The olive fruit fly, Bactrocera oleae, (Rossi) (Diptera:Tephritidae), is a key pest of olive crops. The sterile insect technique (SIT) is an environmentally benign and species-specific method of pest control, aiming to reduce the reproductive potential of a wild population through the mass-release of sterile insects. Previous olive fly SIT trials, involving the release of gamma-ray sterilised mixed-sex populations, achieved limited success. Key problems included altered diurnal mating rhythms of the laboratory-reared insects, leading to assortative mating between released and wild populations, and low competitiveness of the radiation sterilised mass-reared flies. Consequently, the production of competitive, male-only release cohorts is seen as essential. The RIDL (Release of Insects carrying a Dominant Lethal) system is a transgene-based derivative of SIT, one version of which involves the mass release of insects carrying a female specific lethal transgene (fsRIDL). This thesis describes: 1) the development of fsRIDL olive fly strains and the molecular analysis of transgene insertion and function; 2) the analysis of strain life-history parameters; 3) studies into sexual selection and mating compatibility; 4) a caged proof-of-principle population suppression trial; and, 5) selection dynamics on the fsRIDL trait in caged populations. Olive fly fsRIDL strains were developed with full female-lethal penetrance and repressibility. The lead strain displayed similar life-history and sexual competitiveness traits to those of the wild-type strain from which they were derived. In addition, transgenic males showed photoperiod compatibility and strong sexual competitiveness with field-collected wild olive flies. The feasibility of the fsRIDL approach was demonstrated when repeated male releases caused eradication of caged olive fly populations. Although needing field confirmation, these results suggest that fsRIDL olive fly strains may help to mitigate key problems experienced in previous olive fly SIT trials, and could help form the basis of a renewed effort towards olive fly SIT control.

In the light of increasing pesticides resistance in agricultural pests and in insect vectors of human diseases, leading to the rise in occurrence of mosquito-borne diseases, new, efficient and environmentally friendly methods of pest control are needed. Sterile Insect Technique (SIT), relying on mass releases of radiation sterilised males to reduce reproductive potential of target pest populations, although not new, offers an alternative to the use of pesticides and is an environmentally non-polluting method of insect control. Many insect species, however, are not very amenable to classical SIT, due to detrimental side-effects of radiation treatment. We propose a new method, a genetically engineered modification of classical SIT, replacing radiation with genetically induced sterility. Based on conditional expression of male-germline targeted nucleases which introduce double strand breaks into the male germline DNA to render males sterile, this method emulates SIT mechanism, at the same time eliminating radiation and associated detrimental side-effects. Different variants of such a system were investigated in this project, eventually leading to the creation of functional conditional male-sterility systems in two model organisms – the Yellow fever mosquito, Aedes aegypti and the Mediterranean fruit fly, Ceratitis capitata. Both systems utilise chimeric nuclease composed of protamine and FokI cleavage domain fusion. The sperm-specificity and the conditionality of the sterile phenotype have been achieved through the use of tetracycline repressible expression system driven by the β2-tubulin promoter in Ceratitis capitata and by the Topi promoter in Aedes aegypti.

The Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), a major pest of horticulture in eastern Australia, is a relatively poor coloniser of new habitat. This thesis examines behavioural properties that might limit the ability of B. tryoni to establish new populations. As the potential for B. tryoni to establish an outbreak population may be most directly limited by mechanisms associated with dispersal and mating behaviour, these two factors were the focus of this research project. The relevance of dispersal and mating behaviour for control of outbreak populations was assessed. Dispersal (i) Dispersal patterns of males and females are not different. Dispersal of post-teneral male B. tryoni from a point within an orchard near Richmond, New South Wales, was monitored following temporally replicated releases. Application of sterile insect technique (SIT) requires knowledge of dispersal from a release point so that effective release rates can be determined. In addition, dispersal following introduction to new habitat can lead to low or negative population growth and an Allee effect. In Spring and Autumn, 2001 � 2003, three different strains of B. tryoni were released: (1) wild flies reared from infested fruit collected in the Sydney Basin; (2) a laboratory-reared strain with a colour mutation (white marks); and (3) sterile flies obtained by gamma-irradiation of a mass-reared strain. Dispersal was monitored using a grid of traps baited with the male attractant, cuelure. During the majority of releases, flies were massmarked using a self-marking technique and fluorescent pigment powder to enable identification of recaptured flies. A preliminary study found that fluorescent pigment marks had no effect on adult survival and marks did not fade significantly in the laboratory over a period of five weeks after eclosion. As cuelure repels inseminated sexually mature female B. tryoni, unbaited, coloured flat sticky traps, and black and yellow sticky sphere traps baited with a food lure (protein autolysate solution) were used to supplement traps baited with cuelure. The effectiveness of these two sticky trap types was assessed, and recaptures used to compare patterns of dispersal from a release point by male and female B. tryoni. Fluorescent yellow (chartreuse), green, and clear unbaited flat sticky traps were relatively ineffective for monitoring dispersal of sterile male and female B. tryoni, recapturing only 0.1% of released sterile flies. Monitoring dispersal with sticky ball traps baited with protein autolysate solution was more successful, with yellow spheres and black spheres recapturing 1.7% and 1.5%, respectively. Trap colour had no effect on recaptures on flat sticky traps or sticky spheres. Equal recapture rates on yellow and black sticky sphere traps suggests that the odour of yeast autolysate solution