Development and evaluation of a species diagnostic polymerase chain reaction-restriction fragment-length polymorphism procedure for cryptic members of the Culex sitiens (Diptera : Culicidae) subgroup in Australia and the southwest Pacific

Members of the Culex sitiens subgroup are important vectors of arboviruses, including Japanese encephalitis virus, Murray Valley encephalitis virus and Ross River virus. Of the eight described species, Cx. annulirostris Skuse, Cx. sitiens Wiedemann, and Cx. palpalis Taylor appear to be the most abundant and widespread throughout northern Australia and Papua New Guinea (PNG). Recent investigations using allozymes have shown this subgroup to contain cryptic species that possess overlapping adult morphology. We report the development of a polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) procedure that reliably separates these three species. This procedure utilizes the sequence variation in the ribosomal DNA ITS1 and demonstrates species-specific PCR-RFLP profiles from both colony and field collected material. Assessment of the consistency of this procedure was undertaken on mosquitoes sampled from a wide geographic area including Australia, PNG, and the Solomon Islands. Overlapping adult morphology was observed for Cx. annulirostris and Cx. palpalis in both northern Queensland and PNG and for all three species at one site in northwest Queensland.

Enhancement or modulation of the vector competence of ochlerotatus vigilax (Diptera : Culicidae) for Ross River virus by temperature

Two different doses of Ross River virus (1111) were fed to Ochlerotatus vigilax (Skuse), the primary coastal vector in Australia; and blood engorged females were held at different temperatures up to 35 d. After ingesting 10(4.3) CCID50/Mosquito, mosquitoes reared at 18 and 25degreesC (and held at the same temperature) had higher body remnant and head and salivary gland titers than those held at 32degreesC, although infection rates were comparable. At 18, 25, and 32degreesC, respectively, virus was first detected in the salivary glands on days 3, 2, and 3. Based on a previously demonstrated 98.7% concordance between salivary gland infection and transmission, the extrinsic incubation periods were estimated as 5, 4, and 3 d, respectively, for these three temperatures. When Oc. vigilax reared at 18, 25, or 32degreesC were fed a lower dosage of 10(3.3) CCID50 RR/mosquito, and assayed after 7 d extrinsic incubation at these (or combinations of these) temperatures, infection rates and titers were similar. However, by 14 d, infection rates and titers of those reared and held at 18 and 32degreesC were significantly higher and lower, respectively. However, this process was reversible when the moderate 25degreesC was involved, and intermediate infection rates and titers resulted. These data indicate that for the strains of RR and Oc. vigilax used, rearing temperature is unimportant to vector competence in the field, and that ambient temperature variations will modulate or enhance detectable infection rates only after 7 d: extrinsic incubation. Because of the short duration of extrinsic incubation, however, this will do little to influence RR epidemiology, because by this time some Oc. vigilax could be seeking their third blood meal, the latter two being infectious.

DNA vaccine coding for the full-length infectious Kunjin virus RNA protects mice against the New York strain of West Nile virus

A plasmid DNA directing transcription of the infectious full-length RNA genome of Kunjin (KUN) virus in vivo from a mammalian expression promoter was used to vaccinate mice intramuscularly. The KUN viral cDNA encoded in the plasmid contained the mutation in the NS1 protein (Pro-250 to Leu) previously shown to attenuate KUN virus in weanling mice. KUN virus was isolated from the blood of immunized mice 3-4 days after DNA inoculation, demonstrating that infectious RNA was being transcribed in vivo; however, no symptoms of virus-induced disease were observed. By 19 days postimmunization, neutralizing antibody was detected in the serum of immunized animals. On challenge with lethal doses of the virulent New York strain of West Nile (WN) or wild-type KUN virus intracerebrally or intraperitoneally, mice immunized with as little as 0.1-1 mug of KUN plasmid DNA were solidly protected against disease. This finding correlated with neutralization data in vitro showing that serum from KUN DNA-immunized mice neutralized KUN and WN,viruses with similar efficiencies. The results demonstrate that delivery of an attenuated but replicating KUN virus via a plasmid DNA vector may provide an effective vaccination strategy against virulent strains of WN virus.

