Caracterização genética e evolução dos vírus dengue no Estado do Rio Grande do Norte

Dengue is considered as the most important arthropod-borne viral disease throughout the world due to the high number of people at risk to be infected, mainly in tropical and subtropical regions of the planet. The etiologic agent is Dengue Virus (DENV), it is a single positive-stranded RNA virus of the family Flavivirus, genus Flaviviridae. Four serotypes are known, DENV-1, DENV-2, DENV-3 and DENV-4. One of the most important characteristic of these viruses is the genetic variability, which demands phylogenetic and evolutionary studies to understand key aspects like: epidemiology, virulence, migration patterns and antigenic characteristics. The objective of this study is the genetic characterization of dengue viruses circulating in the state of Rio Grande does Norte from January 2010 to December 2012. The complete E gene (1485 pb) of DENV1, 2 e 4 from Brazilian (Rio Grande do Norte) patients was sequenced. Phylogenetic analysis was performed using MEGA 5.2 software, Tamura-Nei model and Neighbor-Joining trees were inferred for the datasets. In Brazil, there is just one DENV-1 genotype (genotype V), one DENV-2 genotype (Asian/American) and two DENV-4 genotypes (genotypes I and II). Brazilian strains of DENV-1 are subdivided in two different lineages (BR-I and BR-II), the Brazilian strains of DENV-2 are subdivided in four lineages (BRI-IV) and genotype II of DENV-4 is subdivided in three Brazilian lineages (BRI-III). The viruses isolated in RN belong to lineage BR-II (DENV-1), BR-IV (DENV-2) and BR-III (DENV-4).The Caribbean and near Latin American countries are the main source of these viruses to Brazil. Amino acids substitutions were detected in three domains of E protein, this makes clear the necessity of studies that associate epidemiological and molecular data to better understand the effects of these mutations. This is the first study about genetic characterization and evolution of Dengue viruses in Rio Grande do Norte, Brazil

Potencial larvicida de extratos de plantas regionais no controle de larvas de Aedes Aegypti (Diptera:Culicidae)

Dengue, amongst the virus illnesses one can get by vectorial transmission, is the one that causes more impact in the morbidity and mortality of world s population. The resistance to the insecticides has caused difficulties to control of vector insect (Aedes aegypti) and has stimulated a search for vegetables with larvicidal activity. The biodiversity of Caatinga is barely known and it is potential of use even less. Some plants of this biome are commercialized in free fairs northeast of Brazil, because of its phytotherapics properties. The vegetables in this study had been selected by means of a questionnaire applied between grass salesmen and natives of the Serido region from Rio Grande do Norte state; culicids eggs had been acquired with traps and placed in container with water for the larva birth. Thirty larvae had been used in each group (a group control and five experimental groups), with four repetitions four times. The vegetables had been submitted to the processes of decoction, infusion and maceration in the standard concentration of 100g of the vegetable of study in 1l of H2O and analyzed after ½, 1, 2, 4, 8, 12, 24 and 48 hours for verification of the average lethal dose (LD50) from the groups with thirty larva. The LD50 was analyzed in different concentrations (50g/l, 100g/l, 150g/l, 200g/l e 300g/l) of Aspidosperma pyrifolium Mart. 48 extracts of rind, leaf and stem of the seven vegetal species: Aspidosperma pyrifolium Mart., Mimosa verrucosa Benth, Mimosa hostilis (Mart.) Benth., Myracrodruon urundeuva Allemão, Ximenia americana L, Bumelia sartorum Mart Zizyphus joazeiro Mart, had been analyzed. The extracts proceeding from the three methods were submitted to the freezedrying, to evaluate and to quantify substances extracted in each process. The results had shown that Aspidosperma pyrifolium Mart. and Myracrodruon urundeuva Allemão are the species that are more distinguished as larvicidal after 24 hours of experiment, in all used processes of extraction in the assays. The Zizyphus joazeiro Mart species has not shown larvicidal activity in none of the assays. In relation to the extraction method, the decoction was the most efficient method in the mortality tax of the A. aegypti larvae

Estudo teórico-bioclimático da potencialidade de desenvolvimento do Aedes aegypti no estado do Rio Grande do Norte