was more important than colour for attraction of post-teneral flies to traps. Using the results of recaptures on odoriferous black and yellow sticky sphere traps within one week of release, regression equations of male and female recaptures per trap were found to be similar (Figure 4-3). This is the first study to clearly indicate that post-teneral dispersal patterns of male and female B. tryoni released from a point do not differ, enabling the use of existing models to predict density of both sexes of B. tryoni following post-teneral dispersal. (ii) Males disperse further in Spring than in Autumn, but this is not temperature-related. Analysis of replicated recaptures in traps baited with cuelure revealed that dispersal of male B. tryoni in an orchard near Richmond, New South Wales, was higher in Spring than in Autumn (Figure 5-6). As the maximum daily temperature was significantly higher in Spring than in Autumn this result was unexpected, since earlier studies have found that B. tryoni disperse at the onset of cool weather in search of sheltered over-wintering sites. Dispersal of post-teneral B. tryoni may have been affected by habitat suitability; it was found that seasonal trends in dispersal could have been influenced by local habitat variables. Low mean dispersal distances in Autumn may be explained by the presence of fruiting hosts in the orchard, or the availability of resources required by over-wintering flies. There was no significant correlation between temperature and mean dispersal distance, suggesting that higher rates of dispersal cannot be explained by temperature-related increases in activity. Recapture rate per trap was significantly negatively correlated with increasing daily maximum and average temperature. This may have consequences for detection of B. tryoni outbreaks in quarantine areas due to reduced cuelure trap efficiency. (iii) Maturity and source variation affect dispersal and response to cuelure. This research indicated that most male and female B. tryoni do not disperse far from a release point, suggesting that an invading propagule would not spread far in the first generation. However, there is considerable variation in flight capability among individuals. Comparison of wild, laboratory-reared white marks, and gamma-irradiated sterile male B. tryoni indicated that mean dispersal distance and redistribution patterns were not significantly affected by fly origin. Despite no difference in dispersal distance from the release point, recaptures of wild and sterile males per Lynfield trap baited with cuelure were highest within one week after release, whereas recaptures of white marks males per trap increased in the second week. This result may offer evidence to support the hypothesis that sterile male B. tryoni respond to cuelure at an earlier age. Rearing conditions used to produce large quantities of males for sterilisation by gamma-irradiation may select for earlier sexual maturity. Mating Behaviour (i) Density and sex ratio do not affect mating, except at low densities. Demographic stochasticity in the form of sex ratio fluctuations at low population density can lead to an observed Allee effect. The effect of local group density and sex ratio on mating behaviour and male mating success of a laboratory-adapted strain of B. tryoni was examined in laboratory cages. In the laboratory-adapted strain of B. tryoni used in this study, a group of one female and one male was sufficient for a good chance of mating success. The proportion of females mated and male mating success was not significantly affected by density or sex ratio, although variability in male mating success was higher at low density. This could indicate that mating success of B. tryoni can be reduced when local group density is low owing to decreased frequency in encounters between males and females. (ii) Mass-reared males exhibit aberrant mating behaviour, but this does not reduce mating success. Strong artificial selection in mass-rearing facilities may lead to decreased competitiveness of sterile males released in SIT programs as a result of alteration or loss of ecological and behavioural traits required in the field. The effects of domestication and irradiation on the mating behaviour of males of B. tryoni were investigated by caging wild, mass-reared and sterile (mass-reared and gammairradiated) males with wild females. Mating behaviour of mass-reared males was different from that of wild males, but behaviour of wild and sterile males was similar. Mass-reared males were found to engage in mounting of other males much more frequently than wild and sterile males, and began calling significantly earlier before darkness. Male calling did not appear to be associated with female choice of mating partners, although this does not exclude the possibility that calling is a cue used by females to discriminate between mating partners. Conditions used to domesticate and rear large quantities of B. tryoni for SIT may select for an alternative male mating strategy, with mass-reared males calling earlier and exercising less discrimination between potential mating partners. Despite differences in behaviour of wild, mass-reared and sterile males, frequency of successful copulations and mating success were similar. (iii) Pheromone-calling by males was increased in larger aggregations but this did not result in significantly more female visits. Finally, large laboratory cages with artificial leks were used to investigate the importance in B. tryoni of male group size for female visitation at lek sites and initiation of male pheromone-calling. Calling propensity of male B. tryoni was increased by the presence of conspecific males. Females visited the largest lek more frequently than single males, but there was no correlation between lek size and female visitation. Female B. tryoni had a limited capacity to perceive a difference between the number of calling males; female visitation at leks was only weakly associated with male calling, suggesting that lek size and the number of pheromone-calling males may not be the only factor important in locating mates in B. tryoni. The weak, but positive correlation between male calling and female visitation may indicate that passive attraction maintains lek-mating in B. tryoni. Further studies are essential on mating behaviour of B. tryoni, including identification of male mating aggregations in the field, measurement of habitat variables associated with male aggregations, the influence of density on wild B. tryoni mating success, and the role of pheromone-calling, in order to optimise use of SIT for control of this pest.

The Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), a major pest of horticulture in eastern Australia, is a relatively poor coloniser of new habitat. This thesis examines behavioural properties that might limit the ability of B. tryoni to establish new populations. As the potential for B. tryoni to establish an outbreak population may be most directly limited by mechanisms associated with dispersal and mating behaviour, these two factors were the focus of this research project. The relevance of dispersal and mating behaviour for control of outbreak populations was assessed. Dispersal (i) Dispersal patterns of males and females are not different. Dispersal of post-teneral male B. tryoni from a point within an orchard near Richmond, New South Wales, was monitored following temporally replicated releases. Application of sterile insect technique (SIT) requires knowledge of dispersal from a release point so that effective release rates can be determined. In addition, dispersal following introduction to new habitat can lead to low or negative population growth and an Allee effect. In Spring and Autumn, 2001 � 2003, three different strains of B. tryoni were released: (1) wild flies reared from infested fruit collected in the Sydney Basin; (2) a laboratory-reared strain with a colour mutation (white marks); and (3) sterile flies obtained by gamma-irradiation of a mass-reared strain. Dispersal was monitored using a grid of traps baited with the male attractant, cuelure. During the majority of releases, flies were massmarked using a self-marking technique and fluorescent pigment powder to enable identification of recaptured flies. A preliminary study found that fluorescent pigment marks had no effect on adult survival and marks did not fade significantly in the laboratory over a period of five weeks after eclosion. As cuelure repels inseminated sexually mature female B. tryoni, unbaited, coloured flat sticky traps, and black and yellow sticky sphere traps baited with a food lure (protein autolysate solution) were used to supplement traps baited with cuelure. The effectiveness of these two sticky trap types was assessed, and recaptures used to compare patterns of dispersal from a release point by male and female B. tryoni. Fluorescent yellow (chartreuse), green, and clear unbaited flat sticky traps were relatively ineffective for monitoring dispersal of sterile male and female B. tryoni, recapturing only 0.1% of released sterile flies. Monitoring dispersal with sticky ball traps baited with protein autolysate solution was more successful, with yellow spheres and black spheres recapturing 1.7% and 1.5%, respectively. Trap colour had no effect on recaptures on flat sticky traps or sticky spheres. Equal recapture rates on yellow and black sticky sphere traps suggests that the odour of yeast autolysate solution was more important than colour for attraction of post-teneral flies to traps. Using the results of recaptures on odoriferous black and yellow sticky sphere traps within one week of release, regression equations of male and female recaptures per trap were found to be similar (Figure 4-3). This is the first study to clearly indicate that post-teneral dispersal patterns of male and female B. tryoni released from a point do not differ, enabling the use of existing models to predict density of both sexes of B. tryoni following post-teneral dispersal. (ii) Males disperse further in Spring than in Autumn, but this is not temperature-related. Analysis of replicated recaptures in traps baited with cuelure revealed that dispersal of male B. tryoni in an orchard near Richmond, New South Wales, was higher in Spring than in Autumn (Figure 5-6). As the maximum daily temperature was significantly higher in Spring than in Autumn this result was unexpected, since earlier studies have found that B. tryoni disperse at the onset of cool weather in search of sheltered over-wintering sites. Dispersal of post-teneral B. tryoni may have been affected by habitat suitability; it was found that seasonal trends in dispersal could have been influenced by local habitat variables. Low mean dispersal distances in Autumn may be explained by the presence of fruiting hosts in the orchard, or the availability of resources required by over-wintering flies. There was no significant correlation between temperature and mean dispersal distance, suggesting that higher rates of dispersal cannot be explained by temperature-related increases in activity. Recapture rate per trap was significantly negatively correlated with increasing daily maximum and average temperature. This may have consequences for detection of B. tryoni outbreaks in quarantine areas due to reduced cuelure trap efficiency. (iii) Maturity and source variation affect dispersal and response to cuelure. This research indicated that most male and female B. tryoni do not disperse far from a release point, suggesting that an invading propagule would not spread far in the first generation. However, there is considerable variation in flight capability among individuals. Comparison of wild, laboratory-reared white marks, and gamma-irradiated sterile male B. tryoni indicated that mean dispersal distance and redistribution patterns were not significantly affected by fly origin. Despite no