A mistura de hexythiazox com acaricidas de ação adulticida tem sido explorada no controle de Brevipalpus phoenicis (Geijskes, 1939) em pomares de citros. Sendo assim, o presente trabalho foi realizado com o objetivo de subsidiar a recomendação de mistura de acaricidas no manejo da resistência de B. phoenicis a hexythiazox. Para esse propósito, foram realizados estudos para 1) avaliar a relação de resistência cruzada entre hexythiazox e cyhexatin em B. phoenicis;2) 2) entender a estabilidade da resistência de B. phoenicis a hexythiazox mediante a comparação da biologia das linhagens suscetível (S) e resistente (R) ao hexythiazox em condições de laboratório e a avaliação da dinâmica da resistência em condições de campo, por um período de dois anos; e 3) avaliar a estratégia da mistura de hexythiazox com os acaricidas cyhexatin ou dicofol mediante a condução de estudos de persistência da atividade biológica de acaricidas sobre as linhagens S e R, além de experimentos de campo para validar essa estratégia. A estimativa da freqüência de resistência a hexythiazox foi realizada com bioensaio de contato direto sobre ovos, utilizando-se a concentração discriminatória de 18 mg de hexythiazox/L de água. A resistência cruzada entre hexythiazox e cyhexatin foi ausente. Não foram verificadas diferenças significativas entre as linhagens S e R, com relação aos parâmetros biológicos avaliados para a confecção das tabelas de vida e fertilidade em condições de laboratório. No entanto, a resistência de B. phoenicis a hexythiazox foi instável em condições de campo; isto é, foram verificadas reduções significativas na freqüência de resistência na ausência de pressão de seleção, tanto em talhões com baixa (<20%) como em talhões com alta (>60%) freqüência de resistência. Estudos de persistência da atividade biológica de hexythiazox, utilizada na forma isolada ou em mistura com cyhexatin ou dicofol, evidenciaram que há discriminação na taxa instantânea de crescimento (ri) das linhagens S e R. Dicofol apresentou menor persistência do que hexythiazox e cyhexatin. As freqüências de resistência a hexythiazox estimadas após a utilização da mistura de hexythiazox com cyhexatin ou dicofol foram significativamente menores do que a freqüência estimada após a utilização de hexythiazox isoladamente; porém maiores do que as freqüências estimadas após a utilização de cyhexatin ou dicofol isoladamente. Apesar do controle satisfatório de B. phoenicis com a utilização da mistura em pomares com baixa freqüência de resistência a hexythiazox, essa estratégia deve ser recomendada com cautela para o manejo da resistência de B. phoenicis a hexythiazox. / The mixture of hexythiazox with acaricides with adulticidal activity has been exploited for controlling Brevipalpus phoenicis (Geijskes, 1939) in citrus groves. Therefore, this research was conducted to subsidize the recommendation of mixture of acaricides in the management of B. phoenicis resistance to hexythiazox. For this purpose, studies were undertaken 1) to evaluate cross-resistance relationship between hexythiazox and cyhexatin; 2) to understand the stability of B. phoenicis resistance to hexythiazox by comparing the life-history of susceptible (S) and resistant (R) strains of B. phoenicis to hexythiazox under laboratory conditions and by evaluating the dynamics of the resistance under field conditions, during a 2-year period; and 3) to evaluate the strategy of the mixture of hexythiazox with cyhexatin or dicofol acaricides by evaluating the persistence of biological activity of the acaricides on S and R strains and conducting field experiments to validate this strategy. The frequency of resistance to hexythiazox was estimated with direct contact bioassays on eggs with the use of discriminating concentration of 18 mg of hexythiazox/L of water. There is no cross-resistance between hexythiazox and cyhexatin. No significant differences were observed between S and R strains in relation to biological parameters evaluated to built life and fertility tables under laboratory conditions. However, the resistance of B. phoenicis to hexythiazox was unstable under field conditions; that is, significant reductions in the frequency of resistance were observed in the absence of selection pressure, either in fields with low (<20%) or high (>60%) frequency of resistance. Studies on persistence of biological activities of hexythiazox, used by itself or in mixture with cyhexatin or dicofol, showed that there is clear discrimination between S and R strains based on the instantaneous rate of increase (ri). Dicofol was less persistent than hexythiazox and cyhexatin. Frequencies of resistance to hexythiazox estimated following the use of mixture of hexythiazox with cyhexatin or dicofol were significantly lower than that with the use of hexythiazox alone; but higher than the frequencies estimated following the use of cyhexatin or dicofol alone. Despite of good control of B. phoenicis in citrus groves with low frequencies of resistance to hexythiazox, the mixture of acaricides must be recommended with cautious for managing the resistance of B. phoenicis to hexythiazox.

