Serological survey of Rickettsia in equids from Vale do Paraíba, São Paulo, Brazil, and their tick identification and molecular investigation of Rickettsia

A febre maculosa brasileira é uma doença grave e letal para seres humanos causada pela bactéria Rickettsia rickettsii. No estado de São Paulo, SP, Brasil, o agente etiológico desta enfermidade é transmitido pelo carrapato Amblyomma sculptum. Conforme descrito na literatura científica, os cavalos infectados com esta bactéria produzem uma forte resposta imune e podem ser importantes sentinelas para a detecção da doença. A presente investigação realizou um levantamento sorológico em cavalos de cinco fazendas do Vale do Paraíba, São Paulo, Brasil, à procura de anticorpos contra Rickettsia rickettsii, Rickettsia parkeri, Rickettsia amblyommatis, Rickettsia rhipicephali e Rickettsia bellii. Em cada fazenda, também foram coletados carrapatos identificados taxonomicamente e examinados por PCR em tempo real para o DNA de Rickettsia spp. Foram coletadas amostras de sangue de 206 cavalos e coletados 334 carrapatos desses animais entre os meses de janeiro e dezembro de 2017. Oitenta carrapatos foram identificados como A. sculptum e 254 Dermacentor nitens. Das amostras de sangue, 7,3% soroconverteram para Rickettsia spp., sendo que, 0,97% apresentaram soropositividade homóloga para R. bellii. Nenhum dos 80 carrapatos de A. sculptum foi positivo com o emprego de PCR em tempo real para Rickettsia spp. Embora não tenham sido detectados carrapatos infectados por Rickettsia spp em cinco fazendas do Vale do Paraíba, os animais apresentaram reações sorológicas positivas para este agente. Assim, outros estudos abrangentes deverão ser realizados na área investigando hospedeiros e vetores, gerando dados para medidas de controle da transmissão da febre maculosa brasileira.Brazilian spotted fever is a serious and lethal illness for humans and is caused by the Rickettsia rickettsii bacteria. In the state of São Paulo/SP (Brazil), the etiological agent of this disease is transmitted by the Amblyomma sculptum tick. It was already shown that horses infected with this bacteria produce a strong immune response and could be important sentinels for the detection of the disease in a proper region. The present investigation performed a serological survey in horses from five farms of Vale do Paraíba, São Paulo state, Brazil, searching for antibodies against, Rickettsia rickettsii, Rickettsia parkeri, Rickettsia amblyommatis, Rickettsia rhipicephali, and Rickettsia bellii. In each farm, ticks were also collected that were taxonomically identified and examined by real-time PCR for Rickettsia spp DNA. Blood samples were collected from 206 horses, and 334 ticks were picked up from these animals from January to December 2017. Eighty ticks wereA. sculptum and 254 Dermacentor nitens. Of the blood samples, 7.3% seroconverted to Rickettsia spp. Of these, 0.97% had a positive serological response to R. bellii. None of the 80 A. sculptum ticks were positive through real-time PCR for Rickettsia spp. Although there was no detection of ticks infected by Rickettsia spp in five farms of Paraíba Valley, the horses presented serological positive reactions against this agent. Thus, further large studies should be conducted in the area targeting hosts and vectors to generate data for control measures of the transmission of Brazilian spotted fever.&nbsp

Ticks and rickettsiae associated with mammals of endemic and nonendemic areas for Brazilian spotted fever

