Suppression of Mexican bean beetle development by foliar applications of azasterols and reversal by cholesterol

Suppression of growth and molting occurred in Mexican bean beetle (Epilachna varivestis) larvae confined with bean plants treated with 25-azacholesterol (25-A) or 3-methoxy-25-azacholesterol (3-M-25-A). No larvae confined with bean plants dipped into 10 ppm of 3-M-25-A completed development and over half failed to complete the first ecdysis. Three larvae treated with 25-A developed everted wing pads on the last instar. Addition of an equal weight of cholesterol to the dipping solution completely nullified the inhibitive effects of a dose of 25-A 100 x greater than that required for inhibitive effects. Treatment of the bean plants with a combination of 25-A and a JH mimic resulted in additive and possibly synergistic pathological developmental effects in confined larvae.

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Résumé Des larves d'Epilachna varivestis élevées sur des plants de haricots trempés dans des solutions d'acétone et eau contenant soit du 25-azacholesterol (25-A) ou du 3-methoxy-25-azacholesterol (3-M-25A), cessent de muer et leur croissance est stoppée. Avec des plants de haricots trempés dans une solution à 10 p.p.m. de. 3 M-25A, aucune larve n'acheva son développement et plus de la moitié des 100 larves expérimentées ne purent achever leur lère mue. Les pupes obtenues ne pesaient fréquemment que la moitié du poids des témoins. Trois larves traitées avec 25-A développèrent des ébauches alaires inversées au cours du dernier stade.Il a été reconnu que ces azasterols inhibent les D²² et D^22,24 sterolreductases dans le tissu larvaire de l'intestin moyen des larves de Manduca sexta (Tobacco Hornworm). Ces enzymes interviennent dans la transformation des principaux phytostérols des feuilles de haricots (stigmasterol, -sitosterol et campesterol) en cholesterol. Comme il y a moins de 1% de cholesterol dans les feuilles de haricots, nous avons essayé de compenser l'effet inhibiteur du 25-A en ajoutant du cholesterol aux solutions du bain. L'addition de 1000 ppm de cholesterol annule complètement les effets inhibiteurs de 1000 ppm de 25-A (100 fois plus que la dose nécessaire pour provoquer des effets inhibiteurs).Des larves de Manduca sexta élevées sur plants de tabac à l'azasterol ne sont pas affectées, bien que leur développement soit grandement retardé par de petites quantités de ces azasterols ajoutés à un aliment synthétique dépourvu de cholestérol. Sans doute, la haute teneur en cholesterol des feuilles de tabac (9% des sterols totaux) est suffisante pour satisfaire les besoins en cholesterol des larves. Des traitements de plants de haricots avec une combinaison de 25-A plus la substance mimétique de l'hormone juvénile (E)-4-[(6,7-epoxy-3-ethyl-7-methyl-2-nonenyl) oxy]-1,2-(methylenedioxy) benzène, ont provoqué des effets pathologiques surjoutés et peut être synergiques sur le développement des larves d'Epilachna.

     

Insect responses to plant water deficits. II. Effect of water deficits in soybean plants on the growth and survival of Mexican bean beetle larvae

Abstract. 1. We examined the effect of water deficits in soybean, Glycine max [L.] Merr., on the growth and survival of Mexican bean beetle larvae, Epilachna varivestis Mulsant (Coleoptera: Coccinellidae).
2. Larvae were reared under growth chamber, glasshouse, and field conditions on foliage from plants that were either well-watered or subjected to water deficits.
3. Larval survival, growth rate and pupal weight were reduced, and development time was increased when larvae were reared on foliage from plants subjected to water deficits. These results are contrary to White's (1974) hypothesis that water deficits in plants cause increased growth and survival of herbivorous insects.
4. Changes in foliage chemistry caused by water deficits, possibly in the concentration of free amino acids, are the most likely causes of the observed effects on growth rate and development time. However, under glasshouse and field conditions, physical changes in foliage and/or the foliage's environment that accompany water deficits are also important.     