A mistura de hexythiazox com acaricidas de ação adulticida tem sido explorada no controle de Brevipalpus phoenicis (Geijskes, 1939) em pomares de citros. Sendo assim, o presente trabalho foi realizado com o objetivo de subsidiar a recomendação de mistura de acaricidas no manejo da resistência de B. phoenicis a hexythiazox. Para esse propósito, foram realizados estudos para 1) avaliar a relação de resistência cruzada entre hexythiazox e cyhexatin em B. phoenicis;2) 2) entender a estabilidade da resistência de B. phoenicis a hexythiazox mediante a comparação da biologia das linhagens suscetível (S) e resistente (R) ao hexythiazox em condições de laboratório e a avaliação da dinâmica da resistência em condições de campo, por um período de dois anos; e 3) avaliar a estratégia da mistura de hexythiazox com os acaricidas cyhexatin ou dicofol mediante a condução de estudos de persistência da atividade biológica de acaricidas sobre as linhagens S e R, além de experimentos de campo para validar essa estratégia. A estimativa da freqüência de resistência a hexythiazox foi realizada com bioensaio de contato direto sobre ovos, utilizando-se a concentração discriminatória de 18 mg de hexythiazox/L de água. A resistência cruzada entre hexythiazox e cyhexatin foi ausente. Não foram verificadas diferenças significativas entre as linhagens S e R, com relação aos parâmetros biológicos avaliados para a confecção das tabelas de vida e fertilidade em condições de laboratório. No entanto, a resistência de B. phoenicis a hexythiazox foi instável em condições de campo; isto é, foram verificadas reduções significativas na freqüência de resistência na ausência de pressão de seleção, tanto em talhões com baixa (<20%) como em talhões com alta (>60%) freqüência de resistência. Estudos de persistência da atividade biológica de hexythiazox, utilizada na forma isolada ou em mistura com cyhexatin ou dicofol, evidenciaram que há discriminação na taxa instantânea de crescimento (ri) das linhagens S e R. Dicofol apresentou menor persistência do que hexythiazox e cyhexatin. As freqüências de resistência a hexythiazox estimadas após a utilização da mistura de hexythiazox com cyhexatin ou dicofol foram significativamente menores do que a freqüência estimada após a utilização de hexythiazox isoladamente; porém maiores do que as freqüências estimadas após a utilização de cyhexatin ou dicofol isoladamente. Apesar do controle satisfatório de B. phoenicis com a utilização da mistura em pomares com baixa freqüência de resistência a hexythiazox, essa estratégia deve ser recomendada com cautela para o manejo da resistência de B. phoenicis a hexythiazox. / The mixture of hexythiazox with acaricides with adulticidal activity has been exploited for controlling Brevipalpus phoenicis (Geijskes, 1939) in citrus groves. Therefore, this research was conducted to subsidize the recommendation of mixture of acaricides in the management of B. phoenicis resistance to hexythiazox. For this purpose, studies were undertaken 1) to evaluate cross-resistance relationship between hexythiazox and cyhexatin; 2) to understand the stability of B. phoenicis resistance to hexythiazox by comparing the life-history of susceptible (S) and resistant (R) strains of B. phoenicis to hexythiazox under laboratory conditions and by evaluating the dynamics of the resistance under field conditions, during a 2-year period; and 3) to evaluate the strategy of the mixture of hexythiazox with cyhexatin or dicofol acaricides by evaluating the persistence of biological activity of the acaricides on S and R strains and conducting field experiments to validate this strategy. The frequency of resistance to hexythiazox was estimated with direct contact bioassays on eggs with the use of discriminating concentration of 18 mg of hexythiazox/L of water. There is no cross-resistance between hexythiazox and cyhexatin. No significant differences were observed between S and R strains in relation to biological parameters evaluated to built life and fertility tables under laboratory conditions. However, the resistance of B. phoenicis to hexythiazox was unstable under field conditions; that is, significant reductions in the frequency of resistance were observed in the absence of selection pressure, either in fields with low (<20%) or high (>60%) frequency of resistance. Studies on persistence of biological activities of hexythiazox, used by itself or in mixture with cyhexatin or dicofol, showed that there is clear discrimination between S and R strains based on the instantaneous rate of increase (ri). Dicofol was less persistent than hexythiazox and cyhexatin. Frequencies of resistance to hexythiazox estimated following the use of mixture of hexythiazox with cyhexatin or dicofol were significantly lower than that with the use of hexythiazox alone; but higher than the frequencies estimated following the use of cyhexatin or dicofol alone. Despite of good control of B. phoenicis in citrus groves with low frequencies of resistance to hexythiazox, the mixture of acaricides must be recommended with cautious for managing the resistance of B. phoenicis to hexythiazox.

