In vitro rearing ofTrissolcus basalis [Hym., Scelionidae] an egg parasitoid ofNezara viridula [Hem., Pentatomidae]

In vitro rearing of the egg parasitoidTrissolcus basalis (WOLL.) from eggs collected on the natural hostNezara viridula (L.) was initiated. Several oligidic diets containing insect material (Manduca sexta hemolymph or host egg content) were tested. Our initial medium with 50% hemolymph induced a high egg mortality, but by decreasing the hemolymph concentration, increasing the hen egg yolk concentration and adding 15% of free amino acids mixture, a hatching rate of 85% of the parasitoid eggs was obtained with 39% reaching the second instar and 33% the third instar. In a medium without hemolymph, but with 18% liquid from parasitized host eggs we obtained 90% to 100% hatching, 25 to 27% reaching the second instar and 8% the third instar. We did not obtain pupation from eggsin vitro, but did get pupae and adults from larvae rearedin vivo to second instar and transfered to anin vitro system.      

Analysis of morphogenetic movements in the development of the notum anlage of Drosophila melanogaster

Summary A comparison of the morphogenetic maps of the notum anlage of Drosophila melanogaster derived from the gynandromorph data and mosaics induced by somatic crossing-over during the first instar larval stage revealed that practically no major morphogenetic movements occur in the development of the anlage between the blastoderm and first instar larval stages and the adult stage. By comparing the morphogenetic map derived from gynandromorphs and the fate map derived from data on the transplantation of fragments of the mature wing imaginal disc, it was observed that no major morphogenetic movements occur in the notum anlage between the stages of the allocation of the disc and the mature disc. The results are consistent with the observations of other authors concerning the larval development of eye-antenna, wing and leg discs.     

Preliminary results on in vitro rearing of the endoparasitoid Cardiochiles nigriceps from egg to second instar

The composition of an artificial medium and technical procedures used for in vitro rearing of the endophagous larval parasitoid Cardiochiles nigriceps Viereck (Hymenoptera, Braconidae), from post-germ band egg to the 2nd instar larva, are described. Amino acids, carbohydrates, salts, and vitamins were supplied in defined amounts as an aqueous solution which, when supplemented with 20 mg/ml of bovine albumin, 5 mg/ml of lactalbumin (enzymatic hydrolysate), 20% (v/v) fetal bovine serum, 20% (v/v) milk and 10% (v/v) chicken egg yolk, allowed for parasitoid growth and molting to the 2nd instar. Molting to the final instar was never observed.     

Susceptibility of Delia radicum to steinernematid nematodes

Isolates of different Steinernema species (S. affine, S. bicornutum, S. feltiae and Steinernema C1) were used in mortality assays with third instar larvae of Delia radicum (L.) (Diptera: Anthomyiidae). The nematode isolates had been obtained by baiting soil regularly grown with cabbage. One isolate (S. feltiae) was the result of a natural infection of a D. radicum puparium. The highest mortality (77%) was obtained with an isolate of S. feltiae (DK1). The isolate DK1 was also used in tests with all larval stages of D. radicum. Mortality around 60% was observed for second and third instar larvae, while first instar larvae showed very low or no susceptibility. Maximum mortality of second and third instar larvae was reached applying only 25 nematodes per larva. Observations of larvae that pupated revealed that some of these puparia contained nematodes. Experiments with hatching puparia showed that a high proportion was infected by nematodes if the flies were prevented from leaving nematode-containing soil. In addition to mortality, the ability of the nematodes to successfully reproduce in the insects was studied. It was found that the species S. feltiae and S. bicornutum reproduced in D. radicum larvae and adults with S. feltiae being the most successful.     

Toxicity of the limonoid allelochemical cedrelone to noctuid larvae

The limonoid cedrelone, a constituent of Cedrela toona and C. odorata (Meliaceae) was evaluated as a larval growth inhibitor for the polyphagous noctuids Peridroma saucia and Mamestra configurata by examining its effects on development and feeding behaviour. Cedrelone significantly inhibited growth of different larval instars when administered orally (in artificial diet), topically or via injection. Nutritional analyses revealed that both growth inhibition and reduced consumption are a consequence of postingestive malaise rather than a peripherally-mediated antifeedant effect. Cedrelone administered via injection to sixth instar P. saucia larvae also inhibited growth, delayed development, and resulted in considerable larval mortality. However, all larvae which survived treatment pupated successfully. The present study suggests that the toxicity of cedrelone does not likely involve the endocrine system.     

