RESUMO

Studies investigating oviposition preference of Ceratitis capitata (Wiedemann, 1824) in table grapes (Vitis vinifera L.) cultivated in the São Francisco River Valley can provide important information to guide control measures, therefore minimizing damage caused by this species in the region. The aim of this study was to evaluate the oviposition preference of C. capitata females for berries cv. 'Italia' collected at five different times (60, 70, 80, 90, and 100 d after production pruning [DAPP]), representing five grapevine growth stages. We also assessed the performance of immature stages of C. capitata regarding the physiological development of the berry. The total soluble solid content was significantly higher in berries at more advanced physiological developmental stages (90 and 100 DAPP). Conversely, these berries showed the lowest values of firmness and titratable acidity. With the onset of physiological development, the average number of punctures per berry increased and reached 5.2 per berry in 100 DAPP berries. The infestation level and pupal weight showed a positive correlation with the growth stage. The highest recovery of pupae was observed in 100 DAPP berries (1.54). Pupal viability values ranged from 50.9 to 64.7% independent of berry maturity stage. The physiological developmental stage of the berry did not affect offspring sex ratio. Results obtained suggest that regardless of the female preference for laying eggs on berries in a more advanced physiological developmental stage, females can initiate the attack to bunches of this cultivar at 60 DAPP, even if the berries have unfavorable physicochemical parameters for oviposition and development of larvae.

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Abstract Fruit flies (Diptera: Tephritidae) are considered the main fruit pests worldwide. In Brazil, two species are predominant: the South American fruit fly, Anastrepha fraterculus and the Mediterranean fruit fly, Ceratitis capitata. In this study, we evaluated the effect of artificial diets with variable pH in their larval development and adult performance. The experiments were carried out in the laboratory at 25 ± 2 °C, 70 ± 10% RH and 12:12h (L:D) photoperiod. Semisolid diets with pH values of 6.0, 5.0, 4.0, 3.0, 2.0, 1.5, and 1.0, adjusted by adding hydrochloric acid were tested. Results indicated that the diet with pH 6.0 did not support larval development of both species of fruit fly. Diets with greater acidic pH values did not allow egg, larvae or pupae development and adult reproduction of A. fraterculus. For C. capitata , the pH of artificial diet exerts greater influence compared to A. fraterculus on the duration and viability of the larval stage, number of pupae, sex ratio and longevity of males.

Resumo As moscas-das-frutas (Diptera: Tephritidae) são consideradas as principais pragas da fruticultura mundial. No Brasil, duas espécies são predomindantes: a mosca-das-frutas Sul-americana, Anastrepha fraterculus e a mosca-do-Mediterrâneo, Ceratitis capitata. Neste estudo avaliamos o efeito de dietas artificiais com pH variável no seu desenvolvimento larval e performance de adultos. Os experimentos foram realizados em laboratório a 25 ± 2 °C, 70 ± 10% UR e fotoperíodo de 12:12 horas. Foram testadas dietas semi-sólidas com valores de pH de 6,0; 5,0; 4,0; 3,0; 2,0; 1,5 e 1,0, ajustados pela adição de ácido clorídrico. Os resultados indicaram que a dieta com pH 6,0 não suportou o desenvolvimento larval de ambas as espécies de mosca-das-frutas. As dietas com pH ácido não permitiram o desenvolvimento de ovos, larvas ou pupas e a reprodução de adultos de A. fraterculus. Para C. capitata o pH da dieta artificial exerceu maior influência do que para A. fraterculus nos parâmetros de duração e viabilidade do estágio larval, número de pupas, razão sexual e longevidade de machos.