A Febre Maculosa Brasileira (FMB) é a enfermidade transmitida por carrapatos de maior importância médica da América Latina. Capivaras são hospedeiros primários para o carrapato vetor Amblyomma sculptum, e amplificadoras da bactéria Rickettsia rickettsii, causadora da FMB. Entretanto, o ciclo epidemiológico da doença pode envolver outros componentes. Nesse sentido, pequenos mamíferos podem atuar como hospedeiros alternativos para carrapatos e para a manutenção do agente. O objetivo do presente estudo foi comparar ixodofauna e riquétsias associadas a pequenos mamíferos de áreas endêmicas e não endêmicas para FMB. Para isso pequenos mamíferos foram capturados (2015-2018) em três áreas endêmicas (E) e três áreas não endêmicas (NE) com alto grau de antropização no estado de São Paulo, e em duas áreas não endêmicas com baixo grau de antropização (BIO), uma no estado do Mato Grosso e outra no estado do Mato Grosso do Sul. Todos os carrapatos em parasitismo nos pequenos mamíferos capturados foram coletados. Soro sanguíneo foi testado para anticorpos anti-riquétsias por Reação de Imunofluorescência Indireta; carrapatos e tecidos dos pequenos mamíferos tiveram DNA extraído e testado na PCR para Rickettsia. Foi realizada uma análise comparativa de fatores ambientais que pudessem alterar a população de carrapatos e pequenos mamíferos do local entre as áreas E e NE. Foram capturados 629 animais de 27 espécies diferentes. A prevalência de parasitismo nesses animais foi de 45,4% nas E, 39% nas NE e 22% nas BIO. Nas E foram coletados 2795 carrapatos (A. sculptum, Amblyomma dubitatum, Amblyomma ovale e Ixodes loricatus); nas NE 1283 (Ixodes schulzei, I. loricatus, A. ovale, A. sculptum e A. dubitatum) e nas BIO 342 (A.sculptum, Amblyomma parvum, A. ovale, Ornithodoros mimon e I. loricatus). A. sculptum correspondeu a 81% dos carrapatos nas E, 10,7% nas NE e 43,8% nas BIO, ocorrendo majoritariamente no marsupial Didelphis sp. nas E e NE. Foram soropositivos 33,3% dos roedores e 19,2% dos marsupiais nas E, 15,7% e 10,3% nas NE e 12,6% e 11,2% nas BIO respectivamente. Nas E a taxa de infecção dos carrapatos testados pela PCR foi de 13,8%, nas NE 24,1% e nas BIO 48,6%. Nenhuma amostra de tecido foi positiva. As áreas NE apresentaram diferenças significativas na variável complexidade de sub-bosque, sendo mais complexo nessas áreas. A composição na comunidade de hospedeiros e carrapatos diferiu entre as três áreas estudadas. Evidencia-se maior circulação de riquétsias e maior número de A. sculptum nas E, apesar da similaridade na abundância do principal pequeno mamífero associado a ambos, Didelphis sp., entre as E e NE.Brazilian spotted fever (BSF) is the most important vector-borne disease in Latin America. Capybaras are primary hosts for the tick vector Amblyomma sculptum, and amplifiers of the bacterium Rickettsia rickettsii, which causes BSF. However, the epidemiological cycle of the disease may involve other components. In this sense, small mammals can act as alternative hosts for ticks and for the maintenance of the agent. The objective of the present study was to compare ixodofauna and rickettsiae associated to small mammals from endemic and nonendemic areas for BSF. For this purpose, small mammals were caught (2015-2018) in three endemic areas (E) and in three non-endemic (NE) areas with a high degree of anthropogenic impact in the state of São Paulo, and in two non-endemic areas with a low degree of anthropogenic impact (BIO ), one in the state of Mato Grosso and another in the state of Mato Grosso do Sul. All ticks on parasitism in the captured small mammals were collected. Blood serum samples were tested for anti-Rickettsia spp. antibodies by Indirect Immunofluorescence Reaction. Ticks and small mammalian tissues had DNA extracted and tested in PCR for Rickettsia. A comparative analysis of environmental factors that could influence the tick and small mammal populations of the site between E and NE areas was carried out. We recorded 629 specimens of animals, belonging to 27 small mammal species. The prevalence of parasitism in these animals was 45.4% in E, 39% in NE and 22% in BIO. In the E, 2795 ticks were collected (A. sculptum, Amblyomma dubitatum, Amblyomma ovale and Ixodes loricatus); in NE 1283 ticks (Ixodes schulzei, I. loricatus, A. ovale, A. sculptum and A. dubitatum) and in BIO 342 ticks (A.sculptum, Amblyomma parvum, A. ovale, Ornithodoros mimon and I. loricatus). Amblyomma sculptum corresponded to 81% of the ticks in the E, 10.7% in the NE and 43.8% in the BIO, occurring mainly in the marsupial Didelphis spp. in E and NE. As much as 33.3% of the rodents and 19.2% of the marsupials in the E, 15.7% and 10.3% in the NE and 12.6%, and 11.2% in the BIO, respectively, were seropositive for Rickettsia spp. In the E, the infection rate of the ticks tested by PCR was 13.8%, in NE 24.1% and in BIO 48.6%. No vertebrate tissue sample was positive for rickettsiae. The NE areas presented significant differences in the variable sub-forest complexity, being more complex in these areas. The composition in the host and tick community differed between the three areas studied. There is evidence of increased circulation of rickettsiae and greater numbers of A. sculptum in E, despite the similarity in the abundance of the main small mammal associated with both, Didelphis spp., between E and NE

