Nyssorhynchus darlingi genome-wide studies related to microgeographic dispersion and blood-seeking behavior.

BACKGROUND: In Brazil, malaria is concentrated in the Amazon Basin, where more than 99% of the annual cases are reported. The main goal of this study was to investigate the population structure and genetic association of the biting behavior of Nyssorhynchus (also known as Anopheles) darlingi, the major malaria vector in the Amazon region of Brazil, using low-coverage genomic sequencing data. METHODS: Samples were collected in the municipality of Mâncio Lima, Acre state, Brazil between 2016 and 2017. Different approaches using genotype imputation and no gene imputation for data treatment and low-coverage sequencing genotyping were performed. After the samples were genotyped, population stratification analysis was performed. RESULTS: Weak but statistically significant stratification signatures were identified between subpopulations separated by distances of approximately 2-3 km. Genome-wide association studies (GWAS) were performed to compare indoor/outdoor biting behavior and blood-seeking at dusk/dawn. A statistically significant association was observed between biting behavior and single nucleotide polymorphism (SNP) markers adjacent to the gene associated with cytochrome P450 (CYP) 4H14, which is associated with insecticide resistance. A statistically significant association between blood-seeking periodicity and SNP markers adjacent to genes associated with the circadian cycle was also observed. CONCLUSION: The data presented here suggest that low-coverage whole-genome sequencing with adequate processing is a powerful tool to genetically characterize vector populations at a microgeographic scale in malaria transmission areas, as well as for use in GWAS. Female mosquitoes entering houses to take a blood meal may be related to a specific CYP4H14 allele, and female timing of blood-seeking is related to circadian rhythm genes.

Evidence of co-circulation of multiple arboviruses transmitted by Aedes species based on laboratory syndromic surveillance at a health unit in a slum of the Federal District, Brazil.

BACKGROUND: Vector-borne diseases, especially arboviruses transmitted by Aedes sp. mosquitos, should be a health policy priority in Brazil. Despite this urgency, there are significant limitations in the traditional surveillance system, mainly in vulnerable areas. This study aimed to investigate the circulation of dengue (DENV), Zika (ZIKV), and chikungunya viruses (CHIKV) by laboratory syndromic surveillance (LSS) in a slum area of the Federal District of Brazil, comparing the results with traditional surveillance data. METHODS: LSS for acute febrile and/or exanthematous symptoms was developed at a health unit of Cidade Estrutural, in order to identify the circulation of arboviruses transmitted by Aedes sp. mosquitos. Between June 2019 and March 2020, 131 valid participants were identified and sera tested by reverse transcription polymerase chain reaction (RT-PCR) for DENV (by serotype), ZIKV, and CHIKV acute infection and by immunoglobulin M enzyme-inked immunosorbent assay (ELISA-IgM) for DENV and CHIKV 15-21 days after symptom onset, when the participant reported no respiratory signs (cough and/or coryza). The results obtained were compared with traditional surveillance data for the study area and period. RESULTS: At least three DENV-1 (2.3%), four DENV-2 (3%), and one CHIKV (0.7%) cases were confirmed in the laboratory, showing evidence of hyperendemicity even though LSS had not reached the historic peak dengue fever months in the Federal District (April-May). When the results obtained here were compared with traditional surveillance, a significant discrepancy was observed, including underreporting of CHIKV infection. CONCLUSIONS: In addition to the risks posed to the study population, the area investigated with its respective socio-environmental profile may be a potential site for spread of the virus, given the cosmopolitan presence of Aedes sp. and human mobility in the Federal District. It is also suggested that traditional epidemiological surveillance may be reporting acute viral infections other than DENV as dengue fever, while underreporting other arboviruses transmitted by Aedes sp. mosquitos in the Federal District.

Sensitivity of Guillain-Barre Syndrome Surveillance in the Brazilian Federal District, using the Capture-Recapture Method.

