Resumo: A malária é um problema de saúde pública e os casos diagnosticados na capital de Roraima, Brasil, têm potencial para caracterizar a carga da doença em todo o estado. O objetivo deste estudo foi descrever os aspectos epidemiológicos, clínicos e laboratoriais dos casos de malária diagnosticados em Boa Vista. Para tanto, foi realizado um estudo descritivo e transversal em duas unidades de saúde do município, com indivíduos diagnosticados com malária e que concordaram em responder ao questionário. De 206 participantes, caracterizados como homens, pardos e jovens, 96% (198) relataram atividade de garimpo ilegal. Entre garimpeiros, 66% (131) vieram de outros estados do Brasil ou de outros países. As minas estavam localizadas principalmente no território Yanomami, em Roraima. A infecção por Plasmodium vivax ocorreu em 74% (153) dos participantes. Entre garimpeiros, houve relatos de internações por malária grave, ataques prévios de malária e atrasos no tratamento após o início dos sintomas. Embora 73% (145) dos garimpeiros tenham relatado saber como a malária era transmitida, apenas 54% (107) usavam mosquiteiros ou repelentes. O uso de Artecom e cloroquina pelos garimpeiros não se destina ao tratamento completo, mas apenas ao alívio dos sintomas, permitindo o retorno às minas de ouro. Isso ressalta a importância da vigilância molecular para detectar a resistência antimalárica. Os indígenas são considerados uma população vulnerável à malária, e os garimpeiros promovem o aumento da malária nas Terras Indígenas de Roraima. Portanto, o acesso ao diagnóstico e tratamento em áreas indígenas invadidas por garimpeiros é imperativo para o enfrentamento dessa doença que assola as comunidades indígenas e ameaça a saúde pública em larga escala na meta de erradicar a malária no estado.

Resumen: La malaria es un problema de salud pública y los casos diagnosticados en la capital de Roraima, Brasil, tienen el potencial de caracterizar la carga de la enfermedad en todo el estado. El objetivo de este estudio fue describir los aspectos epidemiológicos, clínicos y de laboratorio de los casos de malaria diagnosticados en Boa Vista. Para ello, se realizó un estudio descriptivo y transversal en dos unidades de salud del municipio, con personas diagnosticadas con malaria y que aceptaron responder el cuestionario. De 206 participantes, caracterizados como hombres, pardos y jóvenes, el 96% (198) reportó actividad minera ilegal. Entre los mineros, el 66% (131) procedían de otros estados de Brasil o de otros países. Las minas estaban ubicadas principalmente en el territorio Yanomami, en Roraima. La infección por Plasmodium vivax se produjo en el 74% (153) de los participantes. Entre los mineros, hubo relatos de hospitalizaciones por malaria grave, ataques previos de malaria y retrasos en el tratamiento tras de la aparición de los síntomas. Aunque el 73% (145) de los mineros informó saber cómo se transmitía la malaria, solo el 54% (107) usaba mosquiteros o repelentes. El uso de Artecom y cloroquina por parte de los mineros no está destinado a un tratamiento completo, sino al alivio de los síntomas para permitir el regreso a las minas de oro. Esto resalta la importancia de la vigilancia molecular para detectar la resistencia a los antipalúdicos. Los indígenas son considerados una población vulnerable a la malaria, y los mineros promueven el aumento de la malaria en las Tierras Indígenas de Roraima. Por lo tanto, el acceso al diagnóstico y tratamiento en zonas indígenas invadidas por mineros es imperativo para combatir esta enfermedad que asola a las comunidades indígenas y amenaza la salud pública a gran escala en el objetivo de erradicar la malaria en el estado.

