ROSELY DOS SANTOS MALAFRONTE
Projetos de Pesquisa
Unidades Organizacionais
SCPROTOZ-83, Instituto de Medicina Tropical
LIM/49 - Laboratório de Protozoologia, Hospital das Clínicas, Faculdade de Medicina - Líder
LIM/49 - Laboratório de Protozoologia, Hospital das Clínicas, Faculdade de Medicina - Líder
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article 25 Citação(ões) na Scopus Intragenomic variation in the second internal transcribed spacer of the ribosomal DNA of species of the genera Culex and Lutzia (Diptera: Culicidae)(2011) VESGUEIRO, Fabiana Tavares; DEMARI-SILVA, Bruna; MALAFRONTE, Rosely dos Santos; SALLUM, Maria Anice Mureb; MARRELLI, Mauro ToledoCulex is the largest genus of Culicini and includes vectors of several arboviruses and filarial worms. Many species of Culex are morphologically similar, which makes their identification difficult, particularly when using female specimens. To aid evolutionary studies and species distinction, molecular techniques are often used. Sequences of the second internal transcribed spacer (ITS2) of ribosomal DNA (rDNA) from 16 species of the genus Culex and one of Lutzia were used to assess their genomic variability and to verify their applicability in the phylogenetic analysis of the group. The distance matrix (uncorrected p-distance) that was obtained revealed intragenomic and intraspecific variation. Because of the intragenomic variability, we selected ITS2 copies for use in distance analyses based on their secondary structures. Neighbour-joining topology was obtained with an uncorrected p-distance. Despite the heterogeneity observed, individuals of the same species were grouped together and correlated with the current, morphology-based classification, thereby showing that ITS2 is an appropriate marker to be used in the taxonomy of Culex.- Assessment of asymptomatic Plasmodium spp. infection by detection of parasite DNA in residents of an extra-Amazonian region of Brazil(2018) ALENCAR, Filomena E. C. de; MALAFRONTE, Rosely dos Santos; CERUTTI JUNIOR, Crispim; FERNANDES, Licia Natal; BUERY, Julyana Cerqueira; FUX, Blima; REZENDE, Helder Ricas; DUARTE, Ana Maria Ribeiro de Castro; MEDEIROS-SOUSA, Antonio Ralph; MIRANDA, Angelica EspinosaBackground: The hypotheses put forward to explain the malaria transmission cycle in extra-Amazonian Brazil, an area of very low malaria incidence, are based on either a zoonotic scenario involving simian malaria, or a scenario in which asymptomatic carriers play an important role. Objectives: To determine the incidence of asymptomatic infection by detecting Plasmodium spp. DNA and its role in residual malaria transmission in a non-Amazonian region of Brazil. Methods: Upon the report of the first malaria case in 2010 in the Atlantic Forest region of the state of Espirito Santo, inhabitants within a 2 km radius were invited to participate in a follow-up study. After providing signed informed consent forms, inhabitants filled out a questionnaire and gave blood samples for PCR, and thick and thin smears. Followup visits were performed every 3 months over a 21 month period, when new samples were collected and information was updated. Results: Ninety-two individuals were initially included for follow-up. At the first collection, all of them were clearly asymptomatic. One individual was positive for Plasmodium vivax, one for Plasmodium malariae and one for both P. vivax and P. malariae, corresponding to a prevalence of 3.4% (2.3% for each species). During follow-up, four new PCR-positive cases (two for each species) were recorded, corresponding to an incidence of 2.5 infections per 100 personyears or 1.25 infections per 100 person-years for each species. A mathematical transmission model was applied, using a low frequency of human carriers and the vector density in the region, and calculated based on previous studies in the same locality whose results were subjected to a linear regression. This analysis suggests that the transmission chain is unlikely to be based solely on human carriers, regardless of whether they are symptomatic or not. Conclusion: The low incidence of cases and the low frequency of asymptomatic malaria carriers investigated make it unlikely that the transmission chain in the region is based solely on human hosts, as cases are isolated one from another by hundreds of kilometers and frequently by long periods of time, reinforcing instead the hypothesis of zoonotic transmission.
