Bt Proteins Have No Detrimental Effects on Larvae of the Green Lacewing, <Emphasis Type="Italic">Chrysopa pallens</Emphasis> (Rambur) (Neuroptera: Chrysopidae)

Biosafety of a genetically modified crop is required to be assessed prior to its commercialization. For this, a suitable artificial diet was developed and used to establish a dietary exposure test for assessing the toxicity of midgut-active Bt insecticidal proteins on Chrysopa pallens (Rambur). Subsequently, this dietary exposure test was used to evaluate the toxicity of the proteins Cry1Ab, Cry1Ac, Cry1Ah, Cry1Ca, Cry1F, Cry2Aa, Cry2Ab, and Vip3Aa on C. pallens larvae. Temporal stability, bioactivity, and the intake of the insecticidal proteins were confirmed by enzyme-linked immunosorbent assay and a sensitive-insect bioassay. The life history characteristics, such as survival, pupation, adult emergence, 7-day larval weight, larval developmental time, and emerged male and female fresh weights remained unaffected, when C. pallens were fed the pure artificial diet (negative control) and the artificial diets containing 200 μg/g of each purified protein: Cry1Ab, Cry1Ac, Cry1Ah, Cry1Ca, Cry1F, Cry2Aa, Cry2Ab, or Vip3Aa. On the contrary, all of the life history characteristics of C. pallens larvae were adversely affected when fed artificial diet containing boric acid (positive control). The results demonstrate that diets containing the tested concentrations of Cry1Ab, Cry1Ac, Cry1Ah, Cry1Ca, Cry1F, Cry2Aa, Cry2Ab, and Vip3Aa have null effects on C. pallens larvae. The outcome indicates that genetically modified crops expressing the tested Bt proteins are safe for the lacewing, C. pallens.     

Chromosome number variability in central european members of the<Emphasis Type="Italic">Festuca ovina</Emphasis> and<Emphasis Type="Italic">F. pallens</Emphasis> groups (sect.<Emphasis Type="Italic">Festuca</Emphasis>)

Chromosome numbers for 98 plants ofF. pallens, 19 ofF. psammophila, F. belensis andF. vaginata, and 44 ofF. ovina (originating from Austria, the Czech Republic, Germany, Slovakia and Latvia) are given. In addition to theF. ovina andF. pallens groups, chromosome counts for the following taxa are also reported:F. alpestris (2n=14) reported for the first time in this work,F. amethystina subsp.amethystina (2n=28),F. brevipila (2n=42),F. cinerea (2n=28),F. rupicola subsp.rupicola (2n=42) andF. versicolor subsp.versicolor (2n=14).InF. pallens, two ploidy levels (2n=2x=14+0-1B, 2n=4x=28+0-1B) as well as two natural triploid plants (2n=21+0-1B), were found. In addition to the fourF. pallens types that have been distinguished in Austria, one new tetraploid type (F. pallens “scabrifolia”) from the Czech Republic and Germany is reported and its taxonomy is discussed. The distributions of the Oberösterreich-Niederösterreich and Pannonisches-HügellandF. pallens types outside of Austria are documented.Only the diploid chromosome number (2n=14) was found inF. psammophila andF. vaginata. Chromosome numbers forF. psammophila subsp.muellerstollii andF. belensis (both 2n=14) were determined here for the first time. Two ploidy levels, 2n=14+0-5B corresponding toF. ovina subsp.ovina and 2n=28 corresponding toF. ovina subsp.guestphalica andF. cf.duernsteinensis were confirmed inF. ovina. Differences in chromosome structure (simple and multiple secondary constrictions) betweenF. pallens as opposed toF. psammophila andF. vaginata are discussed. A complete survey of published chromosome counts for Central European species from theF. ovina andF. pallens groups is included.     

