Direct selection of mutants of Drosophila resistant to juvenile hormone analogues

A direct screening technique was devised in which applications of a juvenile hormone analogue (JHA) (RO-20-3600; ENT No. 70357; 6,7-Epoxy-3-methyl-7-ethyl-1-(3,4-[methyl-enedioxy]-(phenoxy)-2-cis/trans-octene), Hoffman-LaRoche) to the food resulted in the death of all the F-1 descendants of mutagenized Drosophila melanogaster males with the exception of those animals which possessed sexlinked mutations affecting the metabolism of JHA. These exceptions would comprise (a), those mutant animals which possess any resistance to the lethal effects of the JHA (JH^R mutants) and/or (b), those mutant animals which have a defective JH metabolism and would normally die but are rescued by the exogenous application of JHA (JH^? mutants). Approximately 95, 850 mutagenized chromosomes were tested in this screen, 99,6% of which were killed by the JHA. Of the surviving 181 fertile males, 6 were found to be JH^R mutants. The statistically significant resistance factors ranged from 1.4X to 21.5X. The analysis of the most resistant mutant strain indicates that it may possibly be a behavioral mutant. The implications of such JH^R mutants are discussed in terms of ‘third generation pesticides’.     

Penetration and persistence of an insect growth regulator in the pupa of the stable fly, Stomoxys calcitrans.

An insect growth regulator, (E)-6,7-epoxy-1-(p-ethylphenoxy)-3,7-dimethyl-2-octene (Stauffer R-20458) with insect juvenile hormone activity readily penetrated the puparium of white pharate pupae of Stomoxys calcitrans. The quantity and the persistence of the chemical in the puparium and in the developing pharate adult were determined over a 168 hr post-treatment period. The loss of chemical from the pharate stage was almost total within 168 hr, but over 20 per cent of the applied dose was still present in the puparium after the same interval.     

The activity of juvenile hormone mimics for the eggs of Rhodnius prolixus

Twenty four juvenile hormone (JH) mimics of several different chemical classes were applied to freshly laid eggs of Rhodnius prolixus. Activity of the JH mimics was assessed in terms of their ability to inhibit eclosion. All chemical classes tested except one of the natural JHs and two very closely related long chain compounds contained members which inhibited eclosion; the most active chemicals were derivatives of geranyl para-aminobenzoic acid esters and aryl methylene dioxyphenyl ethers. In general, those chemicals of a particular class which had highest morphogenetic activity when applied to last instar larvae, were also most active in inhibiting eclosion. However, not all structure/activity relationships of JH mimics for last instar larvae were applicable to eggs; in particular the presence of a terminal epoxide group increased activity greatly for eggs but only slightly for last instar larvae. The dose per unit weight to inhibit eclosion is far greater than the equivalent dose to disrupt normal metamorphosis.     

Protection of potato crop against Phthorimaea operculella (Zeller) infestation using frass extract of two noctuid insect pests under laboratory and storage simulation conditions

The oviposition deterrent effect of water extract of Spodoptera littoralis and Agrotis ipsilon larval frass on Phthorimaea operculella adult females was studied using two types of larval food “Natural host and Semi-artificial diet” under laboratory and storage simulation (semi-field) conditions. Extracted frass of fed larvae on semi-artificial diet showed complete oviposition deterrent effect at treatments with 4th, 5th and 6th instars of S. littoralis, also at treatments with 1st–3rd and 6th instars of A. ipsilon, while the same effect was observed when the larvae fed on castor oil leaves as a natural host only at treatment with frass extract of A. ipsilon 6th instar larvae. Presence of low amounts of phenols and flavonoids in water extract of A. ipsilon larval frass resulted in relatively more effect as oviposition deterrent to fertile adult females on treated oviposition sites, while the opposite effect was obtained in S. littoralis larval frass experiments. At semi-field experiments, the percentage reduction of laid eggs reached 100% after two?days at treatments with frass extracts of 4th and 5th S. littoralis larval instars and A. ipsilon 6th instar larvae fed on semi-artificial diet and/or castor oil leaves. Percentage reduction of laid eggs for untreated sacks reached 93.24 and 48.95% after 2 and 30?days, respectively, when placed between treated sacks, in comparison with the mean number of laid eggs for isolated control.     