- Mitochondrial genome of Plasmodium vivax/simium detected in an endemic region for malaria in the Atlantic Forest of Espirito Santo state, Brazil: do mosquitoes, simians and humans harbour the same parasite?(2017) BUERY, Julyana Cerqueira; RODRIGUES, Priscila Thihara; NATAL, Licia; SALLA, Lais Camoese; LOSS, Ana Carolina; VICENTE, Creuza Rachel; REZENDE, Helder Ricas; DUARTE, Ana Maria Ribeiro de Castro; FUX, Blima; MALAFRONTE, Rosely dos Santos; FALQUETO, Aloisio; CERUTTI JR., CrispimBackground: The transmission of malaria in the extra-Amazonian regions of Brazil, although interrupted in the 1960s, has persisted to the present time in some areas of dense Atlantic Forest, with reports of cases characterized by particular transmission cycles and clinical presentations. Bromeliad-malaria, as it is named, is particularly frequent in the state of Espirito Santo, with Plasmodium vivax being the parasite commonly recognized as the aetiologic agent of human infections. With regard to the spatial and temporal distances between cases reported in this region, the transmission cycle does not fit the traditional malaria cycle. The existence of a zoonosis, with infected simians participating in the epidemiology, is therefore hypothesized. In the present study, transmission of bromeliad-malaria in Espirito Santo is investigated, based on the complete mitochondrial genome of DNA extracted from isolates of Plasmodium species, which had infected humans, a simian from the genus Allouata, and Anopheles mosquitoes. Plasmodium vivax/simium was identified in the samples by both nested PCR and real-time PCR. After amplification, the mitochondrial genome was completely sequenced and compared with a haplotype network which included all sequences of P. vivax/simium mitochondrial genomes sampled from humans and simians from all regions in Brazil. Results: The haplotype network indicates that humans and simians from the Atlantic Forest become infected by the same haplotype, but some isolates from humans are not identical to the simian isolate. In addition, the plasmodial DNA extracted from mosquitoes revealed sequences different from those obtained from simians, but similar to two isolates from humans. Conclusions: These findings strengthen support for the hypothesis that in the Atlantic Forest, and especially in the state with the highest frequency of bromeliad-malaria in Brazil, parasites with similar molecular backgrounds are shared by humans and simians. The recognized identity between P. vivax and P. simium at the species level, the sharing of haplotypes, and the participation of the same vector in transmitting the infection to both host species indicate interspecies transference of the parasites. However, the intensity, frequency and direction of this transfer remain to be clarified.
- Unexpected detection of Plasmodium vivax and Plasmodium falciparum DNA in asymptomatic blood donors: fact or artifact?(2014) MENDRONE JR., Alfredo; CERUTTI JR., Crispim; LEVI, Jose Eduardo; BOULOS, Marcos; SANCHEZ, Maria Carmen Arroyo; MALAFRONTE, Rosely dos Santos; SANTI, Silvia Maria Di; ODONE JR., VicenteA study searching for Plasmodium vivax and Plasmodium falciparum DNA among blood donors from the non-endemic area in Brazil reported a rate of 7.41%. This number is at least three times higher than what has been observed in blood donors from the Amazon, an endemic area concentrating >99% of all malaria cases in Brazil. Moreover, the majority of the donors were supposedly infected by P. falciparum, a rare finding both in men and anophelines from the Atlantic forest. These findings shall be taken with caution since they disagree with several publications in the literature and possibly overestimate the actual risk of malaria transmission by blood transfusion in Sao Paulo city.