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Fruit flies (Diptera: Tephritidae) are considered the main fruit pests worldwide. In Brazil, two species are predominant: the South American fruit fly, Anastrepha fraterculus and the Mediterranean fruit fly, Ceratitis capitata. In this study, we evaluated the effect of artificial diets with variable pH in their larval development and adult performance. The experiments were carried out in the laboratory at 25 ± 2 °C, 70 ± 10% RH and 12:12h (L:D) photoperiod. Semisolid diets with pH values of 6.0, 5.0, 4.0, 3.0, 2.0, 1.5, and 1.0, adjusted by adding hydrochloric acid were tested. Results indicated that the diet with pH 6.0 did not support larval development of both species of fruit fly. Diets with greater acidic pH values did not allow egg, larvae or pupae development and adult reproduction of A. fraterculus. For C. capitata , the pH of artificial diet exerts greater influence compared to A. fraterculus on the duration and viability of the larval stage, number of pupae, sex ratio and longevity of males.

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Tetrastichus giffardianus Silvestri (Hymenoptera: Eulophidae) is a gregarious koinobiont endoparasitoid of frugivorous dipterans. In recent years, T. giffardianus has been detected parasitizing Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) in different Brazilian semiarid sites. Thus, T. giffardianus has potential to be used in the biological control of C. capitata in semi-arid conditions. However, one of the factors limiting the use of this parasitoid is the lack of detailed information on rearing methods and bioecological aspects. Thus, the main objectives of this study were to demonstrate a rearing technique for T. giffardianus and to determine its population increase potential in laboratory. For this specimens of T. giffardianus were obtained in the field through of the collection of chestnut fruits (Terminalia catappa L. - Combretaceae) infested with C. capitata. The specimens obtained were used to adapt a rearing methodology for T. giffardianus under laboratory conditions (temperature: 25 ± 2ºC; relative humidity: 60 ± 10%; 12 h photophase). Information on demographic parameters for a T. giffardianus population was obtained using the same methodology. Based on the parameters evaluated, we found that the intrinsic rate of population increase of T. giffardianus was 0.34, the average time of one generation was 19.7 days, the population doubling time was 2.03 days and the finite rate of population increase (λ) was 1.41. The demographic parameters obtained demonstrated that, with the rearing technique used, it is possible to maintain populations of T. giffardianus and enable them to multiply in laboratory.

Tetrastichus giffardianus Silvestri (Hymenoptera: Eulophidae) é um endoparasitoide coinobionte gregário de dípteros frugívoros. Nos últimos anos, T. giffardianus tem sido detectado parasitando Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) em diferentes locais do semiárido brasileiro. Assim, T. giffardianus tem potencial para ser utilizado no controle biológico de C. capitata nas condições do semiárido. Entretanto, um dos fatores limitantes para uso deste parasitoide é a falta de informações detalhadas sobre sua metodologia de criação e seus aspectos bioecológicos. Portanto, o principal objetivo deste estudo foi demonstrar uma técnica de criação para T. giffardianus e determinar seu potencial de crescimento populacional, em laboratório. Para isto, espécimens de T. giffardianus foram obtidos em campo, através da coleta de frutos de castanhola (Terminalia catappa - Combretaceae) infestados com C. capitata. Os exemplares obtidos foram utilizados para adaptar uma metodologia de criação para T. giffardianus em condições de laboratório (temperatura de 25 ± 2ºC, umidade relativa de 60 ± 10% e fotofase de 12 horas). As informações sobre os parâmetros demográficos para uma populacional de T. giffardianus foram obtidas usando a mesma metodologia. Com base nos parâmetros avaliados foi constatado que a taxa intrínseca de aumento populacional de T. giffardianus foi de 0,34, o tempo médio de uma geração foi de 19,7 dias, o tempo de duplicação da população foi de 2,03 dias e a taxa finita de aumento populacional (λ) foi de 1,41. Os parâmetros demográficos obtidos demonstraram que, com a técnica de criação utilizada, é possível manter populações de T. giffardianus e multiplicá-las em laboratório.