difference in dispersal distance from the release point, recaptures of wild and sterile males per Lynfield trap baited with cuelure were highest within one week after release, whereas recaptures of white marks males per trap increased in the second week. This result may offer evidence to support the hypothesis that sterile male B. tryoni respond to cuelure at an earlier age. Rearing conditions used to produce large quantities of males for sterilisation by gamma-irradiation may select for earlier sexual maturity. Mating Behaviour (i) Density and sex ratio do not affect mating, except at low densities. Demographic stochasticity in the form of sex ratio fluctuations at low population density can lead to an observed Allee effect. The effect of local group density and sex ratio on mating behaviour and male mating success of a laboratory-adapted strain of B. tryoni was examined in laboratory cages. In the laboratory-adapted strain of B. tryoni used in this study, a group of one female and one male was sufficient for a good chance of mating success. The proportion of females mated and male mating success was not significantly affected by density or sex ratio, although variability in male mating success was higher at low density. This could indicate that mating success of B. tryoni can be reduced when local group density is low owing to decreased frequency in encounters between males and females. (ii) Mass-reared males exhibit aberrant mating behaviour, but this does not reduce mating success. Strong artificial selection in mass-rearing facilities may lead to decreased competitiveness of sterile males released in SIT programs as a result of alteration or loss of ecological and behavioural traits required in the field. The effects of domestication and irradiation on the mating behaviour of males of B. tryoni were investigated by caging wild, mass-reared and sterile (mass-reared and gammairradiated) males with wild females. Mating behaviour of mass-reared males was different from that of wild males, but behaviour of wild and sterile males was similar. Mass-reared males were found to engage in mounting of other males much more frequently than wild and sterile males, and began calling significantly earlier before darkness. Male calling did not appear to be associated with female choice of mating partners, although this does not exclude the possibility that calling is a cue used by females to discriminate between mating partners. Conditions used to domesticate and rear large quantities of B. tryoni for SIT may select for an alternative male mating strategy, with mass-reared males calling earlier and exercising less discrimination between potential mating partners. Despite differences in behaviour of wild, mass-reared and sterile males, frequency of successful copulations and mating success were similar. (iii) Pheromone-calling by males was increased in larger aggregations but this did not result in significantly more female visits. Finally, large laboratory cages with artificial leks were used to investigate the importance in B. tryoni of male group size for female visitation at lek sites and initiation of male pheromone-calling. Calling propensity of male B. tryoni was increased by the presence of conspecific males. Females visited the largest lek more frequently than single males, but there was no correlation between lek size and female visitation. Female B. tryoni had a limited capacity to perceive a difference between the number of calling males; female visitation at leks was only weakly associated with male calling, suggesting that lek size and the number of pheromone-calling males may not be the only factor important in locating mates in B. tryoni. The weak, but positive correlation between male calling and female visitation may indicate that passive attraction maintains lek-mating in B. tryoni. Further studies are essential on mating behaviour of B. tryoni, including identification of male mating aggregations in the field, measurement of habitat variables associated with male aggregations, the influence of density on wild B. tryoni mating success, and the role of pheromone-calling, in order to optimise use of SIT for control of this pest.

No description available.

Thesis (MSc)--Stellenbosch University, 2011. / ENGLISH ABSTRACT: The Mediterranean fruit fly (medfly), Ceratitis capitata, is a highly invasive species throughout the world and considered as one of the most successful agricultural and economical pests. The increase of global trade in fruit and human travel combined with the biology of the medfly has allowed the species to spread from its proposed Afrotropical origin, to a number of locations throughout the world. In the Western Cape various control strategies have been implemented to control medfly populations, including insecticides and more environmentally-friendly techniques such as the Sterile Insect Technique (SIT). In order to be effective, however, an SIT program requires some knowledge of the population structure and the movement of individuals between pest-occupied sites. The identification of sites from which re-invasion is most likely to occur and knowledge regarding the primary routes through which pests are likely to re-establish are critical to ensure successful SIT programmes. To provide