Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear un derstanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5–7 vast areas of the tropics remain understudied.8–11 In the American tropics, Amazonia stands out as the world’s most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepre sented in biodiversity databases.13–15 To worsen this situation, human-induced modifications16,17 may elim inate pieces of the Amazon’s biodiversity puzzle before we can use them to understand how ecological com munities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple or ganism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region’s vulnerability to environmental change. 15%–18% of the most ne glected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lostinfo:eu-repo/semantics/publishedVersio

A social and ecological assessment of tropical land uses at multiple scales: the Sustainable Amazon Network

Science has a critical role to play in guiding more sustainable development trajectories. Here, we present the Sustainable Amazon Network (Rede Amazonia Sustentavel, RAS): a multidisciplinary research initiative involving more than 30 partner organizations working to assess both social and ecological dimensions of land-use sustainability in eastern Brazilian Amazonia. The research approach adopted by RAS offers three advantages for addressing land-use sustainability problems: (i) the collection of synchronized and co-located ecological and socioeconomic data across broad gradients of past and present human use; (ii) a nested sampling design to aid comparison of ecological and socioeconomic conditions associated with different land uses across local, landscape and regional scales; and (iii) a strong engagement with a wide variety of actors and non-research institutions. Here, we elaborate on these key features, and identify the ways in which RAS can help in highlighting those problems in most urgent need of attention, and in guiding improvements in land-use sustainability in Amazonia and elsewhere in the tropics. We also discuss some of the practical lessons, limitations and realities faced during the development of the RAS initiative so far.Keywords: Social–ecological systems, Tropical forests, Land use, Interdisciplinary research, Sustainability, Trade-off

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear understanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5,6,7 vast areas of the tropics remain understudied.8,9,10,11 In the American tropics, Amazonia stands out as the world's most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepresented in biodiversity databases.13,14,15 To worsen this situation, human-induced modifications16,17 may eliminate pieces of the Amazon's biodiversity puzzle before we can use them to understand how ecological communities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple organism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region's vulnerability to environmental change. 15%–18% of the most neglected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lost

Comparación entre dos métodos de obtención de DNA a ser usados como protocolos alternativos para la detección de parásitos humanos causadores de malaria por nested PCR

Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisas Básicas em Malária. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisas Básicas em Malária. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisas Básicas em Malária. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisas Básicas em Malária. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisas Básicas em Malária. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisas Básicas em Malária. Ananindeua, PA, Brasil.Um diagnóstico laboratorial correto e preciso de malária humana ainda é considerado um desafio, pois o método de referência, o da gota espessa com colocação pelo Giemsa, apresenta limitações que dificultam o controle da malária. Devido a esses problemas, várias pesquisas têm objetivado desenvolver métodos alternativos para o diagnóstico da malária. Grande parte desses estudos aborda métodos de diagnóstico molecular, que têm acarretado o desenvolvimento de algumas alternativas ao método de coloração pelo Giemsa. No entanto, esses métodos, por sua vez, apresentam suas limitações, que incluem seu alto custo, a complexidade do protocolo e a variação da qualidade das fontes de DNA e de reagentes. Neste aspecto, a técnica de nested PCR tem demonstrado ser um bom método e pode ser melhorado usando uma fonte de DNA de alta qualidade. Neste estudo foram avaliados dois métodos para a obtenção de DNA de amostras de sangue seco colhidas em papel de filtro: 1) lavagem e 2) saponina/Chelex-100. O segundo método apresentou sensibilidade e especificidade mais altas em relação ao primeiro, pois detectou mais infecções, tanto simples como mistas, bem como infecções por Plasmodium malariae. Com base nesses resultados, apresentamos o segundo como o protocolo de escolha para a obtenção de DNA. A técnica de nested PCR usando saponina/Chelex-100 para extração de DNA pode ser um método alternativo ou complementar de diagnóstico de parasitas da malária humana, mas não é considerado adequado para o uso de rotina.The correct and precise laboratory diagnosis of human malaria is still a challenge because the reference method, the Giemsa-stained thick blood smear (TS), has limitations that present problems for malaria control. Because of these problems, several studies have attempted to develop alternative methods for malaria diagnosis. Many of these studies focus on molecular diagnosis methods and have led to the development of some alternatives to TS. However, their limitations include high cost, protocol complexity and variable quality of DNA sources and reagents. Nested PCR has been shown to be a good method in this respect and it can be improved by using a high-quality source of DNA. In this study we evaluated two methods for the obtainment of DNA from dried blood samples on filter paper: 1) washing and 2) saponin/chelex-100. The second method showed higher sensitivity and specificity compared to the first, as it detected more infections, whether single or mixed, as well as Plasmodium malariae infections. Based on these results, we present this method as the protocol of choice for DNA obtainment. Nested PCR using saponin/chelex-100 for DNA extraction could be an alternative or complementary diagnosis method for human malaria parasites, but it is not appropriate for routine use

