Laboratory Evaluation of Isaria fumosorosea CCM 8367 and Steinernema feltiae Ustinov against Immature Stages of the Colorado Potato Beetle

The Colorado potato beetle, Leptinotarsa decemlineata, has developed resistance to most registered pesticides and has become one of the most difficult insect pests to control. Development of new biopesticides targeting this pest might solve the resistance problem and contribute to sustainable crop production. Laboratory experiments were conducted to assess the efficacy of Isaria fumosorosea (syn. Paecilomyces fumosoroseus) strain CCM 8367 against L. decemlineata when applied alone or combined with the entomopathogenic nematode Steinernema feltiae. The last-instar larvae of the Colorado potato beetle showed the highest susceptibility to I. fumosorosea followed by pre-pupae and pupae. The median lethal concentration (LC₅₀) was estimated to be 1.03×10⁶ blastospores/ml. The strain CCM 8367 was more virulent, causing 92.6% mortality of larvae (LT₅₀ = 5.0 days) compared to the reference strain Apopka 97, which caused 54.5% mortality (LT₅₀ = 7.0 days). The combined application of the fungus with the nematodes increased the mortality up to 98.0%. The best results were obtained when S. feltiae was applied simultaneously with I. fumosorosea (LT₅₀ = 2.0 days); later application negatively affected both the penetration rate and the development of the nematodes. We can conclude that the strain CCM 8367 of I. fumosorosea is a prospective biocontrol agent against immature stages of L. decemlineata. For higher efficacy, application together with an entomopathogenic nematode is recommended.     

Infection and mortality of Microtheca ochroloma (Coleoptera: Chrysomelidae) by Isaria fumosorosea (Hypocreales: Cordycipitaceae) under laboratory conditions

Microtheca ochroloma Stål, the yellowmargined leaf beetle, is a pest in crucifer crops during the late fall and winter months in Florida. On organic farms, it is difficult to control due to the restricted use of insecticides, in addition to the lack of specific natural enemies. The objective of this study was to evaluate a blastospore-formulated product of Isaria fumosorosea (PFR-97^TM 20% WDG) against this beetle. In the first experiment, four of the beetle's life stages were treated with a suspension of 3 × 10⁷ blastospores/ml. Mean corrected mortality of treated insects was significantly higher in 1st and 3rd instars than in the egg, pupal, and adult stages. Larvae infected by I. fumosorosea exhibited reduced growth and unsuccessful molting. The second experiment quantified mortality of first instars of M. ochroloma by four concentrations of PFR-97^TM. Mean corrected larval infection/treatment was significantly (2.6 times) higher with a concentration of 4?g of product per 100?ml of water compared to concentrations of 1–3?g per 100?ml of water. Different factors that might have affected the pathogenicity of I. fumosorosea against M. ochroloma are discussed.     

A possible therapy by juvenile hormone I to Spodoptera littoralis (Boisd.) larvae previously treated by precocene II

An effective constant dose (55 μg) of precocene II (PII) was topically tested against the last three instars of Spodoptera littoralis (Boisd.) larvae. Application of PII induced morphogenetic abnormalities typical of juvenile hormone (JH) excess. The resultant imperfect insects included larval‐pupal mosaic and partial or severe cases of untanned pupae. The sixth‐instar larvae were more sensitive to PII administration than the two preceding instars. However, sensitivity of the last larval instar to PII varied with the timing of dose application relative to the developmental status of the larvae. Whereas the newly ecdysed (0‐day old) larvae were more sensitive, the older larvae of the same sixth‐instar showed sharp decrease in their sensitivity to PII with a concomitant increase of their age. Application of a single dose (5 μg) of JH I to PII pre‐treated larvae significantly (P < 0.001) reduced the production of imperfect insects where many PH‐treated larvae developed successfully to apparent normal pupae. Although a single dose of PII was more effective on S. littoralis larvae than repeated daily doses, the effectiveness of JHI‐therapy to PH pre‐treated larvae by repeated doses was less effective in producing perfect insects than JH‐therapy to PII pre‐treated larvae by single doses. The reversal of any of these by applied JHI is quite interesting but the mechanisms remain to be unraveled.     