this important information to SIT and area-wide pest control programs in South Africa, sampling at two different spatial scales (regional- and fine-scale) in South Africa was undertaken. Regional scale sampling was done at 13 locations in the Western Cape and fine scale sampling was done at 13 locations within the Ceres-valley. All individuals were genotyped at 11 polymorphic microsatellite markers and selected individuals from the regional scale were sequenced for the mitochondrial gene COI. Our results show that populations at regional- and fine-scale in the Western Cape are characterized by high levels of genetic diversity (HEregional = 0.805; HEfine = 0.803). Little or weak population differentiation was detected at the regional- and fine-scales, suggesting overall high levels of gene flow among sampling locations. These findings were supported by coalescent based methods indicating sufficient levels of gene flow to prevent population differentiation between neighbouring (200m) and distant (350km) populations. However, natural dispersal in C. capitata has been shown to rarely exceed 10 km. As such, high levels of gene flow between distant populations are more likely the result of humanmediated dispersal, linked to the movement of fresh produce within South Africa. This high level of gene flow has important implications for pest management practices, as my results suggests that area-wide pest management should be undertaken at a regional scale, rather than on a farm or valley scale. My results are placed within a management framework, and I argue for more stringent control when fruit are transported within South Africa. Of particular interest for future studies is the investigation of gene flow at broader spatial scales (i.e. the whole of South Africa) and a comparison of the genetic diversity, population differentiation and gene flow patterns of C. capitata with that of Ceratitis rosa will be important to establish a successful pest management strategy in South Africa. / AFRIKAANSE OPSOMMING: Die Mediterreense vrugtevlieg (medvlieg), Ceratitis capitata, is ‘n indringerspesie wêreldwyd en word beskou as een van die mees suksesvolle ekonomiese en landbou peste. Die medvlieg het ‘n Afrotropiese oorsprong, maar die toename in wêreldwye handel en reis, gekombineer met die biologie van die medvlieg het gelei tot die verspreiding van die spesie na ‘n groot aantal bestemmings regoor die wêreld. Die Wes-Kaap provinsie van Suid-Afrika implementeer tans verskeie strategieë om medvlieg bevolkings te beheer. Hierdie strategieë sluit in die gebruik van plaagdoders sowel as meer omgewingsvriendelike tegnieke soos die Steriele Insektegniek (SIT). Om ‘n effektiewe SIT program te implementeer vereis basiese kennis ten opsigte van die genetiese struktuur van die bevolking sowel as van die beweging van individue tussen verskillende pesvoorkomsgebiede. Die identifisering van areas van waar herkolonisering mees waarskynlik kan plaasvind en kennis in verband met die primêre roetes waardeur pes spesies hervestig, is van kritiese belang om ‘n suksesvolle SIT program te verseker. Medvlieg individue is op twee verskillende ruimtelike skale (streeks- en plaaslike-skaal) versamel om die nodige inligting aan SIT en area-wye pes beheer programme in Suid-Afrika te verskaf. Streeks-skaal individue is by 13 lokaliteite regoor die Wes-Kaap versamel en plaaslike-skaal individue by 13 lokaliteite in die Ceres-vallei. Alle versamelde individue is vir 11 polimorfiese mikrosatelliet merkers gegenotipeer en DNS volgordebepaling van geselekteerde individue vanuit die streek-skaal is gedoen vir die mitochondriale geen COI. My resultate toon dat bevolkings op beide skale gekarakteriseer word deur hoë vlakke van genetiese diversiteit (HEstreeks = 0.805; HEplaaslik = 0.803) en geen of swak bevolkings differensiasie. Hierdie resultate dui daarop dat daar hoë vlakke van geenvloei tussen bevolkings is. Hierdie bevindinge word verder ondersteun deur metodes gebaseer op die statistiese eienskappe van die genealogiese verhouding tussen allele onder sekere mutasie en demografiese modelle, wat voldoende vlakke van geenvloei aandui tussen nabye (200m) sowel as verafgeleë (350km) bevolkings om bevolkings differensiasie te verhoed. Natuurlike beweging in C. capitata is egter selde meer as 10 km, sodanig kan die hoë vlakke van geenvloei toegeskryf word aan die verspreiding van individue met menslike hulp, spesifiek in die vervoer van varsprodukte in Suid-Afrika. Hierdie hoë vlak van geenvloei het verreikende implikasies vir pes beheer praktyke, omdat my resultate voorstel dat area-wye pes beheer onderneem moet word op ‘n streeks-skaal eerder as op ‘n plaas-tot-plaas of vallei wye area. Ek plaas my resultate in ‘n bestuursraamwerk, waarin ek streng beheer van vrugtevervoer in Suid-Afrika beklemtoon. Verdere navorsing moet fokus op die ondersoek van geenvloei op ‘n landswye skaal (hele Suid-Afrika) in C. capitata, sowel as die vergelyking van die genetiese diversiteit, bevolkings differensiasie en geenvloei patrone van C. capitata met die van Ceratitis rosa om ‘n suksesvolle pes beheer strategie vir Suid-Afrika te formuleer.

Thesis (MSc (Conservation Ecology and Entomology))--University of Stellenbosch, 2011. / Please refer to full text to view abstract.

