Fiocruz in Antarctica - health and environmental surveillance facing the challenges of the 21st century.

FioAntar, FIOCRUZ's research project in Antarctica, is based on the One Health approach. FioAntar aims to generate relevant information that will help reduce the risk of future pandemics and improve the search for chemical compounds and new biological molecules. After four expeditions to Antarctica under the scope of PROANTAR, Fiocruz has identified Influenza H11N2 virus in environmental fecal samples, as well as Histoplasma capsulatum and Bacillus cereus in soil samples. In addition, in a prospective virome analysis from different lakes in the South Shetland Islands, six viral orders were described, supporting future research related to the biodiversity and viral ecology in this extreme ecosystem. Our findings of environmental pathogens of public health importance are a warning about the urgency of establishing a surveillance agenda on zoonoses in Antarctica due to the imminent risks that ongoing environmental and climate changes impose on human health across the planet. FioAntar strives to establish a comprehensive surveillance program across Antarctica, monitoring circulation of pathogens with the potential to transcend continent boundaries, thereby mitigating potential spread. For Fiocruz, Antarctica signifies a new frontier, teeming with opportunities to explore novel techniques, refine established methodologies, and cultivate invaluable knowledge.

Active surveillance for influenza virus and coronavirus infection in Antarctic birds and mammals in environmental fecal samples, South Shetland Islands.

Numerous Antarctic species are recognized as reservoirs for various pathogens, and their migratory behavior allows them to reach the Brazilian coast, potentially contributing to the emergence and circulation of new infectious diseases. To address the potential zoonotic risks, we conducted surveillance of influenza A virus (IAV) and coronaviruses (CoVs) in the Antarctic Peninsula, specifically focusing on different bird and mammal species in the region. During the summer of 2021/2022, as part of the Brazilian Antarctic Expedition, we collected and examined a total of 315 fecal samples to target these respiratory viruses. Although we did not detect the viruses of interest during this particular expedition, previous research conducted by our team has shown the presence of the H11N2 subtype of influenza A virus in penguin fecal samples from the same region. Given the continuous emergence of new viral strains worldwide, it is crucial to maintain active surveillance in the area, contributing to strengthening integrated One Health surveillance efforts.

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.

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.

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.

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.

Socioecological vulnerability and the risk of zoonotic disease emergence in Brazil.

In developing countries, outbreaks of zoonotic diseases (ZDs) result from intertwined ecological, socioeconomic, and demographic processes that shape conditions for (i) increased contact between vulnerable human population and wildlife in areas undergoing environmental degradation and (ii) the rapid geographic spread of infections across socially vulnerable regions. In Brazil, recent increases in environmental and social vulnerabilities, amplified by economic and political crises, are potential triggers for outbreaks. We discuss Brazilian features that favor outbreaks and show a novel quantitative method for zoonotic risk assessment. Using data on nine ZDs from 2001 to 2019, we found that the most significant causal variables were vegetation cover and city remoteness. Furthermore, 8 of 27 states presented low-level risk of ZD outbreaks. Given the ZD-bushmeat connection, we identified central hunted mammals that should be surveilled to prevent spillover events. The current challenge is to coordinate intersectoral collaboration for effective One Health management in megadiverse countries with high social vulnerability and growing environmental degradation like Brazil.

Evaluation of HBV-Like Circulation in Wild and Farm Animals from Brazil and Uruguay.

The origin of the hepatitis B virus is a subject of wide deliberation among researchers. As a result, increasing academic interest has focused on the spread of the virus in different animal species. However, the sources of viral infection for many of these animals are unknown since transmission may occur from animal to animal, human to human, animal to human, and human to animal. The aim of this study was to evaluate hepadnavirus circulation in wild and farm animals (including animals raised under wild or free conditions) from different sites in Brazil and Uruguay using serological and molecular tools. A total of 487 domestic wild and farm animals were screened for hepatitis B virus (HBV) serological markers and tested via quantitative and qualitative polymerase chain reaction (PCR) to detect viral DNA. We report evidence of HBsAg (surface antigen of HBV) and total anti-HBc (HBV core antigen) markers as well as low-copy hepadnavirus DNA among domestic and wild animals. According to our results, which were confirmed by partial genome sequencing, as the proximity between humans and animals increases, the potential for pathogen dispersal also increases. A wider knowledge and understanding of reverse zoonoses should be sought for an effective One Health response.