Activation of the prothoracic gland by juvenile hormone and prothoracicotropic hormone in Mamestra brassicae

The effects of JHA (ZR-515) application or brain implantation on metamorphosis and adult development were examined in the last instar larvae and pupae of Mamestra brassicae. When JHA was applied to neck-ligated 4- or 5-day-old larvae or to the isolated abdomens of 5-day-old larvae containing implanted prothoracic glands taken from 5-day-old larvae, the insects pupated. Dauer pupae and diapausing pupae treated with JHA showed adult development. By contrast, pupation could not be induced by the application of JHA to 2- or 3-day-old neck-ligated larvae or to the isolated abdomens of 5-day-old larvae containing implanted prothoracic glands from 0-day-old larvae. Implantation of a brain into neck-ligated 3- or 5-day-old larvae (at the beginning of gut emptying and wandering) caused pupation of the host. A similar result was obtained when both a brain and the prothoracic glands from 0- or 5-day-old larvae were implanted into the isolated abdomens of 5-day-old larvae. These results indicate that activation of the prothoracic glands by application of JHA is temporally restricted to the last part of the last larval instar and to the pupal stage, while the activation by prothoracicotropic hormone (PTTH) can occur throughout the last larval instar and the pupal stage. In addition, the implantation of brains or application of JHA to neck-ligated 5-day-old larvae 25 days after ligation seldom induced pupation of the hosts, a result which suggests that larval prothoracic glands maintained under juvenile hormone (JH) or PTTH-free conditions for long periods of time may become insensitive to reactivation by both hormones.     

Relationship between an increase of juvenile hormone titer in early instars and the induction of diapause in fully grown larvae of Sesamia nonagrioides

The larvae of Sesamia nonagrioides (Lepidoptera: Noctuidae) grown at 25 degrees C and long photoperiod (16:8h light:dark) pupate in the 5th or 6th (mostly) larval instar, while the larvae reared under a short photoperiod (12:12h) enter diapause during which they consume some food and undergo up to 12 (usually 3-4) stationary larval molts. Diapause programming includes an increase of juvenile hormone (JH) titer in the hemolymph from about 20 to 50 nM in the 4th and 5th instar larvae (titer in earlier instars was not measured). JH I, II, and III are present in approximate ratio 1-2:10:1. The JH titer drops to zero before pupation but remains around 20 nM during diapause. Perfect extra larval molts associated with a body weight increase can be induced in the non-diapausing larvae with a JH analogue (JHA). The weight rise is due to accumulation of reserves and not to a general body growth. The timing of extra molts is similar to the molting pattern of the diapausing larvae only when JHA is present since early larval instars. In the diapausing larvae, JHA application affects neither molting periodicity nor the body weight. It is concluded that (1) Increased JH titer in early larval instars is a part of diapause programming; (2) The extension of larval stage in the diapausing larvae, but not the timing pattern of extra molts, is due to continuously high JH titer; (3) The diapause program includes low food intake, maintenance of a certain body weight, and periodic larval molts.     

A radioimmunoassay for chironomid hemoglobin

A double antibody, competitive radioimmunoassay was developed for the quantitation of stage-specific hemoglobin from the insect Chironomus thummi. The radioimmunoassay will detect as little as 150 picograms of Hb 3, a hemoglobin synthesized and secreted into the hemolymph of larvae during the 4th, but not the 3rd instar. The assay also detects cross-reacting hemoglobins purified from 4th instar larvae and in freshly laid eggs. Application of this radioimmunoassay is discussed in light of the cross-reacting Hbs in the insect.     

Food consumption,utilization and ecological efficiency of Parthenium beetle,Zygogramma bicolorata pallister (Coleoptera: Chrysomelidae)

Food consumption, utilization and ecological efficiency of Zygogramma bicolorata were studied in larvae and adults up to 45 days old. The study revealed that sex had a significant influence on dry food consumption in adults but not in larvae. The other parameters of ecological efficiencies, viz. efficiency of conversion of ingested food (E.C.I.), tissue growth and dry biomass, were highest in the fourth instar and in adult females. The Consumption Index (C.I.) was higher in males than females in both larval and adult stages. Developmental and growth rates were relatively higher in the first and second instars. The fourth instar and adult females were the most efficient consumers and converters of food. Food intake by females was an important determinant of the number of eggs laid.     