Abstract: Malaria is a public health problem and the cases diagnosed in the capital of Roraima, Brazil, show potential to characterize the burden of the disease in the state. This study aimed to describe the epidemiological, clinical, and laboratory aspects of malaria cases diagnosed in Boa Vista. For this purpose, a descriptive, cross-sectional study was conducted in two health units in the city, with individuals diagnosed and who agreed to respond the questionnaire. Of the total of 206 participants, characterized as men, mixed-race, and young, 96% (198) reported participating in illegal mining activity. Among the group of miners, 66% (131) came from other states of Brazil or other countries. The mines were mainly located in the Yanomami territory in Roraima. Plasmodium vivax infection occurred in 74% (153) of participants. In the miner’s group, hospitalizations for severe malaria, previous malaria attacks, and delays in treatment after the onset of symptoms were reported. Although 73% (145) of miners reported knowing how malaria was transmitted, only 54% (107) used mosquito nets or repellents. The use of Artecom and chloroquine by miners is not for the complete treatment but only to relieve symptoms for returning to gold mines, highlighting the importance of molecular surveillance to antimalarial resistance. Indigenous peoples are considered vulnerable to malaria and miners promotes the increase of malaria in Roraima Indigenous Lands. Therefore, access to diagnosis and treatment in Indigenous areas invaded by miners is imperative to confront this disease that ravages Indigenous communities and threatens public health on a large scale to achieve the goal of eliminating malaria in the state.

Resumen: La malaria es un problema de salud pública y los casos diagnosticados en la capital de Roraima, Brasil, tienen el potencial de caracterizar la carga de la enfermedad en todo el estado. El objetivo de este estudio fue describir los aspectos epidemiológicos, clínicos y de laboratorio de los casos de malaria diagnosticados en Boa Vista. Para ello, se realizó un estudio descriptivo y transversal en dos unidades de salud del municipio, con personas diagnosticadas con malaria y que aceptaron responder el cuestionario. De 206 participantes, caracterizados como hombres, pardos y jóvenes, el 96% (198) reportó actividad minera ilegal. Entre los mineros, el 66% (131) procedían de otros estados de Brasil o de otros países. Las minas estaban ubicadas principalmente en el territorio Yanomami, en Roraima. La infección por Plasmodium vivax se produjo en el 74% (153) de los participantes. Entre los mineros, hubo relatos de hospitalizaciones por malaria grave, ataques previos de malaria y retrasos en el tratamiento tras de la aparición de los síntomas. Aunque el 73% (145) de los mineros informó saber cómo se transmitía la malaria, solo el 54% (107) usaba mosquiteros o repelentes. El uso de Artecom y cloroquina por parte de los mineros no está destinado a un tratamiento completo, sino al alivio de los síntomas para permitir el regreso a las minas de oro. Esto resalta la importancia de la vigilancia molecular para detectar la resistencia a los antipalúdicos. Los indígenas son considerados una población vulnerable a la malaria, y los mineros promueven el aumento de la malaria en las Tierras Indígenas de Roraima. Por lo tanto, el acceso al diagnóstico y tratamiento en zonas indígenas invadidas por mineros es imperativo para combatir esta enfermedad que asola a las comunidades indígenas y amenaza la salud pública a gran escala en el objetivo de erradicar la malaria en el estado. Resumen Roraima estado epidemiológicos Vista ello municipio cuestionario 20 participantes hombres jóvenes 96 198 (198 ilegal 66 131 (131 países Yanomami 74 153 (153 grave 73 145 (145 54 107 (107 repelentes completo oro antipalúdicos tanto 2 9 19 (19 6 13 (13 7 15 (15 14 (14 5 10 (10 1 (1 (

