Laboratory evaluation of certain natural compounds against the melon ladybird beetle,Epilachna chrysomelina F. attacking cucurbit plants

Neem Azal-T/S, Neudosan and Spruzit flüssig at different concentrations were evaluated against the second and fourth instar larvae of E. chrysomelina under laboratory conditions. In case of acute effects, the highest mortality was recorded when larvae were offered squash leaves treated with the highest concentration (2%) of Neem Azal-T/S, and the lowest mortality was, however recorded after treating them with the lowest concentration (0.25%). On the other hand, the second instar larvae were more susceptible to any compound at any concentration than the fourth instar one. One per cent Neudosan formulation showed the highest toxicity to both the 2nd and 4th instar larvae. As the concentration of this compound decreased the mortality percentage of treated larvae decreased to reach the minimum at the lowest concentration (0.25%). The same trend could be applied for Spruzit flüssig compound. When Neem Azal-T/S was compared with Neudosan, the later at any concentration was more toxic than the former. Generally, Neudosan and Spruzit flüssig seemed to be active against E. chrysomelina during the first three days after treatment, then, deterioration occurred among this compound. In case of latent effects, the duration of the treated fourth instar larvae of E. chrysomelina with any tested compound, was prolonged. An increase in the concentration caused prolongation of this period to reach the longest at the highest concentration. This prolongation, was more pronounced after Neem Azal-T/S treatment followed by Neudosan and Spruzit flüssig. The same trend could be applied for the duration of the pupal stage as being affected by the different concentrations of the tested compounds which were applied to the fourth instar larvae. However, adults of E. Chrysomelina treated as fourth instar larvae with high concentrations of Neem Azal-T/S showed some adult malformations. The sex ratios of adults produced from fourth instar larvae treated with different concentrations of the tested natural compounds were about 1?:?1, except for two cases; 1% Neudosan where the number of females were double the number of males and 0.5% Spruzit flüssig where the number of males were double the number of females. An increase in the concentration of any tested compound caused an obvious decrease in the percentage of emerged adults. The least emergence percentage was recorded in the case of Spruzit flüssig, followed by Neudosan and Neem Azal-T/S. Treating the fourth instar larvae with the different concentrations of the tested natural compounds significantly shortened the longevity of produced adults and decreased the fecundity.     

Biological studies ofDiadegma semiclausum (Hym., Ichneumonidae), a parasite of diamondback moth

Laboratory studies were conducted on certain aspects of biology ofDiadegma semiclausum Hellén, a larval parasite of a crucifer pest,Plutella xylostella (L.). Within the range of 15°C to 35°C, the higher temperature, the shorter was the duration of larval and adult stages. Egg hatching and adult emergence were high at 15°C to 30°C but were significantly reduced at 35°C. The higher the temperature, the higher was the proportion of males produced. Temperature threshold was 5.74°C for eggs, 3.80°C for larvae, 5.91°C for pupae and 6.60°C for adults.D. semiclausum oviposition in the first threeP. xylostella larval instars produced more parasite males than females but oviposition in the fourth instar produced significantly more females than males. Parasite adults tended to emerge from their pupae from 06∶00 to 09∶00 hours although some emerged at other hours during the photophase. Adult longevity and production of eggs increased when adults were provided with a food source (honey) compared with no food or provision of water alone. Parasite adults survived and laid eggs for 28 days when provided with food but for only three days when deprived of food.     