Encarsia formosa parasitizing the Poinsettia-strain of the cotton whitefly,Bemisia tabaci, on Poinsettia: bionomics in relation to temperature

Adult longevity, developmental time and juvenile mortality ofEncarsia formosa Gahan (Hymenoptera:Aphelinidae) parasitizing the Poinsettia-strain ofBemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) on Poinsettia (Euphorbia pulcherrima Willd.) were investigated in laboratory experiments at three temperatures: 16 °C, 22 °C and 28 °C. Furthermore, the parasitoid's preference for different larval stages of the whitefly was determined at 24.5 °C. The lifespan ofE. formosa decreased with temperature from one month at 16 °C to nine days at 28 °C. A lower temperature threshold of 11 °C for adult development was found. The development of juvenile parasitoids inB. tabaci lasted more than two months at the lowest temperature, but was only 14 days when temperature was 28 °C. The lower temperature threshold for immature development was 13.3 °C, yielding an average of 207 day-degrees for the completion of development into adults. Juvenile mortality was high, varying from about 50% at 16 °C to about 30% at 22 °C and 28 °C.E. formosa preferred to oviposit in the 4th instar and prepupal stages ofB. tabaci followed by the 2nd and 3rd instars. The preference for the pupal stage was low. The parasitoid used all instars of the whitefly for hostfeeding, with no apparent differences between the stages. The average duration of the oviposition posture was four minutes. Demographic parameters were calculated from life tables constructed from the data. The intrinsic rate of increase (r _m) and the net reproductive rate (R ₀) increased with temperature from 0.0279 day⁻¹ at 16 °C to 0.2388 day⁻¹ at 28 °C and from about 12 at 16 °C to about 66 at 28 °C, respectively.     

Pathogenicity of the Entomopathogenic Fungus, <Emphasis Type="Italic">Lecanicillium muscarium</Emphasis>, against the Sweetpotato Whitefly <Emphasis Type="Italic">Bemisia tabaci</Emphasis> under Laboratory and Glasshouse Conditions

The potential for using the entomopathogenic fungus Lecanicillium muscarium to control the sweetpotato whitefly, Bemisia tabaci has been established in the laboratory by other studies. Laboratory studies however frequently overestimate the level of control achieved by biological control agents in the glasshouse. Before full-scale commercial or field development is considered, glasshouse trials are required to confirm laboratory results. Under both controlled laboratory and glasshouse conditions high mortality of second instar B. tabaci was recorded after application of L. muscarium. The potential of incorporating L. muscarium into integrated pest management strategies for the control of B. tabaci is discussed.     

Chronic toxicity of cadmium toChironomus riparius Meigen — effects upon larval development and adult emergence

Chronic cadmium toxicity tests were carried out withChironomus riparius Meigen using a static-with-replacement exposure procedure. Significant reductions in larval development, survival and production were detected at 0.15 mg Cd l^–1 a value approximately 5000 times lower than the 48 h LC50 for fourth instar larvae. The first instar appears to be the most critical stage in terms of sensitivity, with the number of adults eventually emerging reflecting first instar mortality. Male emergence was slightly delayed at 0.15 mg Cd l^–1. The results of this study illustrate the importance of chronic toxicity tests in identifying sensitive life cycle stages and explaining pollutant-induced stress.     

Field Evaluation of Amitus bennetti (Hymenoptera: Platygasteridae), a Parasitoid of Bemisia argentifolii (Hemiptera: Aleyrodidae), in Cotton and Bean

Field cage experiments were conducted in Riverside, California to quantify the impact of releases of the parasitoid Amitus bennetti Viggiani & Evans on mortality of the whitefly Bemisia argentifolii Bellows & Perring. Single-row 50-m-long plots were planted with either cotton or bean. Cages were erected over the plants in each row, and adult whiteflies were released into the cages. Approximately 10 days later, adult parasitoids were released. Marked individual whiteflies were scored every 4 days for 6 weeks. Paired life tables were then constructed from census data from release and control cages over a single whitefly generation. Total whitefly mortality in release cages (71% in bean, 61% in cotton) was significantly greater than in control cages (25% in bean, 34% in cotton). The marginal rate for mortality attributable directly to the parasitism was 0.535 in the bean plots and 0.201 in the cotton plots. In addition, other mortality was greater in the release plots, possibly reflecting death of parasitized hosts before larval parasitoids could complete development. Parasitism was the greatest mortality factor in the study.     