- The genetic diversity of Plasmodium malariae and Plasmodium brasilianum from human, simian and mosquito hosts in Brazil(2012) GUIMARAES, L. O.; BAJAY, M. M.; WUNDERLICH, G.; BUENO, M. G.; ROEHE, F.; CATAO-DIAS, J. L.; NEVES, A.; MALAFRONTE, R. S.; CURADO, I.; KIRCHGATTER, K.Plasmodium malariae is a protozoan parasite that causes malaria in humans and is genetically indistinguishable from Plasmodium brasilianum, a parasite infecting New World monkeys in Central and South America. P. malariae has a wide and patchy global distribution in tropical and subtropical regions, being found in South America, Asia, and Africa. However, little is known regarding the genetics of these parasites and the similarity between them could be because until now there are only a very few genomic sequences available from simian Plasmodium species. This study presents the first molecular epidemiological data for P. malariae and P. brasilianum from Brazil obtained from different hosts and uses them to explore the genetic diversity in relation to geographical origin and hosts. By using microsatellite genotyping, we discovered that of the 14 human samples obtained from areas of the Atlantic forest, 5 different multilocus genotypes were recorded, while in a sample from an infected mosquito from the same region a different haplotype was found. We also analyzed the longitudinal change of circulating plasmodial genetic profile in two untreated non-symptomatic patients during a 12-months interval. The circulating genotypes in the two samples from the same patient presented nearly identical multilocus haplotypes (differing by a single locus). The more frequent haplotype persisted for almost 3 years in the human population. The allele Pm09-299 described previously as a genetic marker for South American P. malariae was not found in our samples. Of the 3 non-human primate samples from the Amazon Region, 3 different multilocus genotypes were recorded indicating a greater diversity among isolates of P. brasilianum compared to P. malariae and thus, P. malariae might in fact derive from P. brasilianum as has been proposed in recent studies. Taken together, our data show that based on the microsatellite data there is a relatively restricted polymorphism of P. malariae parasites as opposed to other geographic locations.
- Epidemiology of Disappearing Plasmodium vivax Malaria: A Case Study in Rural Amazonia(2014) BARBOSA, Susana; GOZZE, Amanda B.; LIMA, Nathalia F.; BATISTA, Camilla L.; BASTOS, Melissa da Silva; NICOLETE, Vanessa C.; FONTOURA, Pablo S.; GONCALVES, Raquel M.; VIANA, Susana Ariane S.; MENEZES, Maria Jose; SCOPEL, Kezia Katiani G.; CAVASINI, Carlos E.; MALAFRONTE, Rosely dos Santos; SILVA-NUNES, Monica da; VINETZ, Joseph M.; CASTRO, Marcia C.; FERREIRA, Marcelo U.Background: New frontier settlements across the Amazon Basin pose a major challenge for malaria elimination in Brazil. Here we describe the epidemiology of malaria during the early phases of occupation of farming settlements in Remansinho area, Brazilian Amazonia. We examine the relative contribution of low-density and asymptomatic parasitemias to the overall Plasmodium vivax burden over a period of declining transmission and discuss potential hurdles for malaria elimination in Remansinho and similar settings. Methods: Eight community-wide cross-sectional surveys, involving 584 subjects, were carried out in Remansinho over 3 years and complemented by active and passive surveillance of febrile illnesses between the surveys. We used quantitative PCR to detect low-density asexual parasitemias and gametocytemias missed by conventional microscopy. Mixed-effects multiple logistic regression models were used to characterize independent risk factors for P. vivax infection and disease. Principal Findings/Conclusions: P. vivax prevalence decreased from 23.8% (March-April 2010) to 3.0% (April-May 2013), with no P. falciparum infections diagnosed after March-April 2011. Although migrants from malaria-free areas were at increased risk of malaria, their odds of having P. vivax infection and disease decreased by 2-3% with each year of residence in Amazonia. Several findings indicate that low-density and asymptomatic P. vivax parasitemias may complicate residual malaria elimination in Remansinho: (a) the proportion of subpatent infections (i. e. missed by microscopy) increased from 43.8% to 73.1% as P. vivax transmission declined; (b) most (56.6%) P. vivax infections were asymptomatic and 32.8% of them were both subpatent and asymptomatic; (c) asymptomatic parasite carriers accounted for 54.4% of the total P. vivax biomass in the host population; (d) over 90% subpatent and asymptomatic P. vivax had PCR-detectable gametocytemias; and (e) few (17.0%) asymptomatic and subpatent P. vivax infections that were left untreated progressed to clinical disease over 6 weeks of follow-up and became detectable by routine malaria surveillance.
