Flavivirus isolations from mosquitoes collected from Saibai Island in the Torres Strait, Australia, during an incursion of Japanese encephalitis virus

Adult mosquitoes (Diptera: Culicidae) were collected in January and February 2000 from Saibai Island in the Torres Strait of northern Australia, and processed for arbovirus isolation during a period of Japanese encephalitis (JE) virus activity on nearby Badu Island. A total of 84 2 10 mosquitoes were processed for virus isolation, yielding six flavivirus isolates. Viruses obtained were single isolates of JE and Kokobera (KOK) and four of Kunjin (KUN). All virus isolates were from members of the Culex sitiens Weidemann subgroup, which comprised 53.1 % of mosquitoes processed. Nucleotide sequencing and phylogenetic analysis of the pre-membrane region of the genome of JE isolate TS5313 indicated that it was closely related to other isolates from a sentinel pig and a pool of Cx. gelidus Theobald from Badu Island during the same period. Also molecular analyses of part of the envelope gene of KUN virus isolates showed that they were closely related to other KUN virus strains from Cape York Peninsula. The results indicate that flaviviruses are dynamic in the area, and suggest patterns of movement south from New Guinea and north from the Australian mainland.

Regulation of antigen processing and presentation molecules in West Nile virus-infected human skin fibroblasts

Infection of humans with the West Nile flavivirus principally occurs via tick and mosquito bites. Here, we document the expression of antigen processing and presentation molecules in West Nile virus (WNV)-infected human skin fibroblast (HFF) cells. Using a new Flavivirus-specific antibody, 4G4, we have analyzed cell surface human leukocyte antigen (HLA) expression on virus-infected cells at a single cell level. Using this approach, we show that West Nile Virus infection alters surface HLA expression on both infected HFF and neighboring uninfected HFF cells. Interestingly, increased surface HLA evident on infected HFF cultures is almost entirely due to virus-induced interferon (IFN)alpha/beta because IFNalpha/beta-neutralizing antibodies completely prevent increased surface HLA expression. In contrast, RT-PCR analysis indicates that WNV infection results in increased mRNAs for HLA-A, -B, and -C genes, and HLA-associated molecules low molecular weight polypeptide-2 (LMP-2) and transporter associated with antigen presentation-1 (TAP-1), but induction of these mRNAs is not diminished in HFF cells cultured with IFNalpha/beta-neutralizing antibodies. Taken together, these data support the idea that that both cytokine-dependent and cytokine-independent mechanisms account for WNV-induced HLA expression in human skin fibroblasts. (C) 2004 Elsevier Inc. All rights reserved.

Spatial and temporal definition of Ochlerotatus camptorhynchus (Thomson) (Diptera : Culicidae) in the Gippsland Lakes system of eastern Victoria

The confirmed vector of Ross River virus, Ochlerotatus camptorhynchus (Thomson), is the dominant mosquito species inhabiting saline marshes in coastal Victoria. This paper re-examines previously published data on Oc. camptorhynchus, plus additional data collected since that time, and provides greater spatial and temporal definition of Oc. camptorhynchus numbers at seven sites across the Gippsland Lakes system of eastern Victoria. A total of 357 672 Oc. camptorhynchus was captured from 1188 trap-nights across the seven trap sites during trapping seasons from 1990 to 2001. The dominance of Oc. camptorhynchus across the seven sites averaged 75%, with significant differences in mean abundance of Oc. camptorhynchus found between all trap sites. Significant differences in monthly abundance of Oc. camptorhynchus were observed for Wellington Shire. Increase in populations of Oc. camptorhynchus was associated with increases in rainfall at all trap sites, higher minimum temperatures at two of the seven trap sites, and wind speed at one trap site. Prioritisation of mosquito control may be applied based on spatial and temporal factors according to the findings of this study.

Ross River virus disease in Australia, 1886-1998, with analysis of risk factors associated with outbreaks

Ross River virus (RE) is a mosquito-borne arbovirus responsible for outbreaks of polyarthritic disease throughout Australia. To better understand human and environmental factors driving such events, 57 historical reports oil RR Outbreaks between 1896 and 1998 were examined collectively. The magnitude, regularity, seasonality, and locality of outbreaks were found to be wide ranging; however, analysis of climatic and tidal data highlighted that environmental conditions let differently ill tropical, arid, and temperate regions. Overall, rainfall seems to be the single most important risk factor, with over 90% of major outbreak locations receiving higher than average rainfall in preceding mouths. Many temperatures were close to average, particularly in tropical populations; however, in arid regions, below average maximum temperatures predominated, and ill southeast temperate regions, above average minimum temperatures predominated. High spring tides preceded coastal Outbreaks, both in the presence and absence of rainfall, and the relationship between rainfall and the Southern Oscillation Index and Lit Nina episodes suggest they may be useful predictive tools, but only ill southeast temperate regions. Such heterogeneity predisposing outbreaks supports the notion that there are different RE epidemiologies throughout Australia but also Suggests that generic parameters for the prediction and control of outbreaks are of limited use at a local level.