Cross-protective and Infection-Enhancing Immunity in Mice Vaccinated Against Flaviviruses Belonging to the Japanese Encephalitis Virus Serocomplex

The Japanese encephalitis virus serocomplex is a group of mosquito-borne flaviviruses that cause severe encephalitic disease in humans. The recent emergence of several members of this serocomplex in geographic regions where other closely related flaviviruses are endemic has raised urgent human health issues. Thus, the impact of vaccination against one of these neurotropic virus on the outcome of infection with a second, serologically related virus is unknown. We show here that immunity against Murray Valley encephalitis virus in vaccinated mice can cross-protect but also augment disease severity following challenge with Japanese encephalitis virus. Immunepotentiation of heterologous flavivirus disease was apparent in animals immunized with a 'killed' virus preparation when humoral antiviral immunty of low magnitude was elicited. (C) 2002 Elsevier Science Ltd. All rights reserved.

Vector competence of Australian mosquitoes (Diptera : Culicidae) for Japanese encephalitis virus

Australian mosquitoes were evaluated for their ability to become infected with and transmit a Torres Strait strain of Japanese encephalitis virus. Mosquitoes, which were obtained from either laboratory colonies and collected using Centers for Disease Control and Prevention light traps baited with CO2 and octenol or reared from larvae, were infected by feeding on a blood/sucrose solution containing 10(4.5+/-0.1) porcine stable-equine kidney (PS-EK) tissue culture infectious dose(50)/ mosquito of the TS3306 virus strain. After 14 d, infection and transmission rates of 100% and 81%, respectively, were obtained for a southeast Queensland strain of Culex annulirostris Skuse, and 93% and 61%, respectively, for a far north Queensland strain. After 13 or more days, infection and transmission rates of > 90% and greater than or equal to 50%, respectively, were obtained for southeast Queensland strains of Culex sitiens Wiedemann and Culex quinquefasciatus Say, and a far north Queensland strain of Culex gelidus Theobald. Although infection rates were > 55%, only 17% of Ochlerotatus vigilax (Skuse) and no Cx. quinquefasciatus, collected from far north Queensland, transmitted virus. North Queensland strains of Aedes aegypti L., Ochlerotatus kochi (Donitz), and Verrallina funerea (Theobald) were relatively refractory to infection. Vertical transmission was not detected among 673 F, progeny of Oc. vigilax. Results of the current vector competence study, coupled with high field isolation rates, host feeding patterns and widespread distribution, confirm the status of Cx. annulirostris as the major vector of Japanese encephalitis virus in northern Australia. The relative roles of other species in potential Japanese encephalitis virus transmission cycles in northern Australia are discussed.

Mosquito host-feeding patterns and implications for Japanese encephalitis virus transmission in northern Australia and Papua New Guinea

Japanese encephalitis (JE) virus spread to northern Australia during the 1990s, transmitted by Culex annulirostris Skuse and other mosquitoes (Diptera: Culicidae). To determine the relative importance of various hosts for potential vectors of JE virus, we investigated the host-feeding patterns of mosquitoes in northern Australia and Western Province of Papua New Guinea, with particular attention to pigs, Sus scrofa L. - the main amplifying host of JE virus in South-east Asia. Mosquitoes were collected by CDC light traps baited with dry ice and 1-octen-3-ol, run 16.00-08.00 hours, mostly set away from human habitations, if possible in places frequented by feral pigs. Bloodmeals of 2569 mosquitoes, representing 15 species, were identified by gel diffusion assay. All species had fed mostly on mammals: only 30%) were trapped where domestic pigs were kept close to human habitation. From seven of eight locations on the Australian mainland, the majority of Cx. annulirostris had obtained their bloodmeals from marsupials, probably the Agile wallaby Macropus agilis (Gould). Overall proportions of mosquito bloodmeals identified as marsupial were 60% from the Gulf Plains region of Australia, 78% from the Cape York Peninsula and 64% from the Daru area of Papua New Guinea. Thus, despite the abundance of feral pigs in northern Australia, our findings suggest that marsupials divert host-seeking Cx. annulirostris away from pigs. As marsupials are poor JE virus hosts, the prevalence of marsupials may impede the establishment of JE virus in Australia.