INTRODUCTION: Guillain-Barre Syndrome (GBS) is an acute immune-mediated polyneuropathy that compromises the peripheral and cranial nerves. It is characterized by rapid-onset paresthesia accompanied by progressive weakness in the lower extremities followed by symmetric ascending paralysis. METHODOLOGY: assessment of sensitivity to detect GBS between March 2017 and May 2019 in a public referral hospital, using the capture-recapture method based on the Chapman estimator and comparing three GBS data sources: the hospital-based sentinel surveillance system (VSBH), Human Immunoglobulin Dispensing Records System (RDIH), and Hospital Information System (SIH). RESULTS: A total of 259 possible cases were identified (captured). Of these, 58 were confirmed and most resided in the Federal District. The VSBH showed the greatest sensitivity in case identification. The temporal distribution of cases showed periods with no cases identified, and more were registered during the rainy season from October to May, when high temperatures also occur. CONCLUSIONS: Increased circulation of arboviruses and gastrointestinal infections during the rainy season may explain the greater concentration of GBS cases. It is important to note that one-third of the cases identified in the different data sources do not converge, demonstrating that no single surveillance system is 100% effective. The severity and possible increase in cases related to GBS demonstrates the need for an improved surveillance system capable of monitoring and following-up cases involving neurological syndromes, regardless of the event preceding infection.

Ecology and larval population dynamics of the primary malaria vector Nyssorhynchus darlingi in a high transmission setting dominated by fish farming in western Amazonian Brazil.

Vale do Rio Juruá in western Acre, Brazil, is a persistent malaria transmission hotspot partly due to fish farming development that was encouraged to improve local standards of living. Fish ponds can be productive breeding sites for Amazonian malaria vector species, including Nyssorhynchus darlingi, which, combined with high human density and mobility, add to the local malaria burden.This study reports entomological profile of immature and adult Ny. darlingi at three sites in Mâncio Lima, Acre, during the rainy and dry season (February to September, 2017). From 63 fishponds, 10,859 larvae were collected, including 5,512 first-instar Anophelinae larvae and 4,927 second, third and fourth-instars, of which 8.5% (n = 420) were Ny. darlingi. This species was most abundant in not-abandoned fishponds and in the presence of emerging aquatic vegetation. Seasonal analysis of immatures in urban landscapes found no significant difference in the numbers of Ny. darlingi, corresponding to equivalent population density during the rainy to dry transition period. However, in the rural landscape, significantly higher numbers of Ny. darlingi larvae were collected in August (IRR = 5.80, p = 0.037) and September (IRR = 6.62, p = 0.023) (dry season), compared to February (rainy season), suggesting important role of fishponds for vector population maintenance during the seasonal transition in this landscape type. Adult sampling detected mainly Ny. darlingi (~93%), with similar outdoor feeding behavior, but different abundance according to landscape profile: urban site 1 showed higher peaks of human biting rate in May (46 bites/person/hour), than February (4) and September (15), while rural site 3 shows similar HBR during the same sampling period (22, 24 and 21, respectively). This study contributes to a better understanding of the larvae biology of the main malaria vector in the Vale do Rio Juruá region and, ultimately will support vector control efforts.

How forest data catalysed change in four successful case studies.

This paper presents four case studies in which forest data catalysed shifts in public policy and corporate activities. Brazil greatly reduced deforestation during the period between 2005 and 2014; Cameroon introduced a structured forest concessions regime; Viet Nam achieved their forest transition; and corporate operations around the world invested in supply chain management to alleviate deforestation concerns. We break the problem-solving required for these achievements into four steps: problem recognition, proposal and choice of solution, putting the solution into effect, and monitoring results. At each of these steps, we consider the relevant forest data. Data helped place issues on policymaker agendas, supported reaching sound decisions and enabled quantitative targets. Policy instruments for implementing change were built around available data and forest monitoring helped evaluate progress. The details of these successes can be an inspiration to those interested in improving collection of data on forests that can effectively support decision-making and better policies. There have been impressive recent improvements to many developing countries' national forest monitoring capabilities. The successful examples of data application presented and evaluated here provide insight into how these new data can be effectively leveraged.

The role of forest conversion, degradation, and disturbance in the carbon dynamics of Amazon indigenous territories and protected areas.