Aedes aegypti é o principal vetor envolvido na cadeia de transmissão de arboviroses como dengue, chikungunya, zika e febre amarela. Diante dos desafios para diminuir o nível de infestação do vetor e do recente aumento do número de casos e agravos dessas doenças, torna-se indispensável aderir a novas estratégias para complementar o controle vetorial. O controle autocida é uma estratégia que induz um controle de natalidade sobre a população do inseto e que tem a criação em massa como etapa fundamental. Assim, o objetivo desse estudo foi aprimorar o protocolo de criação em massa, manipulação e transporte de adultos de Aedes aegypti para aplicação em biofábricas. Foram avaliados dados de 59 lotes de produção em massa da linhagem bissexual MBR-001 (Juazeiro, Bahia) e alguns dos seus parâmetros produtivos. A análise da MBR-001, mostrou baixo rendimento na produtividade de machos (<span style=\"text-decoration:overline; font-style: italic;\">X =18&#37;), com reduzida produção de ovos (24,5 ovos/fêmea), em comparação com outra linhagem da mesma espécie. Com relação aos seus parâmetros produtivos, a manutenção de adultos em diferentes proporções sexuais nas gaiolas não afetou a produção de ovos viáveis (<span style=\"text-decoration:overline; font-style: italic;\">X = 60&#37;). A MBR-001 apresentou também um baixo rendimento, revelando que ajustes no protocolo de criação são necessários. Tais adaptações são cruciais para reduzir a demanda de espaço físico e o custo da produção. Atrelado a isso, foram desenvolvidas gaiolas de criação em massa, e determinadas densidades de adultos (DRS: cm2/mosquito) para otimizar a produção de ovos viáveis. Os testes realizados com a linhagem Rockefeller revelaram melhor desempenho produtivo em DRS de 1.7 e 1.4 (cm2/mosquito). No que diz respeito à criação larval, foi verificada que não há ocorrência de protrandria em fase larvária logo após eclosão, e ainda, foram desenvolvidas seis dietas com insumos nacionais de baixo custo, uma delas apresentou maior rendimento acumulado de machos (<span style=\"text-decoration:overline; font-style: italic;\">X = 45&#37;) em relação à dieta controle (<span style=\"text-decoration:overline; font-style: italic;\">X = 37&#37;). Além disso, uma ampla faixa de densidades larvais foi testada para determinar as condições ótimas de criação, sendo evidenciado que densidades entre 1.0 e 2.5 larvas/mL são adequadas para a produção em massa. Esses resultados contribuem para o aprimoramento das condições ideais de produção de machos estéreis de qualidade, com uma melhor relação custo-benefício. Por fim, foram determinadas as condições iniciais para o transporte de adultos entre biofábrica e áreas-alvo de liberação de insetos estéreis. A imobilização de Ae. aegypti à 2-5&#176;C (45 minutos) com subsequente transporte em baixas temperaturas (4-7&#176;C) por até seis horas, não afetou a sobrevivência e habilidade de voo do mosquito. De forma geral, o presente trabalho forneceu informações, produtos e condições que contribuíram para o desenvolvimento de um protocolo padronizado e para o aprimoramento da criação em massa de Ae. aegypti, visando a produção de insetos de qualidade e a redução dos custos operacionais em estratégias de controle autocida. / Aedes aegypti is the main vector involved in the transmission chain of arboviruses such as dengue, chikungunya, zika and yellow fever. Faced with the challenges to reduce the level of vector infestation and the recent increase in the number of cases and diseases, it is essential to adhere to new strategies to complement vector control. Self-control is a strategy that induces a birth control over the insect population and that has mass creation as a fundamental step. Thus, the objective of this study was to improve the mass rearing protocol, handling and transport of adults of Ae. aegypti for application in facilities. Data from 59 mass production batches of the bisexual strain MBR-001 and some of its productive parameters were evaluated. he analysis of MBR-001 showed low yield in males (<span style=\"text-decoration:overline; font-style: italic;\">X = 18&#37;), with reduced egg production (24.5 eggs/female), compared to another line of the same species. Regarding their productive parameters, the maintenance of adults in different sex ratios in the cages did not affect viable egg production (<span style=\"text-decoration:overline; font-style: italic;\">X = 60&#37;). MBR-001 also showed low performance, revealing that settings in the breeding protocol are required. Such adaptations are crucial to reduce the demand for physical space and the cost of production. Linked to this, mass - rearing cages and certain adult densities (DRS: cm2 / mosquito) were developed to optimize viable egg production. Tests performed with the Rockefeller line showed better productive performance in DRS of 1.7 and 1.4 (cm2 / mosquito). Regarding larval breeding, it was verified that there is no occurrence of protrandria in the larval stage shortly after hatching, and six diets with low cost national inputs were developed, one of them presented higher accumulated males yield (<span style=\"text-decoration:overline; font-style: italic;\">X = 45&#37;) in relation to the control diet (<span style=\"text-decoration:overline; font-style: italic;\">X = 37&#37;). In addition, a wide range of larval densities was tested to determine the optimum breeding conditions, showing that densities between 1.0 and 2.5 larvae/mL are adequate for mass production. These results contribute to the improvement of the ideal conditions of production of quality sterile males, with a better cost-benefit ratio. Finally, initial conditions were determined for the transport of adults between biofacility and target areas of sterile insect release. The immobilization of Ae. aegypti at 2-5&#176;C (45 minutes) with subsequent transport at low temperatures (4-7&#176; C) for up to six hours did not affect the survival and flight ability of the mosquito. In general, the present work provided information, products and conditions that contributed to the development of a standardized protocol and to the enhancement of Ae. aegypti, aiming the production of quality insects and the reduction of operational costs in self-control strategies.