Numerical simulations of wind and temperature structure within an Alpine lake basin, Lake Tekapo, New Zealand

High-resolution numerical model simulations have been used to study the local and mesoscale thermal circulations in an Alpine lake basin. The lake (87 km(2)) is situated in the Southern Alps, New Zealand and is located in a glacially excavated rock basin surrounded by mountain ranges that reach 3000 m in height. The mesoscale model used (RAMS) is a three-dimensional non-hydrostatic model with a level 2.5 turbulence closure scheme. The model demonstrates that thermal forcing at local (within the basin) and regional (coast-to-basin inflow) scales drive the observed boundary-layer airflow in the lake basin during clear anticyclonic summertime conditions. The results show that the lake can modify (perturb) both the local and regional wind systems. Following sunrise, local thermal circulations dominate, including a lake breeze component that becomes embedded within the background valley wind system. This results in a more divergent flow in the basin extending across the lake shoreline. However, a closed lake breeze circulation is neither observed nor modelled. Modelling results indicate that in the latter part of the day when the mesoscale (coast-to-basin) inflow occurs, the relatively cold pool of lake air in the basin can cause the intrusion to decouple from the surface. Measured data provide qualitative and quantitative support for the model results.

Flavivirus isolations from mosquitoes collected from western Cape York Peninsula, Australia, 1999-2000

After the 1st appearance of Japanese encephalitis virus (JE) on mainland Australia in 1998, a study was undertaken to investigate whether JE had become established in enzootic transmission cycles on western Cape York Peninsula. Adult mosquitoes were collected during the late wet season from Kowanyama and Pormpuraaw in April 1999, and Pormpuraaw and Barr's Yard in April 2000. Despite processing 269,270 mosquitoes for virus isolation, no isolates of JE were obtained. However, other flaviviruses comprising Murray Valley encephalitis virus, Kunjin virus, Alfuy virus, and Kokobera virus (KOK) were isolated. Isolates of the alphaviruses Ross River virus, Barmah Forest virus (BF), and Sindbis virus (SIN) also were obtained. The majority (88%) of isolates were from members of the Culex sitiens subgroup. Single isolates of KOK, BF and SIN were obtained from Ochlerotatus vigilax, Oc. normanensis, and Anopheles bancroftii, respectively. The isolations of flaviviruses during the late wet season indicate that conditions were suitable for flavivirus activity in the area. No evidence was found to suggest that JE has become established in enzootic transmission cycles on western Cape York, although study sites and field trips were limited.

A marginalização do Vale do Cávado e do Ave

Marginalizar o Cávado e o Ave é marginalizar o próprio Porto dentro de si e, portanto, todo o Norte. Assim como em Lisboa, há quem de dentro do Porto, ainda não entenda bem o que é o Vale do Cávado e do Ave!; Abstract: marginalize the Cávado and Ave is to marginalize the very Oporto within itself and therefore the entire North. As in Lisbon, some from within the Oporto, still do not understand well what is the Valley of Cávado and Ave!

Agentes ou Jogadores? Os monstros estranhamente reais dos jogos de computadores

Pretendemos mostrar como os sistemas de motion capture permitem transpor para avatares movimentos de pessoas reais para, com o auxílio do processamento gráfico e dos incrementos em Inteligência artificial, criar as planícies de Masahiro Mori, o uncanny valley. Acreditamos que, num futuro próximo, será possível emocionar jogadores se o realismo das interacções no jogo apresentar consistência semelhante às experiências no mundo real.