DIVERSIDADE DE CARRAPATOS E INFECÇÃO NATURAL POR RICKETTSIA EM REMANESCENTE PRIMÁRIO DE MATA ATLÂNTICA NO ESTADO DE SÃO PAULO, BRASIL

Introdução: Os carrapatos são ectoparasitas hematófagos obrigatórios que parasitam diversos vertebrados, incluindo o ser humano. São vetores de diversos patógenos e o gênero Amblyomma possui grande importância em saúde pública por transmitirem as diferentes espécies do gênero Rickettsia. As riquétsias são bactérias causadoras de enfermidades graves em humanos. O objetivo do estudo foi mapear a área de Mata Atlântica primária na Reserva Legado das Águas – Votorantim no Vale do Ribeira – SP, para carrapatos vetores e riquétsias, com a finalidade epidemiológica. Métodos: As capturas foram realizadas de 01/2018 a 12/2021. Os carrapatos foram coletados de pequenos mamíferos capturados em armadilhas do tipo Shermann e Tomahawk. Os carrapatos obtidos de humanos foram retirados dos humanos após o retorno das trilhas. Os espécimes de vida livre foram coletados através de inspeção de folhagens, e todos os indivíduos foram identificados através de chaves dicotômicas. A extração de DNA foi feita utilizando a técnica de isotiocianato de guanidina e a detecção de riquétsias através de qPCR e cPCR utilizando como alvo os genes gltA e ompA. Após esta etapa os amplicons foram sequenciados e submetidos a análise de similaridade pelo BLAST. Resultados: Foram coletados 2.669 espécimes de três gêneros e 13 espécies, e a única espécie encontrada parasitando humanos foi Amblyomma incisum. Dos carrapatos coletados, 6.8% foram positivos para Rickettsia nos genes testados. A análise de similaridade identificou três espécies infectando carrapatos: Rickettsia bellii, em Amblyomma ovale, Amblyomma brasiliense, Ixodes loricatus e Haemaphysalis juxtakochi; Rickettsia parkeri cepa Mata Atlântica em A. ovale e Rickettsia rhipicephali em Há. Juxtakochi. Conclusão: A área de estudo é constituída por mata primária com grande densidade e diversidade de mamíferos refletindo diretamente na densidade e diversidade de carrapatos identificados. A espécie mais abundante foi A. incisum, única encontrada parasitando humanos, negativa para a infecção por Rickettsia. R. parkeri cepa Mata Atlântica causa Febre Maculosa branda em humanos e foi detectada em A. ovale, única espécie incriminada na transmissão dessa doença. As demais espécies de riquétsias detectadas não apresentam relatos de doença em humanos. As áreas preservadas possuem menor circulação de riquétsias quando comparado a áreas com antropizadas, estando a doença intimamente relacionada ao desmatamento e urbanização de ambiente naturais

Reply: Genetic polymorphisms and cerebrovascular disease in children with sickle cell anemia from Rio de Janeiro, Brazil

The aim of the present work was to examine possible genetic risk factors related to the occurrence of cerebrovascular disease (CVD) in Brazilian population, the frequency of βS-globin gene haplotypes and co-inheritance with α-thalassemia (-α3.7kb) and single nucleotide polymorphism of methylenetetrahydrofolate reductase (MTHFR-C677T), Factor V Leiden (FV-G1691A) and prothrombin (PT-G20210A) genes in children from Rio de Janeiro. Ninety four children with sickle cell anemia (SCA) were included, 24 patients with cerebrovascular involvement and 70 patients without CVD as control group. The mean age of children at the time of the cerebrovascular event was similar to the control group. The frequency of -α3.7kb thalassemia was similar in both groups (p=0.751). Children with Bantu/Atypical βS-globin gene haplotype presented 15 times more chance (OR=15.4 CI 95% 2.9-81.6) of CVD than the other βS-globin gene haplotypes. The C677T polymorphism of MTHFR gene was similar in both groups (p=0.085). No mutation in the FV Leiden or PT genes was found. A large study seems necessary to establish the role of these genetic polymorphisms in Brazilian miscegenated population

ATLANTIC BIRD TRAITS: a data set of bird morphological traits from the Atlantic forests of South America