Nested PCR utilizando a gota espessa como fonte do DNA de Plasmodium: uma alternativa para esfregaços de sangue arquivados

Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.The study aimed to evaluate a protocol of nested PCR using archival Giemsa-stained thick blood smears (GTS)as source of Plasmodium DNA. A total of 138 GTS from patients of five municipalities from Pará State (Amazon Region, Brazil) was included in this survey. These samples were classified in three groups (group 1: 85 Plasmodium positive and negative GTS stored in plastic box during five years; group 2: 28 Plasmodium positive and negative GTS stored in wooden box during 10 years; and group 3: 25 Trypanosoma cruzi GTS negative for Plasmodium stored in plastic box during a month) and were submitted to DNA extraction with Chelex-100. Subsequently, extracted DNA samples were quantified and the integrity was verified by electrophoresis. Nested PCR protocol was performed to detect Plasmodium species. The results of nested PCR were compared to microscopy and statistic parameters were calculated by screening test. DNA samples from all groups had acceptable quantity and purity level, but the evaluation of integrity showed 19 degraded samples from group 2. By nested PCR, this group showed very low sensitivity (29.63%) and accuracy (32.14%), while nested PCR for samples from group 1 showed 100% of sensitivity and 97.65% of accuracy. The results of this research showed that samples stored until five years can be useful as Plasmodium DNA source for nested PCR to identify Plasmodium species, being an important alternative to support retrospective studies.O estudo teve como objetivo avaliar um protocolo de nested PCR, utilizando o teste de gota espessa corada por Giemsa (GTS) como fonte de DNA de Plasmodium. Um total de 138 amostras de GTS de pacientes de cinco municípios do Estado do Pará (Região Amazônica, Brasil) foi incluído neste estudo. Essas amostras foram classificadas em três grupos (grupo 1: 85 Plasmodium GTS positivos e negativos armazenados em uma caixa de plástico durante cinco anos; grupo 2: 28 Plasmodium GTS positivos e negativos armazenados na caixa de madeira durante 10 anos; e grupo 3: 25 Trypanosoma cruzi GTS negativos para Plasmodium armazenados em caixa de plástico durante um mês) e foram submetidas à extração de DNA com Chelex-100. Subsequentemente, as amostras de DNA extraídas foram quantificadas e sua integridade foi verificada pela eletroforese. O protocolo de nested PCR foi realizado para detectar resultados das espécies de Plasmodium. Os resultados do nested PCR foram comparados com os de microscopia e os parâmetros estatísticos foram calculados pelo teste de triagem. As amostras de DNA de todos os grupos obtiveram nível, quantidade e pureza aceitáveis, mas a avaliação da integridade mostrou 19 amostras degradadas do grupo 2. Pelo nested PCR, esse grupo mostrou baixa sensibilidade (29,63%) e precisão (32,14%), enquanto que as amostras do grupo 1 apresentaram 100% de sensibilidade e 97,65% de precisão. Os resultados desta pesquisa apontam que as amostras armazenadas até cinco anos podem ser úteis como fonte de DNA de Plasmodium para que o nested PCR identifique as espécies de Plasmodium, sendo uma alternativa importante para apoiar estudos retrospectivos

Frequência de genótipos da proteína circunsporozoíta de Plasmodium vivax em seres humanos e mosquitos anofelinos em área endêmica da região sudeste do Estado do Pará, Brasil