Resumo: A malária é um problema de saúde pública e os casos diagnosticados na capital de Roraima, Brasil, têm potencial para caracterizar a carga da doença em todo o estado. O objetivo deste estudo foi descrever os aspectos epidemiológicos, clínicos e laboratoriais dos casos de malária diagnosticados em Boa Vista. Para tanto, foi realizado um estudo descritivo e transversal em duas unidades de saúde do município, com indivíduos diagnosticados com malária e que concordaram em responder ao questionário. De 206 participantes, caracterizados como homens, pardos e jovens, 96% (198) relataram atividade de garimpo ilegal. Entre garimpeiros, 66% (131) vieram de outros estados do Brasil ou de outros países. As minas estavam localizadas principalmente no território Yanomami, em Roraima. A infecção por Plasmodium vivax ocorreu em 74% (153) dos participantes. Entre garimpeiros, houve relatos de internações por malária grave, ataques prévios de malária e atrasos no tratamento após o início dos sintomas. Embora 73% (145) dos garimpeiros tenham relatado saber como a malária era transmitida, apenas 54% (107) usavam mosquiteiros ou repelentes. O uso de Artecom e cloroquina pelos garimpeiros não se destina ao tratamento completo, mas apenas ao alívio dos sintomas, permitindo o retorno às minas de ouro. Isso ressalta a importância da vigilância molecular para detectar a resistência antimalárica. Os indígenas são considerados uma população vulnerável à malária, e os garimpeiros promovem o aumento da malária nas Terras Indígenas de Roraima. Portanto, o acesso ao diagnóstico e tratamento em áreas indígenas invadidas por garimpeiros é imperativo para o enfrentamento dessa doença que assola as comunidades indígenas e ameaça a saúde pública em larga escala na meta de erradicar a malária no estado. Resumo Roraima estado epidemiológicos Vista tanto município questionário 20 participantes homens jovens 96 198 (198 ilegal 66 131 (131 países Yanomami 74 153 (153 grave sintomas 73 145 (145 transmitida 54 107 (107 repelentes completo ouro antimalárica Portanto 2 9 19 (19 6 13 (13 7 15 (15 14 (14 5 10 (10 1 (1 (

Abstract: Malaria is a public health problem and the cases diagnosed in the capital of Roraima, Brazil, show potential to characterize the burden of the disease in the state. This study aimed to describe the epidemiological, clinical, and laboratory aspects of malaria cases diagnosed in Boa Vista. For this purpose, a descriptive, cross-sectional study was conducted in two health units in the city, with individuals diagnosed and who agreed to respond the questionnaire. Of the total of 206 participants, characterized as men, mixed-race, and young, 96% (198) reported participating in illegal mining activity. Among the group of miners, 66% (131) came from other states of Brazil or other countries. The mines were mainly located in the Yanomami territory in Roraima. Plasmodium vivax infection occurred in 74% (153) of participants. In the miner’s group, hospitalizations for severe malaria, previous malaria attacks, and delays in treatment after the onset of symptoms were reported. Although 73% (145) of miners reported knowing how malaria was transmitted, only 54% (107) used mosquito nets or repellents. The use of Artecom and chloroquine by miners is not for the complete treatment but only to relieve symptoms for returning to gold mines, highlighting the importance of molecular surveillance to antimalarial resistance. Indigenous peoples are considered vulnerable to malaria and miners promotes the increase of malaria in Roraima Indigenous Lands. Therefore, access to diagnosis and treatment in Indigenous areas invaded by miners is imperative to confront this disease that ravages Indigenous communities and threatens public health on a large scale to achieve the goal of eliminating malaria in the state. Abstract state epidemiological clinical Vista purpose descriptive crosssectional cross sectional city questionnaire 20 participants men mixedrace, mixedrace mixed race, race mixed-race young 96 198 (198 activity 66 131 (131 countries 74 153 (153 miner s attacks 73 145 (145 transmitted 54 107 (107 repellents resistance Lands Therefore 2 9 19 (19 6 13 (13 7 15 (15 14 (14 5 10 (10 1 (1 (

BACKGROUND Although blood transfusion is an essential therapeutic procedure, it can present risks, including transmitting infectious diseases, such as malaria. In Acre, the thick blood smear microscopic examination (TBS) is used to screen infected malaria blood donors. However, TBS has low sensitivity for detecting Plasmodium in situations of low parasitaemia, such as those presented by asymptomatic clinically healthy individuals. OBJECTIVES To investigate the pertinence of using polymerase chain reaction (PCR) to detect malarial infection for screening blood donors in Cruzeiro do Sul, Acre, an endemic high-risk malaria area in the Legal Amazon. METHODS A cross-sectional study was conducted among individuals eligible and ineligible to be blood donors, according to clinical and epidemiological criteria. Besides the mandatory screening of HCV, HBV, and HIV tests, malaria PCR and TBS were also carried out on all blood donor candidates who attended the Cruzeiro do Sul Blood Centre from July to September 2022. FINDINGS Of the 230 participants, 209 (91%) were eligible for blood donation by clinical-epidemiological screening. Surprisingly, no blood donor candidate reported a history of malaria. All TBS microscopic tests were negative at the time of recruitment. However, samples from four blood donor candidates (two eligible by clinical and epidemiological malaria criteria and two ineligible by hypertension and recent tattoo) were positive by Plasmodium and P. vivax molecular tests. MAIN CONCLUSIONS Malaria molecular techniques for screening blood donors should be introduced in the Brazilian Blood Centres to maximise recipient safety. Furthermore, selecting zero-risk donors could pave the way to build a transmissible malaria-free environment in the blood bank context in the near future. procedure risks diseases Acre (TBS However parasitaemia (PCR highrisk high risk Amazon crosssectional cross sectional HCV HBV 2022 23 participants 20 91% 91 (91% clinicalepidemiological Surprisingly recruitment tattoo P safety Furthermore zerorisk zero malariafree free future 202 2 9 (91 (9 (