Zebrafish (Danio rerio) fed vitamin E-deficient diets produce embryos with increased morphologic abnormalities and mortality

Vitamin E (α-tocopherol) is required to prevent fetal resorption in rodents. To study α-tocopherol's role in fetal development, a nonplacental model is required. Therefore, the zebrafish, an established developmental model organism, was studied by feeding the fish a defined diet with or without added α-tocopherol. Zebrafish (age, 4-6 weeks) were fed the deficient (E-), sufficient (E+) or lab diet up to 1 years. All groups showed similar growth rates. The exponential rate of α-tocopherol depletion up to ~80 day in E- zebrafish was 0.029±0.006 nmol/g, equivalent to a depletion half-life of 25±5 days. From age ~80 days, the E- fish (5±3 nmol/g) contained ~50 times less α-tocopherol than the E+ or lab diet fish (369±131 or 362±107, respectively; P<.05). E-depleted adults demonstrated decreased startle response suggesting neurologic deficits. Expression of selected oxidative stress and apoptosis genes from livers isolated from the zebrafish fed the three diets were evaluated by quantitative polymerase chain reaction and were not found to vary with vitamin E status. When E-depleted adults were spawned, they produced viable embryos with depleted α-tocopherol concentrations. The E- embryos exhibited a higher mortality (P<.05) at 24 h post-fertillization and a higher combination of malformations and mortality (P<.05) at 120 h post-fertillization than embryos from parents fed E+ or lab diets. This study documents for the first time that vitamin E is essential for normal zebrafish embryonic development.     

A dynamic model of water gain and loss in larval Mexican bean beetle

Abstract. Quantitative effects of temperature, vapour pressure deficit, host, and larval body size were experimentally determined. A simulation model for dynamic water balance in the Mexican bean beetle, Epilachna varivestis Mulsant, is presented and parameters are estimated from laboratory data for water gain/loss equations. The model is based on water loss through the cuticle, spiracles and frass, and water gain through ingestion.     

Sexually differentiated flight responses of the Mexican bean beetle to larval and adult nutrition

Summary Flight of male and female Mexican bean beetle adults was examined in laboratory tests. The experimental design made it possible to examine flight behavior not only with respect to different types of hosts (young vs senescent common bean foliage) but also with respect to effects due to their utilization during particular stages of beetle development. The median flight time of males was significantly affected by the adult host, but not by the juvenile host; whereas, the median flight time of females tended to be more affected by the juvenile than by the adult host. These different effects of hosts on the flight times of males and females resulted in sexual dimorphism in flight when the sexes were fed senescent foliage as adults. Although age significantly affected the flight time of both males and females, the reproductive status of females did not affect their flight times. The significance of these results are discussed with respect to the influence of the nutritional complexity of habitats on life history strategies and population dynamics.     

Insect responses to plant water deficits. I. Effect of water deficits in soybean plants on the feeding preference of Mexican bean beetle larvae

Abstract. 1. We examined the preference of larvae of the Mexican bean beetle, Epilachna varivestis Mulsant (Coleoptera: Coccinellidae), for foliage of soybean ( Glycine max [L.] Merr.) grown under several levels of water deficit. Third instar larvae were exposed simultaneously to excised foliage from plants that were either well-watered (control) or had experienced water deficits (treatment).
2. Water-deficient plants were re-watered 12 h prior to initiating feeding trials to eliminate physical differences between control and treatment foliage such as leaf water potential, diffusive resistance, relative water content, and foliage toughness.
3. Examination of the free amino acid contents of re-watered and excised foliage indicated that amino acid concentrations increased markedly in foliage grown under water deficits, and that this increase persisted during the preference tests.
4. Larvae preferred control foliage, but shifted preference to treatment foliage under mild water deficits. When the leaf water potential of water-deficient treatment foliage was lower than – 1.13 MPa or when it was more than 0.5 MPa lower than that of control foliage, larvae preferred to feed on foliage from well-watered control plants.
5. The expression of preference for well-watered control foliage was coincident with increases in the concentrations of total free amino acids and individual free amino acids in the water-deficient treatment foliage.
6. These results are inconsistent with White's (1974) hypothesis because Mexican bean beetle larvae avoid plants grown under water deficits that have increased concentrations of free amino acids.     

