A randomised double-blind clinical trial of two yellow fever vaccines prepared with substrains 17DD and 17D-213/77 in children nine-23 months old

This randomised, double-blind, multicentre study with children nine-23 months old evaluated the immunogenicity of yellow fever (YF) vaccines prepared with substrains 17DD and 17D-213/77. YF antibodies were tittered before and 30 or more days after vaccination. Seropositivity and seroconversion were analysed according to the maternal serological status and the collaborating centre. A total of 1,966 children were randomised in the municipalities of the states of Mato Grosso do Sul, Minas Gerais and São Paulo and blood samples were collected from 1,714 mothers. Seropositivity was observed in 78.6% of mothers and 8.9% of children before vaccination. After vaccination, seropositivity rates of 81.9% and 83.2%, seroconversion rates of 84.8% and 85.8% and rates of a four-fold increase over the pre-vaccination titre of 77.6% and 81.8% were observed in the 17D-213/77 and 17DD subgroups, respectively. There was no association with maternal immunity. Among children aged 12 months or older, the seroconversion rates of 69% were associated with concomitant vaccination against measles, mumps and rubella. The data were not conclusive regarding the interference of maternal immunity in the immune response to the YF vaccine, but they suggest interference from other vaccines. The failures in seroconversion after vaccination support the recommendation of a booster dose in children within 10 years of the first dose.

Yellow fever impact on brown howler monkeys (Alouatta guariba clamitans) in Argentina: a metamodelling approach based on population viability analysis and epidemiological dynamics

In South America, yellow fever (YF) is an established infectious disease that has been identified outside of its traditional endemic areas, affecting human and nonhuman primate (NHP) populations. In the epidemics that occurred in Argentina between 2007-2009, several outbreaks affecting humans and howler monkeys (Alouatta spp) were reported, highlighting the importance of this disease in the context of conservation medicine and public health policies. Considering the lack of information about YF dynamics in New World NHP, our main goal was to apply modelling tools to better understand YF transmission dynamics among endangered brown howler monkey (Alouatta guariba clamitans) populations in northeastern Argentina. Two complementary modelling tools were used to evaluate brown howler population dynamics in the presence of the disease: Vortex, a stochastic demographic simulation model, and Outbreak, a stochastic disease epidemiology simulation. The baseline model of YF disease epidemiology predicted a very high probability of population decline over the next 100 years. We believe the modelling approach discussed here is a reasonable description of the disease and its effects on the howler monkey population and can be useful to support evidence-based decision-making to guide actions at a regional level.

Characterization of Tomato yellow spot virus, a novel tomato-infecting begomovirus in Brazil

The objective of this work was the biological and molecular characterization of a begomovirus detected in São Joaquim de Bicas, Minas Gerais, Brazil, named TGV-[Bi2], by determining its host range, complete nucleotide sequence and phylogenetic relationships with other begomoviruses. Biological characterization consisted of a host range study using either sap inoculation or particle bombardment as inoculation methods. The yellow spot virus can infect plants in Solanaceae and Amaranthaceae, including economically importat crops as sweet pepper, and weeds as Datura stramonium and Nicotiana silvestris. For the molecular characterization, the full-length genome (DNA-A and DNA-B) was amplified, cloned and completely sequenced. Sequence comparisons and phylogenetic analyses indicated that TGV-[Bi2] constitutes a novel begomovirus species named Tomato yellow spot virus (ToYSV), closely related to Sida mottle virus (SiMoV).

Watermelon transformation with Zucchini yellow mosaic virus coat protein gene and comparison with parental cultivar

The objective of this work was to transfer Zucchini yellow mosaic virus coat protein (ZYMV-CP) and neomycin phosphotransferase II (NPT II) genes to the watermelon 'Crimson Sweet'(CS) genome, and to compare the transgenic progenies T1 and T2 with the nontransformed parental cultivar for morphological, pomological, growth and yield characteristics. The ZYMV-CP gene was transferred by Agrobacterium tumefaciens. The presence of the gene in transgenic T0, T1 and T2 plants was determined by polymerase chain reaction, and the results were confirmed by Southern blot. Two experiments were performed, one in the winter-spring and the other in the summer-autumn. In both experiments, the hypocotyl length of transgenic seedlings was significantly higher than that of nontransgenic parental ones. In the second experiment, the differences between transgenic and nontransgenic individuals were significant concerning fruit rind thickness, flesh firmness, fruit peduncle length, size of pistil scar, and a* values for fruit stripe or flesh color. Transferring ZYMV-CP gene to CS genome affected only a few characteristics from the 80 evaluated ones. The changes in rind thickness, flesh firmness and flesh color a* values are favorable, while the increase in the size of pistil scar is undesirable. The transgenic watermelon line having ZYMV-CP gene and the parental cultivar CS are very similar.