In the current climate of global public health there is the emergence of urban dengue, a disease regarded as acute infectious fever. The disease annually, has affected millions of people worldwide, mostly in the range of the intertropical globe. The disease's main vector in urban areas, the Aedes aegypti mosquito. Recent studies indicate that the distribution of the insect vector of dengue in the geographical area is directly tied to the behavior of environmental restrictions that area, especially among those, the air temperature and relative humidity. From that context, the work aims to estimate and spatializing, monthly, for each municipality in the state of Rio Grande do Norte, the potential of biophysical conditions conducive to the development of Aedes aegypti. Yet, made use of the following methodology: collection of epidemiological data and climatological, Normal climatological, descriptive statistics (measures of central tendency and scatter), uniform distribution, estimation geostatistics and sufer program, version 8.0. The results flagged for a behavior very heterogeneous, both in space and in time, in the case of the potential of biophysical conditions conducive to the development of Aedes aegypti in the state of Rio Grande do Norte. Still, he noted that there is a tendency for lifting the potential of development for the entire state, from the month of January, ending in the month of April mainly in central and western portions of the state. By contrast, there is the permanence of increased potential for development in the eastern portion of the state. The latter record maximum potential in the month of July, resulting probability of greater than 70% have been favorable conditions for the development of Aedes aegypti in that area. In the period between the months of August to December, it is small potential for development of Aedes aegypti in all parts of the state

Atividades biológicas de extratos salinos de sementes de plantas da caatinga contra Aedes Aegypti e investigação da participação de proteínas bioativas

Dengue fever, currently the most important arbovirus, is transmitted by the bite of the Aedes aegypti mosquito. Given the absence of a prophylactic vaccine, the disease can only be controlled by combating the vector insect. However, increasing reports of resistance and environmental damage caused by insecticides have led to the urgent search for new safer alternatives. Twenty - um plant s eed extracts from the Caatinga were prepared , tested and characterized . Sodium phosphate ( 50 mM pH 8.0) was used as extractor. All extracts showed larvicidal and ovipositional deterrence activity . Extracts of D. grandiflora, E. contortisiliquum, A. cearenses , C. ferrea and C. retusa were able to attract females for posture when in low co ncentration . In the attractive concentrations, the CE of E. contortisiliquum and A. cearenses were able to kill 52% and 100% of the larvae respectively . The extracts of A. cearenses , P. viridiflora, E. velutina, M. urundeuva and S. brasiliensis were also pupicides, while extracts of P. viridiflora, E. velutina, E. contortisiliquum , A. cearenses, A. colubrina, D. grandiflora , B. cheilantha , S. spectabilis, C. pyramidalis, M. regnelli e G. americana displayed adulticidal activity. All extracts were toxic to C. dubia zooplankton . The EB of E. velutina and E. contortisiliquum did not affect the viability of fibroblasts . In all extracts were identified at least two potential insecticidal proteins such as enzyme inhibitors, lectins and chitin - binding proteins and components of secondary metabolism . Considering all bioassays , the extracts from A. cearenses, P. viridiflora, E. contortisiliquum , S. brasiliensis, E. velutina and M. urundeuva were considered the most promising . The E. contortisiliquum extracts was the only one who did not show pupicida activity, indicating that its mechanism of action larvicide and adulticidal is related only to the ingesti on of toxic compounds by insect , so it was selected to be fragmenting. As observed for the CE , th e protein fractions of E. contortisiliquum also showed larvicidal activity, highlighting that F2 showed higher larvicidal activity and lower en vironmental toxicity than the CE source. The reduction in the proteolytic activity of larvae fed with crude extra ct and fractions of E. contortisiliquum suggest ed that the trypsin inhibitors ( ITEc) would be resp onsible for larvicidal activity . However the increase in the purification of this inhibitor resulted in loss of larvicidal activity , but the absence of trypsin inhibitor reduced the effectiveness of the fractions , indicating that the ITEC contributes to the larvicidal activity of this extract. Not been observed larvicidal activity and adulticide in rich fraction vicilin, nor evidence of the contribution o f this molecule for the larvicidal activity of the extract. The results show the potential of seeds from plant extracts of Caatinga as a source of active molecules against insects A. aegypti at different stages of its development cycle, since they are comp osed of different active compounds, including protein nature, which act on different mechanisms should result in the death of insec