Combined action of the entomopathogenic fungus <Emphasis Type="Italic">Metarhizium robertsii</Emphasis> and avermectins on the larvae of the colorado potato beetle <Emphasis Type="Italic">Leptinotarsa decemlineata</Emphasis> (Say) (Coleoptera,Chrysomelidae)

Synergy between the entomopathogenic fungus Metarhizium robertsii J.F. Bisch., S.A. Rehner et Humber and a complex of natural avermectins was observed after combined treatment of the Leptinotarsa decemlineata (Say) larvae. This effect was also recorded at the larval stage most resistant to fungi (5 days after molting to the IV instar). Intoxication with avermectins led to a 2.7-fold decrease in food consumption and arrest of larval development. Synergy between the fungus and avermectins was revealed in field experiments conducted in the steppe zone of Western Siberia under strong fluctuations of temperature and humidity. In particular, the median lethal time (LT₅₀) decreased 2.8-fold after combined treatment as compared to treatment with M. robertsii alone. The mechanisms of synergy and the prospects of using the combined treatment for Colorado potato beetle control are discussed.     

Dynamics of host plant selection and host-switching by silver-spotted skipper caterpillars

Investigations of host plant selection in herbivorous arthropods have emphasized the importance of oviposition site selection by adults; however, a more complete picture of this process requires additional consideration of the factors influencing host plant choice during the immature feeding stages. We conducted a series of larval choice experiments to examine both the innate and induced preferences of larvae of the Silver-spotted skipper (Epargyreus clarus L.) on three commonly used hosts (Wisteria, Robinia, and Pueraria). Late instar E. clarus larvae reared on each of the three host plants displayed an overall pattern of innate preferences that correlated well with larval performance measures and reflected differences in foliar nutrient concentrations. Larval preferences were also influenced by rearing host species, indicating a role for feeding-induced preferences. When larvae reared on low-quality Wisteria for the first four instars were switched to higher quality Pueraria for the final instar, they developed more quickly and attained significantly higher pupal mass than larvae maintained on Wisteria throughout development. Similarly, larvae switched from Pueraria to Wisteria for the final instar suffered increased development time and produced significantly smaller pupae than those maintained on Pueraria throughout. Thus host-switching, particularly during the more mobile final instars, appears to offer larvae an opportunity to recoup fitness losses associated with early development on a low-quality host. For an equal amount of consumption, larvae feeding on Pueraria gained 50% more mass than those feeding on Wisteria, reflecting measured differences in foliar nitrogen concentration; despite these overall differences in quality, larval growth efficiency was similar among hosts. Especially in the age of common exotic plant introductions, a full understanding of the behavioral component of host selection by herbivorous insects requires appreciation of the dynamic role that immatures can play in host selection and use.     

Seasonal Abundance and Predatory Potential of <Emphasis Type="Italic">Stethorus aptus</Emphasis> Kapur (Coleoptera: Coccinellidae): A Biocontrol Agent of Tea Red Spider Mite <Emphasis Type="Italic">Oligonychus coffeae</Emphasis> Nietner (Acarina: Tetranychidae)

The tea red spider mite, Oligonychus coffeae Nietner is one of the major pests of tea plants in North-east India. The ladybird beetle, Stethorus aptus Kapur, is a newly reported predator of O. coffeae. Predatory efficiency study of S. aptus under laboratory conditions revealed that adult of S. aptus consumed significantly more mites than larvae. In free choice condition, the predator consumed within a range of 48–56 adults and 82–90 larvae of O. coffeae whereas the 3rd and 4th instar larvae of S. aptus consumed 20–26 adults and 50–60 larvae of O. coffeae per day respectively. Population dynamics of S. aptus was observed for 1 year under field conditions. The maximum density of the predator was recorded during January to March and it gradually declined from September onwards. Population of S. aptus showed positive correlation with its prey O. coffeae and relative humidity while effect of other factors was insignificant.     