Distinct features of Pepper yellow mosaic virus isolates from tomato and sweetpepper

Determination of virus diversity in the field is vital to support a sustainable breeding program for virus resistance of horticultural crops. The present study aimed to characterize four field potyvirus isolates found naturally infecting sweet pepper (Capsicum annuum) (Sa66 and Sa115) and tomato (Lycopersicon esculentum) (IAC3 and Sa21) plants. Their biological characteristics revealed differences among the isolates in their ability to infect distinct Capsicum spp. and tomato genotypes, and in the severity of symptoms caused by these isolates compared to the infection caused by an isolate of Pepper yellow mosaic virus (PepYMV). Absence of cross-reaction was found among the studied isolates with antiserum against Potato virus Y (PVY). However, all isolates reacted, at different intensities, with antiserum against PepYMV. All isolates showed high identity percentage (97 to 99%) of the amino acid sequence of the coat protein with PepYMV (accession AF348610) and low (69 to 80%) with other potyvirus species. The comparison of the 3' untranslated region also confirmed this finding with 97 to 98% identity with PepYMV, and of 47 to 71% with other potyviruses. The results showed that PepYMV isolates were easily differentiated from PVY by serology and that the host response of each isolate could be variable. In addition, the nucleotide sequence of the coat protein and 3' untranslated region was highly conserved among the isolates.

Sugarcane yellow leaf virus infection leads to alterations in photosynthetic efficiency and carbohydrate accumulation in sugarcane leaves

Infection by Sugarcane yellow leaf virus (ScYLV) causes severe leaf symptoms in sugarcane (Saccharum spp.) hybrids, which indicate alterations in its photosynthetic apparatus. To gain an overview of the physiological status of infected plants, we evaluated chlorophyll a fluorescence and gas exchange assays, correlating the results with leaf metabolic surveys, i.e., photosynthetic pigments and carbohydrate contents. When compared to healthy plants, infected plants showed a reduction in potential quantum efficiency for photochemistry of photosystem (PSII) and alterations in the filling up of the plastoquinone (PQ) pool. They also showed reduction in the CO2 net exchange rates, probably as a consequence of impaired quantum yield. In addition, reductions were found in the contents of photosynthetic leaf pigments and in the ratio chlorophyll a/chlorophyll b (chla/chlb). Carbohydrate content in the leaves was increased as a secondary effect of the ScYLV infection. This article discusses the relation of virus replication and host defense responses with general alterations in the photosynthetic apparatus and in the metabolism of infected plants.

Avaliação de danos causados pelo Barley yellow dwarf virus: PAV em cultivares de trigo no Brasil

Com o objetivo de avaliar os danos causados pelo Barley yellow dwarf virus - PAV (BYDV-PAV), em cinco cultivares de trigo (BRS 177, BRS 179, BRS 194, BRS Camboatá e BRS Angico), foi conduzido um experimento no telado da Embrapa-Trigo (Passo Fundo RS) no ano de 2005. Os danos induzidos pelo BYDV-PAV foram determinados por meio de análise de características agronômicas (estatura das plantas e massa de matéria seca) e do rendimento (número de afilhos, espigas e grãos por planta; massa de mil grãos). Os dados foram submetidos à análise de variância e as médias ao teste de Tukey a 5%. Danos significativos em função da infecção viral foram observados em todas as variáveis avaliadas. A característica agronômica mais afetada foi a massa de matéria seca, que variou de 26,1% (BRS 177) a 51,4% (BRS 179). Para estatura de plantas foram observadas reduções de 12,5% (BRS 177) a 15,5% (BRS Camboatá). O rendimento total de grãos foi o mais afetado pela infecção viral, sendo que, danos significativos foram observados em todas as cultivares, cuja redução variou de 34,2% (BRS Camboatá) a 60,8% (BRS 179). No número médio de afilhos por planta, apenas as cultivares BRS Angico e BRS 179 apresentaram reduções de 20% e 26,6%, respectivamente. A redução do número médio de grãos variou de 26,1% (BRS Camboatá) a 54,3% (BRS 179). Também ocorreu diminuição no peso de mil grãos com redução que variou de 16,9% (BRS Camboatá) a 38,4% (BRS 194).