敲减神经肽F基因(npf)对亚洲玉米螟取食、生长发育和繁殖的影响

【目的】神经肽F(neuropeptide F, NPF)是无脊椎动物特有的一类神经肽,因其C末端是苯丙氨酸(F)而命名,参与昆虫的取食、生物节律、学习记忆等多种生理功能的调控。本研究旨在明确NPF对亚洲玉米螟Ostrinia furnacalis生长发育的影响,为害虫防治提供重要依据。【方法】采用一种基于工程菌高效合成靶向昆虫基因的dsRNA的方法经济有效地敲降npf,用低浓度(0.01%)和高浓度(0.02%)dsNPF和dsGFP(对照)分别饲喂亚洲玉米螟1龄初、3龄初和5龄初幼虫直至化蛹,检测5龄幼虫平均取食量、体重、体长、存活率和化蛹率,蛹羽化率和成虫产卵量,以及幼虫各龄期、蛹发育历期和成虫寿命。【结果】从亚洲玉米螟1, 3和5龄初幼虫开始饲喂0.01%和0.02% dsNPF时,与饲喂相应浓度dsGFP的对照相比,除个别点外,5龄幼虫的取食量、体重、体长、存活率和化蛹率,蛹羽化率和成虫单雌产卵量均显著降低,幼虫各龄期、蛹发育历期均显著延长,成虫寿命显著缩短。且dsNPF处理幼虫的龄期越早对发育的影响越大。其中0.01% dsNPF处理的1龄幼虫和0.02% dsNPF处理的3龄幼虫有90%的个体在蛹期死亡,而0.02%dsNPF处理的1龄幼虫有90%的个体在幼虫期死亡。【结论】结果提示NPF对亚洲玉米螟的发育和取食具有调控作用,这为探索新型绿色的害虫防治提供了依据。     

Azadirachtin potentiates the action of ecdysteroid agonist RH-2485 in Spodoptera littoralis

Edysteroid agonist RH-2485 induces an immediate and fatal molt in Spodoptera littoralis when added to the diet of the 2nd and 4th instar larvae at 1 ppm, and to that of the 6th instar larvae at 0.001 ppm concentration. Ten times lower doses fed to the larvae continuously allow an apparently normal larval development that is terminated by a supernumerary larval molt. The other effects of RH-2485 include death during metamorphosis and impaired fertility of emerged adults. The number of progeny is reduced even with low RH-2485 doses that do not interfere with moltings; e.g., insects fed 0.0001 ppm since the 2nd, 4th, and 6th instar produce 72%, 62%, and 22%, respectively, less progeny than the controls. Feeding larvae with 10-1000 ppm Suneem oil (containing about 0.1-10 ppm azadirachtin) causes, in a stage- and dose-dependent manner, a cessation or reduction of feeding, delay of molts, death of larvae and pupae, and sterility of emerged adults; with 10 ppm Suneem oil, the number of progeny is reduced by 20-32%. Presence of Suneem oil in the diet does not influence the potential of RH-2485 to induce a prompt molt, but it increases ten times the potency to elicit a supernumerary larval molt. Certain combinations of RH-2485 with Suneem oil provoke up to 3 extra larval molts. Lethal developmental derangements and sterility are more frequent, and the response of larvae of different age is more uniform, when Suneem oil and RH-2485 are combined than when each of these agents is administered alone.     

Biology and performance of two indigenous larval parasitoids on Tuta absoluta (Lepidoptera: Gelechiidae) in Sudan

Tuta absoluta is an alien invasive pest in Sudan. Since it was detected in the country, the pest continues to cause major tomato losses. Alarmed by its devastating nature and the speed of spread, tomato growers resorted to the indiscriminate use of broad-spectrum insecticides. Promising indigenous parasitoid(s) may contribute to suppression of this pest. The biology and performance of the native parasitoids, Bracon nigricans Szépligeti (Braconidae) and Dolichogenidea appellator (Telenga) (Braconidae), was therefore evaluated against different immature stages of T. absoluta under laboratory conditions. A significantly higher number of fourth instar larvae was accepted for oviposition by B. nigricans. Similarly, fourth instar larvae yielded a considerably higher number of parasitoid offspring compared to third instar larvae. The male ratio of B. nigricans offspring which emerged from both larval instars was high, 0.96 and 0.66 for third and fourth instar larvae of T. absoluta, respectively. There was, however, no significant difference in the number of males and females that emerged from fourth instar larvae. Conversely, the number of hosts killed by stinging behaviour or host feeding was significantly higher on third instar larvae. B. nigricans potential fecundity varied with the age of the wasp with the lowest fecundity on the day of eclosion. The performance of D. appellator in terms of the total number of offspring produced and female progeny was similar for second and third instar larvae of T. absoluta. The preimaginal developmental time for both parasitoid species did not vary with either host stage or sex of the parasitoid. B. nigricans adult longevity was similar for both sexes, while that of D. appellator significantly varied with females living longer than males. Prospects for conservative biological control of T. absoluta are discussed in the light of the results of this study.     