Contrasting egg and larval performances help explain polyphagy in a florivorous butterfly

We examined both egg and larval performances in the polyphagous butterfly Parrhasius polibetes (Lycaenidae) using two host plants differing in morphological and ecological traits. Oviposition on mixed and pure patches of Schefflera vinosa (Araliaceae) and Pyrostegia venusta (Bignoniaceae), as well as the fate of eggs laid on both hosts, was assessed. To disentangle the effects of egg origin and host quality on larval performance, eggs were collected from Schefflera and Pyrostegia, and the corresponding newly hatched larvae were reared either on the natal (control) or non-natal (experimental) host. Lastly, we evaluated whether early and late instars are able to switch to alternative hosts. In both mixed and pure patches, parasitism was significantly lower, and oviposition and hatching rates were significantly higher for eggs laid on Schefflera than on Pyrostegia. Survivorship did not differ among treatments. Larvae fed with Pyrostegia were heavier than those fed with Schefflera, regardless of egg origin. Only early instars fed with Schefflera switched to Pyrostegia in the tests; in the remaining cases, larvae fed on the alternative hosts significantly less than on the controls. Our results help to explain why the use of multiple hosts by P. polibetes is maintained in nature, as the host conferring superior egg survival may incur poor larval performance and vice versa. Oviposition pattern is better understood from a tri-trophic rather than a bi-trophic perspective. Our study also highlights the monophagous condition of individual P. polibetes larvae; the constraint for switching to novel hosts is dependent on both larval instar and host plant species.     

The role of juvenile hormone during larval-pupal transformation of Spodoptera littoralis: Switchover in the sensitivity of the prothoracic gland to juvenile hormone

Application of JHA to the final instar of Spodoptera littoralis larvae before they attain maximal body weight causes delay to or completely blocks the onset of metamorphosis, as indicated by the dorsal vessel exposure. When the same dose of JHA is administered to the larvae just after their attaining maximal body weight, at the time of the onset of gut-emptying, metamorphosis is substantially accelerated.By means of ligature experiments, it is shown that JHA has a direct effect on the prothoracic glands. It inhibits them at the beginning of the final instar and then stimulates them shortly before pupation. It seems that under normal conditions JH is necessary for pupal cuticle formation; those larvae which pupate without JH show adult characteristics.     

Responses of Episyrphus balteatus DeGeer (Diptera: Syrphidae) in relation to prey density and predator size

Laboratory experiments were conducted to determine the functional and numerical responses of the aphidophagous hover fly Episyrphus balteatus DeGeer (Diptera: Syrphidae) to different densities of 4th instar black bean aphids, Aphis fabae Scopoli (Homoptera: Aphididae), on broad bean, Vicia faba L. (Fabaceae). Two different-sized larvae of predators were tested in different densities of similar-sized prey to determine whether functional response parameters depended on the body sizes of predator and prey. In numerical response experiments, gravid E. balteatus females were exposed individually to different densities of 4th instars of A. fabae on cut sections of the broad bean plant, V. faba L. Logistic regression suggested a type II functional response for both larval sizes of E. balteatus. The searching efficiency (a) of the larger larvae was higher than that of the smaller ones. Prey consumption was higher, and handling time (T_h) was lower for larger larvae than smaller ones. The theoretical maximum number of A. fabae nymphs eaten by the different-sized larvae was 125 and 269 nymphs per day. Larger E. balteatus larvae are more efficient predators for aphid management strategies. The reproductive numerical response, in terms of the number of eggs laid, increased curvilinearly with increasing prey density, but the proportion of eggs laid (egg number/prey density) decreased as the initial density of prey increased.     