As autarquias e a educação. Centro e periferia na construção das políticas educativas (1998-2008)

RESUMO: Com o actual quadro de descentralização de atribuições e competências da administração central para as autarquias locais, na área da educação, os municípios passam a investir cada vez mais na acção educativa ao liderarem e planearem políticas educativas locais mais ou menos explícitas, e, nalguns casos, tentando superar carências que o sistema educativo apresenta. Esta pesquisa tem como problemática compreender o papel do Estado na (re)configuração das políticas de educação, quando a tendência para a mudança, de um Estado-educador para um Estado-regulador, tem por pressuposto o discurso neoliberal de que com ‗menos‘ Estado mas maior accountability se obtêm melhores resultados. Este processo origina uma redefinição no papel e funções do Estado no plano social e económico, provocando constrangimentos e conflitos de poder no que respeita ao seu controlo político, com a redistribuição de poderes entre o Estado e a comunidade, entre o central e o local. É neste contexto de mudança que a presente investigação, que se situa no âmbito da análise das políticas educativas, procura averiguar como e com que meios as autarquias locais concretizam as suas competências na área da educação. A estratégia de investigação concentra-se em uma metodologia qualitativa, com a utilização de um estudo exploratório, em três municípios da Região de Lisboa e Vale do Tejo. As actuais políticas educativas derivam da nova visão na gestão da coisa pública – res publica –, como resultado da nova concepção para o próprio Estado, e dos processos de elaboração das decisões político-educativas. Nesta perspectiva, a descentralização passa a ser um instrumento do poder local que favorece o aumento da autoridade democrática dos actores. Todavia, a governação – governance – supõe uma dinâmica de negociação, até mesmo de regulação entre o Estado, a região, o local, a escola e o mercado, feita para atender à construção do interesse geral, que já não é totalmente definido pelo Estado, mas construído em conjunto com as diversas forças políticas, económicas, educativas e sociais. O estudo permitiu evidenciar que a descentralização é posta em causa pelo Estado central, quando este ‗recentraliza‘ decisões e condiciona o poder local, com o fecho da maioria das escolas do primeiro ciclo e a verticalização dos agrupamentos escolares. Por sua vez, algumas políticas educativas como a ‗Escola a Tempo Inteiro‘ fomentam a desregulação dos vínculos laborais, forçando os municípios a aumentar os seus meios técnicos e humanos e a construírem novas infra-estruturas educativas. As políticas educativas passaram a ser concebidas segundo uma matriz híbrida, que visam a municipalização da educação – do pré-escolar e de todo o ensino básico –, por um lado; e fomentam a situação de ‗quase-mercado‘ com a privatização de sectores e o financiamento de várias instituições – que fornecem serviços na área da educação –, por outro lado. ABSTRACT: With the current framework of decentralization of functions and powers from central government to local authorities, in education, the municipalities are investing each more in educational work in leading educational policies and planning places more or less explicit and in some cases, trying to overcome shortcomings that education system. This research aims to understand the role of the state in the (re) configuration of education policies, when the tendency for the change in a State-Educator for a State-regulator, is the assumption that neo-liberal speech that with 'less' State but with more accountability we achieve better results. This process leads to a redefinition of the role and State functions in socio-economic constraints, resulting in power struggles with regard to its political control, with the redistribution of powers between the state and community, between the central and local. It is in this changing context that the present investigation, which lies in the examination of education policy addresses the question how and by what means the local, materialized their skills in education. The strategy focuses on a qualitative methodology, with the use of an exploratory study in three municipalities of Lisbon and Tagus Valley. The current education policies come from the new vision in the management of public affairs - res publica - as a result of the new design for the State itself, and the process of preparation of educational policy decisions. In this perspective, decentralization becomes an instrument of local government that favours the increase of democratic authority of the actors. However, the governance assumes a dynamic negotiation, even in regulation between the State, region, local authorities, school and market, made to suit the construction of general interest, which is not anymore fully defined by the State, but constructed together with the various political, economic, educational and social forces. The study indicates that decentralization is undermined by the central government when it ‗re-centralize‘ decisions and the local conditions, with the closure of most primary schools and with vertical groupings of schools. In turn, some educational policies such as 'Full Time School' forced the municipalities to increase their technical and human resources, to build new educational infrastructure. The educative policies began to be designed according to a hybrid matrix, which aims the decentralization of education - from pre-school and all the primary school - on one hand, and promote the situation of 'quasi-market' with privatization of sectors and the financing of several institutions - that provide services in education -, on the other hand. RÉSUMÉ: Avec le cadre actuel de décentralisation des fonctions et pouvoirs du gouvernement central aux autorités locales, dans l'éducation, les municipalités investissent de plus en plus dans le travail éducatif dans la conduite des politiques éducatives en mener et en faisant la planification des lieux plus ou moins explicites et, dans certains cas, essayer de remédier aux lacunes que présente l'éducation. Donc, nous voulons avec cette recherche comprendre le rôle de l'Etat dans la (re) configuration des politiques d'éducation, alors que la tendance au changement d‘un État-éducateur pour un État-régulateur, a comme l'hypothèse le discours néo-libéral de que avec «moins» État, mais plus d‘accountability on a des meilleurs résultats. Ce processus conduit à une redéfinition du rôle et des fonctions de l'Etat au plan social et économique, en donnant lieu à des luttes de pouvoir à l'égard de son contrôle politique, avec la redistribution des compétences entre l'État et la collectivité, entre les niveaux central et local. C‘est dans ce contexte changeant que la présente enquête, qui réside dans l‘examen de la politique de l‘éducation aborde la question de savoir comment et par quels moyens le local matérialisé leurs compétences dans l‘éducation. La stratégie est axée sur une méthodologie qualitative, avec l'utilisation d'une étude exploratoire dans trois municipalités de Lisbonne et Vallée du Tage. Les politiques actuelles d'éducation sont tirées de la nouvelle vision dans la gestion des affaires publiques - res publica – à la suite de la nouvelle conception de l'État lui-même, et le processus de préparation des décisions politique-éducatives. Dans cette perspective, la décentralisation devient un instrument de gouvernement local qui favorise l'augmentation de l'autorité démocratique des acteurs. Toutefois, la gouvernance assume une dynamique de négociation, même en matière de réglementation entre l'État, la région, le local, l'école et le marché, faite pour répondre à la construction d'intérêt général, qui n'est pas plus entièrement défini par l'Etat, mais construit en ensemble avec les divers forces politiques, économiques, éducatives et sociales. L‘étude indique que la décentralisation est minée par le gouvernement central quand il ‗re-centralise‘ les décisions et les conditions locales, avec la fermeture de la plupart des écoles du premier cycle et avec des groupements verticaux d‘écoles. À leur tour, certaines politiques éducatives telles que ‗l'école à temps plein‘ forcé les municipalités à accroître leurs ressources techniques et humaines, de construire de nouvelles infrastructures éducatives. Les politiques éducatives ont commencé à être conçues selon une matrice hybride, qui vise à la municipalisation de l'éducation - de l'école maternelle et de toute l'école basique - d'une part ; et de promouvoir la situation de «quasi-marché» avec la privatisation de secteurs et le financement de plusieurs institutions - qui offrent des services dans l'éducation -, d‘autre part.