Production of acetylcholine in rat brain following thiamine deprivation and treatment with thiamine antagonists

Abstract— The effects of thiamine deprivation and of treatment with the thiamine antagonists, oxythiamine and pyrithiamine, on the storage and synthesis of acetylcholine were studied in rats. Rats treated with pyrithiamine always developed ataxia and convulsions, and they died in an average of 36 ± 5.0 hr after onset of convulsions. Injections of sublethal doses of eserine after onset of convulsions had no effect or shortened survival time. If injections were started before the onset of convulsions, the survival time was increased to 56 ± 3.3 hr. The content of total acetylcholine-like compounds, measured by bioassay, in the brain was decreased in all three types of thiamine deficiency. On the other hand, the amount of parenterally administered [¹⁴C]pyruvate converted to [¹⁴C]acetylcholine in vivo was affected only by treatment with pyrithiamine. The increase found was probably due to an increased permeability of the blood-brain barrier to the pyruvate. Conversion of [¹⁴C]pyruvate to [¹⁴C]acetylcholine in vitro was decreased significantly in homogenates of brains from both oxythiamine and pyrithiamine-treated animals.     

Effects of vitamin D supplementation on calcium balance and bone growth in young rats fed normal or low calcium diet

OBJECTIVE: We examined the effect of vitamin D supplementation on bone growth in young rats fed a normal or low calcium diet. METHODS: Fifty female Sprague-Dawley rats, 6 weeks of age, were randomized by stratified weight method into five groups with 10 rats in each group: baseline control, 0.5% (normal) or 0.1% (low) calcium diet, and 0.5 or 0.1% calcium diet + vitamin D (25 microg/100 g, food intake). Duration of the experiment was 10 weeks. RESULTS: Vitamin D supplementation stimulated intestinal calcium absorption and increased urinary calcium excretion in rats fed a low or normal calcium diet. Vitamin D supplementation prevented the reduction in periosteal bone gain but enhanced enlargement of the marrow cavity and reduced the maturation-related cancellous bone gain in rats fed a low calcium diet, and increased the maturation-related cancellous and cortical bone gains in rats fed a normal calcium diet. CONCLUSION: This study shows the differential effects of vitamin D supplementation on born growth in young rats fed a normal or low calcium diet.     

Synergistic effect of fumigation with ethyl formate and methyl salicylate on mortality of life stages of adzuki bean beetle,Callosobruchus chinensis (L.)

Ethyl formate (EF) is becoming an insecticide of choice for protecting stored commodities. Along with EF, methyl salicylate (MS) is also naturally occurring, and both are generally regarded as safe fumigants for their higher threshold limit values (TLV) and residual status below minimum residue limit (MRL). Sub-lethal doses of EF and MS were evaluated to determine if there was a synergistic effect on fumigation against life stages of the adzuki bean beetle, Callosobruchus chinensis (L.) infesting adzuki bean grains. Based on lethal dose (LD) values, the C. chinensis eggs and adults were equally susceptible to EF followed by larvae and pupae. Adults were the most MS-sensitive followed by eggs and larvae, and pupae were the most tolerant stage. The mortalities of all the life stages of C. chinensis by combined fumigation with sub-lethal EF and MS doses were significantly higher compared to the sum of their separate fumigations revealing a notable synergistic effect. Moreover, the maximum doses of EF and MS tested for synergistic fumigation test did not hamper the germination potential of adzuki been seeds. The synergistic effect between EF and MS can significantly reduce the dose rate of flammable EF against C. chinensis thereby offering a good prospect for EF + MS formulation as an alternative to safeguard stored adzuki bean.     

Inhibition of immunoglobulin E production in allergic model mice by supplementation with vitamin E and beta-carotene

A diet containing different amounts of vitamin E (alpha-tocopherol; 0.5 mg, 5 mg, 10 mg or 50 mg per 100 g diet) was supplemented to BALB/c mice for 6 weeks. These mice were subcutaneously immunized twice with ovalbumin (OVA). A passive cutaneous anaphylaxis (PCA) analysis demonstrated that the mice fed on the diet containing 5 mg of vitamin E produced the highest level of the OVA-specific immunoglobulin E (IgE) antibody. A lower level of serum IgE was found in the mice supplemented with 0.5 mg, 10 mg and 50 mg of vitamin E. A sandwich ELISA analysis showed that the pattern of the total IgE antibody level among these four groups was the same as that of the allergen-specific IgE. In a separate experiment, 5 mg of vitamin E and/or 50 mg of beta-carotene was supplemented to the basal diet containing vitamin E as alpha-tocopherol acetate (5 mg) in order to evaluate the effect of their combination on OVA-specific and total IgE production in the mice. The supplementation with beta-carotene alone had no effect on OVA-specific or total IgE production. In contrast, supplementation with vitamin E plus beta-carotene effectively suppressed both the antigen-specific and total IgE antibodies. The serum vitamin E and beta-carotene levels were increased by supplementation with the respective compounds. These results strongly suggest that the combination of dietary vitamin E and beta-carotene suppressed IgE production and would therefore help to prevent the type-I allergic reaction.     