Universidade Federal do Pará. Belém, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Universidade Estadual Paulista. Botucatu, SP, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil / Universidade Estadual Paulista. Botucatu, SP, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil / Universidade Federal do Pará. Belém, PA, Brasil.O objetivo deste estudo foi investigar a frequência de genótipos da proteína circunsporozoíta (CSP) em sangue humano e sua correlação com a parasitemia, bem como avaliar a presença desses genótipos em Anopheles no Município de Goianésia do Pará, uma área endêmica do sudeste do Estado do Pará, Brasil, de 2012 a 2013. Amostras de sangue foram coletadas de 118 pacientes com Plasmodium vivax e 369 mosquitos anofelinos. O gene da CSP foi genotipado usando-se a reação em cadeia da polimerase/polimorfismo de comprimento de fragmento de restrição, e a infectividade dos anofelinos foi determinada pelo ELISA. A parasitemia variou de 5-70.000 parasitas/mm³, e os três genótipos (VK210, VK247 e P. vivax-like) foram detectados tanto em infecções simples quanto em mistas. Nenhuma amostra apresentou infecção mista com todos os três genótipos. O genótipo mais frequente foi o VK210, seguido pelo VK247 e o último associado com os valores mais altos de parasitemia (p < 0,0001). Entre os mosquitos identificados, somente 11 espécimes foram infectados; de sete espécimes Anopheles darlingi, quatro foram infectados por Plasmodium falciparum, dois por VK210 e um por VK247. Os três Anopheles albitarsis foram infectados por VK247 e um Anopheles nuneztovari por VK210. O genótipo VK210 continua sendo o mais prevalente no sudeste do Pará; entretanto, novas evidências indicam a adaptação do VK247. Os espécimes An. darlingi, An. albitarsis e An. nuneztovari desempenham um importante papel na transmissão dos genótipos CSP na área de estudo. Essa descoberta pode ser um problema de saúde pública devido à possibilidade de ressurgimento de epidemias de malária por P. vivax em comunidades suscetíveis.The objective of this study was therefore to investigate the frequency of circumsporozoite protein (CSP) genotypes in human blood and their correlation with parasitemia, as well as to evaluate the presence of these genotypes in Anopheles in the Municipality of Goianésia do Pará, an endemic area of southeastern Pará State, Brazil from 2012-2013. Blood samples were collected from 118 patients with Plasmodium vivax and 369 anopheline mosquitoes. The CSP gene was genotyped using the polymerase chain reaction/restriction fragment length polymorphism technique, and the infectivity of the anophelines was determined using ELISA. Parasitemia ranged from 5-70,000 parasites/mm³, and the three genotypes (VK210, VK247, and P. vivax-like) were detected both in single and mixed infections. No sample exhibited mixed infection with all three genotypes. The most frequent genotype was VK210 followed by VK247 and the latter associated with the highest parasitemia values (p < 0.0001). Among the identified mosquitoes, only 11 specimens were infected; of the seven Anopheles darlingi specimens four were infected with Plasmodium falciparum, two with VK210, and one with VK247. The three Anopheles albitarsis specimens were infected with VK247, and one Anopheles nuneztovari specimen was infected with VK210. The VK210 genotype continues to be the most prevalent in southeastern Pará; however, a new evidence shows the adaptation of VK247. The species An. darlingi, An. albitarsis, and An. nuneztovari play an important role in the transmission of CSP genotypes in the study area. This finding may be a public health concern due to the possibility of resurgence of P. vivax malaria epidemics in susceptible communities

Immunogenetic markers associated with a naturally acquired humoral immune response against an N-terminal antigen of Plasmodium vivax merozoite surface protein 1 (PvMSP-1)