Maintaining the abundance of carbon stored aboveground in Amazon forests is central to any comprehensive climate stabilization strategy. Growing evidence points to indigenous peoples and local communities (IPLCs) as buffers against large-scale carbon emissions across a nine-nation network of indigenous territories (ITs) and protected natural areas (PNAs). Previous studies have demonstrated a link between indigenous land management and avoided deforestation, yet few have accounted for forest degradation and natural disturbances-processes that occur without forest clearing but are increasingly important drivers of biomass loss. Here we provide a comprehensive accounting of aboveground carbon dynamics inside and outside Amazon protected lands. Using published data on changes in aboveground carbon density and forest cover, we track gains and losses in carbon density from forest conversion and degradation/disturbance. We find that ITs and PNAs stored more than one-half (58%; 41,991 MtC) of the region's carbon in 2016 but were responsible for just 10% (-130 MtC) of the net change (-1,290 MtC). Nevertheless, nearly one-half billion tons of carbon were lost from both ITs and PNAs (-434 MtC and -423 MtC, respectively), with degradation/disturbance accounting for >75% of the losses in 7 countries. With deforestation increasing, and degradation/disturbance a neglected but significant source of region-wide emissions (47%), our results suggest that sustained support for IPLC stewardship of Amazon forests is critical. IPLCs provide a global environmental service that merits increased political protection and financial support, particularly if Amazon Basin countries are to achieve their commitments under the Paris Climate Agreement.

Larval habitats of Anopheles species in a rural settlement on the malaria frontier of southwest Amazon, Brazil.

Rural settlements are social arrangements expanding in the Amazon region, which generate disturbances in the natural environment, thus affecting the ecology of the species of Anopheles and thus the malaria transmission. Larval habitats are important sources for maintenance of mosquito vector populations, and holding back a natural watercourse is a usual process in the establishment of rural settlements, since the formation of micro-dams represents a water resource for the new settlers. Identifying characteristics of the larval habitats that may be associated with both the presence and abundance of Anopheles vectors species in an environment under ecological transition is background for planning vector control strategies in rural areas in the Amazon. Anopheles larvae collections were performed in two major types of habitats: natural and flow-limited water collections that were constructed by holding back the original watercourse. A total of 3123 Anopheles spp. larvae were captured in three field-sampling collections. The majority of the larvae identified were taken from flow-limited water collections belonged to species of the Nyssorhynchus subgenus (92%), whereas in the natural larval habitats a fewer number of individuals belonged to the Stethomyia (5%) and Anopheles (3%) subgenera. The total of Nyssorhynchus identified (1818), 501 specimens belonged to An. darlingi, 750 to An. triannulatus and 567 for others remaining species. In addition, 1152 could not be identified to subgenus/species level, because they were either in the first-instar or damaged. The primary vector in areas of the Amazon river basin, An. darlingi, was found exclusively in man-made habitats. Statistical analysis display An. triannulatus with specialist behavior for characteristics of man-made habitats. Modifications in the natural environment facilitate the rise of larval habitats for species with epidemiological importance for malaria in the region. This study showed that man-made habitats flow-limited water collections from dry lands could be a factor associated with the increase of An. darlingi and An. triannulatus populations, and other Nyssorhynchus species as well in endemic areas of the Amazon Region.

Perturbações no ambiente natural, e emergência de habitats larvais ocupados por espécies de Anopheles Meigen (Diptera: Culicidae) em assentamento rural no sudoeste da Amazônia brasileira / Disturbances in the natural environment, and emergency of larval habitats occupied by species of Anopheles Meigen (Diptera: Culicidae) in rural settlement in Brazilian Amazon southwestern