Efeito do Soursop yellow blotch virus no desenvolvimento vegetativo e na produção da gravioleira

Os danos causados no desenvolvimento vegetativo e na produção de frutos da gravioleira pelo vírus da mancha-amarela da gravioleira (Soursop yellow blotch virus, SYBV), foram estudados durante os anos de 2000 a 2004 em um experimento com dois tratamentos: plantas sadias e plantas doentes, dispostos em blocos ao acaso, com oito repetições e quatro plantas por parcela. Foram avaliados, anualmente, a altura da planta, diâmetro do caule, número e peso de frutos, sendo que a produção foi monitorada a partir do segundo ano de plantio. As médias relativas à altura de planta, diâmetro do caule, número e peso de frutos das parcelas foram computadas, analisadas estatisticamente e comparadas pelo teste F. As plantas de ambos tratamentos foram originadas de mudas enxertadas, sendo as plantas doentes obtidas por meio de enxertias com propágulos de plantas infectadas com o SYBV. A doença reduziu em 65,11% e 46,72% a altura e o diâmetro do caule, respectivamente, e em 94,7 % e 99,2 % o número e o peso de frutos em relação às plantas sadias.

Danos na produção da abobrinha de moita causados pelo Papaya ringspot virus type W e Zucchini yellow mosaic virus

Este trabalho teve por objetivo avaliar, em condições de casa de vegetação e de campo, os danos causados pelo PRSV-W e ZYMV em abobrinha-de-moita (Cucurbita pepo cv. Caserta). As plantas em casa de vegetação foram inoculadas com os vírus individualmente e em mistura aos 12 e 22 dias após emergência (DAE) e aos 5, 15 e 25 DAE no campo. Em casa de vegetação, as infecções com PRSV-W + ZYMV, PRSV-W e ZYMV, na primeira época de inoculação, ocasionaram reduções de área foliar de 39,6%, 36,8% e 12,1%, respectivamente. As massas fresca e seca também foram significativamente afetadas na primeira época de inoculação. No campo, as plantas com infecções individuais ou mistas dos potyvírus produziram frutos não comerciais em quantidades que variaram de 14 a 861 g/planta, dependendo da idade que foram inoculadas. As plantas tratadas com tampão fosfato aos 5, 15 e 25 DAE produziram em média 573 g, 937 g e 1172 g de frutos comerciais e 282 g, 221 g e 192 g de frutos não comerciais, respectivamente. A redução na massa fresca das plantas foi diretamente relacionada com a época de inoculação, com médias de 60,7% para aquelas inoculadas aos 5 DAE e de 22,7% para aquelas inoculadas aos 15 DAE. Na terceira época de inoculação não houve diferença significativa de massa fresca entre os tratamentos.

Cytokine and nitric oxide production by mouse macrophages infected with brazilian flaviviruses

The Flaviviridae family, Flavivirus genus includes viruses that are transmitted to vertebrates by infected mosquitoes or ticks. The genus Flavivirus includes a variety of viruses that cause diseases such as acute febrile illness, encephalitis, and hemorrhagic fever. Flaviviruses primarily infect blood monocytes and tissue macrophages, which have been shown to be permissive, supporting viral replication and serving as virus reservoirs. On the other hand, these cells may have an important antiviral activity related to modulation by cytokine production and by the capacity of these cells to synthesize reactive free radicals such as nitric oxide (NO) which can have a microbicidal effect. The present study was performed in order to determine the production of cytokines interleukin-1beta (IL-1β), tumor necrosis factor -alpha (TNF-α), transforming growth factor- beta (TGF-β) and interferon -alpha (IFN-α) and NO by macrophages infected with one of four Brazilian flaviviruses, Bussuquara virus (BUSV), Yellow Fever virus (YFV), Rocio virus (ROCV) and Encephalitis Saint Louis virus (SLEV), and to verify the possible antiviral effect of NO during macrophage infection with ROCV. Moreover, we asked if the different viruses were able to regulate bacterial lipopolysaccharide (LPS) induced cytokine production. Our results showed that YFV and SLEV reduced the production of IL-1β and TGF-β by LPS-stimulated macrophages, while ROCV only diminished LPS-stimulated TGF-β synthesis. On the other hand, BUSV more likely favored an enhancement of the LPS-induced production of IL-1β by macrophages. Additionally, while most of the viruses stimulated the production of IFN-α, none of them altered the production of TNF-α by murine macrophages. Interestingly, all viruses induced synthesis of NO that was not correlated with antiviral activity for ROCV.