Scientists have long been trying to understand why the Neotropical region holds the highest diversity of birds on Earth. Recently, there has been increased interest in morphological variation between and within species, and in how climate, topography, and anthropogenic pressures may explain and affect phenotypic variation. Because morphological data are not always available for many species at the local or regional scale, we are limited in our understanding of intra- and interspecies spatial morphological variation. Here, we present the ATLANTIC BIRD TRAITS, a data set that includes measurements of up to 44 morphological traits in 67,197 bird records from 2,790 populations distributed throughout the Atlantic forests of South America. This data set comprises information, compiled over two centuries (1820–2018), for 711 bird species, which represent 80% of all known bird diversity in the Atlantic Forest. Among the most commonly reported traits are sex (n = 65,717), age (n = 63,852), body mass (n = 58,768), flight molt presence (n = 44,941), molt presence (n = 44,847), body molt presence (n = 44,606), tail length (n = 43,005), reproductive stage (n = 42,588), bill length (n = 37,409), body length (n = 28,394), right wing length (n = 21,950), tarsus length (n = 20,342), and wing length (n = 18,071). The most frequently recorded species are Chiroxiphia caudata (n = 1,837), Turdus albicollis (n = 1,658), Trichothraupis melanops (n = 1,468), Turdus leucomelas (n = 1,436), and Basileuterus culicivorus (n = 1,384). The species recorded in the greatest number of sampling localities are Basileuterus culicivorus (n = 243), Trichothraupis melanops (n = 242), Chiroxiphia caudata (n = 210), Platyrinchus mystaceus (n = 208), and Turdus rufiventris (n = 191). ATLANTIC BIRD TRAITS (ABT) is the most comprehensive data set on measurements of bird morphological traits found in a biodiversity hotspot; it provides data for basic and applied research at multiple scales, from individual to community, and from the local to the macroecological perspectives. No copyright or proprietary restrictions are associated with the use of this data set. Please cite this data paper when the data are used in publications or teaching and educational activities. © 2019 The Authors. Ecology © 2019 The Ecological Society of Americ

ATLANTIC BIRD TRAITS

Scientists have long been trying to understand why the Neotropical region holds the highest diversity of birds on Earth. Recently, there has been increased interest in morphological variation between and within species, and in how climate, topography, and anthropogenic pressures may explain and affect phenotypic variation. Because morphological data are not always available for many species at the local or regional scale, we are limited in our understanding of intra- and interspecies spatial morphological variation. Here, we present the ATLANTIC BIRD TRAITS, a data set that includes measurements of up to 44 morphological traits in 67,197 bird records from 2,790 populations distributed throughout the Atlantic forests of South America. This data set comprises information, compiled over two centuries (1820–2018), for 711 bird species, which represent 80% of all known bird diversity in the Atlantic Forest. Among the most commonly reported traits are sex (n = 65,717), age (n = 63,852), body mass (n = 58,768), flight molt presence (n = 44,941), molt presence (n = 44,847), body molt presence (n = 44,606), tail length (n = 43,005), reproductive stage (n = 42,588), bill length (n = 37,409), body length (n = 28,394), right wing length (n = 21,950), tarsus length (n = 20,342), and wing length (n = 18,071). The most frequently recorded species are Chiroxiphia caudata (n = 1,837), Turdus albicollis (n = 1,658), Trichothraupis melanops (n = 1,468), Turdus leucomelas (n = 1,436), and Basileuterus culicivorus (n = 1,384). The species recorded in the greatest number of sampling localities are Basileuterus culicivorus (n = 243), Trichothraupis melanops (n = 242), Chiroxiphia caudata (n = 210), Platyrinchus mystaceus (n = 208), and Turdus rufiventris (n = 191). ATLANTIC BIRD TRAITS (ABT) is the most comprehensive data set on measurements of bird morphological traits found in a biodiversity hotspot; it provides data for basic and applied research at multiple scales, from individual to community, and from the local to the macroecological perspectives. No copyright or proprietary restrictions are associated with the use of this data set. Please cite this data paper when the data are used in publications or teaching and educational activities. © 2019 The Authors. Ecology © 2019 The Ecological Society of Americ

Effect of lung recruitment and titrated Positive End-Expiratory Pressure (PEEP) vs low PEEP on mortality in patients with acute respiratory distress syndrome - A randomized clinical trial