Untersuchungen zur Veränderung der Unkrautflora bei mehrmaligem Getreidenachbau auf einer Tonschwarzerde des Thüringer Beckens

The latent effects of precocenes I and II (PI and PH) and juvenile hormone I (JHI) when topically applied to the last three instars of Spodoptera littoralis (Boisd.) larvae have been studied. Application of both PI or PII resulted in morphogenetic abnormalities resemble some effects induced by administration of JHI, e.g., larval‐pupal intermediate, partial or severe cases of untanned pupae and imperfect moths. In PII‐treatments, the effect was instar‐dose‐dependent. The intermediate dose (55 μg) was more effective on S. littoralis larvae than other doses. The effectiveness of both doses of 40 or 70 μg in production of deformed larvae and pupae decreased when applied as repeated doses instead of single ones. In Pi‐treatments, the lower dose (25 μg) was more harmful to Spodoptera larvae than the higher dose (70 μg). Repeated application by either lower or higher doses did not enhance the production of imperfect insects. Application of JHI induced symptoms ranging from supernumerary instars, larval‐pupal intermediate, untanned pupae and deformed adults. The effect was dose‐dependent. In all tested compounds, there apparently was a latent or delayed effects. Although the insects were treated while they were larvae the complete effects were not apparent until after the insect had become a pupa or an adult. More efforts will be needed to understand how does precocene interferes with the process of tanning or sclerotization?     

Insecticidal activity of entomopathogenic fungi (Hypocreales) for potato psyllid,Bactericera cockerelli (Hemiptera: Triozidae): Development of bioassay techniques,effect of fungal species and stage of the psyllid

The potato psyllid, Bactericera cockerelli (?ulc), is a pest of potato, tomato, and some other solanaceous vegetables and has also been incriminated in the transmission of a bacterial pathogen, Candidatus Liberibacter solanacearum, resulting in a serious disease known as ‘zebra chip’. Although there are several reports of fungal pathogens in psyllids, there are none from B. cockerelli, nor have any fungi been evaluated against it. Five isolates of fungi, one Beauveria bassiana, two Metarhizium anisopliae and two Isaria fumosorosea, were bioassayed against B. cockerelli on potato leaves under ideal conditions for the fungi. All applications were made with a Potter spray tower. With the exception of concentration-effect studies, all other applications were made using 10⁷ conidia/mL in a 2-mL aqueous suspension. All isolates except B. bassiana, produced 95–99% mortality, corrected for control mortality, in adults 2–3 days after application of conidia and 91–99% in nymphs 4 days after application. The corrected mortalities for adults and nymphs treated with B. bassiana were 53 and 78%, respectively, 4 days after application. I. fumosorosea Pfr 97 produced 95% corrected mortality in both first and late third instar nymphs. M. anisopliae (F 52) produced 96% corrected mortality in first and third instar nymphs. Pfr 97 and F 52 were evaluated for insecticidal activity against third instar B. cockerelli using 10⁵, 10⁶, and 10⁷ conidia per mL. Mortality produced by I. fumosorosea Pfr 97 ranged from 83 to 97% and that of M. anisopliae F 52 was 88 to 95% at these concentrations.     

Biosteres longicaudatus: Developmental dependence on host (Anastrepha suspensa) physiology

Larvae of Anastrepha suspensa that were in the first day of the third instar were parasitized by females of the solitary endoparasitoid, Biosteres longicaudatus. At the end of the 6-hr oviposition period, larvae were ligated posterior to the ring gland so that some larvae had parasitoids anterior to the ligature while in others, the parasitoids were in the abdomen, posterior to the ligature. Ninety-two percent of the parasitoids anterior to the ligature hatched to the first through third instars. Parasitoids posterior to the ligature had a 75% egg hatch to the first instar only. No larval molts to the second or subsequent instars occurred in these parasitoids. Upon parabiosis to 3-day-old, unparasitized host pupae, the ligated larvae pupated and 97% of the first-instar parasitoids in these parabiosed larval abdomens molted to the second instar. Newly laid parasitoid eggs transplanted to 3-day-old pupal hosts had less than one-third of the egg hatch of those transplanted to first-day third-instar hosts. The data implicate the physiological state of the host (vis-a-vis pupation and associated events) as being an important factor in the development of the endoparasitoid.     

Ectopically expressed leaf and bulb lectins from garlic (<Emphasis Type="Italic">Allium sativum</Emphasis> L.) protect transgenic tobacco plants against cotton leafworm (<Emphasis Type="Italic">Spodoptera littoralis</Emphasis>)