Introdução: Assentamentos rurais são recorrentes arranjos sociais em expansão na Região Amazônica, que geram perturbações no ambiente natural e na respectiva ecologia de mosquitos vetores envolvidos na transmissão de malária. Objetivos: Elaborar revisão das espécies do gênero Anopheles que foram registradas na Região Amazônica. Para a pesquisa quantitativa, estabelecer potenciais fatores dos ambientes e das características das águas de diversos habitats larvais que possam estar associadas com a presença de espécies de anofelíneos, através de coletas em criadouros de diferentes graus de perturbação ambiental. Métodos: para a revisão das espécies que foram registradas na Amazônia, foram empregados termos de busca, utilizando sete bases bibliográficas de referência. Para os estudos em campo, foram realizadas capturas de larvas juntamente com estimação e mensuração de algumas condições dos habitats larvais, para comparação dos parâmetros analisados entre ambientes naturais e artificiais. Resultados: a revisão mostrou maior quantidade de artigos que notificaram a ocorrência do subgênero Nyssorhynchus, seguido pelos subgêneros: Anopheles, Stethomyia, Kerteszia e Lophopodomyia. O Anopheles darlingi se mostrou a espécie com maior ocorrência nos artigos levantados, muito pelo seu caráter epidemiológico. Importantes vetores auxiliares pertencentes ao subgênero 6 Nyssorhynchus também foram registrados em grande quantidade. Os resultados das análises conduzidas com os dados das capturas em campo demonstraram que as forças de transformação do ambiente natural favorecem o desenvolvimento de habitats larvais de espécies do subgênero Nyssorhynchus, incluindo o An. darlingi, além de um importante vetor secundário, o An. triannulatus. Discussão e Conclusão: projetos de assentamentos na Amazônia brasileira registram índices significativos de casos de malária, devido à ocupação de áreas naturais pelo homem. Este estudo mostrou que o subgênero Nyssorhynchus possui amplo registro na literatura de inquéritos entomológicos do gênero na floresta tropical equatorial, indicando que o agrupamento pode ser considerado endêmico às fronteiras de transformação na região, tendo assim importante papel na manutenção da transmissão da malária. Os estudos de campo apresentaram evidências de que formação de habitats artificiais favorecem formas iniciais de vida dos mosquitos com histórico vetorial para a região. Esses habitats larvais podem ser respostas ao uso do solo e das consequentes alterações provocadas no ambiente natural pelas atividades humanas. As modificações do ambiente natural selecionam e propiciam a formação de habitats para as espécies do Nyssorhynchus. Este agrupamento taxonômico engloba espécies que são consideradas vetores para a malária na região Neotropical.

Introduction: Rural settlements are recurrent social arrangements expanding in the Amazon region, which generate disturbances in the natural environment, affecting the mosquitoes ecology involved in malaria transmission. Objective: A systematic review of Anopheles species were recorded in the Amazon region. For the qualitative research, were establish potential factors of the environment and the characteristics of the waters of several larval habitats that may be associated with the presence of species of Anopheles mosquitoes, by surveys of the samples presents in larval habitats within environmental disturbance gradient. Methods: for the review of the species, were used search terms, in seven bibliographic databases of reference. For studies in the field, larvae were trapped along estimation and measurement of some conditions of the larval habitats, to compare the parameters analyzed between natural and artificial environments. Results: The review showed greater amount of articles that reported the occurrence of Nyssorhynchus subgenre, followed by: Anopheles, Stethomyia, Kerteszia and Lophopodomyia. The Anopheles darlingi proved the species with the highest occurrence in the papers showed, due to its epidemiological importance. Important auxiliary vectors belonging to the subgenus Nyssorhynchus were also recorded in large scale. The results of the qualitative shown that the natural 8 environment transformation forces favor the development of larval habitats of the species of subgenus Nyssorhynchus, including An. darlingi and an important secondary vector, An. Triannulatus. Discussion and Conclusion: settlement projects in the Brazilian Amazon recorded significant rates of malaria cases. This study showed that, the Amazon basin an important determinant of this phenomenon is the formation of artificial habitats that are occupied by early forms of life of mosquito vectors. These larval habitats may be answers to land use and consequent changes in the natural environment caused by human activities. Modifications to the natural environment select and provide the formation of habitats for species in Nyssorhynchus. This taxonomic group includes species that are considered vectors for malaria in the Neotropics. Discussion and Conclusion: settlement projects in the brazilian Amazon recorded significant rates of malaria cases. This study showed that the review of the literature shows Nyssorhynchus subgenre ample record in entomological surveys in the equatorial rainforest, indicating that the group may be considered endemic to the borders of transformation in the region, and thus important role in the maintenance of malaria transmission. Field studies showed evidence that formation of artificial habitats favor early forms of the mosquito vector, with history for malaria in the region. These larval habitats may be answers to land use and consequent changes in the natural environment caused by human activities. Modifications to the natural environment select and provide the formation of habitats for species in Nyssorhynchus.