DENGUE OUTBREAK IN MATO GROSSO STATE, MIDWESTERN BRAZIL

Dengue virus (DENV) is the most frequent arbovirus worldwide. In this study, we report a large outbreak in Mato Grosso State (MT). Serum samples from 604 patients with acute febrile illness for less than five days were inoculated in C6/36 cells, then infected cells were subjected to an indirect immunofluorescence test for DENV serotypes and yellow fever virus. Serum samples were submitted to a multiplex-semi-nested-RT-PCR for 11 flaviviruses. DENV-4 was isolated in 150/604 (24.8%) and DENV-1 in 19/604 (3.1%) specimens. By RT-PCR, 331 (54.8%) samples tested positive for DENV; 321 had single infections (DENV-4 n = 305; DENV-1 n = 15; DENV-3 n = 1), nine had co-infections of DENV-1/DENV-4, and one of DENV-2/DENV-4. DENV-4 was detected in 315/331 (95.2%) positive patients from 17 municipalities, and DENV-1 in 24/331 (7.2%) patients from five cities in north-central MT and the city of Cuiaba. The incidence of infection was higher in patients aged 20-39 (142/331; 42.9%). The NS5 partial nucleotide sequence of DENV-1 was most similar to that of genotype V, DENV-2 to Southeast Asian/American, DENV-3 to genotype III, and DENV-4 to genotype II strains, considered the most frequent strains in Brazil. This outbreak coincided with the introduction of DENV-4 in the state. Cuiaba was hyperendemic for the four DENV serotypes, highlighting the necessity for arbovirus surveillance in MT.

Arboviral diseases in the Western Brazilian Amazon: a perspective and analysis from a tertiary health & research center in Manaus, State of Amazonas

The Fundação de Medicina Tropical Dr. Heitor Vieira Dourado (FMT-HVD), located in Manaus, the capital of the State of Amazonas (Western Brazilian Amazon), is a pioneering institution in this region regarding the syndromic surveillance of acute febrile illness, including arboviral infections. Based on the data from patients at the FMT-HVD, we have detected recurrent outbreaks in Manaus by the four dengue serotypes in the past 15 years, with increasing severity of the disease. This endemicity has culminated in the simultaneous circulation of all four serotypes in 2011, the first time this has been reported in Brazil. Between 1996 and 2009, 42 cases of yellow fever (YF) were registered in the State of Amazonas, and 71.4% (30/42) were fatal. Since 2010, no cases have been reported. Because the introduction of the yellow fever virus into a large city such as Manaus, which is widely infested by Aedes mosquitoes, may pose a real risk of a yellow fever outbreak, efforts to maintain an appropriate immunization policy for the populace are critical. Manaus has also suffered silent outbreaks of Mayaro and Oropouche fevers lately, most of which were misdiagnosed as dengue fever. The tropical conditions of the State of Amazonas favor the existence of other arboviruses capable of producing human disease. Under this real threat, represented by at least 4 arboviruses producing human infections in Manaus and in other neighboring countries, it is important to develop an efficient public health surveillance strategy, including laboratories that are able to make proper diagnoses of arboviruses.