The insecticidal activity of the leaf (ASAL) and bulb (ASAII) agglutinins from Allium sativum L. (garlic) against the cotton leafworm, Spodoptera littoralis Boisd. (Lepidoptera: Noctuidae) was studied using transgenic tobacco plants expressing the lectins under the control of the constitutive CaMV35S promoter. PCR analysis confirmed that the garlic lectin genes were integrated into the plant genome. Western blots and semi-quantitative agglutination assays revealed lectin expression at various levels in the transgenic lines. Biochemical analyses indicated that the recombinant ASAL and ASAII are indistinguishable from the native garlic lectins. Insect bioassays using detached leaves from transgenic tobacco plants demonstrated that the ectopically expressed ASAL and ASAII significantly (P < 0.05) reduced the weight gain of 4th instar larvae of S. littoralis. Further on, the lectins retarded the development of the larvae and their metamorphosis, and were detrimental to the pupal stage resulting in weight reduction and lethal abnormalities. Total mortality was scored with ASAL compared to 60% mortality with ASAII. These findings suggest that garlic lectins are suitable candidate insect resistance proteins for the control of S. littoralis through a transgenic approach.     

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.     

Effect of mating combinations on the host parasitisation and sex allocation in solitary endoparasitoid,Aenasius arizonensis (Hymenoptera: Encyrtidae)

Aenasius arizonensis (Girault) is an important solitary endoparasitoid of Phenacoccus solenopsis Tinsley. To optimise the mass production of high-quality females, it is important to assess the influence of mating regimes on the progeny fitness and sex allocation. We, therefore, hypothesise that mating combinations in A. arizonensis adults emerged from different host instars may influence parasitism and sex allocation in the subsequent generation. Therefore, we compared three nymphal instars (1st, 2nd and 3rd) and adults host stages of P. solenopsis for parasitism and sex allocation by A. arizonensis. Further, F₁ female progeny of the parasitoid emerged from different host instars was henceforth evaluated for its fitness in six mating combinations. A. arizonensis females parasitised all the host stages except the 1st instar nymphs. The parasitised 2nd instar nymphs yielded only males, while the sex ratio in the later host instars was strongly female-biased. The parasitoid females preferred 3rd instar nymphs with respect to higher parasitism (74.0–84.0%) and produced more females in the F₁ progeny as compared to other host stages. F₁ females that emerged from 3rd instar nymphs produced significantly higher parasitism (74.0–79.0%). These mating combinations also yielded more female progeny in the F₂ generation. However, parasitism by F₁ females was significantly lower (9.0–12.0%) when mated with males that emerged from 2nd instar P. solenopsis nymphs. Moreover, latter combinations yielded only male progeny in F₂ generation. These findings can be used in laboratory mass rearing of this parasitoid vis-à-vis biological control of P. solenopsis.     

Effect of dietary selenium supplementation on resistance to baculovirus infection

We have examined the effects of dietary selenium (Se) supplementation on larval growth and immunocompetence of the lepidopteran pest, the cabbage looper, Trichoplusia ni. Supplementation of the diet of T. ni larvae with 10–20 ppm Se resulted in a 1 day delay in pupation. The effects of the addition and/or removal of dietary Se on total Se bioaccumulation and sequestration were determined by neutron activation analysis of pupae. Early penultimate instar larvae moved from selenium containing diet to basal diet lost total pupal Se content down to the level of those fed basal diet. Conversely, larvae moved from basal diet to diet containing additional Se rapidly attained pupal Se levels comparable to larvae fed Se throughout larval development. Therefore, dietary Se is rapidly accumulated or lost during larval development, but significant amounts are sequestered from diet into pupae. Larvae were reared on diet supplemented with 5 or 10 ppm Se until the onset of the penultimate instar then infected per os with increasing concentrations of the fatal baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV). Larvae fed Se in the penultimate and ultimate instars were more resistant to viral infection than larvae not fed Se in the final instars. This study indicates that dietary Se levels rapidly impact Se assimilation and sequestration and that tissue Se levels are an important factor in resistance to AcMNPV infection in larval T. ni.     

Evaluation of Isaria fumosorosea (Hypocreales: Cordycipitaceae) for control of the Asian citrus psyllid,Diaphorina citri (Hemiptera: Psyllidae)