Life cycle of a giant carnivorous caddisfly, <Emphasis Type="Italic">Himalopsyche japonica</Emphasis> (Morton) (Trichoptera: Rhyacophilidae), in the mountain streams of Nagano,Central Japan

 The life cycle of a giant carnivorous caddisfly, Himalopsyche japonica (Morton), was studied in two mountain streams in Nagano Prefecture, Central Japan. Field surveys and rearing experiments in the laboratory were conducted from October 1997 to September 2001. The life cycle of H. japonica was estimated to be a complex univoltine cycle that partly includes bivoltine populations. The adults had a long flight period, from April to September, with three distinct peaks of emergence. First to third instar larvae were collected from June to February, and the last (fifth) instar larvae and pupae appeared throughout the year. In autumn, the larvae belonging to all instars were found, and younger ones overwintered in the fourth instar stage and others in the fifth instar stage. On the other hand, fifth instar larvae and pupae ceased developing in autumn even though the water temperature was higher than the developmental zero temperature. The overwintered pupae emerged as adults in April, and the overwintered fifth instar larvae pupated in May and emerged in June. The larvae which overwintered in the fourth instar stage probably emerged after June. Received: March 19, 2002 / Accepted: January 10, 2003 Present address: United Graduate School of Agricultural Science, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan Tel. +81-265-77-1401; Fax +81-265-74-7496 e-mail: himalo@f8.dion.ne.jp Acknowledgments The author thanks Prof. T. Yoshida, Prof. H. Nakamura, and Associate professor K. Soma, Shinshu University; Mr. T. Nozaki, Kanagawa Environmental Research Center; and Mr. N. Kubota, Environmental Assessment Center in Matsumoto laboratory for their advice and help in accomplishing this research. The author is also grateful to Prof. K. Tanida, Osaka Prefecture University; Dr. T. Ito, Hokkaido Fish Hatchery; Mr. K. Okazaki, Kutchan City Museum; and Mrs. Y. Isobe, Nara Women's University, for suggesting references. Miss. T. Ishiyama, Mr. H. Kojima, Mr. M. Yagyu, and the students of the Forest Animals Laboratory in Shinshu University kindly provided field samples. Correspondence to:T. Tsuruishi     

Long-term effects of Bacillus thuringiensis subsp. israelensis on Aedes aegypti.

Immediate (24 hours exposure) and long-term (until the emergence of the adults) effects of different doses of a primary powder of Bacillus thuringiensis subsp. israelensis (B.t.i.) against first and second instar Aedes aegypti larvae were monitored. The long-term effect was dose-dependent and was materialized by a prolongation of the preimaginal development and continuous cumulative mortality until the emergence of the adults. Mortality values were higher during the fourth larval instar and the pupal stage. Some of the larvae reaching the fourth instar were smaller in size and remained in this state a 2-4 times longer period than in the control and finally died as larvae or very small pupae. The long-term effect was more intense as the treatment was applied earlier during the larval development. The correlation of the immediate lethal effect with the late effect allows the evaluation of the total impact of a larvicidal treatment.     

Biology of the House Fly Predator <Emphasis Type="Italic">Carcinops pumilio</Emphasis> (Erichson) (Coleoptera: Histeridae)

Two larval instars of Carcinops pumilio (Erichson) were identified from a frequency distribution of the head capsule measurements. The total developmental time from egg to adult emergence averaged 20.5 ± 0.1 days at 30 °C. The 2nd instar was the longest, accounting for 39% of the total development time. The 1st instar larvae sustained the highest mortality of 26% whilst there was no mortality in the pupal stage. The immature stages sustained about 50% mortality before adult emergence. The adult females of C. pumilio can live for 140 days and the rate of survival was high in the young adults but decreased as the beetles aged. Newly emerged adults survived for 25.5 ± 0.4 days without feeding. The sex ratio was 0.48 females: 0.52 males. The intrinsic rate of natural increase (r_m) was 0.065, the generation time (T) was 50.019 days and the net reproduction rate (R_o) was 20.191.     

