Lack of fitness costs in dsRNA-resistant Leptinotarsa decemlineata ([Coleoptera]: [Chrysomelidae]).

The Colorado potato beetle, Leptinotarsa decemlineata (Say) ([Coleoptera]: [Chrysomelidae]), is the most important defoliator of solanaceous plants worldwide. This insect displays a notorious ability in adapting to biological and synthetic insecticides, although in some cases this adaptation carries relevant fitness costs. Insecticidal gene silencing by RNA interference is a novel mode of action pesticide against L. decemlineata that is activated by ingestion of a double stranded RNA (dsRNA) targeting a vital L. decemlineata gene. We previously reported laboratory selection of a > 11,000-fold resistant strain of L. decemlineata to a dsRNA delivered topically to potato leaves. In this work, we tested the existence of fitness costs in this dsRNA-resistant colony by comparing biological parameters to the parental strain and an additional susceptible reference strain. Biological parameters included length of egg incubation period, number of eggs per clutch, egg viability, larval viability, length of larval and pupal periods, adult emergence, number of eggs laid per day, sex ratio, and adult longevity. Comparisons between the 3 beetle strains detected no fitness costs associated with resistance to dsRNA. This information is important to guide effective insect resistance management plans for dsRNA insecticides against L. decemlineata applied topically to potato leaves.

Parasitism-Mediated Interactions Between the Ring-Legged Earwig and Sugarcane Borer Larvae.

Sugarcane is one of the most economically important crops in Brazil. The damage caused by Diatraea saccharalis (F.) (Lepidoptera: Crambidae) results in high costs for its production. Cotesia flavipes Cameron (Hymenoptera: Braconidae) is the most efficient means of controlling this pest. The prohibition of burning and the preservation of soil cover through non-tillage practices increased many population densities of natural enemies of the sugarcane borer (SCB) in sugarcane fields. Euborellia annulipes (Lucas) (Dermaptera: Anisolabididae) is a predator found in sugarcane fields and is associated with predation of SCB. This study aimed to evaluate the influence of parasitism of D. saccharalis by C. flavipes on predation by E. annulipes adult females. For this purpose, fourth instar sugarcane borer larvae were exposed to parasitism over a 5-day period or were not parasitized. Subsequently, the prey was supplied to the predator in arenas under choice and no-choice conditions and under different densities to evaluate the feeding preference over a 12-h period and functional response of the predator over a 24-h period. E. annulipes killed 2-3 fourth instar larvae of the sugarcane borer over a 12-h period. SCB parasitism did not affect the feeding preference of the predator and did not alter the type II functional response. However, the handling time of the predator was lower (1.943 h) and there was a higher T/Th ratio on the non-parasitized larvae (12.352). Our findings indicate E. annulipes as a promising biological control agent of D. saccharalis, with a potential to assist in the sugarcane borer management approaches and could to reduce the costs of parasitoid release in the field.

Phytophagy of the predator Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae) fed on prey and Brassicaceae / Fitofagia do predador Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae) em diferentes brassicáceas

Abstract The purpose of this study was to investigate the development and reproduction of the zoophytophagous predator Podisus nigrispinus (Dallas) (Heteroptera: Pentatomidae) fed kale, broccoli and cabbage affects its. Nymphs and adults of this predator were fed on larvae of Plutella xylostella (L.) (Lepidoptera: Plutellidae) as prey with kale, cabbage, or broccoli. In the nymph period, the duration and prey consumption were similar with all the Brassicacea cultivar. However, nymph viability was higher for predators with broccoli leaves. The mean weight of 5th-instar nymphs, newly emerged females and the sex ratio were similar among the Brassicacea cultivars, while newly emerged males were heavier with kale and broccoli leaves. The supply of broccoli leaves resulted in greater oviposition, higher number of eggs per egg mass and longer longevity of P. nigrispinus males and females. Furthermore, the consumption of P. xylostella larvae by adult predators was higher with these cultivars. The net reproductive rate (R0) and mean generation time (T) were highest for predators with prey and broccoli leaves. The reproductive parameters of P. nigrispinus were enhanced when fed on P. xylostella larvae with and broccoli leaves, which can be an alternative diet in laboratory rearing of this predator.