Mosquitoes are the deadliest animal on the planet killing about a million people a year. These insects are competent vectors of multiple pathogens (e.g., Plasmodium sp, filarial worms, and arboviruses). In most species, females are blood feeders, and must consume a blood meal to complete a gonotrophic cycle. Extensive research has been conducted on hematophagy and host-seeking behaviors, but relatively little is known about phytophagy. Sugar feeding is an essential aspect of mosquito biology. Both male and female mosquitoes must consume sugar as a primary fuel source. Mosquitoes use olfactory and visual cues among other cues to find suitable food sources. Abiotic factors, such as temperature and humidity, have been shown to impact mosquito behaviors, including sugar feeding. Recently, sugar feeding has been identified as a promising control target for multiple species of mosquitoes. Attractive toxic sugar baits (ATSBs) attract both male and females through the use of plant derived volatiles. In this work, we first examined the effects of temperature and humidity on the survival of sugar fed Aedes aegypti. We showed that sugar feeding greatly increases longevity in optimal conditions and that humidity impacted survival while temperature less so. Second, we aimed at developing an ATSB for controlling Aedes j. japonicus mosquitoes. Overall, this work sheds light on the importance of abiotic factors and sugar feeding on mosquito survival and lays the groundwork for controlling an invasive mosquito species. / Master of Science in Life Sciences / Mosquitoes are the deadliest organisms on the planet killing about a million people each year due to the multitude of pathogens they may transmit (e.g., malaria parasites, Zika virus, dengue virus). Female mosquitoes transmit pathogens by taking blood meals to obtain essential nutrients needed to develop eggs. If knowledge on host-seeking behavior and blood-feeding is extensive, comparatively less is known about sugar feeding. Sugar feeding is an important behavior displayed by both male and female mosquitoes to obtain energy for flying, reproduction, and survival. Mosquitoes feed on a variety of sugar sources including plant nectar and decaying fruits. They use multiple different cues in order to locate suitable meals including olfaction and vision. Environmental factors such as temperature and humidity affect mosquito activity, dehydration, and sugar feeding. In the present work, we first examined the effects of temperature and humidity on survival in the major disease vector species, Aedes aegypti. As the global temperatures are increasing, it is essential to better understand how mosquitoes adapt and deal with environmental stressors in a changing world. We then aimed at exploiting sugar feeding behavior by developing a novel method of control for another invasive mosquito species, Aedes j. japonicus. Together, these results help us have a better understanding of mosquito biology and ecology which is crucial for predicting future distribution of invasive species and designing new control strategies.