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Tetrastichus giffardianus Silvestri (Hymenoptera: Eulophidae) is a gregarious koinobiont endoparasitoid of frugivorous dipterans. In recent years, T. giffardianus has been detected parasitizing Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) in different Brazilian semiarid sites. Thus, T. giffardianus has potential to be used in the biological control of C. capitata in semi-arid conditions. However, one of the factors limiting the use of this parasitoid is the lack of detailed information on rearing methods and bioecological aspects. Thus, the main objectives of this study were to demonstrate a rearing technique for T. giffardianus and to determine its population increase potential in laboratory. For this specimens of T. giffardianus were obtained in the field through of the collection of chestnut fruits (Terminalia catappa L. - Combretaceae) infested with C. capitata. The specimens obtained were used to adapt a rearing methodology for T. giffardianus under laboratory conditions (temperature: 25 ± 2ºC; relative humidity: 60 ± 10%; 12 h photophase). Information on demographic parameters for a T. giffardianus population was obtained using the same methodology. Based on the parameters evaluated, we found that the intrinsic rate of population increase of T. giffardianus was 0.34, the average time of one generation was 19.7 days, the population doubling time was 2.03 days and the finite rate of population increase (λ) was 1.41. The demographic parameters obtained demonstrated that, with the rearing technique used, it is possible to maintain populations of T. giffardianus and enable them to multiply in laboratory.(AU)

Tetrastichus giffardianus Silvestri (Hymenoptera: Eulophidae) é um endoparasitoide coinobionte gregário de dípteros frugívoros. Nos últimos anos, T. giffardianus tem sido detectado parasitando Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) em diferentes locais do semiárido brasileiro. Assim, T. giffardianus tem potencial para ser utilizado no controle biológico de C. capitata nas condições do semiárido. Entretanto, um dos fatores limitantes para uso deste parasitoide é a falta de informações detalhadas sobre sua metodologia de criação e seus aspectos bioecológicos. Portanto, o principal objetivo deste estudo foi demonstrar uma técnica de criação para T. giffardianus e determinar seu potencial de crescimento populacional, em laboratório. Para isto, espécimens de T. giffardianus foram obtidos em campo, através da coleta de frutos de castanhola (Terminalia catappa - Combretaceae) infestados com C. capitata. Os exemplares obtidos foram utilizados para adaptar uma metodologia de criação para T. giffardianus em condições de laboratório (temperatura de 25 ± 2ºC, umidade relativa de 60 ± 10% e fotofase de 12 horas). As informações sobre os parâmetros demográficos para uma populacional de T. giffardianus foram obtidas usando a mesma metodologia. Com base nos parâmetros avaliados foi constatado que a taxa intrínseca de aumento populacional de T. giffardianus foi de 0,34, o tempo médio de uma geração foi de 19,7 dias, o tempo de duplicação da população foi de 2,03 dias e a taxa finita de aumento populacional (λ) foi de 1,41. Os parâmetros demográficos obtidos demonstraram que, com a técnica de criação utilizada, é possível manter populações de T. giffardianus e multiplicá-las em laboratório.(AU)

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Fopius arisanus (Sonan, 1932) (Hymenoptera: Braconidae) is an egg-larval parasitoid used in control programs of Bactrocera dorsalis (Hendel) and Ceratitis capitata (Wiedemann). In Brazil, C. capitata and Anastrepha fraterculus (Wiedemann) are considered the main tephritid pests of exotic and indigenous fruits. The objective of this study was to study the biology of F. arisanus in C. capitata and A. fraterculus Eggs of the two fruit fly species were used to determine the parasitism rate, number of offspring, emergence rate, sex ratio, adult weight and longevity of male and female F. arisanus These biological parameters were used to develop a fertility life table. We observed higher parasitism and emergence rates of adults, a shorter duration of the egg-adult period and a sex ratio biased to females when F. arisanus was reared in eggs of C. capitata than in those of A. fraterculus However, adults of F. arisanus from eggs of A. fraterculus were heavier and had greater longevity than those obtained from C. capitata eggs. The fertility life table showed better biological and reproductive performance for F. arisanus reared in eggs of C. capitata, although eggs of A. fraterculus also provided positive values for population increase.