Resumo O objetivo deste estudo foi verificar o desenvolvimento e reprodução do zoofitófago Podisus nigrispinus (Dallas) (Heteroptera: Pentatomidae) em couve, brócolis e repolho. Ninfas e adultos deste predador foram alimentados com lagartas de Plutella xylostella (L.) (Lepidoptera: Plutellidae) como presa e receberam folhas de couve, repolho ou brócolis. Durante o período ninfal, a duração do período e o consumo de presas foram semelhantes com as diferentes cultivares de brassicácea. Porém, a viabilidade ninfal foi maior para predadores com folhas de brócolis. O peso de ninfas de quinto instar e de fêmeas recém-emergidas e a razão sexual de P. nigrispinus foram semelhantes entre as cultivares de brassicáceas, enquanto que o peso de machos recém-emergidos foi maior com folhas de couve e brócolis. Folhas de brócolis proporcionaram maiores número de oviposições, ovos por postura e longevidade de machos e fêmeas de P. nigrispinus. Além disso, o consumo de lagartas de P. xylostella por adultos desse predador fora maiores com esta cultivar. A taxa líquida de reprodução (R0) e o tempo médio de geração (T) foram maiores para predadores com presa e folhas de brócolis. Podisus nigrispinus alimentados com lagartas de P. xylostella e folhas de brócolis apresentaram melhores parâmetros reprodutivos, podendo ser uma alternativa para a criação deste predador em laboratório.

Phytophagy of the predator Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae) fed on prey and Brassicaceae.

The purpose of this study was to investigate the development and reproduction of the zoophytophagous predator Podisus nigrispinus (Dallas) (Heteroptera: Pentatomidae) fed kale, broccoli and cabbage affects its. Nymphs and adults of this predator were fed on larvae of Plutella xylostella (L.) (Lepidoptera: Plutellidae) as prey with kale, cabbage, or broccoli. In the nymph period, the duration and prey consumption were similar with all the Brassicacea cultivar. However, nymph viability was higher for predators with broccoli leaves. The mean weight of 5th-instar nymphs, newly emerged females and the sex ratio were similar among the Brassicacea cultivars, while newly emerged males were heavier with kale and broccoli leaves. The supply of broccoli leaves resulted in greater oviposition, higher number of eggs per egg mass and longer longevity of P. nigrispinus males and females. Furthermore, the consumption of P. xylostella larvae by adult predators was higher with these cultivars. The net reproductive rate (R0) and mean generation time (T) were highest for predators with prey and broccoli leaves. The reproductive parameters of P. nigrispinus were enhanced when fed on P. xylostella larvae with and broccoli leaves, which can be an alternative diet in laboratory rearing of this predator.

Selection of prey to improve biological parameters of the predator Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae) in laboratory conditions / Seleção de presas para melhorar parâmetros biológicos do predador Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae) em condições de laboratório

Abstract Mass production of predatory stinkbugs in the laboratory is prioritized to release them into the field as part of IPM programs. Therefore, the aim of this study was to identify the best prey for rearing the predator Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae) among five different species of insect (three of Lepidoptera, one of Coleoptera, and one of Diptera). Second-instar P. nigrispinus nymphs were conditioned in transparent 1000-mL plastic pots, adults were placed in Petri dishes for mating, and both stages were maintained under controlled conditions (25 ± 1°C, 12 hours of photophase, 70 ± 10% RH). Nymphs and adults of P. nigrispinus consumed more Musca domestica (Linnaeus, 1758) (Diptera: Muscidae) larvae than the other tested prey. The consumption of fly larvae was 1.5 larvae/day/nymph and adults 1.7 larvae/day/adult. However, the number of eggs per female was less when the predator consumed M. domestica larvae (407.8 eggs/female) and most when consumed the Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) larvae (797.7 eggs/female). Furthermore, the percentage of hatched eggs was greater when the predator females consumed D. saccharalis larvae (90.0%). D. saccharalis larvae is the best prey to rearing P. nigrispinus.

Resumo A produção massal de percevejos predadores no laboratório é priorizada para liberá-los no campo, como parte de programas de MIP. Portanto, o objetivo deste estudo foi identificar a melhor presa para a criação do predador Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae) entre cinco espécies diferentes de insetos (três de Lepidoptera, uma de Coleoptera e uma de Diptera). Ninfas de segundo ínstar de P. nigrispinus foram acondicionadas em recipientes plásticos transparentes de 1000 ml, adultos foram colocados em placas de Petri para o acasalamento e, ambos os estágios foram mantidos sob condições controladas (25 ± 1°C, fotofase de 12 horas e 70 ± 10% UR). Ninfas e adultos de P. nigrispinus consumiram mais larvas de Musca domestica (Linnaeus, 1758) (Diptera: Muscidae) do que as outras presas testadas. O consumo de larvas de mosca foi de 1,5 larvas/dia/ninfa e adultos 1,7 larvas/dia/adulto. No entanto, o número de ovos por fêmea foi menor quando o predador consumiu larvas de M. domestica (407,8 ovos/fêmea) e maior quando consumiu larvas de Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) (797,7 ovos/fêmea). Além disso, a percentagem de eclosão de ninfas foi maior quando as fêmeas do predador consumiram larvas de D. saccharalis larvas (90,0%). Larvas de D. saccharalis é a melhor presa para a criação de P. nigrispinus.