Epidemiologia e caracterização molecular dos vírus Dengue circulantes no Rio Grande do Norte, 2013-2014

Dengue is an acute infectious disease, usually transmitted by Aedes aegypti mosquitoes. The etiologic agents belong to the family Flaviviridae, genus Flavivirus, and occur as four antigenically related but distinct serotypes designated DENV-1, 2, 3, and 4. In Brazil, the disease represents a national public health problem. The purpose of the present study was to describe epidemiological aspects of dengue in the State of Rio Grande do Norte, from 2013 to 2014. A total of 483 blood or serum samples, collected from January 2013 to December 2014, were studied by RT-PCR for viral detection and typing. The infection was confirmed in 36.44% (176/483) of the cases studied. This study detected the circulation of three serotypes of dengue virus in Rio Grande do Norte, DENV-1, 2, and 4. The predominant serotype in 2013-2014 was the DENV-4, representing 83.51% (81/97) and 68.35% (54/79) of the positive cases, respectively. Regarding the spatial distribution, most of the cases occurred in Natal and Caicó, with 9.28% (9/97) and 18.99% (15/79), respectively. The months with the biggest viral circulation in RN were March 2013 and May 2014. The female gender was the most affected, representing 69.07% (67/97) in 2013 and 54.43% (43/79) in 2014. The most affected age groups were 21-30 years (2013) and 11-20 years (2014) with 25.77% (25/97) and 20.25% (16/79) positive cases, respectively. Phylogenetic analysis indicated that genotype V (DENV-1) and genotype II (DENV-4) circulated in the State. Our results provide information about the dynamics of DENV in the Rio Grande do Norte, important for the development of disease control strategies.

Potencial larvicida de extratos de plantas regionais no controle de larvas de Aedes aegypti

Dengue, amongst the virus illnesses one can get by vectorial transmission, is the one that causes more impact in the morbidity and mortality of world s population. The resistance to the insecticides has caused difficulties to control of vector insect (Aedes aegypti) and has stimulated a search for vegetables with larvicidal activity. The biodiversity of Caatinga is barely known and it is potential of use even less. Some plants of this biome are commercialized in free fairs northeast of Brazil, because of its phytotherapics properties. The vegetables in this study had been selected by means of a questionnaire applied between grass salesmen and natives of the Serido region from Rio Grande do Norte state; culicids eggs had been acquired with traps and placed in container with water for the larva birth. Thirty larvae had been used in each group (a group control and five experimental groups), with four repetitions four times. The vegetables had been submitted to the processes of decoction, infusion and maceration in the standard concentration of 100g of the vegetable of study in 1l of H2O and analyzed after ½, 1, 2, 4, 8, 12, 24 and 48 hours for verification of the average lethal dose (LD50) from the groups with thirty larva. The LD50 was analyzed in different concentrations (50g/l, 100g/l, 150g/l, 200g/l e 300g/l) of Aspidosperma pyrifolium Mart. 48 extracts of rind, leaf and stem of the seven vegetal species: Aspidosperma pyrifolium Mart., Mimosa verrucosa Benth, Mimosa hostilis (Mart.) Benth., Myracrodruon urundeuva Allemão, Ximenia americana L, Bumelia sartorum Mart Zizyphus joazeiro Mart, had been analyzed. The extracts proceeding from the three methods were submitted to the freezedrying, to evaluate and to quantify substances extracted in each process. The results had shown that Aspidosperma pyrifolium Mart. and Myracrodruon urundeuva Allemão are the species that are more distinguished as larvicidal after 24 hours of experiment, in all used processes of extraction in the assays. The Zizyphus joazeiro Mart species has not shown larvicidal activity in none of the assays. In relation to the extraction method, the decoction was the most efficient method in the mortality tax of the A. aegypti larvae

A ocupação urbana do entorno de zonasde proteção ambiental de natal, suas condições de sustentabilidade e a relação com a ocorrência de insetos vetores