Biological Data on <Emphasis Type="Italic">Anovia punica</Emphasis> Gordon (Coleoptera: Coccinellidae), a Predator of <Emphasis Type="Italic">Crypticerya multicicatrices</Emphasis> Kondo &amp; Unruh (Hemiptera: Monophlebidae)

The coccinellid beetle Anovia punica Gordon (Coleoptera: Coccinellidae: Noviini) is an important predator of the Colombian fluted scale, Crypticerya multicicatrices Kondo & Unruh (Hemiptera: Monophlebidae). In order to gather information on the biological traits of A. punica, we conducted a series of studies, including of the developmental time of each life history stage, estimation of life table parameters, and predation rates under laboratory conditions [25.1 ± 1.6°C, with 70.5 ± 7.3% RH, and natural light regime, approx. 12:12 (L:D) h]. Developmental stages of A. punica were categorized as follows: egg stage, four larval instars, prepupal instar, pupal instar, and adult. Developmental time from egg to adult emergence averaged 29.41 ± 1.85 days, and 47.6% of the eggs developed to adulthood. Female and male survival was 94.42 and 90 days, respectively. Life table parameters show that one female of A. punica is replaced by 86 females (R ₀), the intrinsic growth rate (r _m ) was 0.1115, the average generation time (T) was 40 days, and the doubling time (D _t ) was 6.2 days. The life table parameters suggest that A. punica can be used as a potential predator of C. multicicatrices and, more importantly, provided baseline information for a mass-rearing protocol. This is the first detailed study on the biology of A. punica that reports the potential of this predator as a biological control agent for scale insects of the tribe Iceryini.     

Discovery of a Highly Virulent Strain of <Emphasis Type="Italic">Photorhabdus luminescens</Emphasis> ssp<Emphasis Type="Italic">. akhurstii</Emphasis> from Meghalaya,India

Photorhabdus is an insect-pathogenic Gram negative enterobacterium found in the gut of Heterorhabditis nematodes. Photorhabdus is highly virulent to insects, and can kill insects rapidly upon injection at very low concentrations of one to few cells. We characterized the virulence of Photorhabdus symbionts isolated from the Heterorhabditis nematodes collected from various parts of India by injecting different concentrations of bacterial cells into fourth instar larval stage of insect Galleria mellonella. Photorhabdus luminescens ssp. akhurstii strain IARI-SGMG3 from Meghalaya was identified as the most virulent of all the tested strains on the basis of LT₅₀ and LC₅₀ values. This study forms a basis for further investigations on the genetic basis of virulence in Photorhabdus bacteria.     

Selection and Characterization of <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> (Berliner) (Eubacteriales: Bacillaceae) Strains for <Emphasis Type="Italic">Ecdytolopha aurantiana</Emphasis> (Lima) (Lepidoptera: Tortricidae) Control

The citrus fruit borer, Ecdytolopha aurantiana (Lima, 1927) (Lepidoptera: Tortricidae), is responsible for major losses to the citrus industry because it causes rot and drop of fruits. The current study aimed to select and characterize Bacillus thuringiensis (Berliner, 1911) strains toxic to E. aurantiana. For this purpose, 47 B. thuringiensis strains were evaluated in selective bioassays using first instar larvae of E. aurantiana. The lethal concentration (LC₅₀) of the most toxic strains was estimated, and the strains were characterized by morphological, biochemical, and molecular methods. Of the 47 strains tested, 10 caused mortality above 85% and showed mean lethal concentrations between 1.05E+7 and 1.54E+8 spores mL^?1. The lowest LC₅₀ values were obtained for the HD-1 standard strain and the BR145, BR83, BR52, and BR09 strains. The protein profile showed the presence of Cry proteins of 60, 65, 70, 80, and 130 kDa. The molecular characterization showed the presence of cry1, cry2, cry3, and cry11 genes. The morphological analysis identified three different crystalline inclusions: bipyramidal, round, and cuboidal. The cry1 and cry2 genes were the most frequent among the B. thuringiensis strains evaluated and encode Cry proteins toxic to insects of the order Lepidoptera, which agree with the toxicity results obtained by the selective bioassays against E. aurantiana. The results showed four different B. thuringiensis strains toxic to E. aurantiana at the same level as the HD-1 standard strain, and these strains have biotechnological potential for E. aurantiana control through the production of transgenic plants or the formulation of biopesticides.     