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A 4-yr study was done to analyze seasonal patterns underlying host plant-fruit fly-parasitoid interactions in a secondary forest in the Argentinean Yunga and its importance for the implementation of conservation and augmentative biological control. Larval-pupal hymenopteran parasitoids associated with all host plants and fruit fly species were identified and the seasonal occurrence of fruit, infestation levels, parasitism percentage, and relative parasitoid abundance were determined. Three fruit fly species in two genera were found in association with surveyed plants, two of which (Ceratitis capitata (Wiedemann) and Anastrepha fraterculus (Wiedemann)) are of major economic importance. Infestation levels were strongly influenced by environmental factors and peak fruit availability. Five fruit fly parasitoid species were recovered from fly pupae, four braconid species, and one figitid. Time windows for fruit fly population growth were pinpointed. Based on results, the present analysis proposes an effective fruit fly biological control strategy tailored for the northwestern Argentinean citrus-producing area.

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Population fluctuations of Ceratitis capitata (Wiedemann) were evaluated over a period of 12 mo in four altitudinal strata (400-750, 750-1,100, 1,100-1,450, and 1,450-1,800 meters above sea level, masl) in Eastern Guatemala. Within each altitudinal range, sampling plots were established in coffee plantations and adjacent areas, in which Jackson traps were set and baited with Trimedlure. Coffee berries and other host fruits were collected. Population density was lowest at the 400-750 masl stratum and highest at 1,450-1,800 masl. At every altitudinal range, the fluctuations of the pest were associated mainly with the availability of ripe coffee berries as a primary host. From 750-1,450 masl, the pest was also associated with the availability of sweet orange and mandarins in commercial and backyard orchards. The highest densities of the pest were recorded in the dry season. Citrus were the main alternate host where ripe coffee berries were not available. This knowledge on population dynamics of C. capitata will contribute to develop more effective area-wide pest management strategies including the use of sterile insects, natural enemies, and bait sprays.

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The toxicity of essential oils from the citrus peel has been proposed as the major resistance mechanism offered by citrus to fruit fly infestation. We evaluated the insecticidal activity of the ether extracts from the lemon (Citrus limon [L.] Burm.) and grapefruit (C. paradisi Macfadyen) peel as well as from limonene and citral against Anastrepha fraterculus (Wiedemann) and Ceratitis capitata (Wiedemann) immature stages. We also evaluated the toxicity of the extracts at two ripening stages. Extracts proved toxic to A. fraterculus egg and larvae. The lemon and grapefruit extracts showed the same toxicity in both fruit fly species. For A. fraterculus eggs, citral was more toxic than limonene; for larvae, they showed equal toxicity. Anastrepha fraterculus eggs were more sensitive than C. capitata eggs. In conclusion, we provide evidence of chemical resistance mechanisms that could account for the nonhost condition of lemon for A. fraterculus.

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Objetivou-se avaliar a influência da idade (tamanho) das larvas de Ceratitis capitata utilizadas na criação de Diachasmimorpha longicaudata sobre o parasitismo, a razão sexual e a longevidade das fêmeas da progênie, quando as fêmeas mães foram mantidas sozinhas ou competindo pelo mesmo hospedeiro. Para isso, foram oferecidas larvas de cinco, seis, sete e oito dias de idade às fêmeas do parasitoide em "unidades de parasitismo", no interior de gaiolas adaptadas, contendo uma ou duas fêmeas do parasitoide de sete dias de idade. O número de fêmeas do parasitoide dividindo a mesma "unidade de parasitismo" não afetou nenhum aspecto da qualidade do parasitoide. O tamanho do hospedeiro influenciou significativamente no número de parasitoides emergidos. Larvas hospedeiras menores (cinco dias) provocaram uma redução no parasitismo em relação às larvas de seis, sete e oito dias de idade. Houve maior emergência de parasitoides machos de hospedeiros com cinco e seis dias de idade, enquanto mais fêmeas emergiram de larvas com sete e oito dias de idade. As fêmeas de D. longicaudata provenientes de larvas com seis, sete e oito dias viveram mais do que as fêmeas obtidas de larvas com cinco dias de idade. Larvas hospedeiras maiores, quando utilizadas na criação de parasitoides de moscas-das-frutas, aumentam a porcentagem de parasitismo, o número de fêmeas na progênie e a longevidade das fêmeas da progênie, podendo proporcionar vantagens econômicas quando utilizadas para liberação aumentativa no campo.(AU)