IMPORTANCE: The effects of recruitment maneuvers and positive end-expiratory pressure (PEEP) titration on clinical outcomes in patients with acute respiratory distress syndrome (ARDS) remain uncertain. OBJECTIVE: To determine if lung recruitment associated with PEEP titration according to the best respiratory-system compliance decreases 28-day mortality of patients with moderate to severe ARDS compared with a conventional low-PEEP strategy. DESIGN, SETTING, AND PARTICIPANTS: Multicenter, randomized trial conducted at 120 intensive care units (ICUs) from 9 countries from November 17, 2011, through April 25, 2017, enrolling adults with moderate to severe ARDS. INTERVENTIONS: An experimental strategy with a lung recruitment maneuver and PEEP titration according to the best respiratory-system compliance (n = 501; experimental group) or a control strategy of low PEEP (n = 509). All patients received volume-assist control mode until weaning. MAIN OUTCOMES AND MEASURES: The primary outcome was all-cause mortality until 28 days. Secondary outcomes were length of ICU and hospital stay; ventilator-free days through day 28; pneumothorax requiring drainage within 7 days; barotrauma within 7 days; and ICU, in-hospital, and 6-month mortality. RESULTS: A total of 1010 patients (37.5% female; mean [SD] age, 50.9 [17.4] years) were enrolled and followed up. At 28 days, 277 of 501 patients (55.3%) in the experimental group and 251 of 509 patients (49.3%) in the control group had died (hazard ratio [HR], 1.20; 95% CI, 1.01 to 1.42; P = .041). Compared with the control group, the experimental group strategy increased 6-month mortality (65.3% vs 59.9%; HR, 1.18; 95% CI, 1.01 to 1.38; P = .04), decreased the number of mean ventilator-free days (5.3 vs 6.4; difference, −1.1; 95% CI, −2.1 to −0.1; P = .03), increased the risk of pneumothorax requiring drainage (3.2% vs 1.2%; difference, 2.0%; 95% CI, 0.0% to 4.0%; P = .03), and the risk of barotrauma (5.6% vs 1.6%; difference, 4.0%; 95% CI, 1.5% to 6.5%; P = .001). There were no significant differences in the length of ICU stay, length of hospital stay, ICU mortality, and in-hospital mortality. CONCLUSIONS AND RELEVANCE: In patients with moderate to severe ARDS, a strategy with lung recruitment and titrated PEEP compared with low PEEP increased 28-day all-cause mortality. These findings do not support the routine use of lung recruitment maneuver and PEEP titration in these patients. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT01374022

Geoeconomic variations in epidemiology, ventilation management, and outcomes in invasively ventilated intensive care unit patients without acute respiratory distress syndrome: a pooled analysis of four observational studies

Background: Geoeconomic variations in epidemiology, the practice of ventilation, and outcome in invasively ventilated intensive care unit (ICU) patients without acute respiratory distress syndrome (ARDS) remain unexplored. In this analysis we aim to address these gaps using individual patient data of four large observational studies. Methods: In this pooled analysis we harmonised individual patient data from the ERICC, LUNG SAFE, PRoVENT, and PRoVENT-iMiC prospective observational studies, which were conducted from June, 2011, to December, 2018, in 534 ICUs in 54 countries. We used the 2016 World Bank classification to define two geoeconomic regions: middle-income countries (MICs) and high-income countries (HICs). ARDS was defined according to the Berlin criteria. Descriptive statistics were used to compare patients in MICs versus HICs. The primary outcome was the use of low tidal volume ventilation (LTVV) for the first 3 days of mechanical ventilation. Secondary outcomes were key ventilation parameters (tidal volume size, positive end-expiratory pressure, fraction of inspired oxygen, peak pressure, plateau pressure, driving pressure, and respiratory rate), patient characteristics, the risk for and actual development of acute respiratory distress syndrome after the first day of ventilation, duration of ventilation, ICU length of stay, and ICU mortality. Findings: Of the 7608 patients included in the original studies, this analysis included 3852 patients without ARDS, of whom 2345 were from MICs and 1507 were from HICs. Patients in MICs were younger, shorter and with a slightly lower body-mass index, more often had diabetes and active cancer, but less often chronic obstructive pulmonary disease and heart failure than patients from HICs. Sequential organ failure assessment scores were similar in MICs and HICs. Use of LTVV in MICs and HICs was comparable (42·4% vs 44·2%; absolute difference -1·69 [-9·58 to 6·11] p=0·67; data available in 3174 [82%] of 3852 patients). The median applied positive end expiratory pressure was lower in MICs than in HICs (5 [IQR 5-8] vs 6 [5-8] cm H2O; p=0·0011). ICU mortality was higher in MICs than in HICs (30·5% vs 19·9%; p=0·0004; adjusted effect 16·41% [95% CI 9·52-23·52]; p<0·0001) and was inversely associated with gross domestic product (adjusted odds ratio for a US$10 000 increase per capita 0·80 [95% CI 0·75-0·86]; p<0·0001). Interpretation: Despite similar disease severity and ventilation management, ICU mortality in patients without ARDS is higher in MICs than in HICs, with a strong association with country-level economic status