Coxiella and Bartonella spp. in bats (Chiroptera) captured in the Brazilian Atlantic Forest biome.

BACKGROUND: The role of bats as reservoirs of zoonotic agents, especially pathogenic bacteria such as Bartonella and Coxiella, has been discussed around the world. Recent studies have identified bats as potential hosts of species from the proteobacteria phylum. In Brazil, however, the role of bats in the natural cycle of these agents is poorly investigated and generally neglected. In order to analyze the participation of bats in the epidemiology of diseases caused by Bartonella, Coxiella, Rickettsia, Anaplasma and Ehrlichia, we conducted a descriptive epidemiological study in three biogeographic regions of the Brazilian Atlantic Forest. RESULTS: Tissues of 119 bats captured in preserved areas in the states of Rio de Janeiro, Bahia and Santa Catarina from 2014 to 2015 were submitted to molecular analysis using specific primers. Bartonella spp. was detected in 22 spleen samples (18.5%, 95% CI: 11.9-26.6), whose phylogenetic analysis revealed the generation of at least two independent clusters, suggesting that these may be new unique genotypes of Bartonella species. In addition, four samples (3.4%, 95% CI: 0.9-8.3) were positive for the htpAB gene of C. burnetii [spleen (2), liver (1) and heart (1)]. Rickettsia spp., Anaplasma and Ehrlichia were not identified. This is the first study reporting C. burnetii and Bartonella spp. infections in bats from the Atlantic Forest biome. CONCLUSIONS: These findings shed light on potential host range for these bacteria, which are characterized as important zoonotic pathogens.

From mammals back to birds: Host-switch of the acanthocephalan Corynosoma australe from pinnipeds to the Magellanic penguin Spheniscus magellanicus.

Trophically-transmitted parasites are regularly exposed to potential new hosts through food web interactions. Successful colonization, or switching, to novel hosts, occur readily when 'donor' and 'target' hosts are phylogenetically related, whereas switching between distantly related hosts is rare and may result from stochastic factors (i.e. rare favourable mutations). This study investigates a host-switching event between a marine acanthocephalan specific to pinnipeds that is apparently able to reproduce in Magellanic penguins Spheniscus magellanicus from Brazil. Detailed analysis of morphological and morphometrical data from acanthocephalans from penguins indicates that they belong to Corynosoma australe Johnston, 1937. Partial fragments of the 28S rRNA and mitochondrial cox1 genes were amplified from isolates from penguins and two pinniped species (i.e. South American sea lion Otaria flavescens and South American fur seal Arctocephalus australis) to confirm this identification. Infection parameters clearly differ between penguins and the two pinniped species, which were significantly lower in S. magellanicus. The sex ratio of C. australe also differed between penguins and pinnipeds; in S. magellanicus was strongly biased against males, while in pinnipeds it was close to 1:1. Females of C. australe from O. flavescens were smaller than those from S. magellanicus and A. australis. However, fecundity (i.e. the proportion of fully developed eggs) was lower and more variable in females collected from S. magellanicus. At first glance, the occurrence of reproductive individuals of C. australe in Magellanic penguins could be interpreted as an adaptive colonization of a novel avian host through favourable mutations. However, it could also be considered, perhaps more likely, as an example of ecological fitting through the use of a plesimorphic (host) resource, since the ancestors of Corynosoma infected aquatic birds.

Molecular and morphological characterization of Contracaecum pelagicum (Nematoda) parasitizing Spheniscus magellanicus (Chordata) from Brazilian waters.

Three new sequences of Mitochondrial cytochrome c-oxidase subunit 2 (mtDNA cox-2) from C. pelagicum parasite of Spheniscus magellanicus, the Magelanicus penguin, were determined from Brazilian waters. The sequences presented 99 and 98% of similarity with C. pelagicum sequences from Argentina, deposited on GenBank for the same genetic region and with a strong statistical support inferred from the phylogenetic tree. The morphological and ultrastructural studies that were carried out confirmed the genetic analysis.