Influence of induced resistance in soybean on the development and nutrition of the soybean looper and the Mexican bean beetle

Standard gravimetric technique was used to assess the effect of induced resistance in soybean on the consumption and utilization of food by the Mexican bean beetle (MBB), Epilachna varivestis, and the soybean looper (SBL), Pseudoplusia includens. Induced resistance had significant retarding effects on the development and growth of both SBL and MBB. SBL fed leaves of previously injured soybean plants had a 8.5% longer developmental time through the entire larval stage and a 10.4% lower pupal weight than larvae fed leaves from uninjured plants. MBB fed on leaves from previously injured plants through the first three instars had a 9.1% longer developmental time and a 16.5% lower final weight than those fed on leaves from unijured plants. Induced resistance showed no significant effect on total food consumed by either SBL or MBB. Nutritional indices including relative consumption rate (RCR), relative growth rate (RGR), efficiency of conversion of ingested food (ECI), approximate digestibility (AD), and efficiency of conversion of digested food (ECD) were calculated on dry weight measurements. In general, induced resistance had greater impacts on the performance of MBB than on that of SBL.

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Résumé Une technique gravimétrique uniforme a été utilisée pour mesurer l'effet de la résistance induite du soja et la consommation et l'utilisation d'aliments par E. varivestis (Coléop.: Coccinellidae) et P. includens (Lepid.: Noctuidae). La résistance induite a significativement retardé la croissance et le développement des 2 insectes. La consommation de feuilles déjà abîmées a prolongé de 8.5% la durée du développement de P. includens et a réduit de 10.4% le poids des chrysalides. La consommation de feuilles de soja abîmées pendant les 3 premiers stades d'E. varivestis a prolongé le développement de 9.1% et a réduit de 16.5% le poids des nymphes. La résistance induite n'a pas provoqué une modification significative de la quantité de feuilles de soja abîmées chez les 2 insectes. Les indices de nutrition, comprenant le taux de consommation relative (RCR), le taux de croissance relative (RGR), l'efficacité de transformation de l'aliment ingéré (ECI), la digestibilité approchée (AD) et l'efficacité de conversion de l'aliment digéré (ECD), ont été calculés à partir des poids secs. En général, la résistance induite a eu des effets plus nets sur les performances d'E. varivestis que de P. includens.

     

Pattern and process in insect feeding behaviour: A quantitative analysis of the Mexican bean beetle, Epilachna varivestis

No-choice feeding trials and automatic feeding detector studies were used in a quantitative analysis of the feeding behaviour of adult Mexican bean beetles on their host-plant. Different chemical stimuli and physiological factors gave clearly distinguishable patterns in the feeding process. Compared to controls: age had no marked effect; starvation synchronised and reduced the time of initiating feeding, increased consumption rates, increased feeding duration, but did not alter the number of meals; sucrose increased consumption rates over the same feeding duration as controls, and reduced the number of meals; phenolic compounds decreased consumption rates, feeding durations and number of meals. The methodology produces quantitative indices of specific feeding behaviours from the measurement of cumulative consumption/time data, while being sufficiently rapid for screening plant materials and chemicals. The methods are considered generally applicable to laboratory bioassays with phytophagous insects, and are being used to generate a predictive model for the Mexican bean beetle.