Influências de Atta spp. (Hymenoptera: Formicidae) na recuperação da vegetação pós-fogo em floresta de transição amazônica / Influences of leafcutter ant Atta spp. (Hymenoptera: Formicidae) on vegetation recovery after fire in Amazonian transitional forest

Este estudo investigou o papel das saúvas na recuperação da vegetação pós-fogo. Foi hipotetizado que a perturbação do fogo aumenta a abundância de ninhos de saúva (1) e as atividades de remoção de sementes (2) e desfolha (3), especialmente no ambiente de borda. Os ninhos de saúva foram inventariados e mapeados e após 17 meses, checados para o registro de sua atividade e de novas colônias emergindo na área. Realizou-se experimentos comparativos de remoção de sementes e desfolha de plântulas em duas parcelas de 50 ha, uma submetida a fogo anual (tratamento) e outra sem fogo (controle). A abundância de ninhos de saúva foi maior na parcela tratamento que na parcela controle, especialmente na borda da floresta. As espécies encontradas foram: Atta cephalotes, A. laevigata e A. sexdens , sendo esta última a espécie mais abundante e que mostrou aumento em número de colônias ativas após 17 meses. O aumento na abundância de ninhos na parcela tratamento foi acompanhado por um aumento na atividade dessas formigas. Enquanto mais de duas folhas foram arrancadas por plântula nessa parcela, menos de uma foi registrada na parcela controle. Também a abundância média de galhos desfolhados e de sementes removidas por saúvas foi maior na parcela tratamento que na parcela controle. As saúvas tanto podem diminuir ou retardar o processo de regeneração florestal pós-fogo, quanto acelerá-lo, devido à elevada predação seletiva que favorece as espécies menos palatáveis. Desta forma, suas atividades podem modificador apenas a composição da vegetação recuperando-se do fogo.

In this study we investigated the role of leaf-cutting ants in the post-fire vegetation recovery. We hypothesized that a forest plot submitted to annual fire presents: (1) higher abundance of leaf-cutting ant nests; (2) higher removal of seeds; and (3) higher herbivory rates of leaf-cutting ants, when compared to the forest plots without fire (control). The leaf-cutting ant nests were sampled, mapped, and checked up (after 17 months) to register their activity and new colonies emerging from the plot. We made comparative experiments of seeds removal and seedlings herbivory in two 50 ha plots, one submitted to annual fire and another without fire. The abundance of leaf-cutting nests was higher in the plot submitted to fire than in the control plot. The species found were: Atta cephalotes, A. laevigata, and A. sexdens, being the latter the most abundant and the one that showed an increase of active colonies after 17 months. The plot submitted to fire showed a higher abundance of seeds removed by leaf-cutting ants than the control. While more than two leaves were ripped per seedling in the plot under fire, less than one was registered in the control plot. It was also observed that the average abundance of seedlings attacked by leaf-cutting ant in the fire plot was higher than in the plot without fire. This study shows that the leaf-cutting ants may reduce seed germination and seedling recruitment, affecting the reproductive success and the composition of vegetation recovering from fire. Alternatively, the leaf-cutting ants may also speed up the regeneration process of unpalatable plant species. In this sense, we may conclude that the presence of leaf-cutting may interfere in the species composition of post fire forest.

Population dynamics, structure and behavior of Anopheles darlingi in a rural settlement in the Amazon rainforest of Acre, Brazil.