- Ecological characterisation and infection of Anophelines (Diptera: Culicidae) of the Atlantic Forest in the southeast of Brazil over a 10 year period: has the behaviour of the autochthonous malaria vector changed?(2018) BUERY, Julyana Cerqueira; REZENDE, Helder Ricas; NATAL, Licia; SILVA, Leonardo Santana da; MENEZES, Regiane Maria Tironi de; FUX, Blima; MALAFRONTE, Rosely dos Santos; FALQUETO, Aloisio; CERUTTI JUNIOR, CrispimBACKGROUND In southeastern Brazil, autochthonous cases of malaria can be found near Atlantic Forest fragments. Because the transmission cycle has not been completely clarified, the behaviour of the possible vectors in those regions must be observed. A study concerning the entomological aspects and natural infection of anophelines (Diptera: Culicidae) captured in the municipalities of the mountainous region of Espirito Santo state was performed in 2004 and 2005. Similarly, between 2014 and 2015, 12 monthly collections were performed at the same area of the study mentioned above. METHODS Center for Disease Control (CDC) light traps with CO2 were set in open areas, at the edge and inside of the forest (canopy and ground), whereas Shannon traps were set on the edge. FINDINGS A total of 1,414 anophelines were collected from 13 species. Anopheles (Kerteszia) cruzii Dyar and Knab remained the most frequently captured species in the CDC traps set in the forest canopy, as well as being the vector with the highest prevalence of Plasmodium vivax/simium infection, according to molecular polymerase chain reaction techniques. CONCLUSIONS P. vivax/simium was found only in abdomens of the mosquitoes of the subgenus Nyssorhynchus, weakening the hypothesis that this subgenus also plays a role in malaria transmission in this specific region.
- Canine visceral leishmaniasis in the metropolitan area of Sao Paulo: Pintomyia fischeri as potential vector of Leishmania infantum(2017) GALVIS-OVALLOS, Fredy; SILVA, Mariana Dantas da; BISPO, Giulia Baldaconi da Silva; OLIVEIRA, Alessandra Gutierrez de; GONCALVES NETO, Jose Rodriguez; MALAFRONTE, Rosely dos Santos; GALATI, Eunice Aparecida BianchiAmerican visceral leishmaniasis is a zoonosis caused by Leishmania infantum and transmitted mainly by Lutzomyia longipalpis. However, canine cases have been reported in the absence of this species in the Greater Sao Paulo region, where Pintomyia fischeri and Migonemyia migonei are the predominant species. This raises the suspicion that they could be acting as vectors. Therefore, this study sought to investigate specific vector capacity parameters of these species and to compare them with those of Lu. longipalpis s.l. Among these parameters the blood feeding rate, the survival, and the susceptibility to the development of Le. infantum were evaluated for the three species, and the attractiveness of dogs to Pi. fischeri and Mg. migonei was evaluated. The estimated interval between blood meals was shorter for Lu. longipalpis s.l, followed by Pi. fischeri and Mg. migonei. The infection rate with Le. infantum flagellates in Lu. longipalpis was 9.8%, in Pi. fischeri 4.8%, and in Mg. migonei nil. The respective infective life expectancies (days) of Lu. longipalpis, Mg. migonei, and Pi. fischeri were 2.4, 1.94, and 1.68. Both Pi. fischeri and Mg. migonei were captured in the kennel with a predominance (95%) of Pi. fischeri. Considering the great attractiveness of dogs to Pi. fischeri, its susceptibility to infection by Le. infantum, infective life expectancies, and predominance in Greater Sao Paulo, this study presents evidence of Pi. fischeri as a potential vector of this parasite in the region.