Solano-violeta (Solanum violaefolium) é uma ornamental rasteira usada pra cobrir solos de áreas sombreadas. Um vírus que induz manchas anelares nas folhas, tentativamente designado de mancha anular do S. violaefoliumm (S. violaefolium ringspot vírus - SvRSV), transmitido pelo ácaro Brevipalpus phoenicis foi encontrado nesta planta em jardins de Piracicaba, SP. Trata-se de um vírus baciliforme que se acumula no lúmen do retículo endoplasmático induzindo viroplasma citoplasmático, assemelhando-se a outros vírus do tipo citoplasmático, dos transmitidos por ácaros Brevipalpus (Acari: Tenuipalpidea). Este trabalho relata algumas de suas propriedades biológicas e a caracterização molecular parcial. SvRSV foi ser transmitido mecanicamente e pelo B. phoenicis a várias outras espécies botânicas, sempre causando lesões localizadas. Destas, Datura stramonium mostrou-se a melhor como hospedeira experimental. As propriedades físicas do SvRSV in vitro foram: temperatura de inativação - 40-45 ºC; ponto final de diluição - 10-3-10-4; longevidade in vitro- 12 dias. Posteriormente, observou-se também infestação destas plantas por B. obovatus que em ensaios preliminares transmitiu o SvRSV. Em secções ultrafinas, as partículas do SvRSV mostraram-se ligeiramente mais delgadas que as de outros vírus do tipo citoplasmático, transmitidos por Brevipalpus, e por outro lado formavam eventualmente partículas mais longas, às vezes de ca. 1 µm. Como os demais vírus, do tipo citoplasmático, transmitido por Brevipalpus, induz a formação de um viroplasma denso e vacuolado no citoplasma. Em casos favoráveis foram observadas fases do processo de morfogênese por "brotação" a partir do material do viroplasma. Dada sua labilidade não foi possível conseguir sua purificação apesar das inúmeras tentativas, usando diferentes protocolos. Logrou-se a extração de dsRNA a partir de D. stramonium e a partir dele, obter-se dois fragmentos do genoma viral, identificados como parte da proteína de movimento e da replicase, após seu sequenciamento. Foram produzidos pares de "primer" baseado nestas seqüências que amplificaram especificamente, por RT-PCR, fragmentos de DNA de tamanho esperado, a partir do RNA total extraído de lesões foliares de S. violaefolium e D. stramonium infetados. Sondas baseadas nas seqüências obtidas hibridizaram com ss- e dsRNA de lesões de D. stramonium. Ensaios preliminares de RT-PCR e hibridização não resultaram em reação com alguns outros vírus transmitidos por Brevipalpus, do tipo citoplasmático, inclusive o da leprose dos citros (CiLV-C). / Solanum violaefolium is an ornamental Solanaceae, with prostrate, trailing growth cultivated in shaded areas. Plants exhibiting necrotic ringlike spots on the leaves have been found in several gardens and parks at Piracicaba - SP. The ringspot symptoms on the leaves is caused by a vírus, named S. violaefolium ringspot virus (SvRSV), and is transmisible by mite Brevipalpus phoenicis (Acari: Tenuipalpidae). Short bacilliform particles are present within the cisternae of endoplasmic reticulum (ER) and often electron dense viroplasm is present in the cytoplasm, characterist of the cytoplasmatic type of Brevipalpus-borne virus. The present study reports some of its biological properties and partial molecular caracterization. SvRSV is easily transmitted to many plant species either by viruliferous B. phoenicis or mechanically, always causing local lesions. Datura stramonium was proved to be better as experimental host. Its physical properties in vitro were: inactivation thermal point - 40-45 ºC; final diluition point - 10-3-10-4; longevity in vitro - 12 days. Afterwards, its was observed that S. violaefolium plants were infested by B. obovatus that transmitted SvRSV in preliminary assays. Thin sections revealed that SvRSV particles are slightly thinner and sometimes appear very long. In some favorable sections intermediate steps of viral particle morphogenesis by a budding process of the dense material of the viroplasm toward the lumen of ER could be seen. Due to the fragility of the particles, several attempts to purify the virus have failed, despite many protocols tried. It was possible, however, to extract dsRNA from infected tissue of D. stramonium, and two segments of viral genome, respectively with homology to movement protein (mp) and replicase (rep) of some known viruses were obtained. Primers were designed based in these sequences, which amplified by RT-PCR, fragments of DNA of expected size from total RNA extracts from leaves lesions of infected S. violaefolium and D. stramonium. Probes based on obtained sequences hibridizated with ss- and dsRNA from lesions of S. violaefolium and D. stramonium. Preliminary assays of RT-PCR and hybridization did not result in positive reaction with other cytoplasmatic type of Brevipalpus-borne viruses, including citrus leprosis (CiLVC).