This study started from the hypothesis of the existence of a relation between the type of the urban occupation concerning to the sustainability conditions at the proximity of Environment Protected Zones and the occurrence of vectors insects in Natal, Rio Grande do Norte, Brazil. This research, which used data available by the City Administration Health and Urbanization Secretaries (respectively SMS and SEMURB), in the time period of 2006 to 2008, aimed to characterize the study site in terms of urban occupation, relating it to social environmental aspects of land occupation and the occurrence of vectors insects. This study is presented in two papers, the first one linking the occurrence of vectors insects and sustainable development indicators and the second relating the incidence of reported cases of Dengue and Dengue Hemorrhagic Fever (DHF) and the occurrence of larvae infection indexes of Aedes aegypti, in Natal, Rio Grande do Norte State. In the first paper, was made a correlation between Dengue Fever vectors and Visceral and Tegumentar Leishmaniasis vectors and sustainable development indicators, selected from IDS Brasil- 2008. Through factorial analysis a Sustainability Index (SI) was acquired for each region, the northern region of the municipality obtained lower numbers than southern region, which, in its turn, presented better sustainability conditions. Linking this index to vector infestation parameters shows a high significant correlation between the SI and the Breteau Index of Aedes aegypti (p=0,028) as well as with SI and sand flies infestation index (p=0,01). Higher rates in vectors infestation in regions with a lower Sustainable Development Index demonstrates that this index can be used to determine the increasing of probability of Aedes and sand flies occurrence in urban environment. The second paper analyzed the occurrence of the main vector of Dengue and DHF, the Aedes aegypti mosquito, and the relation between larvae infection indexes of this insect and reported cases of the diseases. This study revealed unexpected relation where areas with higher Breteau s Indexes showed lower infection rates of Dengue Fever, although showing high incidence of DHF

Co-circulação dos vírus dengue tipo 1, 2 e 4 nos Estados do Rio Grande do Norte e Paraíba, 2013

Dengue is a viral disease transmitted by female mosquitoes from genus Aedes, the principal urban vector is Aedes aegypti. Actually dengue has caused, in global scale, substantial morbidity and mortality. Four serotypes (antigenically distinct) are known: DENV-1, DENV-2, DENV-3 and DENV-4. The objective of this study was described the epidemiological profile dengue in the states of Rio Grande do Norte (RN) and Paraíba (PB), 2013. For that, suspected cases of dengue were studied, received for Laboratory of Molecular Biology of infectious disease and cancer (LADIC-UFRN) from different Health Units from RN and PB between January and December of 2013. The viral RNA was obtained from serum samples of patient from health units from RN and PB. It were studied 478 suspected cases of dengue , 252 (52,7%) from Rio Grande do Norte and 226 (47,3%) from Paraíba, showeds a global rate of infection global prevalence of 29,7% (142/478). The co-circulation of three serotypes was observed: DENV-1 (9,8% [14/142]), DENV-2 (3,5% [5/142]) and DENV-4 (86,7% [123/142]). People between 21-30 years old were the most affected by the disease during all the period of the study, representing 63,7% of the cases in both states. The genus most affected was female, representing 63,3% of cases in both states. Pau dos Ferros, Rio Grande do Norte, had the highest circulation of disease, with 8,2% (8/97) of cases. In Paraíba, the city most affected was João Pessoa, with (80% (36/45) of cases. The months with the biggest viral circulation in RN and PB were March and August, respectively. These results are very important to understanding the dengue viral activity in RN and PB, providing data that can guide control actions of this disease in support to local control programs

Isolamento de uma quitinase extraída do cefalotórax do camarão marinho Litopenaeus schmitti (BURKENROAD, 1936) : avaliação de suas atividades antimicrobiana e larvicida