A laboratory bioassay was developed to evaluate strains of Isaria fumosorosea Wize, against Diaphorina citri. Up to 100% of adult psyllids were killed at concentrations between 10⁶ and 10⁷ blastospores/ml after 12 days, with derived LC₅₀ values (at 7 days post treatment) between 1.4 × 10⁵ and 2.0 × 10⁶ blastospores/ml for strains ARSEF 3581, FE 9901 and Apopka-97. A significantly higher value (1.5 × 10⁷) was obtained with a conidial formulation of Apopka-97. Average survival times were dosage dependent, i.e. between 10.2 days at 10³ blastospores/ml and 3.5 days at 10⁸ blastospores/ml. Rates of mycosis were also dosage dependent, with up to 100% sporulation on cadavers at 10⁸ blastospores/ml but declining at lower concentrations. The Apopka-97 strain (commercially available as PFR-97) was tested against established D. citri infestations in potted citrus in greenhouse cages. Treatments at label rates reduced psyllid populations by approximately 50% over 3 weeks. The combination of PFR-97 with emulsifiable oils (0.25% v/v) did not increase psyllid mortality compared with either agent alone. Imidacloprid applied as a drench killed 100% of psyllids within 3 weeks. Subsequent greenhouse tests during humid conditions were hampered by natural dissemination of I. fumosorosea to untreated psyllids, suggesting that this fungus is spread by air movement and may be highly effective under very humid conditions. In later tests, a Cladosporium sp. rapidly colonised psyllid cadavers and leaf surfaces, but was not pathogenic in laboratory tests. Our studies confirm the potential of I. fumosorosea to be used in IPM strategies for D. citri that rely on other tactics, such as insecticidal oils and native or introduced biological control agents.     

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.     

Entomopathogenic fungi disturbed the larval growth and feeding performance of Ocinara varians (Lepidoptera: Bombycidae) larvae

Abstract Feeding experiments using three strains of entomopathogenic fungi, Beauveria bassiana, Metarhizium anisopliae and Isaria fumosorosea were conducted with newly moulted 3rd–5th instar Ocinara varians Walker larvae in the laboratory. The mortality of larvae immersed individually in spore suspension (1 × 10⁷ spores/mL) of all the strains was ≥ 80% except 5th instar larvae treated with M. anisopliae which transformed into pupae, but did not result in adult emergence. The growth (total body mass), consumption, relative consumption rate and relative growth rate, were reduced at all three larval stages, while developmental time was extended in infected larvae with concurrent significant increase in approximate digestibility in infected larvae. Conversion of digested food (ECD) and ingested food (ECI) values declined in infected larvae as compared to the healthy larvae (control). The 5th instar larvae treated with M. anisopliae showed higher ECD and ECI values than control. Based on mortality and growth inhibition it can be suggested that all the studied fungal strains have a high potential for biocontrol and could be developed into biocontrol agents against O. varians.     

Effects of gamma radiation on different developmental stages of the oriental tobacco budworm,Helicoverpa assulta (Lepidoptera: Noctuidae)

Ionizing radiation is increasingly used as an alternative to post‐harvest crop fumigation by methyl bromide. We studied the effects of gamma irradiation on Helicoverpa assulta (Lepidoptera: Noctuidae) at different stages of development to determine the minimal dose for the prevention of normal emergence of adults. We selected five doses of gamma rays (100, 200, 300, 400 and 500 Gy) based on preliminary experiments and irradiated eggs, larvae, pupae and adults. A dose of 100 Gy to eggs allowed 21.83% of larvae to pupate, but these all died during the pupal stage. A dose of 100 Gy to last‐instar larvae caused larval or pupal death, or the emergence of abnormal adults; no normal adults developed. Irradiation of pupae with doses of 300 Gy and above resulted either in their death or emergence of abnormal adults; however, after 100 or 200 Gy, normal adults emerged and F₁ eggs were produced, but no eggs hatched. Following irradiation of adults, eggs were produced at all doses, although the numbers were significantly decreased compared to untreated controls (P < 0.05; 69.45–125.50 vs. 475.05 eggs per female); however, none of the eggs hatched. As prevention of normal emergence is a key outcome for measuring the effectiveness of radiation, then the 100 Gy dose was effective for irradiation of eggs and larvae, and 300 Gy for pupae.     

Comparative effect of various powder diets on growth of big-eyed bug Geocoris ochropterus (Fieber) (Hemiptera: Geocoridae)

Application of powder diets may promote development of the predator Geocoris ochropterus (Fieber) when densities are low in greenhouses or fields. Development and nymphal survival ratio of G. ochropterus fed on different powder diets including silkworm pupae (A), crickets (B), silkworm pupae and sunflower seed (C) and crickets and sunflower seed (D) were examined. Total average development period was 30.14 days for instars fed on silkworm pupae, 34.43 days for instars fed on crickets, 26.28 days for instars fed on silkworm pupae and sunflower seed and 29.67 days for instars fed on crickets and sunflower seed, and they were significantly different. Head width, body length, forewing length, dry body weight of adults and sex ratio were not affected by the different diets. There were significantly different found in survival rates among the diets when the nymph reached 5th instar. Results indicate that the combination powder of silkworm pupae and sunflower seed will promise as an alternative diet in the greenhouses to maintain the population of G. ochropterus.     