Biting activity of Aedes scapularis (Rondani) and Haemagogus mosquitoes in Southern Brazil (Diptera: Culicidae)

The biting activity of a population of Aedes scapularis (Rondani), Haemagogus capricornii Lutz and Hg. leucocelaenus (Dyar and Shannon) in Southern Brazil was studied between March 1980 and April 1983. Data were obtained with 25-hour human bait catches in three areas with patchy residual forests, named "Jacaré-Pepira", "Lupo" Farm, and "Sta. Helena" Farm, in the highland region of S. Paulo State (Brazil). Data obtained on Ae. scapularis were compared with those formerly gathered in the "Ribeira'' Valley lowlands, and were similar, except in the "Lupo" Farm study area, where a precrepuscular peak was observed, not recorded at the "Jacaré-Pepira" site or in the "Ribeira" Valley. In all the areas this mosquito showed diurnal and nocturnal activity, but was most active during the evening crepuscular period. These observations support the hypothesis about the successful adaptation of Ae. scapularis to man-made environments and have epidemiological implications that arise from it. As for Haemagogus, results obtained on the "Lupo" and "Sta. Helena" regions agree with previous data obtained in several other regions and show its diurnal activity. The proximity of "Lupo" Farm, where Hg. capricornii and Hg. leucocelaenus showed considerable activity, to "Araraquara" city where Aedes aegypti was recently found, raises some epidemiological considerations about the possibility of urban yellow fever resurgence.