A dengue é atualmente uma das doenças tropicais com maior rapidez de propagação e de grande alcance mundial (OMS, 2013). No Brasil, o governo federal, bem como os estados e os municípios gastam montantes significativos em sua prevenção e controle. O presente trabalho analisa aspectos econômicos e sociais da implementação de uma nova técnica de combate ao mosquito Aedes aegypti, a TIE, vis-à-vis os métodos convencionais. Essa técnica é avaliada com base no estudo de caso da Biofábrica Moscamed, responsável pelo desenvolvimento da mesma no Brasil. A TIE consiste na esterilização dos insetos e posterior soltura no ambiente, com a finalidade de diminuir a população de insetos selvagens, estes vetores do vírus da dengue. Dois cenários são analisados: cenário atual de medidas de prevenção e controle adotadas pelas prefeituras e o cenário de adoção da TIE, em substituição aos métodos atuais. São tomados como estudos de caso a aplicação da TIE em Jacobina e Juazeiro, na Bahia, cujos dados foram extrapolados para uma simulação também para o município de Piracicaba, em São Paulo. Calcula-se o VPL para o cenário da TIE, considerando um horizonte temporal de quatro anos e a discussão é conduzida no escopo simplificado de uma Análise de Impacto Regulatório (AIR). Foram analisados ainda os gastos das prefeituras para o controle da dengue e de seu vetor, com base no ano de 2013. Realizou-se uma análise dos aspectos sociais e econômicos concernentes à dengue, que apontou como impactos relevantes as mortes, o afastamento dos doentes de suas atividades e o alto número de notificações mesmo após aplicação dos métodos de controle vigentes. Por se tratar de uma questão de saúde pública a análise tem foco majoritariamente social, uma vez que para se implementar projetos com benefícios sociais a intervenção e os investimentos públicos são de fundamental importância, suplantando a visão econômica de viabilidade do projeto. A mensuração dos benefícios da TIE foi limitada pelo curto período em que os experimentos de campo estão sendo aplicados, não havendo ainda resultados publicados sobre sua eficácia. Do mesmo modo, não se tem dados que permitam calcular os benefícios dos atuais métodos de controle do vetor e da doença. Assim, não foi possível conduzir uma comparação de eficácia e uma estimativa da possibilidade de associação entre os métodos tradicionais e a TIE. Em suma, mediante análise fundamentada em um roteiro de referência proposto para análise de impacto regulatório conclui-se que projetos com avaliação positiva do ponto de vista social, mesmo com indicadores econômicos negativos, são geralmente financiados pela esfera pública e, no caso aqui analisado da dengue, devem buscar melhorar os resultados dos atuais programas que não têm sido capazes de evitar os prejuízos sociais e econômicos dessa doença no Brasil. / Dengue has been one of the most important tropical diseases that has showed a fast and worldwide propagation according to the WHO (2013). In Brazil, the federal government as well as states and municipalities are spending significant amounts of resources of all kinds on its prevention and control. Yet it seems worthless. This paper examines the economic feasibility of implementing a new technique to fight the mosquito Aedes aegypti, the TIE (Técnica do Inseto Estéril-Sterile Insect Technique), vis-à-vis the conventional methods employed. The TIE is evaluated through the case study of the Moscamed, a biofirm which is responsible for its development and propagation in Brazil. The TIE consists on the sterilization of insects and their release into the environment, aiming to reducing the population of susceptible wild insects, which can transmit the virus of dengue among population. Two scenarios are analyzed: 1) the current scenario of prevention and control adopted by municipalities, and 2) the scenario that describes the adoption of the TIE, replacing the current methods. The cities of Jacobina and Juazeiro in Bahia are used as case\'s study, and whose data were extrapolated for a simulation also for the city of Piracicaba, located in the state of São Paulo. The Net Present Value (NPV) is calculated for the scenario of TIE, considering a four years\' timeline, and additionally a discussion is conducted in the scope of a simplified Regulatory Impact Analysis (RIA). It was also analyzed the spending of municipalities for the control of dengue and its vector, based on the year of 2013. It was conducted an analysis of the social and economic aspects related to dengue, which identified as relevant impacts: deaths, absence of patients of their labor activities and the high number of notifications even after implementation of the existing control methods. Because it is a public health problem, this analysis had mostly a social focus. It is well known that to implement projects with social benefits, the government intervention and public investment are of fundamental importance, despite the economic view of the project feasibility. The measurement of the benefits from the TIE was limited by the short period in where the field experiments were taken place. In fact, there are no enough published results about its effectiveness. Similarly, there is no data allowing calculation of the benefits of the current vector control and disease. Thus, it was not possible to conduct a comparison of efficiency and an estimation of a possible association between traditional methods and TIE. In summary, this analysis, based on a proposed reference roadmap for Regulatory Impact Analysis, concludes that projects with positive assessment from a social point of view, even if they have negative economic indicators, are usually funded by the public sphere. In the specific dengue case, the government should seek to improve the results of current programs that have not been able to avoid the social and economic losses of this disease in Brazil.