Radiation biology and inherited sterility in false codling moth (Lepidoptera: Tortricidae)

False codling moth, Cryptophlebia leucotreta (Meyrick), male and female mature pupae and newly emerged adults were treated with increasing doses of gamma radiation and either inbred or out-crossed with fertile counterparts. For newly emerged adults, there was no significant relationship between dose of radiation and insect fecundity when untreated females were mated to treated males (N female by T male). However, fecundity of treated females mated to either untreated (T female by N male) or treated males (T female by T male) declined as the dose of radiation increased. A similar trend was observed when mature pupae were treated. The dose at which 100% sterility was achieved in treated females mated to untreated males (T female by N male) for both adults and pupae was 200 Gy. In contrast, newly emerged adult males treated with 350 Gy still had a residual fertility of 5.2% when mated to untreated females, and newly emerged adult males that were treated as pupae had a residual fertility of 3.3%. Inherited effects resulting from irradiation of parental (P1) males with selected doses of radiation were recorded for the F1 generation. Decreased F1 fecundity and fertility, increased F1 mortality during development, and a significant shift in the F1 sex ratio in favor of males was observed when increasing doses of radiation were applied to the P1 males.     

Life history studies of <Emphasis Type="Italic">Prorops nasuta</Emphasis>, a parasitoid of the coffee berry borer

Life history studies were conducted in the laboratory on the African parasitoid Prorops nasuta Waterston (Hymenoptera: Bethylidae), a parasitoid of the coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Scolytidae). The female wasp enters an infested coffee berry, kills the adult borer and seals the entrance of the berry with the body of the borer, impeding the entry of other organisms into the berry. The preoviposition period ranges from 3 to 14 days (mean 5.42 ± 0.37 SE). During this time females feed on the immature stages and paralyse fully grown larvae and pupae of the CBB. P. nasuta is an idiobiont solitary ectoparasitoid. Eggs are laid externally on the last instar larvae and pupae. Mean development time (egg to adult) for males and females was 27.7 (±0.37 SE) and 30 (±0.12 SE) days, respectively. Median survival for wasps fed on final instar CBB larvae was 27.7 days, significantly longer than any other treatment, while for females without food it was 2.5 days. In culture, females produced an average of 4.3 (±0.39 SE) progeny during their lifetimes. Adults began emerging at 30.6 days (±0.28 SE) after cultures were started and peak production was reached at 36 days, declining thereafter. Males normally emerged from coffee beans 2–3 days before females. Males usually emerged from 07:00 to 09:00h and females from 10:00 to 14:00 h. The culture sex ratio (proportion of males) was 0.21. Virgin females produced only male offspring.     

Immunocompetence of the red turpentine beetle, Dendroctonus valens LeConte (Coleoptera: Curculionidae, Scolytinae): Variation between developmental stages and sexes in populations in China

Immune defense imposes fitness costs as well as benefits, so organisms should optimize, not maximize, their immune function through their life cycle. We investigated this issue in the red turpentine beetle, Dendroctonus valens LeConte (Coleoptera: Curculionidae, Scolytinae), which is a pine-killing invasive beetle in China, though it is usually considered as a secondary pest in its native range of North America. We hypothesized that pathogen pressure may affect these beetles differently throughout their life history. We measured the insect's immunocompetence throughout life, determining encapsulation ability and phenoloxidase activity in larval stages, pupae and adults. Pupae had the highest encapsulation ability, but encapsulation was not different between final instar larvae and adults. Phenoloxidase (PO) activity was highest in final instar larvae and pupae, followed by the second instar larvae and adults. Total phenoloxidase activity increased significantly from the second instar larval stage to pupae, and then decreased in adults. Although the second instar larvae had the lowest phenoloxidase activity, more than 90% of total PO existed in the hemolymph in the form of the active enzyme, as compared with pupae, in which over 60% of PO occurred as a proenzyme. Both active PO and total PO were much higher in females than in males, though no significant differences were detected between the encapsulation ability of male and female adults. This result suggests the existence of a sexual dimorphism of immunocompetence in D. valens adults. Variations in immunocompetence across developmental stages suggest that D. valens adopts diverse investment strategies in immunocompetence during different stages. Potential reasons for variation in immunocompetence among developmental stages and between the sexes of D. valens are discussed.     