Biological control of tobacco cutworm,Spodoptera litura Fabricius with entomopathogenic nematodes

The efficacies of several entomopathogenic nematodes ofSteinernema andHeterorhabditis spp. were examined against tobacco cutworm,Spodoptera litura Fabricius.H. bacteriophora HY showed 100% mortality after 20 h against 2nd instar of tobacco cutworm. In the case of 3–4th instar,S. carpocapsae PC.,H. bacteriophora HY andS. monticola CR showed 100% mortality after 47 h. In the case of 5–6th instar,S. carpocapsae PC proved more effective than the others. Generally, the number of nematodes harvested increased as their size decreased. Also, the highest number of nematodes was obtained in the 5–6th instar ofS. litura byH. bacteriophora HY, showing about 1.3×10⁶ nematodes per larva.In vitro culturedS. carpocapsae PG showed 100% mortality after 73 h against 5–6th instar tobacco cutworm, indicating that nematodes producedin vitro can be potentially used for the biological control ofS. litura instead of nematodesin vivo.     

Selection of Beauveria bassiana isolates for control of the whiteflies Bemisia tabaci and Trialeurodes vaporariorum on the basis of their virulence, thermal requirements, and toxicogenic activity

As part of a 3-fold approach to select potential mycoinsecticides for whitefly control, we evaluated infectivity, thermal requirements, and toxicogenic activity of the entomopathogenic fungus Beauveria bassiana (Ascomycota: Clavicipitaceae) under laboratory conditions. Twenty-five native B. bassiana isolates and a commercially available mycoinsecticide (based on B. bassiana) were evaluated for virulence to fourth instar nymphs of sweetpotato whitefly, Bemisia tabaci, and greenhouse whitefly, Trialeurodes vaporariorum, at a concentration of 1 × 10⁷ conidia/ml. All isolates were pathogenic for both whitefly species, whereas mortality rates varied from 3 to 85%. A second series of bioassays was conducted on 10 selected isolates using four 10-fold concentrations ranging from 1 × 10⁵ to 1 × 10⁸ conidia/ml. Median lethal concentrations (LC₅₀) of the four most virulent isolates varied from 1.1 × 10⁵ to 6.2 × 10⁶ conidia/ml and average survival time (AST) of treated nymphs from 5.9 to 7.4 days. T. vaporariorum were significantly more susceptible to all B. bassiana isolates than B. tabaci. The thermal biology of the eight most virulent isolates to both whitefly species was investigated at six temperatures (10–35 °C). The colony radial growth rate was estimated from the slope of the linear regression of colony radius on time and data were then fitted to a modified generalized β function that accounted for 90.5–99.3% of the data variance. Optimum temperatures for extension rate ranged from 23.1 to 27.1 °C, whereas maximum temperatures for fungal growth varied from 31.8 to 36.6 °C. On the basis of their virulence and thermal requirements, three isolates showed promise as candidates for whitefly management in Mediterranean greenhouses. Whilst in vitro production of macromolecular compounds toxic to Galleria mellonella larvae was not a requisite for virulence, ASTs of larvae injected with Sephadex G-25 fractions from candidate isolates ranged from 1.4 to 3.7 days compared with 5–6 days for non-toxic G-25 fractions. In addition, proteinase K treatment significantly reduced their toxic activity suggesting that they were proteins and revealing the potential of these isolates to be further improved through biotechnology to kill the pest more quickly.     