Solano-violeta (Solanum violaefolium) é uma ornamental rasteira usada pra cobrir solos de áreas sombreadas. Um vírus que induz manchas anelares nas folhas, tentativamente designado de mancha anular do S. violaefoliumm (S. violaefolium ringspot vírus - SvRSV), transmitido pelo ácaro Brevipalpus phoenicis foi encontrado nesta planta em jardins de Piracicaba, SP. Trata-se de um vírus baciliforme que se acumula no lúmen do retículo endoplasmático induzindo viroplasma citoplasmático, assemelhando-se a outros vírus do tipo citoplasmático, dos transmitidos por ácaros Brevipalpus (Acari: Tenuipalpidea). Este trabalho relata algumas de suas propriedades biológicas e a caracterização molecular parcial. SvRSV foi ser transmitido mecanicamente e pelo B. phoenicis a várias outras espécies botânicas, sempre causando lesões localizadas. Destas, Datura stramonium mostrou-se a melhor como hospedeira experimental. As propriedades físicas do SvRSV in vitro foram: temperatura de inativação - 40-45 ºC; ponto final de diluição - 10-3-10-4; longevidade in vitro- 12 dias. Posteriormente, observou-se também infestação destas plantas por B. obovatus que em ensaios preliminares transmitiu o SvRSV. Em secções ultrafinas, as partículas do SvRSV mostraram-se ligeiramente mais delgadas que as de outros vírus do tipo citoplasmático, transmitidos por Brevipalpus, e por outro lado formavam eventualmente partículas mais longas, às vezes de ca. 1 µm. Como os demais vírus, do tipo citoplasmático, transmitido por Brevipalpus, induz a formação de um viroplasma denso e vacuolado no citoplasma. Em casos favoráveis foram observadas fases do processo de morfogênese por “brotação” a partir do material do viroplasma. Dada sua labilidade não foi possível conseguir sua purificação apesar das inúmeras tentativas, usando diferentes protocolos. Logrou-se a extração de dsRNA a partir de D. stramonium e a partir dele, obter-se dois fragmentos do genoma viral, identificados como parte da proteína de movimento e da replicase, após seu sequenciamento. Foram produzidos pares de “primer” baseado nestas seqüências que amplificaram especificamente, por RT-PCR, fragmentos de DNA de tamanho esperado, a partir do RNA total extraído de lesões foliares de S. violaefolium e D. stramonium infetados. Sondas baseadas nas seqüências obtidas hibridizaram com ss- e dsRNA de lesões de D. stramonium. Ensaios preliminares de RT-PCR e hibridização não resultaram em reação com alguns outros vírus transmitidos por Brevipalpus, do tipo citoplasmático, inclusive o da leprose dos citros (CiLV-C). / Solanum violaefolium is an ornamental Solanaceae, with prostrate, trailing growth cultivated in shaded areas. Plants exhibiting necrotic ringlike spots on the leaves have been found in several gardens and parks at Piracicaba – SP. The ringspot symptoms on the leaves is caused by a vírus, named S. violaefolium ringspot virus (SvRSV), and is transmisible by mite Brevipalpus phoenicis (Acari: Tenuipalpidae). Short bacilliform particles are present within the cisternae of endoplasmic reticulum (ER) and often electron dense viroplasm is present in the cytoplasm, characterist of the cytoplasmatic type of Brevipalpus-borne virus. The present study reports some of its biological properties and partial molecular caracterization. SvRSV is easily transmitted to many plant species either by viruliferous B. phoenicis or mechanically, always causing local lesions. Datura stramonium was proved to be better as experimental host. Its physical properties in vitro were: inactivation thermal point – 40-45 ºC; final diluition point - 10-3-10-4; longevity in vitro – 12 days. Afterwards, its was observed that S. violaefolium plants were infested by B. obovatus that transmitted SvRSV in preliminary assays. Thin sections revealed that SvRSV particles are slightly thinner and sometimes appear very long. In some favorable sections intermediate steps of viral particle morphogenesis by a budding process of the dense material of the viroplasm toward the lumen of ER could be seen. Due to the fragility of the particles, several attempts to purify the virus have failed, despite many protocols tried. It was possible, however, to extract dsRNA from infected tissue of D. stramonium, and two segments of viral genome, respectively with homology to movement protein (mp) and replicase (rep) of some known viruses were obtained. Primers were designed based in these sequences, which amplified by RT-PCR, fragments of DNA of expected size from total RNA extracts from leaves lesions of infected S. violaefolium and D. stramonium. Probes based on obtained sequences hibridizated with ss- and dsRNA from lesions of S. violaefolium and D. stramonium. Preliminary assays of RT-PCR and hybridization did not result in positive reaction with other cytoplasmatic type of Brevipalpus-borne viruses, including citrus leprosis (CiLVC).