Unforeseen Plasmodium infections in the Atlantic Forest of Brazilian Extra-Amazonian region could jeopardise malaria elimination. A human malaria case was registered in Três Forquilhas, in the Atlantic Forest biome of Rio Grande do Sul, after a 45 years’ time-lapsed without any malaria autochthonous notification in this southern Brazilian state. This finding represents the expansion of the malaria distribution areas in Brazil and the southernmost human malaria case record in South America in this decade. The coexistence of the bromeliad-breeding vector Anopheles (Kerteszia) cruzii and non-human primates in the Atlantic Forest regularly visited by the patient claimed for the zoonotic origin of this infection. The reemergence of Atlantic Forest human malaria in Rio Grande do Sul was also discussed.

BACKGROUND The prompt diagnosis of plasmodial species for effective treatment prevents worsening of individual health and avoids transmission maintenance or even malaria reintroduction in areas where Plasmodium does not exist. Polymerase chain reaction (PCR) allows for the detection of parasites below the threshold of microscopic examination. OBJECTIVE Our aim was to develop a real-time PCR test to reduce diagnostic errors and increase efficacy. METHODS The lower limit of quantification and the linearity/analytical sensitivity to measure sensitivity or limit of detection (LoD) were determined. Intra-assay variations (repeatability) and alterations between assays, operators, and instruments (reproducibility) were also assessed to set precision. FINDINGS The linearity in SYBR™ Green and TaqMan™ systems was 106 and 102 copies and analytical sensitivity 1.13 and 1.17 copies/μL, respectively. Real-time PCR was more sensitive than conventional PCR, showing a LoD of 0.01 parasite (p)/μL. Reproducibility and repeatability (precision) were 100% for up to 0.1 p/μL in SYBR™ Green and 1 p/μL in TaqMan™ and conventional PCR. CONCLUSION Real-time PCR may replace conventional PCR in reference laboratories for P. vivax detection due to its rapidity. The TaqMan™ system is the most indicated when quantification assays are required. Performing tests in triplicate when diagnosing Plasmodium-infected-asymptomatic individuals is recommended to minimise diagnostic errors.

BACKGROUND AND OBJECTIVE Brazil is responsible for a large number of Plasmodium vivax cases in America. Given the emergence of P. vivax parasites resistant to chloroquine and the effectiveness of antifolates in vivax malaria treatment together with a correlation between mutations in P. vivax dhfr and dhps genes and SP treatment failure, the point mutations in these genes were investigated. METHODS Blood samples from 54 patients experiencing vivax malaria symptomatic episodes in the Amazonian Region were investigated. Genomic DNA was extracted using a DNA extraction kit (QIAGENTM). Nested polymerase chain reaction (PCR) amplification was carried out followed by Sanger sequencing to detect single nucleotide polymorphisms (SNPs). FINDINGS All tested isolates showed non-synonymous mutations in pvdhfr gene: 117N (54/54, 100%) and 58R (25/54, 46%). Double mutant allele 58R/117N (FRTNI, 28%) was the most frequent followed by triple mutant alleles (58R/117N/173L, FRTNL, 11%; 58R/61M/117N, FRMNI, 5% 117N/173L, FSTNL, 4%) and quadruple mutant allele (58R/61M/117N/173L, FRMNL, 2%). A single mutation was observed at codon C383G in pvdhps gene (SGKAV, 48%). CONCLUSION No evidence of molecular signatures associated with P. vivax resistance to SP was observed in the Brazilian samples.