Suitability of Different Prey Aphids on the Growth,Development and Reproduction of <Emphasis Type="Italic">Chrysoperla zastrowi sillemi</Emphasis> (Esben-Petersen) (Chrysopidae: Neuroptera)

The prey preference of polyphagous predator, green lacewing (Chrysoperla zastrowi sillemi (Esben-Petersen)) was evaluated against five prey aphids viz., mustard aphid (Lipaphis erysimi), green peach aphid (Myzus persicae), cabbage aphid (Brevicorynebrassicae), black bean aphid (Aphis craccivora), spirea aphid (Aphis spiraecola) of agriculture importance and compared with eggs of Corcyracephalonica (Stainton). Lacewing larvae preferred Myzus persicae most followed by Brevicorynebrassicae. The highest growth index (8.31), larval survival (94.50 %), larval weight (10.45 mg), pupal weight (8.78 mg), faster multiplication rate (0.051) and fecundity (183.4 per gravid female) of the predator were recorded on M. persicae. However, the chrysopid reared on Corcyra eggs performed best in all biological parameters and fitness, than on aphid preys. This study explores the possibilities of selecting the most suitable prey aphid species for its exploitation as supplement for mass multiplication of chrysopid during off-season or unavailability of Corcyra eggs.     

Evaluation of <Emphasis Type="Italic">Nastus fausti</Emphasis> Reitter (Coleoptera: Curculionidae: Entiminae: Nastini) for biological control of invasive giant hogweeds (<Emphasis Type="Italic">Heracleum</Emphasis> spp.)

The weevil Nastus fausti Reitter (Coleoptera, Curculionidae) was evaluated for its potential in the biological control of invasive giant hogweeds (Heracleum spp.). Quantitative sampling suggested that at a high population density (more that 3–4 mature larvae per plant) damage by N. fausti larvae could have some negative impact on the above-ground part of the plant. However, no-choice laboratory tests showed that N. fausti females were able to feed on a number of Apiaceae genera, including such important cultivated crops as carrot, parsnip, and celeriac. Feeding on these plants did not cause any significant decrease in female survival or fecundity. Moreover, at least part of N. fausti larvae may feed and develop on roots of these plants, and the rate of their growth and development does not differ significantly from that in larvae fed on roots of H. mantegazzianum. N. fausti adult and larval feeding on Angelica purpurascens, representative of related genus of the same tribe, was recorded under natural conditions, too. In combination, these data suggest that N. fausti is an oligophagous species connected with plants from at least several genera of Apiaceae and thus it cannot be considered a potential agent for biological control of invasive Heracleum species.     

The seasonal development cycle of <Emphasis Type="Italic">Aphantopus hyperantus</Emphasis> (L.) (Lepidoptera,Nymphalidae: Satyrinae) in Leningrad Province

The simultaneous influence of the photoperiodic and temperature conditions on pre-diapause and postdiapause larval development of the ringlet Aphantopus hyperantus was studied. At the short day (12 h of light a day) all larvae reached the III instar and entered diapause at all the temperatures tested (18–24°C). At the long day (22 h) the number of diapausing larvae increased with decreasing temperature, and the larvae diapaused at the III and IV instars. The I and II instar larvae (before winter) developed and grew faster under short-day, and the hibernated larvae, under long-day conditions. At the short day the growth rate of the I and II instar larvae did not depend on the temperature, and at the long day it decreased as the temperature increased. After hibernation the growth rate of the IV instar larvae increased with the temperature under long-day conditions, and did not depend on the temperature under short-day ones. Thus, acceleration or deceleration of development, depending on the photoperiodic and temperature conditions, regulated the timing of diapause in the overwintering stage (the III instar larva) and maintained the univoltine seasonal cycle. The males of A. hyperantus developed and grew faster than females while their weight at the larval and pupal stages was smaller. The thermal reaction norms for development (the lower temperature threshold and the coefficient of thermal sensitivity of development) were calculated for the eggs, IV and V instar larvae, and pupae. The coefficient of thermal sensitivity of development for the IV and V instar larvae taken together was lower, and the threshold was higher than the corresponding values for eggs and pupae. The thermal reaction norms for development of the latter stages were the same. The larvae of A. hyperantus retained their sensitivity to the photoperiod after hibernation. Resumption of development after cold reactivation occurred only under long-day conditions, whereas under short-day ones the overwintered larvae entered a repeated diapause. The possibility of photoperiodic reactivation and the absence of spontaneous resumption of development in these larvae were demonstrated.     