Tanto as técnicas de controle biológico quanto a Técnica do inseto estéril (TIE), são utilizadas em vários países para o controle, supressão e até mesmo erradicação de moscas-das-frutas e outras pragas da agricultura, pecuária e saúde publica. O uso de tais técnicas minimiza o emprego contínuo de inseticidas, protege o ambiente e se adequa aos padrões de segurança alimentar. No entanto, é necessário para a implementação de tais programas, tecnologia para produzir milhões de parasitóides e a própria praga em laboratório com qualidade biológica similar aos insetos encontrados na natureza e com custo competitivo com o controle químico. Os objetivos deste trabalho foi estabelecer protocolos para criação artificial de A. sp. 1 aff. fraterculus em dieta larval líquida que permita atingir níveis de criação massal, para uma possível diminuição no custo da criação e determinar a dose de radiação esterilizante para adultos de A. sp. 1 aff. fraterculus atendendo os parâmetros de qualidade exigidos pela Técnica do Inseto Estéril com insetos provenientes da criação do Laboratório de Radioentomologia do CENA/USP. Foram testadas sete dietas em relação à dieta convencional utilizada no laboratório de Radioentomologia do CENA/USP, que foi usada como controle. Todas as sete dietas tiveram em comum a exclusão do ágar na sua formulação. Somente duas das dietas testadas foram adequadas para o desenvolvimento larval da mosca, estas quando comparadas com a dieta padrão, apresentaram resultados inferiores com relação ao volume de larvas recuperadas, emergência e o peso das pupas, porém, não apresentaram diferenças significativas quanto aos períodos de desenvolvimento, recuperação pupal, razão sexual e longevidade sob estresse. Concluí-se que é possível a substituição da dieta com ágar por dietas líquidas para criação artificial de A. sp.1 aff. fraterculus, de custo reduzido e com maior praticidade de manejo,porém devidos aos seus parâmetros de qualidade inferiores aos da dieta padrão, são necessários maiores testes principalmente quanto à adaptabilidade do inseto ao novo meio. Para determinar a dose esterilizante o presente trabalho analisou a fertilidade, fecundidade, emergência, habilidade de vôo e longevidade sob estresse de A. fraterculus irradiadas com doses de 40, 50, 60, 70 e 80 Gy. A fonte de radiação gama foi um irradiador de Co-60 (Gammabeam-650) localizado no mesmo laboratório. Os testes foram realizados sob as seguintes condições ambientais: 26 ± 1 ° C, 70 ± 5% RH, e fotofase de 12:12. A dose de r adiação gama esterilizante para machos de A. sp. 1 aff. fraterculus foi determinada sendo 70 Gy. Fêmeas irradiadas, mesmo com a dose mais baixa de 40 Gy, não ovipositaram, possivelmente pela atrofia dos ovários causada pela radiação. A radiação não afetou os demais parâmetros de qualidade do inseto como emergência, longevidade e habilidade de vôo / Both the biological control techniques as the Sterile Insect Technique (SIT), are used in many countries to control, suppress and even eradicate fruit flies and other pests in agriculture and public health. The use of such techniques minimizes the continuous employment of insecticides, protects the environment and conforms to standards for food safety. However, it is necessary to implement such programs, technology to produce millions of parasitoids and the pest in its own laboratory with biological quality similar to the insects found in nature and cost competitive with chemical control. The objectives of this study was to establish protocols for artificial rearing of A. sp. 1 aff. fraterculus in liquid larval diet that will achieve levels of mass production for a possible reduction in the cost of establishing and determining the dose of radiation sterilization of adult A. sp. 1 aff. fraterculus meeting the quality parameters required by the Sterile Insect Technique with insects from the creation of Radioentomologia Laboratory of CENA / USP. Seven experimental diets compared to the conventional diet used in laboratory Radioentomologia of CENA / USP, which was used as control. All seven diets have in common the exclusion of agar in its formulation. Only two of the diets tested were suitable for larval development of the fly, they compared with the standard diet, showed inferior results with respect to the volume of recovered larvae, pupae and weight of emergency, however, no significant differences regarding the periods of development , pupal recovery, sex ratio and longevity under stress. It is possible to replace the diet with agar for liquid diets for artificial creation of A. sp.1 aff. fraterculus, reduced cost and greater convenience of handling, but due to their quality standards lower than the standard diet, more tests are needed especially regarding the adaptability of the insect to the new environment. To determine the sterilizing dose this study examined the fertility, fecundity, emergence, flight ability and longevity under stress in A. fraterculus irradiated with 40, 50, 60, 70 and 80 Gy. The radiation source was a gamma irradiator with a Co-60 (Gammabeam-650) located in the same laboratory. The tests were performed under the following environmental conditions: 26 ± 1 ° C, 70 ± 5% RH and photoperiod of 12:12. The dose of gamma radiation sterilization for male A. sp. 1 aff. fraterculus was determined with 70 Gy. Irradiates females, even with the lower dose of 40 Gy, layed no eggs, possibly due to atrophy of the ovaries caused by radiation. The radiation did not affect the other quality parameters such as insect emergence, longevity and ability to fly