Healthy Pupae Research Articles

Understanding the differential performance in hygienic behavior against dead brood of drones and workers of Apis mellifera: a chemical approach

Hygienic behavior is a social health mechanism displayed by honey bee workers of Apis mellifera by which dead or diseased brood is removed from the hive. Such behavior is known to be induced by olfactory cues. The efficiency of hygienic behavior toward drone brood cells is significantly lower than that of worker brood cells. A possible explanation for this difference lies in the cell wax cap of drone brood, which may act as a barrier to the diffusion of characteristic odors associated with diseased brood. The aim of this research was to explore the importance of drone cell wax cap as an interfering factor in the diffusion of chemical signals within honey bee colonies. To this end, brood removal percentages were compared among several types of brood: pin-killed worker, pin-killed drone, pin-killed worker with a healthy drone wax cap, healthy worker with a pin-killed drone wax cap, and healthy worker covered with a healthy drone wax cap. Results showed that the removal behavior did not differ between cells with pin-killed worker and pin-killed worker with a healthy drone wax cap, but was significantly higher towards healthy worker pupae covered with dead drone wax cap compared to the negative control (healthy worker pupa covered with a healthy drone cell wax cap). Present evidence suggests that chemical signals associated with the dead drone brood would be less volatile and retained to some extent in the wax cap, and partially explain the differential hygienic behavior between both types of brood.

Cynipid galls on oak leaves are resilient to leaf vein disruption

Oaks serve as host plants for numerous insects, including those forming galls. Galls induced on oaks are completely dependent on leaf resources. Many other folivores damage veins of leaves, which may result in cutting galls off from sources of assimilates, nutrients and water. We hypothesised that the disruption of the continuity of leaf vascular tissues stops gall development, leading to the death of the larva. Leaves of sessile oak (Quercus petraea) with Cynips quercusfolii galls in the initial stage of development were marked. The diameter of the galls was measured, and the vein on which the gall was present was cut. Four experimental treatments were established: control – with no cutting, cutting the vein distal to the gall relative to the petiole, cutting the vein basal to the gall and cutting both sides. The average survival rate (live galls at the end of the experiment including healthy larvae, pupae or imagines inside) – was 28.9%. The rate varied depending on the treatment and was 13.6% in the treatment with the vein cut on both sides and about 30% in the remaining treatments. However, this difference was not statistically significant. The growth dynamics of galls are highly dependent on the experimental treatment. The largest galls grew in the control treatment, and the smallest galls were in the treatments with the veins cut on both sides. Unexpectedly, even cutting veins on both sides did not result in the immediate dieback of the galls. The results suggest that the galls are very strong nutrient and water sinks. The functions of the cut vein are likely taken over by other lower-order veins, allowing nourishment of the gall to complete larva development.

Batch Rearing Aedes aegypti.

Standardized rearing methods for the yellow fever mosquito Aedes aegypti are critical to facilitate controlled laboratory studies. This protocol describes a batch rearing protocol for Aedes aegypti stocks that yields healthy eggs, larvae, pupae, and adults in the laboratory for long-term colony maintenance and experimental manipulation. Foundational principles for the rearing and containment of these life cycle stages, as well as steps for mating and blood feeding Aedes aegypti to yield viable eggs for continuous culture or storage, are detailed.

Fenoxycarb Effects 7th instar Galleria mellonella Larvae by Triggering Extra Larval Stages

Insect growth regulators are specific chemicals with hormonal effects on insects and these types of chemicals target to reduce the nonspecific effects of pesticides on nature and living things. In this study, we investigated the effects of one of the insect growth regulators, fenoxycarb which mimics the juvenile hormone action, on the growth of a harmful lepidopteran species Galleria mellonella, and it’s some developmental parameters. For this purpose, fenoxycarb was applied on day 0 of 7th instar larvae of the greater wax moth, Galleria mellonella. The treatment with 100 ng fenoxycarb had no developmental effect on Galleria mellonella larvae. The treatment with relatively high doses of fenoxycarb on day 0 induced the supernumerary larvae formation of 99% and larvae which molted extra larval instar formed healthy pupae in the ratio of 98-99%. Consequently, the effects of fenoxycarb on Galleria mellonella larvae occurred in a dose-dependent manner. Although fenoxycarb is a potent insect growth regulator, late Galleria mellonella larvae are not susceptible to this chemical, unlike some other Lepidopter insect species.

Codon optimization, expression in Escherichia coli, and immunogenicity analysis of deformed wing virus (DWV) structural protein.

BackgroundDeformed wing virus (DWV) is a serious threat to honey bees (Apis mellifera) and is considered a major cause of elevated losses of honey bee colonies. However, lack of information on the immunogenicity of DWV structural proteins has hindered the development of effective biocontrol drugs.MethodsWe optimized the VP1, VP2 and VP3 codons of DWV surface capsid protein genes on the basis of an Escherichia coli codon bias, and the optimized genes of roVP1, roVP2 and roVP3 were separately expressed in E. coli and purified. Next, the three recombinant proteins of roVP1, roVP2 and roVP3 were intramuscularly injected into BALB/c and the immunogenicity was evaluated by the levels of specific IgG and cytokines. Furthermore, anti-roVP-antisera (roVP1 or roVP2 or roVP3) from the immunized mice was incubated with DWV for injecting healthy white-eyed pupae for the viral challenge test, respectively.ResultsThe optimized genes roVP1, roVP2 and roVP3 achieved the expression in E. coli using SDS-PAGE and Western blotting. Post-immunization, roVP2 and roVP3 exhibited higher immunogenicity than roVP1 and stimulated a stronger humoral immune response in the mice, which showed that the recombinant proteins of roVP3 and roVP2 induced a specific immune response in the mice. In the challenge test, data regarding quantitative real-time RT-PCR (qRT-PCR) from challenged pupae showed that the level of virus copies in the recombinant protein groups was significantly lower than that of the virus-only group at 96 h post-inoculation (P < 0.05). Among them, the degree of neutralization using antibodies raised to the recombinant proteins are between approximately 2-fold and 4-fold and the virus copies of the roVP3 group are the lowest in the three recombinant protein groups, which indicated that specific antibodies against recombinant proteins roVP1, roVP2 and roVP3 of DWV could neutralize DWV to reduce the virus titer in the pupae. Collectively, these results demonstrated that the surface capsid protein of DWV acted as candidates for the development of therapeutic antibodies against the virus.