Gamma irradiation as a quarantine treatment for apples infested by codling moth (Lep., Tortricidae)

Abstract: Effects of gamma radiation on the fifth instar codling moth, Cydia pomonella (L.), larvae were examined. Mature larvae were exposed to a series of gamma radiation doses ranging from 50 to 250 Gy and survival to pupae and adults was examined. The results showed that pupation and adult emergence decreased with increasing radiation dose. The results also showed that diapausing larvae were more sensitive to irradiation treatment than non-diapausing larvae, and females were more sensitive than males. A dose of 150 Gy reduced adult emergence to less than 2% in non-diapausing larvae, while a dose of 200 Gy completely prevented it. Furthermore, none of the emerging moths exposed to a dose of 150 Gy were females; at 100 Gy dose, the percentage of females was less than 14%. Irradiating larvae in apple fruit in a small-scale laboratory experiment produced similar results. Tests in which >100 000 larvae (in the fifth instar) were irradiated in an artificial rearing medium with a dose of 200 Gy resulted in no adult emergence. Similar results were also obtained when >32 000 larvae in the same stage were exposed in apple fruit to the same dose. The results indicate that the use of ionizing radiation as a quarantine treatment for codling moth infested fruits is feasible and requires a relatively low dose.     

Neural inhibition of the corpus allatum in the last larval instar differs from that in adults in the blood-feeding insect, Rhodnius prolixus

An active corpus allatum (CA) in the blood feeding insect, Rhodnius prolixus, releases a substance that inhibits metamorphosis in larvae, and activates egg production in adults. In adults, transecting the nervus corporis cardiacum II's (NCCII), which are attached posteriorly to each protocerebral lobe, greatly increases egg production indicating that the adult CA is activated and receives neural inhibition from cells associated with the NCCII [Chiang, Arch. Insect. Biochem. Physiol. 39:126-131 (1998)]. In the present study, the NCCII's in fifth instar larvae were transected immediately before or after feeding to determine if these nerves normally inhibit CA activity in the last larval instar. Approximately 20 to 25 days following ingestion of a blood meal, L5's with transected NCCII's emerged as fully-formed adults with no larval characteristics. Examination of the brain in these recently emerged adults revealed that the NCCII's were absent. Since fifth instar larvae with transected NCCII's emerged with no juvenile characteristics, cutting the NCCII's did not activate the CA, indicating that the mechanism for inhibition of the CA differs in the last larval instar and adult animals.     

薇甘菊甲醇提取物对二疣犀甲生长发育的影响

【目的】探讨入侵杂草薇甘菊Mikania micrantha的利用价值, 及其在棕榈害虫二疣犀甲Oryctes rhinoceros生态防控中的应用前景。【方法】采用浸渍法和室内饲喂法, 研究了薇甘菊甲醇提取物对二疣犀甲取食量、 卵孵化、 化蛹、 羽化及幼虫发育等的影响。【结果】薇甘菊提取物对二疣犀甲具有很好的生长发育调节活性。拒食活性研究结果表明, 在不同供试浓度下, 其对二疣犀甲3龄幼虫均表现拒食活性, 且拒食率与处理浓度呈正相关。薇甘菊提取物处理二疣犀甲的卵后, 孵化率明显降低, 且孵化期延长, 10 mg/mL提取物处理后孵化率仅达66.66%, 孵化期比对照延迟3 d, 同时初孵幼虫死亡率也高达40.43%。采用添加薇甘菊提取物的饲料饲喂1龄幼虫后, 幼虫体重增长减缓, 在处理浓度为10和5 mg/g时, 处理后90 d体重增加量分别为3.83 g和4.53 g, 而对照组体重增加量达到6.87 g。薇甘菊提取物处理老熟幼虫后, 对其化蛹具有抑制作用, 造成化蛹率降低, 化蛹时间延长及蛹的畸形。经薇甘菊提取物处理二疣犀甲蛹后, 成虫羽化率降低, 羽化时间明显延长, 且畸形成虫数量增加, 主要表现为翅无法正常伸展、 虫体瘦小、 无法正常爬行等。【结论】薇甘菊具有开发为生物源昆虫生长调节剂、 并在二疣犀甲防控中应用的潜力。     