Introduction This thesis concerns the mosquito-borne arbovirus Ross River (RR) virus. The main objectives were to determine the vector associations, the incidence, costs and natural history of disease, and behavioural and environmental risks for infection in tropical Queensland. 1. Literature review On the basis of the literature review there is strong evidence that Aedes vigilax, Ae. camptorhynchus and Culex annulirostris are important vectors in Australia. Aedes camptorhychus does not occur in Queensland. There is evidence that two peri-domestic container-breeding mosquitoes, Ae. notoscriptus and Ae. aegypti, may be vectors of the virus. The virus has been isolated from many other species but the role of most of these is unclear. It is unclear which vertebrate species are the major reservoirs for human infection. Studies are inconsistent with regard to the prevalence, duration and severity of symptoms and debility during RR virus disease. Nearly all epidemiological studies of RR virus have been descriptive. Therefore one can only surmise what the risks for human infection might be. Epidemiological studies to determine the associations between exposures and risk for disease do not exist. 2. Virus isolation from mosquitoes During 1996-1998 61,619 mosquitoes were processed for virus isolation. Thirty-three isolates of RR virus were made. The largest number (14) were from Ae. carmenti. The minimum infection rate (MIR) per 1,000 was 2.4. Isolates were also made from Ae. imprimens (1 isolate, MIR 10.3), Ae. kochi (2, 0.2), Ae. lineatus (1, 0.2), Ae. notoscriptus (1, 1.6), Ae. vigilax (1, 0.3), Cx. annulirostris (9, 0.3), Cx. vicinus (1, 4.0) and Mansonia septempunctata (3, 5.8). Ross River virus has been isolated from Cx. annulirostris, Ae. vigilax, Ae. notoscriputus and Ae. kochi but not from the other species. Ross River virus was not isolated from Ae. aegypti. Twenty-six isolates came from in or near a colony of 15,000 spectacled flying-fox, Pteropus conspicillatus. The proportion of RR virus positive pools from within 1 km. of this colony was significantly greater than elsewhere for all species combined and for Cx. annulirostris but not for Ae. carmenti. 3. The incidence and costs of Ross River virus disease Unpublished data on National, State and Territory notifications was collected. Crude incidence rates using census figures for denominator data were calculated. The same was done for the areas in which the other studies described in the thesis were carried out. An estimate of the cost of Ross River virus disease in Australia was made. During the period 1991-1998 of the States and Territories the Northern Territory (NT) had the highest and the Australian Capital Territory (ACT) the lowest notification incidences. These were 62-281 and 0-3 per 100,000 per annum, respectively. During this period the notification incidence for Queensland ranged from 70 to 149 per 100,000 per annum. For the local government areas of Cairns and Mareeba, where the majority of cases of RR virus disease for the studies described in this thesis were recruited, the notification incidences were between 74 and 267, and 28 and 200, respectively. On the basis of an average of 4,800 cases per annum in Australia the cost of serological testing and medical consultations were estimated at $443,520 and $105,600. Lost earnings were estimated at $1,798,560. The total cost for medical consultations, serological testing and lost earnings will therefore be over $2 million in an average year. 4. The natural history of Ross River virus disease In 1998 incident cases of RR virus disease were ascertained. Fifty-seven eligible cases were recruited but only 47 could be reviewed on 3 occasions and data on these were analysed. Cases were followed for up to 197 days. Review included history, examination and the administration of the Clinical Health Assessment Questionnaire (CLINHAQ) and Short Form-36 (SF-36) . On initial review the 3 most common symptoms were arthralgia, joint stiffness and myalgia affecting 97.9, 89.4 and 59.6% of cases, respectively. The joint types most commonly affected by pain at the initial review were the ankles, wrists, interphalangeal joints of the fingers, knees and metacarpophalangeal joints. Objective signs of joint inflammation were rare. The prevalence of signs of inflammation decreased and the prevalence