Selection of insect parasitic nematodes for biological control of the garden chafer,Phyllopertha horticola

Larvae ofPhyllopertha horticola L. (Coleoptera: Scarabaeidae) cause increasing problems on sports fields and lawns in NW-Europe. A biological control programme using insect parasitic nematodes is being developed. This paper contains the results of bioassays with various species and isolates of the nematode generaHeterorhabditis andSteinernema. In bioassays in small pots with moist sand, most of the nematode isolates gave 30–60% mortality against each of the three larval stages. The susceptibility of the grubs for nematode infection generally increased with larval development.H. bacteriophora, H. heliothidis, H. megidis, a DutchHeterorhabditis isolate NLH-E87.3 andS. glaseri 326 showed the highest mortality rates, with nearly 100% mortality of third instar grubs. The DutchHeterorhabditis isolate NLH-E87.3 andS. glaseri 326 were selected as candidates for further studies on their potential as biological control agents forP. horticola grubs in the field.     

Effects of azadirachtin after systemic uptake intoBrassica oleracea on larvae ofPieris brassicae

Petioles of intact cabbage leaves were placed in azadirachtin solutions for 24 h. Azadirachtin or active metabolites of this compound appeared to be translocated systemically as performance of first instar larvae ofPieris brassicae L. placed on disks from those leaves was affected. The position within a leaf from which disks were excised did not affect the extent of these effects, indicating a homogeneous distribution of active compounds in the laminar tissue. At doses ranging from 1–10 ppm of azadirachtin in the solution taken up by the leaves, gain of larval fresh weight and leaf area consumed were reduced. These effects were dose dependent up to 60–90 ppm. At 10 ppm and higher doses no larva reached the second instar within 72 h. Mortality during the 72 h period was highest at 10 ppm, due to the coincidence of lethal action with ecdysis in combination with the effect on developmental rate. Azadirachtin solutions applied directly to the surface of leaf disks in order to produce known doses on a leaf fresh weight basis gave results similar to the systemic treatment for weight gain, leaf area consumed and developmental rate, while mortality was higher in the latter treatment.     

Activité,in vivo, d''inhibiteurs de la biosynthèse de l''ecdysone chezLocusta migratoria

Summary Among about 50 compounds synthesized to inhibit enzymes involved in the biosynthesis pathway of ecdysone we selected seven molecules which showed a strong effect on ecdysone production byLocusta migratoria prothoracic glands incubatedin vitro. These molecules mostly possess a specific activity on ecdysone biosynthesis which is irreversible. The compounds were administered in one or several injections of aqueous or oily solutions at different times in the course of the two last larval instars. Three inhibitors led in a 10% ratio to a prolongation (sometimes more than 3 times the standard length) of the instar, pointing out a decrease in the ecdysone biosynthesis. Two other inhibitors induced some morphogenetic modifications in the adults, as size reduction or wing alterations, and metamorphosis difficulties. Thein vivo low activity compared with the strong onein vitro could be due to difficulties for the compounds to reach the prothoracic glands without degradation. The variation of inhibitory activity which appearsin vivo between the seven compounds studied is not linked either with the chemical structures of the molecules (which are very near one another) or with theirin vitro activity.

     

Influence of Clavicipitaceous Endophyte Infection in Ryegrass on Development of the ParasitoidMicroctonus hyperodaeLoan (Hymenoptera: Braconidae) inListronotus bonariensis(Kuschel) (Coleoptera: Curculionidae)

Laboratory experiments were conducted to examine the effect of ryegrass infection by the endophytic fungusAcremonium loliiLatch, Christensen and Samuels onMicroctonus hyperodaeLoan, a parasitoid ofListronotus bonariensis(Kuschel). Progression of parasitoids through the larval instar stages was shown to depend on adequate nutrition of the weevil host. Compared to confinement on endophyte-free ryegrass, parasitized weevils held on nonpreferred diets comprising leaf segments from endophyte-infected ryegrass and switchgrass contained parasitoid larvae with retarded development. Similarly, development of parasitoid larvae was retarded in hosts feeding on artificial diet containing diterpenes and alkaloids ofA. loliiorigin. Several diterpenes incorporated into the diet reduced survival of the parasitoid larvae. Attack rate of parasitoids was reduced when the quality of potential host weevils was compromised by confinement on nonpreferredA. lolii-infected ryegrass or without food for 14 days.     