Entomological investigations in a focus of dengue transmission in Cairns, Queensland, Australia, by using the sticky ovitraps

Sticky ovitraps (patent pending) were used to sample female Aedes aegypti (L.) weekly in a focus of dengue activity in Cairns, Queensland, Australia. In February 2003, transmission of dengue virus serotype 2 began in the suburb of Parramatta Park, peaking in mid-March 2003. This suburb features many older, unscreened houses with high populations of Ae. aegypti. Highest densities (2-3.5 females per trap per week) were obtained during peak dengue transmission (January and February) before mosquito control was initiated. Beginning in late March, female Ae. aegypti collected in sticky ovitraps were tested for dengue viral RNA by using a TaqMan reverse transcription-polymerase chain reaction assay. Dengue viral RNA was detected in six pools of Ae. aegypti collected in late March. The highest minimum infection rate was 116/1000 mosquitoes. After the initiation of larval control (containers treated with S-methoprene or lambda-cyhalothrin) and adult control (interior harborage sites sprayed with lambda-cyhalothrin) in early March, trap collections dropped to

Differences in climatic factors between Ross River virus disease outbreak and nonoutbreak years

Ross River virus is a common mosquito-borne arbovirus responsible for outbreaks of polyarthritic disease throughout Australia. To better understand climatic factors preceding outbreaks, we compared seasonal and monthly rainfall and temperature trends in outbreak and nonoutbreak years at four epidemic-prone locations. Our analyses showed that rainfall in outbreak years tended to be above average and higher than rainfall in nonoutbreak years. Overall temperatures were warmer during outbreak years. However, there were a number of distinct deviations in temperature, which seem to play a role in either promoting or inhibiting outbreaks. These preliminary findings show that climatic differences occur between outbreak and nonoutbreak years; however, seasonal and monthly trends differed across geo-climatic regions of the country. More detailed research is imperative if we are to optimize the surveillance and control of epidemic polyarthritic disease in Australia.

Risks for ross river virus disease in tropical Australia

Background There are no analytical studies of individual risks for Ross River virus (RRV) disease. Therefore, we set out to determine individual risk and protective factors for RRV disease in a high incidence area and to assess the utility of the case-control design applied for this purpose to an arbovirus disease. Methods We used a prospective matched case-control study of new community cases of RRV disease in the local government areas of Cairns, Mareeba, Douglas, and Atherton, in tropical Queensland, from January I to May 31, 1998. Results Protective measures against mosquitoes reduced the risk for disease. Mosquito coils, repellents, and citronella candles each decreased risk by at least 2-fold, with a dose-response for the number of protective measures used. Light-coloured clothing decreased risk 3-fold. Camping increased the risk 8-fold. Conclusions These risks were substantial and statistically significant, and provide a basis for educational programs on individual protection against RRV disease in Australia. Our study demonstrates the utility of the case-control method for investigating arbovirus risks. Such a risk analysis has not been done before for RRV infection, and is infrequently reported for other arbovirus infections.

Evaluation of Melanotaenia duboulayi (Atheriniformes : Melanotaeniidae), Hypseleotris galii (Perciformes : Eleotridae), and larvicide Vectolex (R) WG (Bacillus sphaericus) for integrated control of Culex annulirostris

Australian freshwater fish species Melanotaenia duboulayi and Hypseleotris galii were selected for a small plot field evaluation of an integrated pest management strategy using native fish and VectoLex® WG (Bacillus sphaericus) for the control of Culex annulirostris Skuse, the principal freshwater vector of arbovirus Ross River virus in Australia. When tested alone, the level of control afforded by M. duboulayi and H. galii was highly dependent on the prerelease density of mosquito larvae; and even when stocking rates as high as 10 g per pond (>30 kg/ha) were used, larval abundance was too high to attain adequate control from fish alone. In contrast, treatment with VectoLex WG at 500 g/ha resulted in 100% mortality of Cx. annulirostris immatures, but no residual activity was evident. The delayed reduction of Cx. annulirostris immatures in ponds stocked with fish alone, and the recolonization by Cx. annulirostris in ponds after treatment with B. sphaericus, did not occur when both treatments were combined.

Laboratory evaluation of the effect of alternative prey and vegetation on predation of Culex annulirostris immatures by Australian native fish species

The impact of alternative prey and simulated vegetation on Culex annulirostris Skuse predation efficacy by Australian smelt, Retropinna semoni (Retropinnidae); crimson-spotted rainbowfish, Melanotaenia duboulayi (Melanotaeniidae); empire gudgeon, Hypseleotris compressa (Eleotridae); estuary perchlet, Ambassis marianus (Ambassidae); firetail gudgeon, Hypseleotris galii (Eleotridae); fly-specked hardyhead, Craterocephalus stercusmuscarum (Atherinidae); and Pacific blue-eye, Pseudomugil signifer (Atherinidae), was evaluated in Queensland, Australia. The presence of chironomid midge larvae and tusked frog, Adelotus brevis (Leptodactylidae), tadpoles did not have a significant negative impact on the predation rates of Cx. annulirostris by these 7 fish species. Hypseleotris galii, M. duboulayi, and R. semoni demonstrated strong preference for larvae of Cx. annulirostris over both alternative prey species. In the presence of alternative prey species, the mean predation rate of M. duboulayi on larvae of Cx. annulirostris remained greater than that of other fish species tested. When evaluated at varying densities of simulated vegetation, predation rates of all fish species were similar to those reported in open conditions.