- Natural infection in anopheline species and its implications for autochthonous malaria in the Atlantic forest in Brazil(2013) DUARTE, Ana Maria R. C.; PEREIRA, Diego M.; PAULA, Marcia B. de; FERNANDES, Aristides; URBINATTI, Paulo R.; RIBEIRO, Andressa F.; MELLO, Maria Helena S. H.; MATOS JR., Marco O.; MUCCI, Luis F.; FERNANDES, Licia N.; NATAL, Delsio; MALAFRONTE, Rosely S.Background: A descriptive study was carried out in an area of the Atlantic Forest with autochthonous malaria in the Parelheiros subdistrict on the periphery of the municipality of Sao Paulo to identify anopheline fauna and anophelines naturally infected with Plasmodium as well as to discuss their role in this peculiar epidemiological context. Methods: Entomological captures were made from May 2009 to April 2011 using Shannon traps and automatic CDC traps in four areas chosen for their different patterns of human presence and incidences of malaria (anthropic zone 1, anthropic zone 2, transition zone and sylvatic zone). Natural Plasmodium infection was detected by nested PCR based on amplification of the 18S rRNA gene. Results: In total, 6,073 anophelines were collected from May 2009 to April 2011, and six species were identified in the four zones. Anopheles cruzii was the predominant species in the three environments but was more abundant in the sylvatic zone. Anopheles (Kerteszia) cruzii specimens from the anthropic and sylvatic zones were positive for P. vivax and P. malariae. An. (Ker.) bellator, An. (Nys.) triannulatus, An. (Nys.) strodei, An. (Nys.) lutzi and An. (Ano) maculipes were found in small numbers. Of these, An. (Nys.) triannulatus and An. (Nys.) lutzi, which were collected in the anthropic zone, were naturally infected with P. vivax while An. (Nys.) triannulatus from the anthropic zones and An. (Nys.) strodei from the transition zone were positive for P. malariae. Conclusion: These results confirm that Anopheles (Kerteszia) cruzii plays an important role as a major Plasmodium vector. However, the finding of other naturally infected species may indicate that secondary vectors are also involved in the transmission of malaria in the study areas. These findings can be expected to help in the implementation of new measures to control autochthonous malaria in areas of the Atlantic Forest.
- Malaria outside the Amazon region: Natural Plasmodium infection in anophelines collected near an indigenous village in the Vale do Rio Branco, Itanhaem, SP, Brazil(2013) NEVES, Amanda; URBINATTI, Paulo Roberto; MALAFRONTE, Rosely dos Santos; FERNANDES, Aristides; PAGANINI, Wanderley da Silva; NATAL, DelsioA few cases of Plasmodium vivax malaria in which anophelines of subgenus Kerteszia were incriminated as vectors have been reported outside the Amazon region, in the Atlantic Forest. This study was carried out near an indigenous Guarani village in the Curucutu reserve, an environmental protection area in the municipality of Itanhaem in the state of Sao Paulo, Brazil, on November 30, 2009, February 18, 2010, April 29, 2010 and May 26, 2010. Mosquitoes were collected along the route to the Guarani village where the edge of the Branco river floodplain meets the forests on the mountain slopes. Adult forms were collected with CO2-baited CDC traps and Shannon traps from twilight to 10:00 P.M. Anopheles cruzii predominated in both traps. The other species collected in the CDC traps were An. pseudomaculipes/maculipes, An. fluminensis and An. mediopunctatus/forattinii/costai. In addition to the latter three species, An. apicimacula/intermedius and An. strodei were also found in the Shannon traps. All but An. cruzii and An. strodei belong to subgenus Anopheles. A total of 506 mosquitoes were assayed by PCR to detect natural infection by Plasmodium species. In the CDC traps, An. fluminensis and An. pseudomaculipes/maculipes were positive for Plasmodium malariae, while in the Shannon traps An. pseudomaculipes/maculipes was positive for Plasmodium vivax and Plasmodium malariae and An. cruzii was positive for P. malariae, resulting in a minimum infection rate of 0.24%. Our findings suggest that An. cruzii may be incriminated in the transmission of malaria between monkeys and humans, as this species was found to be infected by P. malariae. They also highlight the need for an understanding of the role of anophelines from outside subgenus Kerteszia in the transmission of malaria in the Atlantic Forest, as these were also found to be naturally infected by P. vivax and P. malariae.