Morphology and development of <Emphasis Type="Italic">Pteromalus cerealellae</Emphasis> (Ashmead) (Hymenoptera: Pteromalidae) on <Emphasis Type="Italic">Callosobruchus maculatus</Emphasis> (F.) (Coleoptera: Chrysomelidae)

Pteromalus cerealellae (Ashmead) (Hymenoptera: Pteromalidae) is an ectoparasitoid of several stored-product insect pests. Very little information has been published on its biology and development in host larvae, which typically are concealed within seeds. We documented the development of P. cerealellae within fourth instar larvae of its concealed host, Callosobruchus maculatus (F.) (Coleoptera: Chrysomelidae) infesting cowpea seeds. The preimaginal life stages of the parasitoid were characterized for the first time using morphological structures revealed by microscopic techniques including scanning electron microscopy. Pteromalus cerealellae produces hymenopteriform eggs and larvae. Eggs hatch into 13-segmented first instar larvae with peripneustic condition of spiracles. The larvae have simple, tusk-like mandibles, whereas the mandibles of the pupae and the adults are of the conventional toothed types. Using statistical analyses of the sizes of the larval mandibles and head capsules in conjunction with reliable characters such as the number of exuviae on the body of parasitoid larvae, cuticular folding, and excretion of the meconium, we recorded four larval instars for P. cerealellae. The data showed significant positive correlations between larval mandible lengths and widths of larval head capsules, as well as between mandible lengths and larval instars, suggesting that mandible length is a good predictor of the number of instars in P. cerealellae. Developmental time from egg to adult emergence was ∼12 d for females and ∼11 days for males at 30 ± 1°C, 70 ± 5% r.h. and 12L:12D photoperiod.     

爵床的甲醇提取物对小菜蛾的生物活性作用

为探明爵床(Justicia procumbens)甲醇提取物对小菜蛾的生物活性,采用室内生测法测定了爵床甲醇提取物对小菜蛾的触杀、拒食、胃毒、生长发育抑制和产卵忌避作用。结果表明,爵床甲醇提取物对小菜蛾幼虫具有较强的触杀、拒食、胃毒和生长发育抑制活性,对小菜蛾成虫具有较强的产卵忌避活性。在触杀试验中,药后1、2 d和3 d爵床甲醇提取物对小菜蛾3龄幼虫的致死中浓度(LC50)分别为5.17、4.05和3.06 mg/m L;在拒食试验中,药后1 d和2 d提取物对3龄幼虫的选择性拒食中浓度(AFC50)分别为2.64和3.13 mg/m L,药后1 d和2 d提取物对3龄幼虫的非选择性拒食中浓度(AFC50)分别为3.70、4.54 mg/m L;在胃毒试验中,药后4、5、6 d和7 d提取物对3龄幼虫的致死中浓度(LC50)分别为8.13、3.65、2.88、2.23 mg/m L;在生长发育抑制试验中,药后1 d和2 d提取物对3龄幼虫的抑制中浓度(IC50)分别为2.02、1.40 mg/m L;在产卵忌避试验中,药后1、2 d和3 d提取物对小菜蛾成虫的选择性产卵忌避中浓度(AOC50)分别为2.61、3.66、4.58 mg/m L,药后1、2和3 d提取物对小菜蛾成虫的非选择性产卵忌避中浓度(AOC50)分别为3.19、4.52、5.65 mg/m L。由此证实,爵床提取物对小菜蛾具有显著的毒杀活性,具有开发为新型高效、低毒植物源农药的潜在价值。     

Effectiveness of Heterohabditis bacteriophora strain GPS11 applications targeted against different instars of the Japanese beetle Popillia japonica