Selection of prey to improve biological parameters of the predator Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae) in laboratory conditions.

Mass production of predatory stinkbugs in the laboratory is prioritized to release them into the field as part of IPM programs. Therefore, the aim of this study was to identify the best prey for rearing the predator Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae) among five different species of insect (three of Lepidoptera, one of Coleoptera, and one of Diptera). Second-instar P. nigrispinus nymphs were conditioned in transparent 1000-mL plastic pots, adults were placed in Petri dishes for mating, and both stages were maintained under controlled conditions (25 ± 1°C, 12 hours of photophase, 70 ± 10% RH). Nymphs and adults of P. nigrispinus consumed more Musca domestica (Linnaeus, 1758) (Diptera: Muscidae) larvae than the other tested prey. The consumption of fly larvae was 1.5 larvae/day/nymph and adults 1.7 larvae/day/adult. However, the number of eggs per female was less when the predator consumed M. domestica larvae (407.8 eggs/female) and most when consumed the Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) larvae (797.7 eggs/female). Furthermore, the percentage of hatched eggs was greater when the predator females consumed D. saccharalis larvae (90.0%). D. saccharalis larvae is the best prey to rearing P. nigrispinus.

Potential of Entomopathogenic Fungi as Biological Control Agents of Diamondback Moth (Lepidoptera: Plutellidae) and Compatibility With Chemical Insecticides.

The objectives were to evaluate the efficiency of entomopathogenic fungi against Plutella xylostella (L.) and the compatibility of the most virulent isolates with some of the insecticides registered for use on cabbage crops. Pathogenicity tests used isolates of Beauveria bassiana, Metarhizium rileyi, Isaria fumosorosea, Isaria sinclairii, and Lecanicillium muscarium standardized at a concentration of 10(7) conidia/ml. Cabbage leaf discs were immersed in these suspensions, and after evaporation of the excess water, were placed 10 second-instar larvae of P. xylostella, totaling 10 leaf discs per treatment. Mortality was assessed 7 d after treatment, and the isolates that caused mortality>80% were used to estimate LC50 and LT50. The compatibilities of the most virulent isolates and the insecticides were tested from the mixture of these into the culture medium, and after solidifying, the medium was inoculated with an aliquot of the isolated suspension. The following parameters were evaluated: growth of the colony, number and viability of conidia after 7 d. The isolated IBCB01, IBCB18, IBCB66, and IBCB87 of B. bassiana, LCMAP101 of M. rileyi, and ARSEF7973 of I. sinclairii caused mortality between 80 and 100%, with LC50 and LT50 between 2.504 to 6.775×10(4) conidia/ml and 52.22 to 112.13 h, respectively. The active ingredients thiamethoxam and azadirachtin were compatible with the entomopathogenic fungi. The results suggest that the use of these isolates is an important alternative in the pesticidal management of P. xylostella, with the possible exception of the associated use of chemical controls using the active ingredients thiamethoxam or azadirachtin.

Interactions Between Bt-Bioinsecticides and Podisus nigrispinus (Dallas) (Hemiptera: Pentatomidae), a Predator of Plutella xylostella (L.) (Lepidoptera: Plutellidae).

Bioinsecticides are being increasingly used to protect vegetable crops against herbivores, but data on the side effects of such strategy on the third trophic level are still required. We investigated the influence of the Bacillus thuringiensis subsp. kurstaki HD-1 strain and of the commercial bioinsecticide Agree® on the biological aspects of the predator Podisus nigrispinus (Dallas) (Hemiptera: Pentatomidae) when feeding on Plutella xylostella (L.) (Lepidoptera: Plutellidae)-infected larvae. On average, infected larvae were consumed more often than the non-infected larvae throughout the predator nymphal development, and no effect on nymph survival was observed. Population growth parameters (R 0, r m, and λ) resulting from the fertility life tables did not differ among treatments. The results led to the conclusion that P. nigrispinus can be used in combination with the product Agree® or the strain HD-1 for the control of P. xylostella. The combined use of these control agents helps to maintain the balance of the ecosystem and contributes for the production of food for healthy human consumption.