Spatio-temporal analyses of West Nile virus transmission in Culex mosquitoes in northern Illinois, USA, 2004

After a severe outbreak of West Nile virus (WNV) in Cook County, Illinois, in 2002, detections of WNV in mosquitoes were frequent across the state in the following years despite small numbers of human cases. We conducted a spatio-temporal analysis of Culex (subgenus Culex) mosquitoes collected in 2004 in three mosquito abatement districts (MAD) in Cook County by calculating monthly estimates of mosquito density, prevalence of infected mosquitoes, and exposure intensity, which in turn is a product of mosquito density and infection rates. Mosquito infections were detected early at three sites in late May and were widely detected throughout the three MADs in the summer with infection rates as high as 13 per 1000 in August. Exposure intensities were higher at sites adjacent to the Des Plaines River, especially in August and September. The aggregated pattern of WNV transmission along the river might be related to the existence of substantial forest preserves and wetlands that might produce ecological conditions favorable for mosquito proliferation and interactions between mosquitoes and birds.

Assessment of the occurrence and potential risks of pharmaceuticals and their metabolites in fish and water using liquid chromatography mass spectrometry

A comprehensive method for the analysis of 11 target pharmaceuticals representing multiple therapeutic classes was developed for biological tissues (fish) and water. Water samples were extracted using solid phase extraction (SPE), while fish tissue homogenates were extracted using accelerated solvent extraction (ASE) followed by mixed-mode cation exchange SPE cleanup and analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS). Among the 11 target pharmaceuticals analyzed, trimethoprim, caffeine, sulfamethoxazole, diphenhydramine, diltiazem, carbamazepine, erythromycin and fluoxetine were consistently detected in reclaimed water. On the other hand, caffeine, diphenhydramine and carbamazepine were consistently detected in fish and surface water samples. In order to understand the uptake and depuration of pharmaceuticals as well as bioconcentration factors (BCFs) under the worst-case conditions, mosquito fish were exposed to reclaimed water under static-renewal for 7 days, followed by a 14-day depuration phase in clean water. Characterization of the exposure media revealed the presence of 26 pharmaceuticals while 5 pharmaceuticals including caffeine, diphenhydramine, diltiazem, carbamazepine, and ibuprofen were present in the organisms as early as 5 h from the start of the exposure. Liquid chromatography ultra-high resolution Orbitrap mass spectrometry was explored as a tool to identify and quantify phase II pharmaceutical metabolites in reclaimed water. The resulting data confirmed the presence of acetyl-sulfamethoxazole and sulfamethoxazole glucuronide in reclaimed water. To my knowledge, this is the first known report of sulfamethoxazole glucuronide surviving intact through wastewater treatment plants and occurring in environmental water samples. Finally, five bioaccumulative pharmaceuticals including caffeine, carbamazepine, diltiazem, diphenhydramine and ibuprofen detected in reclaimed water were investigated regarding the acute and chronic risks to aquatic organisms. The results indicated a low potential risk of carbamazepine even under the worst case exposure scenario. Given the dilution factors that affect environmental releases, the risk of exposure to carbamazepine will be even more reduced.

Aedes aegypti pharate first instar aseasonal quiescence cues anticipatory plasticity with implications for urban vector ecology and control

The eggs of the dengue fever vector Aedes aegypti possess the ability to undergo an extended quiescence period hosting a fully developed first instar larvae within its chorion. As a result of this life history stage, pharate larvae can withstand months of dormancy inside the egg where they depend on stored reserves of maternal origin. This adaptation known as pharate first instar quiescence, allows A. aegypti to cope with fluctuations in water availability. An examination of this fundamental adaptation has shown that there are trade-offs associated with it. ^ Aedes aegypti mosquitoes are frequently associated with urban habitats that may contain metal pollution. My research has demonstrated that the duration of this quiescence and the extent of nutritional depletion associated with it affects the physiology and survival of larvae that hatch in a suboptimal habitat; nutrient reserves decrease during pharate first instar quiescence and alter subsequent larval and adult fitness. The duration of quiescence compromises metal tolerance physiology and is coupled to a decrease in metallothionein mRNA levels. My findings also indicate that even low levels of environmentally relevant larval metal stress alter the parameters that determine vector capacity. ^ My research has also demonstrated that extended pharate first instar quiescence can elicit a plastic response resulting in an adult phenotype distinct from adults reared from short quiescence eggs. Extended pharate first instar quiescence affects the performance and reproductive fitness of the adult female mosquito as well as the nutritional status of its progeny via maternal effects in an adaptive manner, i.e., anticipatory phenotypic plasticity results as a consequence of the duration of pharate first instar quiescence and alternative phenotypes may exist for this mosquito with quiescence serving as a cue possibly signaling the environmental conditions that follow a dry period. M findings may explain, in part, A. aegypti's success as a vector and its geographic distribution and have implications for its vector capacity and control.^

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

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.