Field and laboratory tests were conducted from 2001 through 2007 to assess the effectiveness of entomopathogenic nematode Heterorhabditis bacteriophora strain GPS11 applications targeted against different instars of the Japanese beetle, Popillia japonica. During summer flight, P. japonica adults were trapped and caged on turfgrass plots for oviposition. Larval development was monitored for the occurrence of each instar. Nematodes were applied in the field against each developing instar at 2.5 × 10⁹ infective juveniles/ha. In 2001, field data obtained in October resulted in 75%, 53%, and 33% control with the applications targeted against the first, second, and third instars, 69, 28, and 9 days after treatment (DAT), respectively. In 2002 field trial, data obtained in October indicated 97%, 88%, and 0% control when the applications were targeted against the first, second, and third instars at 66, 43, and 14 DAT, respectively. Additional plots established in 2002 to determine efficacy against each instar at 14 DAT showed control of the first, second, and third instars to be 55%, 53%, and 0%, respectively. In laboratory tests conducted in 2002, 2004, and 2007, P. japonica collected from the field at the occurrence of each instar were exposed to H. bacteriophora at concentrations of 0, 10, 33, 100, 330, or 1000 infective juveniles/grub. Probit analysis of the mortality from three of the four sets of tests conducted showed the first instar to be significantly more susceptible to H. bacteriophora than the third instar at the LC₅₀ level and all tests showed the first instar to be significantly more susceptible than the third instar at the LC₉₀ level. In addition to the observed decrease in the third instar susceptibility to H. bacteriophora, soil temperatures in the mid-western United States during late September and October rapidly decline often reaching below 15 °C by the beginning of October when grubs are in the third instar stage of development. Therefore, we conclude that the applications of the nematodes made in August or September will provide higher control than those made in October, due to the more appropriate temperature for nematode activity and the presence of more susceptible larval stages. Early nematode applications may also provide an opportunity for nematodes to recycle and cause secondary infections.     

Temperature and the biology and predation of Ilione albiseta (Diptera: Sciomyzidae) — Potential biological control agent of liver fluke

The effect of temperature on predation by Ilione albiseta (Diptera: Sciomyzidae) on Lymnaea peregra was investigated at 14°, 17°, 20°, 23° and 26°C. The mean dry weight of snail tissue (Lymnaea peregra) attacked and consumed per day by first and second instar I. albiseta larvae was highest at 20°C while for third instar and total larval duration period it was greatest at 23°C. The mean number of snails killed per day during the third instar was also highest at 23°C. The total amount of snail tissue consumed by I. albiseta larvae increased significantly from first to second instar and from second to third instar at each constant temperature. Mean survival period of unfed first instar larvae decreased from 28.4 days at 14°C to 11 days at 26°C and the mean length of the second instar cephalopharyngeal skeleton decreased with increasing temperatures. As temperature increased the rate of consumption of oxygen (dissolved in water) by first and third instar larvae rose.     

Effects of <Emphasis Type="Italic">Paecilomyces fumosoroseus</Emphasis> and <Emphasis Type="Italic">Encarsia formosa</Emphasis> on the control of the greenhouse whitefly: preliminary assessment of a compatability study

The effect of separate and combined activity of Paecilomyces fumosoroseus Wize (Brown and Smith) Trinidadian strain T11 and the parasitoid, Encarsia formosa Gahan, was assessed on populations of the greenhouse whitefly, Trialeurodes vaporariorum (Westwood), infesting Phaseolus vulgaris L. (French bean) and Pelargonium x domesticum (regal geranium) plants in replicate experiments. When infested bean and geranium plants were exposed to E. formosa for 2 days, and 4 days later sprayed with P. fumosoroseus blastospores, whitefly percent mortality was 99.5% and 75.5%, 94.6% and 59.4% for experiments 1 and 2, respectively. Treatment of infested bean plants with either E. formosa or P. fumosoroseus resulted in 87.8% and 78.7%, 73.1% and 97.0% whitefly mortality for experiments 1 and 2, respectively, while similar treatment of infested geranium plants resulted in 9.2% and 52.8%, 34.3% and 64.5% whitefly mortality for experiments 1 and 2, respectively. Our results support the use of E. formosa and P. fumosoroseus in combination in Experiment 1 for the treatment of whitefly infested P. vulgaris plants since a significant difference in mortality is observed than when either E. formosa or P. fumosoroseus is applied alone. However, in experiment 2, the combination treatment on P. vulgaris was no more effective than spraying P. fumosoroseus alone. On P. x domesticum plants, only P. fumosoroseus alone is needed for efficient control of the whitefly compared to the combination treatment. The relative timing of parasitoid oviposition and fungal infection are critical in determining the outcome of the interaction and are plant host dependent.