Chitinases are enzymes involved in degradation of chitin and are present in a range of organisms, including those that do not contain chitin, such as bacteria, viruses, plants and animals, and play important physiological and ecological roles. Chitin is hydrolyzed by a chitinolytic system classified as: endo-chitinases, exo-chitinases and N-acetyl-b-D-glucosaminidases. In this study a Litochitinase1 extracted from the cephalotorax of the shrimp Litopenaeus Schmitt was purified 987.32 times using ionexchange chromatography DEAE-Biogel and molecular exclusion Sephacryl S-200. These enzyme presented a molecular mass of about 28.5 kDa. The results, after kinetic assay with the Litochitinase1 using as substrate p-nitrophenyl-N-acetyl-b-Dglucosaminideo, showed apparent Km of 0.51 mM, optimal activity at pH ranging from 5.0 to 6.0, optimum temperature at 55°C and stability when pre-incubated at temperatures of 25, 37, 45, 50 and 55°C. The enzyme showed a range of stability at pH 4.0 to 5.5. HgCl2 inhibited Litochitinase1 while MgCl2 enhances its activity. Antimicrobial tests showed that Litochitinase1 present activity against gram-negative bacterium Escherichia coli in the 800 μg/mL concentration. The larvicidal activity against Aedes aegypti was investigated using crude extracts, F-III (50-80%) and Litochitinase1 at 24 and 48 hours. The results showed larvicidal activity in all these samples with EC50 values of 6.59 mg/mL for crude extract, 5.36 mg/mL for F-III and 0.71 mg/mL for Litochitinase1 at 24 hours and 3.22 and 0.49 mg/mL for the F-III and Litochitinase1 at 48 hours, respectively. Other experiments confirmed the presence of chitin in the midgut of Aedes aegypti larvae, which may be suffering the action of Litochitinase1 killing the larvae, but also the absence of contaminating proteins as serine proteinase inhibitors and lectins in the crude extract, F-III and Litochitinase1, indicating that the death of the larvae is by action of the Litochitinase1. We also observed that the enzymes extracted from intestinal homogenate of the larvae no have activity on Litochitinase1. These results indicate that the enzyme can be used as an alternative to control of infections caused by Escherichia coli and reducing the infestation of the mosquito vector of dengue.

A eliminação do mosquito do dengue em ambientes residenciais: uma questão de cuidado ambiental?

Dengue is currently considered one of the most relevant public health problems worldwide. Studies indicate the surroundings of the houses as the preferred sites for the proliferation of Aedes aegypti. The residential areas are privileged environments for human development and contribute to the formation of the individual s identity and for the establishment of affective, social and cultural bonds. The purpose of this study was to investigate possible links between psychological indicators of pro-environmentalism and conservation status of residential backyards. Data collection was performed in 147 homes and methodological strategy involved the use of interview, the Scale of Ecocentric and Anthropocentric Environmentalism, Scale of Consideration of Future Consequences and a tool for environmental evaluation. It was found that the participants expressed as environmental practices the garbage recycling, besides they had the knowledge of how the transmission of dengue occurs. These residents showed ecofriendly motivated commitment: pro-environmentalist ecocentric and anthropocentric. In evaluating the backyard it was verified that the conservation conditions, in almost half of the homes, appeared as carelessness on the part of residents and those conditions are conducive to the proliferation of Aedes aegypti. The pro-environmentalists and guidance for the future identified by the scales were not associated with the conservation status of the backyards. However, it was found that the trends of reduction and stability of infestation levels are associated with self-reported environmental care. These results can contribute to the discussion and design of new mosquito control actions and practices of education and health information among the population

A atuação do enfermeiro no combate à dengue e a febre chikungunya: estudo comparativo nos municípios de Parnamirim e Santa Cruz/RN

The research aimed to analyze the role of nurses in the Family Health Strategy (FHS) in the control of Dengue and Chikungunya fever in the cities of Parnamirim and Santa Cruz. It is exploratory and descriptive nature of research, with a qualitative approach, which was developed with nurses of the Family Health Strategy in the municipalities of Parnamirim and Santa Cruz. All research process followed the ethical standards laid out research in Resolution No. 466/2012 of the National Health Council about research involving human beings. Data collection was performed using a semi-structured questionnaire with open and closed questions, organized into three parts: the profile of respondents, knowledge of the disease (transmission, prevention, control) and description of practices. The actions performed by nurses are punctual performed when there is growing disease of the number of cases in order to try to reduce the number of cases. The principal methodology used for them is to talk and guidelines resulting from campaigns and home visits, using leaflets and social networks. Regarding knowledge of nurses in the city of Santa Cruz on Dengue Fever and Chikungunya, is clearly some gaps. Nursing professionals know recognize a suspected case of dengue, but get confused when trying to explain to the Chikungunya fever, exhibiting the same symptoms of dengue. Although everybody had participated in a training on Chikungunya Fever and Dengue, a very limited knowledge of nurses regarding the clinical management. It is suggested that further studies are developed on the ground a few nurses have accepted to participate in the study, so that we can identify strategies, interventions, activities and nursing actions that are consistent with the reality of working in favor of coping Aedes epidemics aegypti that have positive impact on reducing the infestation rates and may be suitable and applied in other regions.