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Résumé Des expériences sans choix et des études avec un enregistreur automatique de l'alimentation ont été utilisées pour une analyse quantitative du comportement alimentaire des adultes d'Epilachna varivestis sur leur plante hôte.La consommation cumulée à partir de données temporelles d'insectes isolés a permis de déterminer les courbes temporellement cumulées de la consommation moyenne, les surfaces recouvertes par ces courbes, un rapport entre les aires des courbes expérimentales et témoin, le moment du début du repas, le nombre et la durée des périodes de jeûne et de repas et les taux de consommation pendant les repas. Les témoins ont été comparés aux résultats avec des stimuli physiologiques et chimiques variés.L'âge n'a pas d'effet sensible, peut être parce qu'il s'agit d'un insecte à longévité élevée. La privation d'aliment synchronise et réduit le temps de latence avant le repas, accroît les taux de consommation et les périodes de repas, mais augmente les taux de consommation pour les périodes d'alimentation identiques par rapport aux témoins; le nombre de périodes de repas et le nombre et la durée des périodes de jeûne diminuent. Quelques composés phénologes augmentent le temps avant le repas tandis que d'autres ne le font pas. Beaucoup de composés phénologes diminuent les taux de consommation, les durées et le nombre de repas, tandis qu'ils augmentent la durée des périodes de jeûne et diminuent leur nombre.Cette méthodologie fournit des indices quantitatifs des comportements alimentaires spécifiques et est suffisamment rapide pour tester des végétaux et des substances chimiques. Ces méthodes paraissent applicables à la majorité des essais en laboratoire avec des insectes phytophages, et ont été utilisées pour construire un modèle prédictif sur Epilachna varivestis.

     

Benzyladenine reversal of abscisic Acid inhibition of growth and RNA synthesis in germinating bean axes

The effect of benzyladenine on growth, ATP pool size and specific radioactivity, and the rate and amount of RNA synthesis in aseptically cultured axes of Phaseolus vulgaris during the first 24 hours of germination were measured in experiments where the duration of benzyladenine application and its concentration were varied. Maximum promotion of growth (25%) occurs at 10(-5)m benzyladenine. Maximum promotion of RNA synthesis (44%) occurs at 10(-5)m benzyladenine. Benzyladenine has little effect on the size or specific radioactivity of the ATP pool. Benzyladenine can completely counteract abscisic acid inhibition of growth and RNA synthesis, and these reversals are measurable in 2 hours. GA(1) and GA(3) do not promote growth or counteract abscisic acid inhibition of growth in germinating bean axes in these experiments.     

The effects of temperature and size on growth and mortality of cod larvae

The optimal temperatures for growth of four groups of hatchery-reared cod larvae (geometric mean weight: 73, 191, 249 and 251 μg), reared on rotifers at four or five constant temperatures between 4 and 16° C for 14, 12, 9 and 16 days were 9.7, 12.3, 12.7 and 13.4° C, respectively. The maximum growth rate also increased with size and was 6.5, 9.6, 11.7 and 11.3% day⁻¹ for the respective size groups. The optimal temperature for survival was 8.5–8.8° C for all size groups. The results indicate an opposite relationship between (1) size and optimal temperature for growth and (2) size and maximum growth rate of cod larvae, to that observed for juvenile and immature cod.     

The effect of late pregnancy supplementation of ewes with vitamin E on lamb vigour

The experiment measured lamb responses to supplementation of the pregnant ewe diet with vitamin E above requirement. Crossbred ewes were mated with either Suffolk or Texel rams. Twin-bearing ewes were randomly allocated (approximately 21 months of age at allocation) to one of four treatment groups (20 ewes per group, 10 mated with Suffolk and 10 with Texel rams). Treatments imposed were 50, 100, 150 or 250 IU supplementary vitamin E per ewe per day to give a four treatment by two sire-type factorial experimental design. Ewes were fed concentrates to meet energy requirements for stage of pregnancy and hay ad libitum. Diets were introduced approximately 6 weeks before lambing. Blood samples were obtained prior to introduction of diets, 17 days after introduction of diets and within 24 h of lambing from a subset of eight ewes per treatment (32 total). Colostrum samples were obtained from 10 ewes per treatment, 12 h after birth of the first lamb. All births were observed and a lamb vigour score was assigned to each lamb 5 min after birth. At 1 and 12 h after birth, rectal temperature, and at 12 h after birth, sex, crown-rump length and BW of each lamb were recorded. Mean ewe plasma α-tocopherol concentration prior to introduction of the diets was 1.5 μg/ml (s.e.m. 0.09) and did not differ between groups. There were positive linear (P < 0.001) effects of dietary vitamin E on plasma (17 days after introduction of diets) and colostrum (12 h after birth) α-tocopherol concentrations. Lamb vigour scores were superior (P < 0.001) for lambs sired by Texel rather than Suffolk rams but there were no differences as a result of vitamin E supplementation. Lamb mortality was low and unrelated to either sire or supplementary vitamin E. Lamb birth and weaning weights were also unaffected by vitamin E supplementation. Supplementing the ewe with vitamin E therefore had no effect on any lamb measurements.