Ação de inseticidas sobre os ovos e lagartas da broca-pequena-dofruto do tomate, em bioensaio de laboratório / Action of insecticides on the eggs and larvae of the borer small-fruit tomato in laboratory bioassay

A broca-pequena-do-fruto é praga-chave na cultura do tomate por causar danos significativos às partes reprodutivas. Devido a isto, foi objetivo deste trabalho avaliar a eficiência de inseticidas sobre ovos e lagartas recém-emergidas, com e sem a adição de óleo vegetal (0,25%), em bioensaios de laboratório. Frutos com ovos foram coletados em cultivo de tomate estaqueado na quinzena posterior à última aplicação de agrotóxicos, sendo selecionados os frutos com ovos de coloração variável de branco a marrom claro, com 1 e 4 dias de incubação, e imersos em 1 L da calda inseticida por 5 segundos. O delineamento estatístico utilizado foi o inteiramente casualizado, com média de 18 frutos nos tratamentos e de, aproximadamente, 4 ovos/fruto. A avaliação da ação inseticida de 24 produtos foi realizada após a imersão na calda inseticida, observando-se, sob microscópio estereoscópico, os ovos quanto à integridade do córion, textura e coloração, lagartas emergidas, bem como os orifícios de entrada e de saída das larvas (aos 7 e 21 dias). Os produtos testados diferiram da testemunha quanto à densidade de lagartas eclodidas, bem como quanto à redução populacional de lagartas, podendo-se destacar Trebon 100 SC (etofenprox; 200 mL do produto comercial/100 L), Lannate BR (methomil; 100 mL), Thiobel 500 (cartap; 250 g) e Vertimec 18 CE (abamectin; 100 mL). A adição de óleo vegetal resultou em incremento na eficiência dos produtos.

Action of insecticides on tomato fruit borer eggs and larvae using laboratory bioassay. The tomato fruit borer is a key tomato pest in light of its damages to the plants’ reproductive parts. Therefore, the present study was aimed to evaluate the effectiveness of insecticides on the eggs and newly hatched larvae, when applied alone or associated with vegetable oil (0.25%), in laboratory bioassays. Fruits with eggs were collected in staked tomato crops fifteen days after the last application of agro-chemicals, selecting the fruits with eggs colored from white to light brown, which had between 1 and 4 days of incubation, followed by immersion of the fruit for 5 seconds in 1 L of the insecticide solutions. The experimental design was completely randomized, with an average of 18 fruits per treatment (24 insecticides and a control) and about 4 eggs per fruit. The evaluations were performed 7 and 21 days after immersion, using a stereomicroscope to observe the corion integrity, egg color, number of larvae emerged, and larvae fruit holes (entry and exit). The insecticides differed significantly from the control, most notably Trebon SC 100 (etofenprox; 200 mL of the commercial product/100 L), Lannate BR (methomil; 100 mL), Thiobel 500 (cartap; 250 g) and Vertimec 18 CE (abamectin; 100 mL). The addition of vegetable oil increased the insecticides’ effectiveness.

Interaction between the predator Podisus nigrispinus (Hemiptera: Pentatomidae) and the entomopathogenic bacteria Bacillus thuringiensis.

Plutella xylostella (L.) is susceptible to both the entomopathogen Bacillus thuringiensis and the predator, Brazilian spined soldier bug [Podisus nigrispinus (Dallas)]. The objective of this study was to measure the interaction between the bacterium B. thuringiensis and the predator P. nigrispinus. We also studied the behavior of P. nigrispinus in relation to its choice between B. thuringiensis-infected and healthy P. xylostellais larvae. In the first treatment, P. nigrispinus nymphs were fed daily with B. thuringiensis-infected P. xylostella larvae and distilled water. In the second treatment, nymphs were fed daily with healthy larvae and a suspension of B. thuringiensis as a source of water. The control nymphs were fed daily with healthy larvae and water. Adult P. nigrispinus were separated by sex, couples were formed, and they were fed daily with P. xylostella larvae derived from the treatments. We followed the development of P. nigrispinus and measured its biological characteristics. On the basis of these data, parameters were determined for the construction of life tables. A choice test was used to compare infected and healthy larvae. The HD1 strain of B. thuringiensis does not affect the biological characteristics of P. nigrispinus when fed infected larvae and water or healthy larvae and B. thuringiensis suspension. Our study shows that integrated management of P. xylostella, a pest of the Brassicaceae, is feasible by using the HD1 strain of B. thuringiensis and the predator P. nigrispinus, because the predator shows no preference for infected or healthy P. xylostella larvae.

Treatment of streptozotocin-induced diabetic rats with vanadate and phlorizin prevents the over-expression of the liver insulin receptor gene.