São Paulo State University. Department of Biology. São José do Rio Preto, SP, Brazil / São José do Rio Preto Medical School. Department of Skin. Infectious and Parasitic Diseases. São José do Rio Preto, SP, Brazil.São José do Rio Preto Medical School. Department of Skin. Infectious and Parasitic Diseases. São José do Rio Preto, SP, Brazil.São José do Rio Preto Medical School. Department of Skin. Infectious and Parasitic Diseases. São José do Rio Preto, SP, Brazil / São Paulo State University. Department of Biology. São José do Rio Preto, SP, Brazil.Federal University of Sergipe. Department of Biology. São Cristóvão, SE, Brazil.Oswaldo Cruz Foundation. Leônidas and Maria Deane Institute. Manaus, AM, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisa Básica de Malária. Belém, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisa Básica de Malária. Belém, PA, Brasil.Oswaldo Cruz Foundation. Leônidas and Maria Deane Institute. Manaus, AM, Brazil.São Paulo State University. Department of Biology. São José do Rio Preto, SP, Brazil / São José do Rio Preto Medical School. Department of Skin. Infectious and Parasitic Diseases. São José do Rio Preto, SP, Brazil / Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisa Básica de Malária. Belém, PA, Brasil.Background: Humoral immune responses against proteins of asexual blood-stage malaria parasites have been associated with clinical immunity. However, variations in the antibody-driven responses may be associated with a genetic component of the human host. The objective of the present study was to evaluate the influence of co-stimulatory molecule gene polymorphisms of the immune system on the magnitude of the humoral immune response against a Plasmodium vivax vaccine candidate antigen. Methods: Polymorphisms in the CD28, CTLA4, ICOS, CD40, CD86 and BLYS genes of 178 subjects infected with P. vivax in an endemic area of the Brazilian Amazon were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The levels of IgM, total IgG and IgG subclasses specific for ICB2-5, i.e., the N-terminal portion of P. vivax merozoite surface protein 1 (PvMSP-1), were determined by enzyme-linked immuno assay. The associations between the polymorphisms and the antibody response were assessed by means of logistic regression models. Results: After correcting for multiple testing, the IgG1 levels were significantly higher in individuals recessive for the single nucleotide polymorphism rs3116496 in CD28 (p = 0.00004). Furthermore, the interaction between CD28 rs35593994 and BLYS rs9514828 had an influence on the IgM levels (p = 0.0009). Conclusions: The results of the present study support the hypothesis that polymorphisms in the genes of co-stimulatory components of the immune system can contribute to a natural antibody-driven response against P. vivax antigens

Knowledge, Attitudes and Practices Regarding Sylvatic Rabies among High-Risk Households in Cear&aacute; State, Brazil

Rabies transmitted by sylvatic populations has become an increasing concern in Brazil. A total of 113 participants with a history of contact with sylvatic populations were interviewed in 27 municipalities of Cear&aacute; State in northeast Brazil. Questionnaires included questions on knowledge, attitudes and practices (KAP) regarding sylvatic rabies. Most of the respondents (92%) knew about rabies and confirmed at least one species that transmitted the disease (79.6%). Of these respondents, 69% mentioned monkeys, and 67.2% mentioned dogs. However, 16% of the respondents listed an incorrect species. In general, knowledge on the symptoms and signs and on prevention measures was weak. The majority raised pets (93.8%), most commonly dogs and cats, and, of all the pets, 85.7% were claimed to be vaccinated against rabies. A total of 67.3% reported the appearance of free-living wild animals around their houses, mostly marmosets and wild canids; 18.3% reported that sylvatic populations had attacked animals or humans. Seventy-three percent had raised or still were raising wild animals as pets, mostly capuchin monkeys (79.5%) and marmosets (24.1%). This is the first KAP study on sylvatic rabies in Brazil. The data indicate important knowledge gaps and risk behavior within a high-risk population. There is a need for strengthening and improving sylvatic rabies surveillance and control, combined with the intensification of education and information campaigns

Morphological atypia and molecular profile of

This study aimed to perform morphological and molecular analyses of parasites isolated from the blood of malaria-infected individuals during an outbreak in the Microregion of Cametá, State of Pará, Brazilian Amazon. A total of 260 positive samples were identified by microscopy as Plasmodium vivax; however, in three samples, forms considered unusual for the species were found and defined as morphological atypia of P. vivax. Single P. vivax infection was confirmed by qPCR in all samples. Among 256 genotyped samples, the VK247 genotype alone was identified in 255 samples, and the VK210 genotype was found in only one. The study showed that this malaria outbreak was caused by the etiological agent P. vivax, and for the first time, morphological atypia was described in isolates circulating in Brazil. Likewise, for the first time, the VK247 genotype was detected predominantly in single infections in an area of the State of Pará, which may suggest a greater circulation of the genotype in the region