Uma estação biológica para o estudo dos mosquitos e dos outros animaes silvestres relacionados com a febre amarela

The outbreak of the jungle or forest yellow fever, through the adapta¬tion, quite recently of the yellow fever virus o the forest mosquitoes, brou¬ght the necessity of ecological researches on hese mosquitoes, as well as on the wild animals they bite, some of them being susceptible to the desease. This has been done by the special yellow fever Service of the State of Sao Paulo, in a special Biological Station in Perús, São Paulo, which has been built in the midst of the jungle. This station was made with plain materials, and covered with straw, but was confortable enough for the technical work, i nthe early months of 1938. During the months in which the investigations were being carried on, the following interesting results were obtained: 1. As we have already pointed out in other places, the forest mosquitoes biting us during daytime, are always new born insects, having not yet sucked blood, as it is the general rule with all mosquitoes, and therefore also, with the anopheles and stegomyia, and this explains why nobody gets malaria or yellow fever, transmitted by anofeles or by aedes aegypti during the day. We think therefore, the jungle yellow fever, got during daytime is not due to the infected jungle or forest mosquito biting, but to infection through the human skin coming into close contact with tre virus, which the forest mosquitoes lay with their dejections, on the leaves of the trees where they remain sitting du¬ring the day. 2. As it is the rule with anopheles, stegomyia and other mosquitoes, the insects once having sucked blood, take nocturnal habits and, therefore, bite us, only during the night, so it happens with the forest mosquito, and insects with developped eggs and blood in stomach have been caught within the sta¬tion house, during the night. During the day, these mosquitoes do not bite, but remain quite still on the leaves of the trees, in the damp parts of the woods. 3. Jungle or forest mosquitoes can easely bite wild animals, some with more avidity then ethers, as it has bee npointed out to the opossum (didei-phis) and other animals. They also bite birds having very thin skin and only exceptionally, cold bloods animals. 5. Is has hot been possible to ascertain how forest mosquitoes are able to live, from onde season to another, through winter, when temperature drops near and even below zero. They have not been found in holes of the terrain, of trees and of animals, as it is the rule in cold countries. During winter, in the forest, it is possible to find larvs in the holes of bambus and trees full of water. As wild animals do not harbour the yellow fever virus for a long time in their body, it is diffcult to explain how the desease lasts from one season to another. Many ecological features on the mosquito, remains yet to be explained and therefore it in necessary to go on with the investigations, in bio¬logical stations, such as that one built up in Perús, São Paulo.

Ecology of Haemagogus sp. and Sabethes sp. (Diptera: Culicidae) in relation to the microclimates of the Caxiuanã National Forest, Pará, Brazil

This study was conducted in a meteorological tower located in the Caxiuanã Forest (municipality of Melgaço, Pará, Brazil) with the aim of assessing the vertical stratification of species of Haemagogus and Sabethes, potential vectors of the yellow fever virus. To investigate the role of microclimates in mosquito stratification, bimonthly collections were conducted at ground level (0 m), 8 m, 16 m and 30 m (canopy level), with the aid of entomological nets and suction tubes, from July 2005-April 2006. A total of 25,498 mosquitoes were collected; specimens of Sabethes sp. and Haemagogus janthinomyswere found mostly at heights of 16 m and 30 m while Hg. leucocelaenus was most frequently observed at ground level. The largest number of vector species was collected during the rainiest months, but this difference between seasons was not statistically significant. However, the number of Hg. janthinomys was positively correlated with variations in temperature and relative humidity.

Biodiversity and influence of climatic factors on mosquitoes (Diptera: Culicidae) around the Peixe Angical hydroelectric scheme in the state of Tocantins, Brazil

The influence of climatic factors on the seasonal frequency of mosquitoes (Diptera: Culicidae) at the Peixe Angical hydroelectric scheme (Tocantins, Brazil) was evaluated in the present paper. Mosquito surveys were conducted in the municipality of Peixe and in areas surrounding the reservoir in the municipalities of Paranã and São Salvador do Tocantins during two daytime periods (10 am-12 noon and 2 pm-4 pm) and two night-time periods (6 pm-8 pm and 6 pm-10 am) over 14 months. In total, 10,840 specimens from 42 species were captured, 84.5% of which belonged to the Culcinae. The most common species were Anopheles darlingi, Psorophora albipes and Sabethes chloropterus. The number of Culicidae specimens was higher in months with higher rainfall and air humidity than during the drier months. The large population of Ps. albipes and the presence of both An. darlingi (primary vector for human malaria parasites) and Haemagogus janthinomys (primary vector for yellow fever virus) are highlighted.