Competence of Macrotermes michaelseni (Isoptera: Macrotermitinae) larvae to differentiate into soldiers under the influence of juvenile hormone analogue (ZR-515, methoprene)

It has been shown that only third instar larvae of Macrotermes michaelseni have the competence to differentiate into presoldiers under the influence of juvenile hormone analogue (JHA). The timing of events leading to presoldier formation was independent of JHA dose above the threshold. Further studies with homogeneous groups of third instar larvae of different ages showed that only larvae of a certain age (0–6 days) could respond to topically applied JHA to produce presoldiers and intercastes (intermediate forms). Older larvae did not respond, hence, 0–6 days interval is the competence period for presoldier differentiation in this species. It seems also that the corpora allata of those individuals which differentiate into presoldiers become activated during the competence period, possibly by the parents or other means.     

Development of Microplitis bicoloratus on Spodoptera litura and implications for biological control

Microplitis bicoloratus Chen (Hymenoptera:Braconidae:Microgastrinae), a new species of Microplitis Förster from China, is a solitary endoparasitoid of the larvae of the cotton leafworm, Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae). This parasitoid is the first to be successfully reared and evaluated in the laboratory as a potential agent for the biological control of S. litura in China. Oviposition, immature development, and the effects of parasitism on the development of S. litura were studied. In long-term oviposition trials, females laid eggs on S. litura larvae for up to 10 days; oviposition was heavily skewed toward the first few days, with approximately one third of the eggs laid on day 1 and over 50% laid by day 3. This rapid oviposition rate increases the potential for biological suppression of host populations because the likelihood of mortality for the parasites from exposure to detrimental environmental factors or generalist predators increases with time. Immature development of the parasitoid in its host only required 7 days: eggs hatched within 24 h, the first instar larva required 2 days, the second instar larva needed 3 days, and the third instar larvae exited the host and pupated in 1 day, at 27±1°C, 60–80% relative humidity and a 12:12-h (long day) photoperiod. The development of the parasitized hosts was disrupted. When the parasitoid larvae finished development, the body weights of host larvae were significantly reduced regardless of which host instar was parasitized. Our results suggest that M. bicoloratus has considerable potential as a biological control agent for S. litura.     

Some aspects of activity regulation of Galleria mellonella PTTH cells

Using the Galleria prothoracicotropic bioassay, five small neurosecretory cells occurring in each dorsolateral part of protocerebrum of Galleria mellonella brain were identified as prothoracicotropic hormone (PTTH) cells. It was found that the critical period for the release of PTTH from a brain implanted in neck-ligated larva lasts up to the third day after implantation. The content of paraldehyde-fuchsin positive neurosecretory material (NSM) in PTTH cells was determined during the penultimate and last larval instar, during pupal instar, and in starved or poststarvation fed or space-deprived last instar larvae. Two peaks of NSM in PTTH cells were found in the penultimate instar (in freshly molted, and 76-h-old larvae), four peaks in the last instar larvae (in freshly molted, and in 67-, 132-, and 174-h-old larvae), and one peak in the pupal instar (in 56-76-h-old pupae). It was also observed that upon starvation NSM accumulated in PTTH cells, while after 3 h of poststarvation feeding it was released. In permanent space-deprived last instar larvae no NSM occurred in PTTH cells. In all investigated larval instars a rapid release of NSM from PTTH cells was found a few hours after molt associated with the beginning of the feeding period. The significance of the NSM content in PTTH cells is discussed in relation to ecdysteroid titer.     

Effects of a juvenile hormone analogue on the duration of the fifth instar in the milkweed bug Oncopeltus fasciatus

Topical application of JHA to fifth instar nymphs of Oncopeltus fasciatus, immediately following ecdysis from the fourth instar, decreases the duration of the fifth instar by approximately 36 hr in addition to inducing a supernumerary larval moult. JHA appears to accelerate the time of subsequent ecdysis in two ways: first, the onset of ecdysone secretion is accelerated, and is accompanied by a similarly premature initiation of mitotic activity in epidermal cells. This is the classical prothoracicotropic action of JH. Second, the period between the onset of mitotic activity and the time of ecdysis itself is shortened. That is, once cellular activities associated with the moulting cycle are triggered by ecdysone, such activities are completed more rapidly in the presence of JHA. It appears that the larval-larval moult induced by JHA requires intrinsically less time to accomplish than a normal metamorphic moult.     