Acaricidal effect of <Emphasis Type="Italic">Schinus molle</Emphasis> (Anacardiaceae) essential oil on unengorged larvae and engorged adult females of <Emphasis Type="Italic">Rhipicephalus sanguineus</Emphasis> (Acari: Ixodidae)

The current concern about resistance to acaricides and the impact of toxic waste on the environment has led to the search of vegetal alternatives in the control of the brown tick of the dog Rhipicephalus sanguineus. Schinus molle L. (Anacardiaceae) derivatives have been associated with insecticidal, antimicrobial and antiprotozoal activities and essential oil showed to be lethal to R. microplus larvae. This study aimed at evaluating the acaricidal effect of essential oil of S. molle (EO_Sm) on engorged adult females and larval stages of R. sanguineus. One-hundred engorged females were obtained from the ears, interdigital spaces, neck, groin and base of the tail of two cross-bred dogs. The larvae package test was accomplished with 21-day-old larvae and five concentrations (v/v) of EO_Sm (0.125, 0.25, 0.50, 1 and 2%) in an anionic detergent, a synthetic acaricide (cypermethrin) and detergent and deionized water as controls. The immersion adult test was carried out with nine concentrations (0.125, 0.25, 0.50, 1, 2, 4, 8, 16, 20%) of the EO_Sm. At the concentration of 2%, EO_Sm caused 99.3% of larval mortality. In adults, inhibition of oviposition, egg hatching (EH) and reproductive efficiency (RE) values were dose-dependent from 4 to 20% EO_Sm; the lowest values of EH (29.62) and RE (22.61) were achieved with 20% EO_Sm. Strong and negative correlations were found between concentration of EO_Sm and EH (r?=???0.948) and between concentration of EO_Sm and RE (r = ??0.985). This study demonstrated for the first time the acaricidal effect of EO_Sm on larvae and reproductive parameters of engorged adult females of R. sanguineus.     

Developmental dynamics of myogenesis in the shipworm <Emphasis Type="Italic">Lyrodus pedicellatus</Emphasis> (Mollusca: Bivalvia)

Background

The shipworm Lyrodus pedicellatus is a wood-boring bivalve with an unusual vermiform body. Although its larvae are brooded, they retain the general appearance of a typical bivalve veliger-type larva. Here, we describe myogenesis of L. pedicellatus revealed by filamentous actin labelling and discuss the data in a comparative framework in order to test for homologous structures that might be part of the bivalve (larval) muscular ground pattern.

Results

Five major muscle systems were identified: a velum retractor, foot retractor, larval retractor, a distinct mantle musculature and an adductor system. For a short period of larval life, an additional ventral larval retractor is present. Early in development, a velum muscle ring and an oral velum musculature emerge. In late stages the lateral and dorsal mantle musculature, paired finger-shaped muscles, an accessory adductor and a pedal plexus are formed. Similar to other bivalve larvae, L. pedicellatus exhibits three velum retractor muscles, but in contrast to other species, one of them disappears in early stages of L. pedicellatus. The remaining two velum retractors are considerably remodelled during late larval development and are most likely incorporated into the elaborate mantle musculature of the adult.

Conclusions

To our knowledge, this is the first account of any larval retractor system that might contribute to the adult bodyplan of a (conchiferan) mollusk. A comparative analysis shows that a pedal plexus, adductors, a larval velum ring, velum retractors and a ventral larval retractor are commonly found among bivalve larvae, and thus most likely belong to the ground pattern of the bivalve larval musculature.