Studies on mosquitoes (Diptera: Culicidae) and anthropic environment: 1- Parity of blood seeking Anopheles (Kerteszia) in South-Eastern Brazil

Populations of Anopheles (Kerteszia) were sampled fortnightly over a one-year period (August 1991 to July 1992) at Ribeira Valley, S. Paulo State, Brazil. Indoor and outdoor collections were made on human bait at evening crepuscular period. The Polovodova technique for age grading was applied to 3,501 females of Anopheles cruzii and to 416 females of An. bellator. That sample represented 34.4% of the total number of mosquitoes collected. The most abundant species found was An. cruzii. However, An. bellator showed an endophagy that was almost three times greater than that of An. cruzii. The overall parous rate was 25.4% and uniparity was practically dominant one. A proportion of 26.9% of An. cruzii and 12.0% of An. bellator were found to be uniparous. Only three outdoor females of the former species (0.1%) showed biparity. Parity of An. cruzii was higher in females caught outdoors than in those caught indoors. Nevertheless, 497 nulliparous females examined (417 cruzii and 80 bellator) had ovaries that had advanced to Christophers and Mer stages III to V. These results imply that these females had already practised hematophagy. Relating these results to those from the parous females, a high statistical significance was found, leading to the conclusion that gonothophic discordance is a common pattern among these anophelines. Further, these results obtained with human bait catches strongly suggest that nearly 38.0% of these host-seeking females had already taken at least one previous blood-meal. So it is possible that enough time could thus be available for the plasmodian development in the vectors.

Studies on mosquitoes (Diptera: Culicidae) and anthropic environment: 2 - Immature stages research at a rice irrigation system location in South-Eastern Brazil

A relation between a rice irrigation system and mosquito breeding was established in a study undertaken at the Ribeira Valley Experimental Station, from January through December 1992. Flooding favoured Anopheles (Nyssorhynchus) and Culex (Melanoconion) species, while empty paddies condition were propitious to Aedes scapularis and Culex (Culex) species. Compared with a more primitive area of the same region, several species showed high a degree of adaptation to the anthropic environment. Among them, Anopheles albitarsis, a potential malaria vector that breeds in the irrigation system, has shown immature stage production thirteen times higher than at the natural breeding sites. In addition, Ae. scapularis, An. oswaldoi, Cx. bastagarius, and Cx. chidesteri presented high levels of synanthropy.

Studies on mosquitoes (Diptera: Culicidae) and anthropic environment: 3- Survey of adult stages at the rice irrigation system and the emergence of Anopheles albitarsis in South-Eastern, Brazil

A study of adult Culicidae ecology was carried out from January 1992 through January 1993 at the rice irrigation system of the Ribeira Valley Experimental Station. The adaptation of Anopheles albitarsis to the anthropic environment became evident through the adult collections made at its various habitats represented by the irrigation system and the edge of the residual pond, as well as at those made within the local patchy residual woods. Other potential disease vectors were prevalent in the irrigated system too. There were Aedes scapularis, Culex nigripalpus and Cx. ribeirensis that were collected at various habitats. Remarkable differences among their prevalences were obtained such as between the natural forest and anthropic environments. In the former An. albitarsis was practically non-existent, thus suggesting that it might be considered as eusynathropic. As the populations of other species seemed to increase in the anthropic environment, they may be regarded as hemisynanthropes. Observations suggest the hypothesis that the development of irrigated land may be a factor in the emergence of An. albitarsis, and some other species, as well as the possibility of an increase in the transmission of mosquito-borne diseases such as malaria.