Administration of vanadate, an insulinomimetic agent, has been shown to normalize the increased number of insulin receptors in the liver of streptozotocin-induced diabetic rats. In the present study, the effects of vanadate on various steps of expression of the liver insulin receptor gene in diabetic rats have been analyzed and compared with those of phlorizin, a glucopenic drug devoid of insulinomimetic properties. Livers of rats killed 23 days after streptozotocin injection showed a 30-40% increase in the number of cell surface and intracellular insulin receptors, a 50-90% increase in the levels of 9.5 and 7.5 kb insulin receptor mRNA species, and a 20% decrease in the relative abundance of the A (exon 11-) insulin receptor mRNA isotype. Daily administration of vanadate or phlorizin from day 5 to day 23 prevented the increase in insulin receptor number and mRNA level, and vanadate treatment also normalized receptor mRNA isotype expression. Unlike observations in vivo, vanadate and phlorizin differentially affected the expression of the insulin receptor gene in Fao hepatoma cells. Vanadate treatment (0.5 mmol/l for 4 h) decreased the levels of the 9.5 and 7.5 kb insulin receptor transcripts by at least twofold, without affecting the relative abundance of the A insulin receptor mRNA isotype. In contrast, phlorizin treatment (5 mmol/l for 4 h) slightly increased or did not affect the levels of the 9.5 and 7.5 kb insulin receptor transcripts respectively, and increased by twofold the relative expression of the A insulin receptor mRNA isotype. It is suggested that, although mediated in part by a reversal of hyperglycemia, normalization of liver insulin receptor gene expression by vanadate treatment in diabetic rats may also involve a direct inhibitory effect of this drug on gene expression.

Vanadate, but not insulin, inhibits insulin receptor gene expression in rat hepatoma cells.

Insulin and vanadate treatments have recently been shown to reverse the overexpression of the hepatic insulin receptor (IR) gene in streptozotocin-induced diabetic rats. To better understand the mechanisms underlying these effects, the abilities of insulin and vanadate to affect IR gene expression have been comparatively examined in Fao hepatoma cells, an insulin-responsive cell line. Exposure of Fao cells to insulin (1 microM) or vanadate (500 microM) for 24 h led to a 2-fold decrease in IR number in total cellular membranes. Insulin treatment did not affect IR messenger RNA (mRNA) level regardless of time of exposure and concentration. In contrast, vanadate treatment caused a time- and dose-dependent decrease in IR mRNA level, which was maximal (4-fold change) after a 24-h exposure to 500 microM vanadate and was fully reversible. Insulin treatment increased from 28 to 39% the relative expression of isotype A IR mRNA, but vanadate treatment did not significantly affect this parameter. Vanadate treatment did not modify mRNA half-life (3.5 h) in 5, 6 dichlorobenzimidazole riboside-treated cells but decreased by 4-fold the transcriptional activity of the IR gene. These data show for the first time that, although both insulin and vanadate decrease total cellular IR number in Fao cells, only vanadate decreases IR mRNA level. It does so by inhibiting transcription of the IR gene, suggesting an action on the gene promoter which could be mediated by changes in the level of expression and/or of phosphorylation of trans-acting factors.

[Regulation of insulin receptor expression and its gene]. / Régulation de l'expression du récepteur de l'insuline et de son gène.

The insulin receptor is a membrane macromolecule whose expression on the cell surface is essential for cell sensitivity to insulin. Current knowledge on the regulation of expression of the insulin receptor and its gene in human and animal cells is presented. Although ubiquitously distributed, the insulin receptor and its messenger RNA (mRNA) are mainly expressed in metabolically active cells such as hepatocytes and adipocytes. Two receptor isoforms, generated by alternative splicing of exon 11, have been identified. Isoform B (exon 11+) predominates in liver and adipocytes, and isoform A (exon 11-) in brain, spleen and leukocytes. In vivo and in several cell models, the expression of the insulin receptor and/or its mRNA is under positive regulation by glucocorticoid hormones and negative regulation by insulin. Glucocorticoid hormones stimulate receptor gene transcription and receptor protein synthesis. Insulin stimulates receptor protein degradation and, in certain cell types, decreases receptor mRNA level. Vanadate (an insulinomimetic agent) corrects, in vivo, the hyperexpression of the liver receptor observed in experimental insulinopenic diabetes, but its effects on receptor expression in vitro are complex and vary with the cell type. In vivo the insulin receptor and/or its mRNA are expressed early in fetal development with a high level, in liver, of isoform A. Maximal expression is reached at the end of gestation and then decreases after birth. In several cell models, receptor protein and/or mRNA expression is affected by cell growth and/or differentiation. Several cis- and trans-acting factors regulating the expression of the human insulin receptor gene and its response to glucocorticoid hormones have been identified.