A atuação do enfermeiro no combate à dengue e a febre chikungunya: estudo comparativo nos municípios de Parnamirim e Santa Cruz/RN

The research aimed to analyze the role of nurses in the Family Health Strategy (FHS) in the control of Dengue and Chikungunya fever in the cities of Parnamirim and Santa Cruz. It is exploratory and descriptive nature of research, with a qualitative approach, which was developed with nurses of the Family Health Strategy in the municipalities of Parnamirim and Santa Cruz. All research process followed the ethical standards laid out research in Resolution No. 466/2012 of the National Health Council about research involving human beings. Data collection was performed using a semi-structured questionnaire with open and closed questions, organized into three parts: the profile of respondents, knowledge of the disease (transmission, prevention, control) and description of practices. The actions performed by nurses are punctual performed when there is growing disease of the number of cases in order to try to reduce the number of cases. The principal methodology used for them is to talk and guidelines resulting from campaigns and home visits, using leaflets and social networks. Regarding knowledge of nurses in the city of Santa Cruz on Dengue Fever and Chikungunya, is clearly some gaps. Nursing professionals know recognize a suspected case of dengue, but get confused when trying to explain to the Chikungunya fever, exhibiting the same symptoms of dengue. Although everybody had participated in a training on Chikungunya Fever and Dengue, a very limited knowledge of nurses regarding the clinical management. It is suggested that further studies are developed on the ground a few nurses have accepted to participate in the study, so that we can identify strategies, interventions, activities and nursing actions that are consistent with the reality of working in favor of coping Aedes epidemics aegypti that have positive impact on reducing the infestation rates and may be suitable and applied in other regions.

Isolamento de uma quitinase extraída do cefalotórax do camarão marinho Litopenaeus schmitti (BURKENROAD, 1936) : avaliação de suas atividades antimicrobiana e larvicida

Chitinases are enzymes involved in degradation of chitin and are present in a range of organisms, including those that do not contain chitin, such as bacteria, viruses, plants and animals, and play important physiological and ecological roles. Chitin is hydrolyzed by a chitinolytic system classified as: endo-chitinases, exo-chitinases and N-acetyl-b-D-glucosaminidases. In this study a Litochitinase1 extracted from the cephalotorax of the shrimp Litopenaeus Schmitt was purified 987.32 times using ionexchange chromatography DEAE-Biogel and molecular exclusion Sephacryl S-200. These enzyme presented a molecular mass of about 28.5 kDa. The results, after kinetic assay with the Litochitinase1 using as substrate p-nitrophenyl-N-acetyl-b-Dglucosaminideo, showed apparent Km of 0.51 mM, optimal activity at pH ranging from 5.0 to 6.0, optimum temperature at 55°C and stability when pre-incubated at temperatures of 25, 37, 45, 50 and 55°C. The enzyme showed a range of stability at pH 4.0 to 5.5. HgCl2 inhibited Litochitinase1 while MgCl2 enhances its activity. Antimicrobial tests showed that Litochitinase1 present activity against gram-negative bacterium Escherichia coli in the 800 μg/mL concentration. The larvicidal activity against Aedes aegypti was investigated using crude extracts, F-III (50-80%) and Litochitinase1 at 24 and 48 hours. The results showed larvicidal activity in all these samples with EC50 values of 6.59 mg/mL for crude extract, 5.36 mg/mL for F-III and 0.71 mg/mL for Litochitinase1 at 24 hours and 3.22 and 0.49 mg/mL for the F-III and Litochitinase1 at 48 hours, respectively. Other experiments confirmed the presence of chitin in the midgut of Aedes aegypti larvae, which may be suffering the action of Litochitinase1 killing the larvae, but also the absence of contaminating proteins as serine proteinase inhibitors and lectins in the crude extract, F-III and Litochitinase1, indicating that the death of the larvae is by action of the Litochitinase1. We also observed that the enzymes extracted from intestinal homogenate of the larvae no have activity on Litochitinase1. These results indicate that the enzyme can be used as an alternative to control of infections caused by Escherichia coli and reducing the infestation of the mosquito vector of dengue.