethal to New World monkey; 13% (37/283) were detected Callitrichine gammaherpesvirus 3 (CalHV-3), responsible for lymphoproliferative disease that can be fatal in NWP. In addition, CalHV-3 / HSV-1 co-infection was in 11.6% (33/283) of the samples. MAIN CONCLUSIONS: Pan-herpesvirus was useful to identify species-specific herpesviruses and virus from human that can infect animals. Furthermore, during an outbreak of YF other infections should be monitored.

Back to basics: Could simple experiments resolve important parasitology enigmas? The rarity of Ascaris eggs compared with Trichuris eggs in archeology and other contexts.

Most parasitologists face a conundrum throughout their careers: 'which parasite eggs are more resistant to environmental stress, Ascaris spp. or Trichuris spp.?' In this analysis, our experimental and statistical analyses demonstrated that Trichuris sp. is more resistant than Ascaris sp. We highlight that desiccation exerts a major effect on the conservation of Ascaris eggs, and this may result in an underestimation of Ascaris eggs in paleoparasitological records. This observation can be extrapolated to more modern scenarios, for example, parasitological research in animal feces from semiarid environments, where whipworms are more common than roundworms. Similarly, this could be a plausible explanation for the higher frequency and abundance of whipworms than roundworms, when other hypotheses are unsupported.

Chagas disease in the context of the 2030 agenda: global warming and vectors

The 2030 Agenda for Sustainable Development is a plan of action for people, planet and prosperity. Thousands of years and centuries of colonisation have passed the precarious housing conditions, food insecurity, lack of sanitation, the limitation of surveillance, health care programs and climate change. Chagas disease continues to be a public health problem. The control programs have been successful in many countries in reducing transmission by T. cruzi; but the results have been variable. WHO makes recommendations for prevention and control with the aim of eliminating Chagas disease as a public health problem. Climate change, deforestation, migration, urbanisation, sylvatic vectors and oral transmission require integrating the economic, social, and environmental dimensions of sustainable development, as well as the links within and between objectives and sectors. While the environment scenarios change around the world, native vector species pose a significant public health threat. The man-made atmosphere change is related to the increase of triatomines' dispersal range, or an increase of the mobility of the vectors from their sylvatic environment to man-made constructions, or humans getting into sylvatic scenarios, leading to an increase of Chagas disease infection. Innovations with the communities and collaborations among municipalities, International cooperation agencies, local governmental agencies, academic partners, developmental agencies, or environmental institutions may present promising solutions, but sustained partnerships, long-term commitment, and strong regional leadership are required. A new world has just opened up for the renewal of surveillance practices, but the lessons learned in the past should be the basis for solutions in the future.

Fiocruz Biological Collections: strengthening Brazil's biodiversity knowledge and scientific applications opportunities.

Biological collections are central in understanding and preserving life on Earth. In Brazil, the most representative collections are kept by natural history museums, whose primary focus is in invertebrates, vertebrates and vascular plants. Only a few institutions keep repositories in different kingdoms. The Oswaldo Cruz Foundation (Fiocruz), established in 1900, is a strategic public health institution of the Ministry of Health of Brazil. As such, Fiocruz is responsible for a wide range of activities, from basic research to the development and production of vaccines, drugs, reagents and diagnostic kits. Its biological collections were soon established in the expeditions made by naturalists and physicians seeking integrated knowledge of the fauna, flora and tropical diseases. Since then, they have been part of the institutional policy. In a few decades, those collections were already in the forefront of basic and applied research on tropical parasitic and infectious diseases. Currently, they comprise thirty-three repositories representing part of the Brazilian diversity of bacteria, fungi, protozoa, helminths, arthropods, molluscs and plants of medical and environmental importance. Different methods of long-term preservation are applied for the conservation of this wide range of organisms represented by about 6 million specimens. Herein, we describe this range of collections and discuss their complementary role as repositories of groups not represented in other biological collections in Brazil. These valuable biological materials have been used in public health and medical research, as well as for technological development and innovation in Brazil. Parallel to this specific usage, Fiocruz biological collections have played and continue to play a unique and important role in understanding and conserving part of Brazil's biodiversity that is currently under-represented in other biological and natural history collections in Brazil and South America.

Are immunoenzymatic tests for intestinal protozoans reliable when used on archaeological material?