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.     

Influence of the entomopathogenic fungus Isaria fumosorosea on Axinoscymnus cardilobus (Coleoptera: Coccinellidae) under laboratory conditions

Effects of entomopathogenic fungus Isaria fumosorosea on biological characteristics and life table parameters of Axinoscymnus cardilobus, a predator of whiteflies, were studied using five different conidial concentrations under laboratory conditions. Non-significant differences were observed among different fungal concentrations on the percent survival of all immature stages. The survival of A. cardilobus on different host plants did not differ significantly. The prey consumption of third instar A. cardilobus on Bemisia tabaci treated with different fungal concentrations differed significantly at different time intervals. The developmental periods for all immature stages (from eggs, first, second, third, fourth instar nymph and pupae up to emergence) among the treatments were not statistically significant when compared to that of control. In the present study, fecundity, longevity, egg viability and life table parameters of females were not statistically significant over the different concentrations. It can be concluded that control strategies tested are compatible to a greater extent and incorporation of these have promising prospects for control of whitefly.     

Assessment of Ceratitis capitata (Diptera,Tephritidae) pupae killed by heat or cold as hosts for rearing Spalangia cameroni (Hymenoptera: Pteromalidae)

In this work, we study the suitability of using dead medfly Ceratitis capitata pupae, killed by heat- or cold-shock, for the mass rearing of Spalangia cameroni, a pupal parasitoid of key pests. 100% mortality of medfly pupae could be accomplished with cold-shock at –20°C for 60 min or with heat-shock at 55°C for 30 min. Neither parasitism percentage nor sex ratio of the offspring differed significantly among heat-shocked, cold-shocked and untreated pupae. In addition, there was no significant difference in the percentage of parasitoids that aborted (♂♂ or ♀♀) among pupal treatments. Some of the pupae were covered with peat because the third larval instar of the medfly buries itself before pupation. However, the buried pupae were not parasitised at a greater or lesser rate than those not covered with peat. The percentage of parasitism was also unaffected by whether the pupae had been killed recently or had been stored at between 4°C and 6°C over 15 or 30 days. The use of dead hosts and later storage permitted the following: (a) the use of hosts over long periods of time; (b) a rapid increase in parasitoid numbers and (c) the availability of pupae killed at the most suitable postpupation times for the production of parasitoids. Furthermore, in biological control projects, the use of dead parasitised pupae in the field avoids the risk of enhancing the pest and allows an increase in parasitism in the field through the use of pupae treated with cold- or heat-shock.     

Efficacy of multiple biocontrol agents against the sweet potato whitefly Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) on tomato

Abstract: Biological control provides an environmentally harmonious and potentially stable management tactic to combat noxious pests such as Bemisia tabaci, notorious for its resistance to synthetic pesticides. Bioassays conducted under control chamber conditions integrating applications of the parasitoid Encarsia formosa, reared for 20 years on Trialeurodes vaporariorum, and the fungus Verticillium lecanii on the third‐fourth instar nymphs of B. tabaci on tomato, showed a comparable effect between the parasitoid‐fungus combined treatment and the fungus treatment alone (70.7% vs. 70.4%). Analysis of our results indicates antagonism between the two biocontrol agents related to the parasitoids’ ability to discriminate between infected and healthy B. tabaci nymphs. The parasitoid treatment alone produced 36.3% mortality, with no mortality in the distilled water controls. The behavioural performance of the parasitoid could have either genetic or environmental causes. Bioassays studying the feeding habit of the imported mirid predator Macrolophus caliginosus (adults) and the indigenous mirid Camptotylus reuteri (nymphs and adults) on eggs, or early second instar nymphs of B. tabaci, and choice preference tests indicated a significant difference in feeding between M. caliginosus and C. reuteri. There was no significant difference in percentage feeding of M. caliginosus on eggs (2.2%) or second instar nymphs (8.0%). There was a significant difference in feeding of M. caliginosus adults (18.6%) when offered eggs and second instars in the same arena compared with eggs or second instars offered separately. These results could be attributed to the biological behaviour of the predator having a type III functional response. Studies with the local C. reuteri species showed no significant difference in adult and nymphal consumption on second instars of B. tabaci compared with nymphs on eggs. However, C. reuteri adults fed less on eggs compared with nymphs. This local predatory species appears to be more efficient than M. caliginosus in feeding on particular stages of B. tabaci without depending on prey density. This is further supported by the low consumption of both adults and nymphs in the choice test (4% and 2.3%, respectively) compared with M. caliginosus adults (18.6%).     

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.