In Brazil, malaria still remains a clinically important febrile syndrome for local populations and travelers, occurring mostly in the Amazon Basin. This review aims to report the main efforts employed to control this disease since the 1940s and the emergence of Plasmodium falciparum and Plasmodium vivax chemoresistance to chloroquine and sulphadoxine-pyrimethamine among other drugs. Additionally, in vivo, in vitro and molecular studies as well as malaria chemoresistance consequences on disease morbidity and policy treatment guidelines were commented.

The relationship between autoimmunity and malaria is not well understood. To determine whether autoimmune responses have a protective role during malaria, we studied the pattern of reactivity to plasmodial antigens of sera from 93 patients with 14 different autoimmune diseases (AID) who were not previously exposed to malaria. Sera from patients with 13 different AID reacted against Plasmodium falciparum by indirect fluorescent antibody test with frequencies varying from 33-100%. In addition, sera from 37 AID patients were tested for reactivity against Plasmodium yoelii 17XNL and the asexual blood stage forms of three different P. falciparum strains. In general, the frequency of reactive sera was higher against young trophozoites than schizonts (p < 0.05 for 2 strains), indicating that the antigenic determinants targeted by the tested AID sera might be more highly expressed by the former stage. The ability of monoclonal auto-antibodies (auto-Ab) to inhibit P. falciparum growth in vitro was also tested. Thirteen of the 18 monoclonal auto-Ab tested (72%), but none of the control monoclonal antibodies, inhibited parasite growth, in some cases by greater than 40%. We conclude that autoimmune responses mediated by auto-Ab may present anti-plasmodial activity.

The glutamate-rich protein (GLURP) is an exoantigen expressed in all stages of the Plasmodium falciparum life cycle in humans. Anti-GLURP antibodies can inhibit parasite growth in the presence of monocytes via antibody-dependent cellular inhibition (ADCI), and a major parasite-inhibitory region has been found in the N-terminal R0 region of the protein. Herein, we describe the antiplasmodial activity of anti-GLURP antibodies present in the sera from individuals naturally exposed to malaria in a Brazilian malaria-endemic area. The anti-R0 antibodies showed a potent inhibitory effect on the growth of P. falciparum in vitro, both in the presence (ADCI) and absence (GI) of monocytes. The inhibitory effect on parasite growth was comparable to the effect of IgGs purified from pooled sera from hyperimmune African individuals. Interestingly, in the ADCI test, higher levels of tumour necrosis factor alpha (TNF-α) were observed in the supernatant from cultures with higher parasitemias. Our data suggest that the antibody response induced by GLURP-R0 in naturally exposed individuals may have an important role in controlling parasitemia because these antibodies are able to inhibit the in vitro growth of P. falciparum with or without the cooperation from monocytes. Our results also indicate that TNF-α may not be relevant for the inhibitory effect on P. falciparum in vitro growth.

INTRODUÇÃO: A letalidade da malária na região extra-amazônica é cerca de 80 vezes maior do que na Amazônia, que concentra 99,8% dos casos do país. Em áreas de transmissão de dengue, como o Rio de Janeiro, o atraso no diagnóstico e tratamento da malária dos pacientes com febre, provenientes de áreas endêmicas de malária, pode ser, entre outros fatores, devido à confusão entre o diagnóstico das duas doenças pelos generalistas da rede de assistência médica. Neste trabalho, apresentamos as consequências do atraso diagnóstico em três pacientes com malária por Plasmodium falciparum; P. malariae e P. vivax, que, após o périplo habitual para tratamento de dengue, procuraram a nossa instituição onde foram corretamente diagnosticados e submetidos aos tratamentos adequados. MÉTODOS: Descrição de três casos de malária diagnosticada tardiamente e encaminhados ao IPEC/ FIOCRUZ, entre os anos de 2007 e 2008. RESULTADOS: uma brasileira proveniente de Moçambique, primo-infectada por P. falciparum, com malária diagnosticada após 6 dias do início da febre, morreu com malária cerebral e choque. Outro paciente com malária por P. malariae teve um curso grave e prolongado, mas ficou curado após o tratamento específico. A terceira paciente diagnosticada tardiamente apresentou malária por P. vivax adquirida na região de Mata Atlântica no Estado do Rio. CONCLUSÕES: Os profissionais de saúde do Rio devem ser treinados para aperfeiçoar a vigilância e o tratamento oportuno da malária e evitar desfechos mórbidos e fatais. Sugere-se que uma investigação de focos de malária autóctone em áreas de mata no estado seja realizada.