Intestinal protozoans found in ancient human samples have been studied primarily by microscopy and immunodiagnostic assays. However, such methods are not suitable for the detection of zoonotic genotypes. The objectives of the present study were to utilize immunoenzimatic assays for coproantigen detection of Cryptosporidium sp., Giardia duodenalis, and Entamoeba histolytica/Entamoeba dispar in sixty ancient human and animal samples collected from 14 archaeological sites in South America, and to carry out a critical analysis of G. duodenalis according to results obtained from three diagnostic methodologies: microscopy, immunodiagnostic tests (immunoenzymatic and immunofluorescence), and molecular biology (PCR and sequencing). More than half (31/60) of the samples analyzed using immunoenzymatic tests were positive for at least one of the intestinal protozoans, with 46.6% (28/60) corresponding to G. duodenalis, 26.6% (16/60) to Cryptosporidium sp., and 5% (3/60) to E. histolytica/E. dispar. Cryptosporidium sp. and G. duodenalis coinfection was observed in 15% (9/60) of the samples, whereas all three protozoans were found in 5% (3/60) of samples. In the Northeast Region of Brazil, by immunoenzymatic tests there is evidence that G. duodenlais and Cryptosporidium sp. have infected humans and rodents for at least 7150 years. However, for G. duodenalis, the results from the three diagnostic tests were discordant. Specifically, despite the efficiency of the molecular biology assay in the experimental models, G. duodenalis DNA could not be amplified from the ancient samples. These results raise the following question: Are all ancient samples positive for coproantigen of G. duodenalis by immunoenzymatic tests truly positive? This scenario highlights the importance of further studies to evaluate the sensitivity and specificity of the immunoenzymatic method in the archaeological context.

SISS-Geo: Leveraging Citizen Science to Monitor Wildlife Health Risks in Brazil.

The well-being of human and wildlife health involves many challenges, such as monitoring the movement of pathogens; expanding health surveillance; collecting data and extracting information to identify and predict risks; integrating specialists from different areas to handle data, species and distinct social and environmental contexts; and the commitment to bringing relevant information to society. In Brazil, there is still the difficulty of building a system that is not impaired by its large territorial extension and its poorly integrated sectoral policies. The Brazilian Wildlife Health Information System, SISS-Geo (SISS-Geo is the abbreviation of "Sistema de Informação em Saúde Silvestre Georreferenciado" (which translates to "Georeferenced Wildlife Health Information System") and can be accessed at http://www.biodiversidade.ciss.fiocruz.br or http://sissgeo.lncc.br (in Portuguese)), is a platform for collaborative monitoring that intends to overcome the challenges in wildlife health. It aims at the integration and participation of various segments of society, encompassing the registration of animals occurrences by citizen scientists; the reliable diagnosis of pathogens from the laboratory and expert networks; and computational and mathematical challenges in analytical and predictive systems, model interpretation, data integration and visualization, and geographic information systems. It has been successfully applied to support decision-making on recent wildlife health events, such as a Yellow Fever epizootic.

Methodological innovations for the study of irreplaceable samples reveal giardiasis in extinct animals (Nothrotherium maquinense and Palaeolama maior).

The use of diagnostic methods that prevent irreplaceable samples (from museum collections, archaeological and paleontological samples) of being consumed or that increase their yield is relevant. For museum collections, archaeological and paleontological samples it is essential to conserve samples, subsamples or portions for future research. We are addressing methods for conservation of irreplaceable samples that could be fully consumed. Innovations in methodologies that are used in studies of Paleoparasitology and Paleomicrobiology will contribute to the preservation of collections. Therefore, to the development of archaeology and paleontology in the future, we evaluated whether the discarded material of the immunochromatography test could be used for molecular diagnosis and vice versa. We used a genotyped experimental coprolite positive for Giardia duodenalis. The diagnosis was positive for giardiasis in both cases. This methodology can be corroborated with the coprolite of a Paleolama maior (extinct llama) previously diagnosed for G. duodenalis with an immunoenzymatic test. The residue of the pre-digestion step of the DNA extraction before adding Proteinase K was confirmed positive with the immunochromatographic test. Also, the DNA extraction residue from a coprolite of Nothrotherium maquinense (ground sloth) was tested positive with immunochromatographic test for G. duodenalis. These are the oldest findings for G. duodenalis confirming that this intestinal parasite occurred among Northeastern Brazilian Megafauna animals from the late Pleistocene period, correlated to human occupation. The relevance of these results will allow the study by different methodological approaches from a small amount of material, reusing discarded materials.

Study of infectious diseases in archaeological bone material - A dataset.

Bones of human and ground sloth remains were analyzed for presence of Trypanosoma cruzi by conventional PCR using primers TC, TC1 and TC2. Sequence results amplified a fragment with the same product size as the primers (300 and 350pb). Amplified PCR product was sequenced and analyzed on GenBank, using Blast. Although these sequences did not match with these parasites they showed high amplification with species of bacteria. This article presents the methodology used and the alignment of the sequences. The display of this dataset will allow further analysis of our results and discussion presented in the manuscript "Finding the unexpected: a critical view on molecular diagnosis of infectious diseases in archaeological samples" (Pucu et al. 2017) [1].