A biological assessment of Apophua simplicipes (Hymenoptera: Ichneumonidae) as a parasitoid of the oblique banded leafroller, Choristoneura rosaceana (Lepidoptera: Tortricidae)

Apophua simplicipes (Cresson) (Hymenoptera: Ichneumonidae) is a common parasitoid of the oblique banded leafroller, Choristoneura rosaceana (Harris) (Lepidoptera: Tortricidae) in organically managed apple orchards in the southern interior of British Columbia, Canada. The biological characteristics of a laboratory colony of A. simplicipes were studied. When held at 15 and 25°C and provided with honey water, individual females survived an average of 60.6±6.1 and 29.8±4.7 days, oviposited 196.7±50.7 and 326.6±51.3 eggs and parasitized a total of 163.4±40.4 and 229.4±35.8 hosts, respectively. Females oviposited into first through fourth instar oblique banded leafrollers, with significantly more parasitism occurring in the first two instars compared to the third and fourth instars. No parasitoid larvae survived past the first larval stage in parasitized fourth instar hosts. Apophua simplicipes did not parasitize larvae of three-lined leafroller, Pandemis limitata (Robinson) (Lepidoptera: Tortricidae) which is sympatric with oblique banded leafrollers in orchards in the southern interior of British Columbia. Female predation and host feeding from wounds on early instars of both leafroller species was observed under laboratory conditions. In addition, early instar hosts exited diet feeding sites in response to the probing activity of the ovipositing wasps. A similar escape reaction in the orchard may cause a leafroller larva to move away from its feeding site, making it more vulnerable to predation or movement off the tree. Apophua simplicipes larvae emerged from fifth and sixth instar hosts. Parasitized oblique banded leafroller hosts consumed significantly less meridic diet than unparasitized female larvae from fifth instar through to parasitoid emergence or leafroller pupation. Our laboratory results suggest that A. simplicipes may reduce field populations of oblique banded leafroller and decrease pest feeding damage.     

The effects of Plagiorchis noblei metacercariae on the development and survival of Aedes aegypti larvae in the laboratory

Plagiorchis noblei infections impair the survival and development of fourth instar Aedes aegypti larvae. Mortality during the larval and pupal stages reached 92%, and 60% of the emerging adults were malformed. The metacercariae interfere with pupation and the emergence of adults. Larvae and pupae that fail to transform to the next developmental stage within the normal time characteristically persist for extended periods, but invariably die without transforming. Whereas 82% of the control larvae gave rise to functional adults, only 4% of infected larvae managed to do so. Such effects may facilitate the transmission of the parasite.     

Horizontal transmission of Parathelohania anophelis to the copepod, Microcyclops varicans, and the mosquito, Anopheles quadrimaculatus

The copepod Microcyclops varicans was infected with Parathelohania anophelis by unincleate meiospores from a field-collected fourth instar Anopheles quadrimaculatus larva. Large numbers of unincleate, pyriform spores developed in the copepod. These spores were fed to early instar A. quadrimaculatus larvae, infecting both males and females, resulting in the production of cylindrical, binucleate spores in the adult. These spores were responsible for vertical transmission, through the eggs, to the larvae. The original spore type collected from the field was found in the male progeny from the infected females. Another P. anophelis-infected mosquito colony was established by feeding spores from a single, field-collected, infected copepod to A. quadrimaculatus larvae. The microsporidium was continuously maintained by vertical transmission in newly established infected colonies.     

Pupal diapause of Helicoverpa armigera (Lepidoptera: Noctuidae): sensitive stage for thermal induction in the Okayama (western Japan) population