Molecular and morphological characterization of Contracaecum pelagicum (Nematoda) parasitizing Spheniscus magellanicus (Chordata) from Brazilian waters / Caracterização molecular e morfológica de Contracaecum pelagicum (Nematoda) parasito de Spheniscus magellanicus (Chordata) em águas brasileiras

Three new sequences of Mitochondrial cytochrome c-oxidase subunit 2 (mtDNA cox-2) from C. pelagicum parasite of Spheniscus magellanicus, the Magelanicus penguin, were determined from Brazilian waters. The sequences presented 99 and 98% of similarity with C. pelagicum sequences from Argentina, deposited on GenBank for the same genetic region and with a strong statistical support inferred from the phylogenetic tree. The morphological and ultrastructural studies that were carried out confirmed the genetic analysis.

Foram determinadas três novas sequências da região do Citocromo c-oxidase da subunidade II do DNA mitocondrial (cox-2 mtDNA) de Contracaecum pelagicum, parasito de Spheniscus magellanicus, pinguim Magalhães, de águas brasileiras. As sequências apresentaram 99 e 98% de similaridade com sequências de C. pelagicum da Argentina depositadas no GenBank da mesma região genética com forte suporte estatístico inferido pela arvore filogenética. Estudos morfológicos e ultraestruturais realizados confirmaram a identidade genética.

New records and descriptions of digeneans from the Magellanic penguin Spheniscus magellanicus (Forster) (Aves: Sphenisciformes) on the coast of Brazil.

Five species of digeneans parasitic in the Magellanic penguin Spheniscus magellanicus (Forster) from off the Brazilian coast of the western Atlantic are reported for the first time from this host and described. These are Mesostephanus odhneri (Travassos, 1924) Lutz, 1935, Posthodiplostomum macrocotyle Dubois, 1937, Stephanoprora uruguayensis Holcman-Spector & Olagüe, 1989, Ascocotyle (Phagicola) longa Ransom, 1920 and Ascocotyle (Phagicola) sp. One other digenean, Cardiocephaloides physalis (Lutz, 1926) Sudarikov, 1959, was also recorded. The taxonomy of the species and available data on their life-cycles are commented upon in relation to the possible origins of digenean infections of the Magellanic penguin.

Marine debris ingestion by Magellanic penguins, Spheniscus magellanicus (Aves: Sphenisciformes), from the Brazilian coastal zone.

Magellanic penguins (Spheniscus magellanicus) are non-breeding winter visitors to the Brazilian coast. In 2008 and 2010, plastic items and other marine debris were found in the stomachs and intestines of 15% of 175 dead penguins collected in the Lagos Region of the state of Rio de Janeiro. One bird had its stomach perforated by a plastic straw, which may have caused its death. There are few records of penguins ingesting plastic litter, but previous studies have found similar levels of debris ingestion among Magellanic penguins stranded on the Brazilian coast (35.8% of 397 birds). The high incidence of marine debris in this species in Brazil may result at least in part from the predominance of juveniles reaching these waters, as juvenile penguins may have a broader diet than adults. It is unclear to what extent plastic ingestion affects the mortality rate in this species and whether the incidence in stranded birds reflects that in the entire population. The present study addresses the increasing impact of plastic debris on marine life.

Degradation and detoxification of three textile Azo dyes by mixed fungal cultures from semi-arid region of Brazilian Northeast

The aim of this work was to study the degradation and detoxification of three textile azo dyes (Reactive Red 198, Reactive Red 141 and Reactive Blue 214) by mixed fungal cultures from semi-arid region of Brazilian Northeast. Sediment samples of twenty water reservoirs in the surroundings of Serra da Capivara National Park, area of environmental preservation in the caatinga in the State of Piauí, with semi-arid climate, were evaluated in order to select the consortia of fungi capable to degrade and detoxify these dyes. The mixed fungal culture from Caldeirão Escuridão (CE) reservoir was the most efficient in the degradation and detoxification of the dyes tested.