     

Laboratory and simulated-field bioassays for assessing mixed cultures of <Emphasis Type="Italic">Lysinibacillus sphaericus</Emphasis> against <Emphasis Type="Italic">Aedes aegypti</Emphasis> (Diptera: Culicidae) larvae resistant to temephos

Aedes aegypti (L.) is the main vector of tropical diseases such as dengue, chikungunya and Zika. Due to the overuse of insecticides, Ae. aegypti resistant populations have increased. Biological control with Lysinibacillus sphaericus (Ahmed) has been used against Culex sp. and Anopheles sp. Although Ae. aegypti is refractory to the binary toxin of L. sphaericus spores, vegetative cells have been shown to be effective against Ae. aegypti larvae. In this work, the effect of L. sphaericus vegetative cells on Ae. aegypti temephos-resistant larvae was assessed under lab and simulated field conditions. L. sphaericus caused about 90% mortality of insecticide-resistant Ae. aegypti larvae under simulated field conditions. Likewise, Ae. aegypti larvae were more sensitive to mixed cultures of L. sphaericus than to individual strains; then, the most effective mixed culture exhibited an LC₅₀ of 1.21 × 10⁵ CFU/mL with Rockefeller larvae and 8.04 × 10⁴ CFU/mL with field-collected larvae. Additionally, we found that mixed cultures composed of two L. sphaericus strains were more effective than a culture formed by the three strains. Our results suggest that mixed cultures comprising L. sphaericus vegetative cells could be useful for controlling temephos-resistant populations of Ae. aegypti, as evidenced by the effectiveness demonstrated under laboratory and simulated field conditions.     

On distribution and ecology of some species of cleonines (Coleoptera,Curculionidae): IV. Genus <Emphasis Type="Italic">Lixus</Emphasis> F., subgenus <Emphasis Type="Italic">Eulixus</Emphasis> Reitt.

Data on habitats, food plants of larvae and adults, feeding, mating, oviposition, larval and pupal development, natural enemies, and distribution of seven weevil species (Lixus canescens F.-W., L. iridis Ol., L. myagri Ol., L. punctirostris Boh., L. subtilis Boh., L. incanescens Boh., and L. brevipes Bris.) are given. New host plants of L. canescens and L. iridis are revealed. Gall induction by L. brevipes is reported for the first time. Distribution of all the species in Ukraine and Russia (the latter based on the literature) are given in more detail. Information on the known and potential economic importance of every species is provided.     

Rate of consumption,biological parameters,and population growth capacity of <Emphasis Type="Italic">Orius laevigatus</Emphasis> fed on <Emphasis Type="Italic">Spodoptera exigua</Emphasis>

Orius laevigatus (Fieber) (Hemiptera: Anthocoridae) is already successfully used to control Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) in protected crops. In the present work, the predatory capacity of O. laevigatus on the eggs and larvae of Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) as well as the population growth capacity of O. laevigatus fed on eggs of the noctuid were determined. Fourth and 5th instar nymphs had a greater consumption of eggs than younger ones. Females consumed more eggs than males. Only 5th instar nymphs and adults preyed on the neonatal larvae of S. exigua. The intrinsic rate of natural increase did not differ between O. laevigatus fed with S. exigua eggs and those offered eggs of the substitute host Ephestia kuehniella (Zeller) (Lepidoptera: Pyralidae). Thus, O. laevigatus is a good candidate for the biological control of S. exigua, a cosmopolitan pest of many crops.     

Parasitism of <Emphasis Type="Italic">Spodoptera litura</Emphasis> (Lepidoptera: Noctuidae) by <Emphasis Type="Italic">Microplitis prodeniae</Emphasis> (Hymenoptera: Braconidae)

Spodoptera litura Fabricius is a major vegetable pest that is widely distributed throughout tropical, subtropical and temperate regions. Microplitis prodeniae Rao and Chandry is a solitary endoparasitoid of S. litura. To assess the potential use of this parasitoid as a biological control agent, the reproductive schedule, fecundity and functional response of M. prodeniae were investigated under conditions of 28 ± 1°C and 70 ± 10% relative humidity with a 14:10-h L:D photoperiod. The parasitoid’s average lifetime fecundity was 171.0 ± 10.4 eggs, of which approximately 50% were laid within the first 3 days. Additionally, M. prodeniae exhibited a Holling type II functional response, and the estimated maximum numbers of the 1st, 2nd and 3rd instar larvae that were parasitized by a single M. prodeniae female were 71.6, 78.4 and 41.5 larvae over a 24-h period, respectively. The results of this study suggest that M. prodeniae has great potential as a candidate for controlling S. litura and can guide efforts in its mass production.