This work was carried out in order to evaluate the influence of age (size) of larvae Ceratitis capitata used in the rearing of the Diachasmimorpha longicaudata on parasitism, sex ratio of the progeny and the longevity of progeny's females, when the mothers were maintained alone or sharing the same host. Larvae at different ages (five, six, seven and eight days old), in "units of parasitism" were offered to female parasitoids, inside adapted cages, containing one or two seven days old female parasitoids. The number of females of the parasitoid sharing the same "unit of parasitism" in the same cage did not affect any aspect of parasitoid quality. However, the size of the host had significant influence on the number of emerged parasitoids. The youngest host larvae (five days old) reduced parasitism more than larvae at six, seven and eight days old. There was higher emergence of male parasitoids of five and six days old hosts, while more females emerged from seven and eight days old larvae. The females of D. longicaudata from larvae with six, seven and eight days old lived more than those obtained from females of five days old larvae. When larger host larvae are used in the rearing of parasitoids of fruit flies, it increases parasitism, the number of female progeny and the longevity of female progeny; it can provide economic advantages when used for augmentative liberation on the field.(AU)

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Objetivou-se avaliar a influência da idade (tamanho) das larvas de Ceratitis capitata utilizadas na criação de Diachasmimorpha longicaudata sobre o parasitismo, a razão sexual e a longevidade das fêmeas da progênie, quando as fêmeas mães foram mantidas sozinhas ou competindo pelo mesmo hospedeiro. Para isso, foram oferecidas larvas de cinco, seis, sete e oito dias de idade às fêmeas do parasitoide em "unidades de parasitismo", no interior de gaiolas adaptadas, contendo uma ou duas fêmeas do parasitoide de sete dias de idade. O número de fêmeas do parasitoide dividindo a mesma "unidade de parasitismo" não afetou nenhum aspecto da qualidade do parasitoide. O tamanho do hospedeiro influenciou significativamente no número de parasitoides emergidos. Larvas hospedeiras menores (cinco dias) provocaram uma redução no parasitismo em relação às larvas de seis, sete e oito dias de idade. Houve maior emergência de parasitoides machos de hospedeiros com cinco e seis dias de idade, enquanto mais fêmeas emergiram de larvas com sete e oito dias de idade. As fêmeas de D. longicaudata provenientes de larvas com seis, sete e oito dias viveram mais do que as fêmeas obtidas de larvas com cinco dias de idade. Larvas hospedeiras maiores, quando utilizadas na criação de parasitoides de moscas-das-frutas, aumentam a porcentagem de parasitismo, o número de fêmeas na progênie e a longevidade das fêmeas da progênie, podendo proporcionar vantagens econômicas quando utilizadas para liberação aumentativa no campo.(AU)

This work was carried out in order to evaluate the influence of age (size) of larvae Ceratitis capitata used in the rearing of the Diachasmimorpha longicaudata on parasitism, sex ratio of the progeny and the longevity of progeny's females, when the mothers were maintained alone or sharing the same host. Larvae at different ages (five, six, seven and eight days old), in "units of parasitism" were offered to female parasitoids, inside adapted cages, containing one or two seven days old female parasitoids. The number of females of the parasitoid sharing the same "unit of parasitism" in the same cage did not affect any aspect of parasitoid quality. However, the size of the host had significant influence on the number of emerged parasitoids. The youngest host larvae (five days old) reduced parasitism more than larvae at six, seven and eight days old. There was higher emergence of male parasitoids of five and six days old hosts, while more females emerged from seven and eight days old larvae. The females of D. longicaudata from larvae with six, seven and eight days old lived more than those obtained from females of five days old larvae. When larger host larvae are used in the rearing of parasitoids of fruit flies, it increases parasitism, the number of female progeny and the longevity of female progeny; it can provide economic advantages when used for augmentative liberation on the field.(AU)