[Diversity of wild and domestic mammal's intestinal helminths from the Caatinga of the Parque Nacional Serra da Capivara, Southeast of Piauí, Brazil]. / Diversidade de helmintos intestinais em mamíferos silvestres e domésticos na Caatinga do Parque Nacional Serra da Capivara, Sudeste do Piauí, Brasil.

Biodiversity studies allow ecosystem assessment and monitoring of environmental changes and impacts. Parasite diversity could reflect the host/ parasite coevolutionary process and the environment changes that permit the loss, gain or maintenance of species. This survey used species/morphotypes of helminths eggs found in feces from seven wild mammal species (the groups Dasypodidae and Large Cats, and Tamandua tetradactyla, Cebus apella, Alouatta caraya, Cerdocyon thous, Pecari tajacu) and from two domestic species (Canis familiaris and Sus scrofa), which occur within the Serra da Capivara National Park (PNSC) and surrounding areas in order to analise the diversity of mammal intestinal helminths. This work used the helminthological fauna findings of wild and domestic mammals, to consider a possible helminth flux between these two host groups using Unweighted Pair Group Method with Arithmetic Mean (UPGMA) of the hosts based on helminthological fauna composition. The results indicate that the region of the PNSC still maintains environmental conditions that still keep wild mammal helminthological fauna composition different from the one found for domestic mammals.

Diversidade de helmintos intestinais em mamíferos silvestres e domésticos na Caatinga do Parque Nacional Serra da Capivara, Sudeste do Piauí, Brasil / Diversity of wild and domestic mammal’s intestinal helminths from the Caatinga of the Parque Nacional Serra da Capivara, Southeast of Piauí, Brazil

Estudos sobre biodiversidade permitem avaliações ecossistêmicas e monitoramentos de impactos e mudanças ambientais. A diversidade parasitária pode refletir o processo coevolutivo entre parasitos e hospedeiros, e mudanças ambientais que permitem a perda, ganho ou a manutenção das espécies nesse processo. O presente trabalho utilizou espécies/morfotipos de ovos de helmintos encontrados em fezes de sete mamíferos silvestres (os grupos Dasypodidae e grandes felinos, e as espécies: Tamandua tetradactyla, Cebus apella, Alouatta caraya, Cerdocyon thous, Pecari tajacu) e dois domésticos (Canis familiaris e Sus scrofa), que cohabitam o Parque Nacional Serra da Capivara (PNSC) e seu entorno, para analisar a diversidade dos helmintos intestinais desses mamíferos. Neste trabalho, utilizou-se achados da fauna helmintológica de mamíferos silvestres e domésticos, para avaliar um possível luxo de helmintos entre estes dois grupos hospedeiros usando Unweighted Pair Group Method with Arithmetic Mean (UPGMA) dos hospedeiros com base na composição de suas faunas helmintológicas. Os resultados indicam que a região do PNSC, ainda mantém condições ambientais que ainda propiciam a manutenção da composição da fauna de helmintos de mamíferos silvestres, diferente da encontrada nos animais domésticos.

Biodiversity studies allow ecosystem assessment and monitoring of environmental changes and impacts. Parasite diversity could relect the host/ parasite coevolutionary process and the environment changes that permit the loss, gain or maintenance of species. his survey used species/morphotypes of helminths eggs found in feces from seven wild mammal species (the groups Dasypodidae and Large Cats, and Tamandua tetradactyla, Cebus apella, Alouatta caraya, Cerdocyon thous, Pecari tajacu) and from two domestic species (Canis familiaris and Sus scrofa), which occur within the Serra da Capivara National Park (PNSC) and surrounding areas in order to analise the diversity of mammal intestinal helminths. his work used the helminthological fauna indings of wild and domestic mammals, to consider a possible helminth lux between these two host groups using Unweighted Pair Group Method with Arithmetic Mean (UPGMA) of the hosts based on helminthological fauna composition. he results indicate that the region of the PNSC still maintains environmental conditions that still keep wild mammal helminthological fauna composition diferent from the one found for domestic mammals.