of normal joints on examination increased through the reviews. The prevalence of use of NSAIDs decreased through the reviews. On the basis of CLINHAQ items regarding work performance functioning at work improved through the course of the reviews. Linear regression with days since symptom onset as the independent variable was performed for some variables. The 8 dimensions of the SF-36 were standardized to the Queensland population and analyzed longitudinally . Analyses of the CLINHAQ functional disability index (FDI) and the visual analogue scales (VASs) for pain, global severity, fatigue, gastrointestinal complaints and sleep, and the depression and anxiety scales were also performed. The slopes of all fitted regressions except the SF-36 general health dimension were significantly different from zero. All measures of disease severity returned to normal by 8 months from onset, many in a shorter period. 5. Behavioural and environmental risks for infection Fifty-five incident cases of RR virus disease were recruited and formed the basis of a case-control study of behavioural and environmental risks. They were matched to 85 controls. In the year prior to symptom onset the only leisure exposure that significantly altered risk was camping [Odds ratio (OR) = 2.15; 95% confidence interval (CI) = 1.07-4.35]. No peri-domestic activities in the year prior to onset significantly altered disease risk. Leisure exposures were also assessed in a 3-week exposure period ending 4 days prior to symptom onset. None significantly altered risk. No peri-domestic activity in this period significantly altered risk and nor did exposure to vertebrates or mosquitoes. Containers and vegetation around the subject's dwelling did not significantly increase risk. The presence of ice cream containers and buckets was significantly protective when assessed by questionnaire, however this was also assessed by inspection and was found to increase risk though not significantly so. It was concluded that the former finding was due to differential misclassification of exposure status. The premise condition index (PCI) was measured. A low PCI for the subject's house was associated with a significantly increased risk (3-4 relative to 7-9 as reference category: OR = 3.15, 95% CI = 1.07-9.25). Window screening did not alter disease risk and air-conditioning in the house or the bedroom decreased risk but not significantly so. Use of protective measures, except bed nets, in the year prior to onset was found to decrease disease risk. Personal repellents, mosquito coils and citronella candles significantly decreased risk. A dose response was shown for the number of protective measures from personal repellents, aerosol and surface sprays, mosquito coils, citronella candles and mosquito "zappers" used in the year prior to symptom onset. Pet ownership and proximity of dwelling to horses did not significantly alter risk. A preference for light coloured clothing was significantly protective (0.37, 0.15-0.89). Stratification by gender, date of symptom onset and geographical area was performed. Stratification by geographical area included a coastal and tablelands stratum. There were differences between the stratum-specific odds ratios for camping in the year prior to symptom onset, the presence of bromeliads in the subject's garden and a preference for light coloured clothing. Multivariate analysis demonstrated confounding by use of personal repellents, mosquito coils and citronella candles. When modeled together these were found to cause confounding among themselves. They also caused significant confounding of camping, outdoor work and the presence of banana trees in the subject's yard. Multivariate analysis of the association between PCI and disease risk failed to demonstrate confounding by use of protective measures or time between symptom onset and review. 6. Synthesis and conclusions The three research Chapters form a coherent body of public health research on the epidemiology (Chapters 5 and 6) and ecology (Chapters 4 and 7) of RR virus, and the natural history of RR virus disease (Chapter 6) in tropical Queensland. Conclusions are drawn from the research in the thesis. A set of priorities for future public health research on RR virus is suggested, and a pilot control program for Ross River virus disease in tropical Queensland is recommended.