外源激素对褐飞虱翅型分化的影响

该文报道外源蜕皮激素(ED)和保幼激素类似物(JHA)对褐飞虱Nilaparvata lugens (Stal)翅型分化及体内酯酶活性、可溶性糖和游离氨基酸含量等的影响。褐飞虱在3、4龄若虫期经JHA处理后,所羽化出的成虫短翅型比例明显升高;而在5龄若虫期处理,所羽化出成虫的短翅型比例无明显变化。ED处理能增加3、4龄若虫体内酯酶活性、可溶性糖及游离氨基酸含量,而JHA处理则能有效地抑制ED的这些生物活性,这可能是JHA处理后褐飞虱成虫短翅型比例提高的生理生化基础。     

Functional response of Chrysoperla carnea (Neuroptera: Chrysopidae) to Helicoverpa armigera (Lepidoptera: Noctuidae): Effect of prey and predator stages

Abstract Understanding predator–prey interactions has a pivotal role in biological control programs. This study evaluated the functional response of three larval instars of the green lacewing, Chrysoperla carnea (Stephens), preying upon eggs and first instar larvae of the cotton bollworm, Helicoverpa armigera Hübner. The first and second instar larvae of C. carnea exhibited type II functional responses against both prey stages. However, the third instar larvae of C. carnea showed a type II functional response to the first instar larvae of H. armigera, but a type III functional response to the eggs. For the first instar larvae of C. carnea, the attack rate on H. armigera eggs was significantly higher than that on the larvae, whereas the attack rate of the second instar C. carnea on H. armigera larvae was significantly higher than that on the eggs. For the third instar larvae of C. carnea, the attack rate on the larvae was 1.015 ± 0.278/h, and the attack coefficient on the eggs was 0.036 ± 0.005. The handling times of the third instar larvae on larvae and eggs were 0.087 ± 0.009 and 0.071 ± 0.001 h, respectively. The highest predation rate was found for the third instar larvae of C. carnea on H. armigera eggs. Results of this study revealed that the larvae of C. carnea, especially the third instar, had a good predation potential in controlling H. armigera eggs and larvae. However, for a comprehensive estimation of the bio‐control abilities of C. carnea toward H. armigera, further field‐based studies are needed.     

Phenology of Parasetigena silvestris (Diptera: Tachinidae), gypsy moth (Lymantria dispar) (Lepidoptera: Lymantriidae) larval parasitoid and its efficiency for parasitisation

Parasetigena silvestris is a univoltine, solitary, larval endoparasitoid which lays its eggs on the surface of gypsy moth larvae. Field collection of the host larvae (2nd through 5th instar) from an artificially established gypsy moth population were made to compare stage specific parasitism between larvae without and with P. silvestris tachinid eggs. The tachinid oviposition rate detected was highest in second instar larvae, and then decreased as larvae developed toward full maturity. The opposite was true for tachinid parasitoid emergence which had no emergence from second through third host instar larvae. Fourth instar gypsy moth larvae, however, experienced significantly higher parasitism by P. silvestris in the larvae with eggs than those without the eggs. The braconid wasp Cotesia melanoscelus caused significantly higher parasitism in early instar larvae with P. silvestris eggs than in those without the eggs. The tachinid prefers to lay more eggs on parasitised larvae by the braconid even though the braconid is a superior competitor to the fly during multiparasitism. Factors influencing parasitism rates by P. silvestris such as host-parasitoid synchronisation and the multiparasitism interaction with C. melanoscelus are discussed.     

Developmental interactions between the army wormLeucania separata [Lep.: Noctuidae] and its parasiteApanteles ruficrus [Hym.: Braconidae]

The developmental interactions between the gregarious endoparasitoidApanteles ruficrus Hal. and the army worm,Leucania separata Walker were investigated. The parasitoid preferred young host larvae and developed in 9.5 days irrespective of host age at the time of parasitization. The growth of parasitized host larvae were depressed. The net maximum weight of the host larva was positively correlated with the number of parasitoid eggs laid when the 2nd instar was parasitized. And when parasitizing in 2nd instar, the weight of parasitoid was negatively correlated with the number of eggs laid. The parasitoid has an ability to regulate the size of the host and the parasitoid itself according to the number of eggs laid when the host larva is very small.