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The Mediterranean fruit fly Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) is one of the principal exotic pests affecting Brazilian production in the northeastern and southeastern regions of Brazil. In the south, it is has potential as a serious threat to temperate-climate fruit farms, since it is already found in urban and suburban communities in this region. We studied the biological characteristics of C. capitata populations from Pelotas-RS (temperate climate), Petrolina-PE (tropical), and Campinas-SP (subtropical). Ceratitis capitata biology was studied under controlled temperature (15, 20, 25, 30, and 35 ± 1 °C), 70 ± 10% RH, and 14:10 L:D photoperiod. The duration and survival rate of the egg, larval, and pupal stages were evaluated and the thermal requirements of these three populations were determined. The duration and survival of these developmental stages varied with temperature, with similar values for the three populations, except for some variation in the egg phase. Egg to adult developmental time for all three populations was inversely proportional to temperature; from 15 to 30 °C developmental time varied from 71.2 to 17.1, 70.2 to 17.1, and 68.5 to 16.9 days, respectively. Survival during development was affected at 15 to 30 °C, and differed significantly from survival at 20 to 25 °C. At 35 °C, immature stages did not develop. The basal temperature and degree-day requirement were similar for all immature stages except for the egg stage. The basal temperatures and thermal constants were 9.30 and 350, 8.47 and 341, and 9.60 °C and 328 degree-days for the Pelotas, Petrolina, and Campinas populations, respectively. Results suggested that survival and thermal requirements are similar for these tropical, subtropical, and temperate populations of C. capitata, and demonstrate the species' capacity to adapt to different climate conditions.

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The present report shows a partial biochemical characterization and life cycle expression of N-ß-alanyldopamine hydrolase (Tan protein) in Ceratitis capitata and Drosophila melanogaster. This enzyme catalyzes the hydrolysis of N-ß-alanyldopamine (NBAD), the main tanning precursor of insect brown cuticles. It also plays an important role in the metabolism of brain neurotransmitters, recycling dopamine and histamine. In contrast to NBAD-synthase, Tan is expressed constitutively in epidermis and does not respond directly to microbial challenge. Immunodetection experiments showed the novel localization of NBAD-hydrolase in the embryo central neural system and in different regions of the adult brain, in addition to optic lobes. We sequenced and characterized Drosophila mutants tan¹ and tan³. The latter appears to be a mutant with normal expression in neural tissue but weak one in epidermis.

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This study aimed to evaluate adult emergence and duration of the pupal stage of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), and emergence of the fruit fly parasitoid, Diachasmimorpha longicaudata (Ashmead), under different moisture conditions in four soil types, using soil water matric potential. Pupal stage duration in C. capitata was influenced differently for males and females. In females, only soil type affected pupal stage duration, which was longer in a clay soil. In males, pupal stage duration was individually influenced by moisture and soil type, with a reduction in pupal stage duration in a heavy clay soil and in a sandy clay, with longer duration in the clay soil. As matric potential decreased, duration of the pupal stage of C. capitata males increased, regardless of soil type. C. capitata emergence was affected by moisture, regardless of soil type, and was higher in drier soils. The emergence of D. longicaudata adults was individually influenced by soil type and moisture factors, and the number of emerged D. longicaudata adults was three times higher in sandy loam and lower in a heavy clay soil. Always, the number of emerged adults was higher at higher moisture conditions. C. capitata and D. longicaudata pupal development was affected by moisture and soil type, which may facilitate pest sampling and allow release areas for the parasitoid to be defined under field conditions.