INTRODUCTION: The mortality of malaria in the extra-Amazon region is about 80 times higher than in the Amazon region, where malaria is concentrated (99.8% of cases). In areas of dengue transmission, delay in the diagnosis and treatment of malaria in patients with fever who reside in areas of malaria transmission can be due to the confusion between the clinical diagnoses of both diseases by nonspecialist doctors, among other factors. This work presents some of the consequences of delayed diagnosis in three patients with malaria by Plasmodium falciparum, P. malariae and P. vivax, who, after following the usual route for Dengue treatment, sought our institution, where they were correctly diagnosed and adequately treated. METHODS: Description of three cases of malaria with delayed diagnosed malaria referred to the Outpatient Clinic for Acute Febrile Diseases, IPEC/FIOCRUZ-RJ, between 2007 and 2008. RESULTS: A Brazilian from Mozambique, primo-infected with P. falciparum was diagnosed with malaria six days after the onset of fever and died of cerebral malaria and shock. Another patient with P.malariae malaria presented a severe and prolonged course, but was cured after specific treatment. A third patient, with delayed diagnosis of P. vivax malaria, acquired it in the Atlantic Forest region in the State of Rio. CONCLUSIONS: Health professionals from non-endemic areas for malaria should be trained to optimize the surveillance and early treatment of malaria and prevent morbid and fatal outcomes. An investigation of outbreaks of autochthonous malaria in the State of Rio de Janeiro is suggested.

Plasmodium vivax control is now being hampered by drug resistance. Orthologous Plasmodium falciparum genes linked to chloroquine or sulfadoxine-pyrimethamine chemoresistance have been identified in P. vivax parasites, but few studies have been performed. The goal of the present work is to characterise pvmdr1 and pvdhfr genes in parasite isolates from a Brazilian endemic area where no molecular investigation had been previously conducted. The pvmdr1 analysis revealed the existence of single (85.7%) and double (14.3%) mutant haplotypes, while the pvdhfr examination showed the presence of double (57.2%) and triple (42.8%) mutant haplotypes. The implications of these findings are discussed.

The antibody response to Plasmodium falciparum parasites of naturally infected population is critical to elucidate the role of polymorphic alleles in malaria. Thus, we evaluated the impact of antigenic diversity of repetitive and family dimorphic domains of the merozoite surface protein 2 (MSP-2) on immune response of 96 individuals living in Peixoto de Azevedo (MT-Brazil), by ELISA using recombinant MSP-2 proteins. The majority of these individuals were carrying FC27-type infections. IgG antibody responses were predominantly directed to FC27 parasites and were correlated to the extension of polymorphism presented by each MSP-2 region. This finding demonstrated the impact of the genetic polymorphism on antibody response and therefore, its importance on malaria vaccine efficacy.

In this work we investigated the frequency of polymorphism in exon II of the gene encoding most of the amino-terminal region of the serine rich antigen (SERA) in Plasmodium falciparum field samples. The blood samples were colleted from P. falciparum infected individuals in three areas of the Brazilian Amazon. Two fragments have been characterized by polymerase chain reaction: one of 175 bp corresponding to the repeat region with 5 octamer units and one other of 199 bp related to the 6 repeat octamer units of SERA protein. The 199 bp fragment was the predominant one in all the studied areas. The higher frequency of this fragment has not been described before and could be explained by an immunological selection of the plasmodial population in the infected individuals under study. Since repeat motifs in the amino-terminal region of SERA contain epitopes recognized by parasite-inhibitor antibodies, data reported here suggest that the analysis of the polymorphism of P. falciparum isolates in different geographical areas is a preliminary stage before the final drawing of an universal vaccine against malaria can be reached.