BACKGROUND: Anopheles darlingi is the major vector of malaria in South America, and its behavior and distribution has epidemiological importance to biomedical research. In Brazil, An. darlingi is found in the northern area of the Amazon basin, where 99.5% of the disease is reported. METHODS: The study area, known as Ramal do Granada, is a rural settlement inside the Amazon basin in the state of Acre. Population variations and density have been analysed by species behaviour, and molecular analysis has been measured by ND4 mitochondrial gene sequencing. RESULTS: The results show higher density in collections near a recent settlement, suggesting that a high level of colonization decreases the vector presence. The biting activity showed higher activity at twilight and major numbers of mosquitos in the remaining hours of the night in months of high density. From a sample of 110 individual mosquitoes, 18 different haplotypes were presented with a diversity index of 0.895, which is higher than that found in other Anopheles studies. CONCLUSIONS: An. darlingi depends on forested regions for their larval and adult survival. In months with higher population density, the presence of mosquitoes persisted in the second part of the night, increasing the vector capacity of the species. Despite the intra-population variation in the transition to rainy season, the seasonal distribution of haplotypes shows no change in the structure population of An. darlingi.

Factors affecting the abundance of leaf-litter arthropods in unburned and thrice-burned seasonally-dry Amazonian forests.

Fire is frequently used as a land management tool for cattle ranching and annual crops in the Amazon. However, these maintenance fires often escape into surrounding forests, with potentially severe impacts for forest biodiversity. We examined the effect of experimental fires on leaf-litter arthropod abundance in a seasonally-dry forest in the Brazilian Amazon. The study plots (50 ha each) included a thrice-burned forest and an unburned control forest. Pitfall-trap samples were collected at 160 randomly selected points in both plots, with sampling stratified across four intra-annual replicates across the dry and wet seasons, corresponding to 6, 8, 10 and 12 months after the most recent fire. Arthropods were identified to the level of order (separating Formicidae). In order to better understand the processes that determine arthropod abundance in thrice-burned forests, we measured canopy openness, understory density and litter depth. All arthropod taxa were significantly affected by fire and season. In addition, the interactions between burn treatment and season were highly significant for all taxa but Isoptera. The burned plot was characterized by a more open canopy, lower understory density and shallower litter depth. Hierarchical partitioning revealed that canopy openness was the most important factor explaining arthropod order abundances in the thrice-burned plot, whereas all three environmental variables were significant in the unburned control plot. These results reveal the marked impact of recurrent wildfires and seasonality on litter arthropods in this transitional forest, and demonstrate the overwhelming importance of canopy-openness in driving post-fire arthropod abundance.

Role of Brazilian Amazon protected areas in climate change mitigation.

Protected areas (PAs) now shelter 54% of the remaining forests of the Brazilian Amazon and contain 56% of its forest carbon. However, the role of these PAs in reducing carbon fluxes to the atmosphere from deforestation and their associated costs are still uncertain. To fill this gap, we analyzed the effect of each of 595 Brazilian Amazon PAs on deforestation using a metric that accounts for differences in probability of deforestation in areas of pairwise comparison. We found that the three major categories of PA (indigenous land, strictly protected, and sustainable use) showed an inhibitory effect, on average, between 1997 and 2008. Of 206 PAs created after the year 1999, 115 showed increased effectiveness after their designation as protected. The recent expansion of PAs in the Brazilian Amazon was responsible for 37% of the region's total reduction in deforestation between 2004 and 2006 without provoking leakage. All PAs, if fully implemented, have the potential to avoid 8.0 +/- 2.8 Pg of carbon emissions by 2050. Effectively implementing PAs in zones under high current or future anthropogenic threat offers high payoffs for reducing carbon emissions, and as a result should receive special attention in planning investments for regional conservation. Nevertheless, this strategy demands prompt and predictable resource streams. The Amazon PA network represents a cost of US$147 +/- 53 billion (net present value) for Brazil in terms of forgone profits and investments needed for their consolidation. These costs could be partially compensated by an international climate accord that includes economic incentives for tropical countries that reduce their carbon emissions from deforestation and forest degradation.

Indigenous lands, protected areas, and slowing climate change.

Recent climate talks in Copenhagen reaffirmed the crucial role of reducing emissions from deforestation and degradation (REDD). Creating and strengthening indigenous lands and other protected areas represents an effective, practical, and immediate REDD strategy that addresses both biodiversity and climate crises at once.