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Chitin-binding vicilin from Erythrina velutina seeds was purified by ammonium sulfate followed by affinity chromatography on a chitin column and gel filtration on Superose-6-10-300-GL. The Erythrina velutina vicilin, called EvV, is a tetrameric glycoprotein composed of 1.85% carbohydrates and M r of 216.6 kDa, consisting of two subunits of M r of 54.8 and two subunits of M r of 50.8 kDa. The EvV homogeneity was confirmed in native PAGE where it was observed to be a unique acid-protein band with slow mobility in this gel. Effect of EvV on C. capitata larvae was examined by bioassay and its mechanism of action was determined by immunodetection techniques and fluorescence localization in chitin structures that are present in C. capitata digestory system. EvV when added to diet caused strong effect on mortality (ED50 of 0.14%) and larval mass (WD50 of 0.12%). These deleterious effects were associated to the binding to chitin structures present in peritrophic membrane and to gut epithelial cells, and its low digestibility in C. capitata digestive tract. These results are the first demonstration of a proteinaceous bioinsecticide from plant origin effective against C. capitata larvae. EvV may be part of the pest management programs or an alternative in plant improvement program.

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A proteinaceous trypsin inhibitor was purified from Crotalaria pallida seeds by ammonium sulfate precipitation, affinity chromatography on immobilized trypsin-Sepharose and TCA precipitation. The trypsin inhibitor, named CpaTI, had M(r) of 32.5 kDa as determined by SDS-PAGE and was composed of two subunits with 27.7 and 5.6 kDa linked by disulfide bridges. CpaTI was stable at 50 degrees C and lost 40% of activity at 100 degrees C. CpaTI was also stable from pH 2 to 12 at 37 degrees C. CpaTI weakly inhibited chymotrypsin and elastase and its inhibition of papain, a cysteine proteinase, were indicative of its bi-functionality. CpaTI inhibited, in different degrees, digestive enzymes from Spodoptera frugiperda, Alabama argillacea, Plodiainterpunctella, Anthonomus grandis and Zabrotes subfasciatus guts. In vitro and in vivo susceptibility of Callosobruchus maculatus and Ceratitis capitata to CpaTI was evaluated. C. maculatus and C. capitata enzymes were strongly susceptible, 74.4+/-15.8% and 100.0+/-7.3%, respectively, to CpaTI. When CpaTI was added to artificial diets and offered to both insect larvae, the results showed that C. maculatus was more susceptible to CpaTI with an LD(50) of 3.0 and ED(50) of 2.17%. C. capitata larvae were more resistant to CpaTI, in disagreement with the in vitro effects. The larvae were more affected at lower concentrations, causing 27% mortality and 44.4% mass decrease. The action was constant at 2-4% (w/w) with 15% mortality and 38% mass decrease.

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A laboratory bioassay was developed to determine both the chemical toxicity and the phototoxicity of the xanthene dye, phloxine B (D&C Red No 28), to the immature stages of the Mediterranean fruit fly, Certitis capitata (Wiedemann). An additional goal was to find out which main tissues are affected first. A low, but significant, level of toxicity was observed when the insects were maintained in the dark: at the point of adult ecdysis, the LC50 was 11.03 mM. As expected, after 8-h exposure of late larva III to light, a high level of mortality was produced (LC50 at ecdysis: 0.45 mM) as a dose-dependent function of dye concentration. At sublethal concentrations of the dye, the surviving insects showed a number of physiological abnormalities. Phloxine B appeared to mainly affect the larval longitudinal muscles as well as the abdominal muscles of ecdysing adults, giving rise to abnormal puparia and failed adult ecdysis, respectively. Moreover, a significant phloxine B-dependent delay in the jumping of surviving larvae for dispersal was documented. This could be attributed to a delay in attaining a threshold weight for jumping and/or to abnormalities in neuromuscular coordination, thus reinforcing the idea of pleiotropic effects of the dye.

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