Helicoverpa armigera (Hübner) exhibits a facultative pupal diapause, which depends on temperature and photoperiod. Pupal diapause is induced at 20 degrees C by short photoperiods and inhibited by long photoperiods during the larval stage. However, in some pupae (35% of males and 57% of females) of a non-selected field population from Okayama Prefecture (34.6 degrees N), diapause is not induced by short photoperiods. In the present experiment, the importance of temperature for diapause induction was studied in the non-diapausing strain, which was selected from such individuals reared at 20 degrees C under a short photoperiod of 10L:14D. Furthermore, the sensitive stage for thermal determination of pupal diapause was determined by transferring larvae of various instars and pupae between 20 degrees C and 15 degrees C. Diapause was induced by 15 degrees C without respect to photoperiod. When larvae or pupae reared from eggs at 20 degrees C under a short or a long photoperiod were transferred to 15 degrees C in the periods of the middle fifth instar to the first three days after pupation, the diapause induction rate was significantly reduced in both males and females, especially in females. In contrast, when larvae or pupae reared at 15 degrees C were transferred to 20 degrees C in the same periods, diapause was induced in males, but not in females. However, the diapause induction rate of pupae transferred to 20 degrees C on the fourth day after pupation was significantly increased in females. The results show that temperature is the major diapause cue in the photoperiod-insensitive strain and the periods of middle fifth larval instar to early pupal stage are the thermal sensitive stages for pupal diapause induction with some different responses to temperatures between males and females in H. armigera.     

Anopheles stephensi and Toxorhynchites amboinensis: aseptic rearing of mosquito larvae on cultured cells

Aseptic larvae of Anopheles stephensi and Toxorhynchites amboinensis were reared on a continuous cell line (RU TAE 12 V) from the mosquito, T. amboinensis, that grew in suspension as multicellular vesicles. Surface-sterilized eggs were hatched in a 24-well plate containing 0.2 ml of Leibovitz's L-15 medium per well and incubated in a humidified atmosphere. Toxorhynchites amboinensis eggs of 36 hr or older were placed singly to assure hatching and avoid cannibalism. Hatching rates were over 80%. All larval instars were maintained in L-15 medium at 28 C with a 12-hr photoperiod. Anopheles stephensi larvae were reared in 25-cm2 tissue culture flasks containing 10 ml of L-15 medium with 30 to 50 first and second instar larvae or 10 third and fourth instar larvae per flask. Toxorhynchites amboinensis larvae remained in the 24-well plate in 1.5 ml of medium through the second instar; third instar larvae were kept in 12-well plates (3 ml of medium per well) and transferred to 25-cm2 flasks (10 ml per flask) when they reached the fourth instar. First and second instar A. stephensi larvae were fed cultured cells once, and third or fourth instar larvae twice a day. Toxorhynchites amboinensis larvae were fed vesicles once during the first 4 days after hatching, and every 1 or 2 days thereafter. Each A. stephensi larva consumed approximately 2 X 10(6) cells, and T. amboinensis larvae 10 times more cells before pupating. Anopheles stephensi pupated after 7 to 8 days and adults emerged during days 9 to 11. Pupation in T. amboinensis began on day 21 after hatching and adults emerged 5 days later. Cell lines isolated from A. stephensi larvae or embryos of the ticks Rhipicephalus sanguineus and Anocentor (Dermacentor) nitens supported only limited growth of A. stephensi larvae. Defibrinated hamster (Mesocricetus auratus) blood, though readily ingested, did not support the growth of A. stephensi whereas larvae reared on blood cells plus T. amboinensis cells showed limited growth.     

Sublethal effects of chronic exposure to tebufenozide on the development, survival, and reproduction of the tufted apple bud moth (Lepidoptera: Tortricidae)

The lethal and sublethal effects of tebufenozide on the survival, development, and reproduction of a field strain of tufted apple bud moth, Platynota idaeusalis (Walker) (Lepidoptera: Tortricidae), were assessed by feeding first and third instars tebufenozide-treated diet until pupation. Larval mortality was 27.4 and 44.7% at 0.1 and 0.2 ppm for first instars and 21.9 and 57.8% at 0.2 and 0.4 ppm for third instars, respectively. Treated larvae exhibited higher pupal mortalities, lower pupal weights, and generally more deformed adults than untreated larvae. Larval development was not affected by tebufenozide when neonates were exposed, but development was accelerated slightly at 0.4 ppm for both males and females when third instars were exposed. All treatments produced sex ratios biased toward males. When paired with either treated or untreated males, females resulting from neonates treated at 0.2 ppm and from third instars treated at both 0.2 and 0.4 ppm laid from 37 to 65% fewer eggs. A reduction in fertility was only found when third instars were treated at the higher 0.4 ppm rate. These results suggest that tebufenozide can exhibit a significant effect on the population dynamics of the tufted apple bud moth.