Response of the honey bee (Apis mellifera) proteome to Israeli acute paralysis virus (IAPV) infection

Recent declines in honey bee (Apis mellifera L., 1758) populations worldwide have spurred significant research into the impact of pathogens on colony health. The role of the Israeli acute paralysis virus (IAPV) on hive mortality has become of particular concern since being correlated with colony losses. However, the molecular interactions between IAPV and its host remain largely unknown. To investigate changes in host protein expression during IAPV infection, mass-spectrometry-based quantitative proteomics was used to compare IAPV-infected and healthy pupae. Proteins whose expression levels changed significantly during infection were identified and functional analysis was performed to determine host systems and pathways perturbed by IAPV infection. Among the A. mellifera proteins most affected by IAPV, those involving translation and the ubiquitin–proteasome pathway were most highly enriched and future investigation of these pathways will be useful in identifying host proteins required for infection. This analysis represents an important first step towards understanding the honey bee host response to IAPV infection through the systems-level analysis of protein expression.

Dynamics of the presence of israeli acute paralysis virus in honey bee colonies with colony collapse disorder.

The determinants of Colony Collapse Disorder (CCD), a particular case of collapse of honey bee colonies, are still unresolved. Viruses including the Israeli acute paralysis virus (IAPV) were associated with CCD. We found an apiary with colonies showing typical CCD characteristics that bore high loads of IAPV, recovered some colonies from collapse and tested the hypothesis if IAPV was actively replicating in them and infectious to healthy bees. We found that IAPV was the dominant pathogen and it replicated actively in the colonies: viral titers decreased from April to September and increased from September to December. IAPV extracted from infected bees was highly infectious to healthy pupae: they showed several-fold amplification of the viral genome and synthesis of the virion protein VP3. The health of recovered colonies was seriously compromised. Interestingly, a rise of IAPV genomic copies in two colonies coincided with their subsequent collapse. Our results do not imply IAPV as the cause of CCD but indicate that once acquired and induced to replication it acts as an infectious factor that affects the health of the colonies and may determine their survival. This is the first follow up outside the US of CCD-colonies bearing IAPV under natural conditions.

Natural and artificial envenomation ofCeratitis capitatabyEupelmus urozonusand the search for new bioinsecticides

This study was designed to indentify novel bioactive molecules in the venom of the parasitoid Eupelmus urozonus Dalman (Hymenoptera: Eupelmidae). Parasitism by E. urozonus induces apparent paralysis in the larvae of the host Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) and it arrests the development of host pupae. Parasitoid eggs were transferred from stung to unparasitized host pupae to determine whether adult female stings or bites from the first instars were responsible for the above effects. The two treatments gave the same results, indicating that both parasitoid adults and larvae produced venoms capable to compromise host development. A protocol was developed to artificially microinject E. urozonus venom into healthy host pupae and adults at known concentrations to study the effects. The microinjection of venom was found to produce the same macroscopic result as natural parasitization, indicating that host developmental arrest was caused by molecules produced by the parasitoid venom glands. One-tenth, one-twentieth, and one-hundredth of the contents of a female venom reservoir was sufficient to compromise the development of 100, 90, and 50% of the microinjected host pupae, respectively. The microinjection of 0.1 female venom equivalents into host adults always caused death within 24 h. Extraction and freezing did not affect the activity of the E. urozonus venom, which facilitates its storage, whereas denaturation treatments demonstrated that the bioactive molecules were proteins. The venom was also found to prevent the hosts from decaying for over 2 weeks and it promoted the accumulation of unknown subspherical granules in the host haemocoel. These results suggest the potential identification of novel molecules with interesting biological activity with various possible applications.

Feasibility Study of silkworm pupa sex identification with pattern matching

Silkworm sex identification is one of the important processes in the sericulture industry because it can assist in effectively separating strong and healthy silkworm pupae from the weak ones. In this paper, we study and show that a desired moderate accuracy and fast response time in silkworm gender identification can be realized by deploying a widely-used and simple normalized cross correlation (NCC)-based pattern matching operation in our optical penetration-based silkworm pupa gender identification structure. Other key features are ease of implementation, adaptive learning ability, and low component counts. Experimental proof of concept is performed by using three 636-nm wavelength light emitting diodes, one 1600×1200-pixel web camera, an 8-bit microcontroller, a notebook computer, and our LabView program. There are 25 female and 20 male silkworm pupae under the study. Experimental results show that male silkworm pupae can be completely identified under the NCC-based pattern matching performed in both the Cartesian and polar coordinate systems. For the female silkworm pupa, the best measured accuracies of 80% and 56% are obtained by performing the NCC-based pattern matching operations in the Cartesian and polar coordinate systems, respectively. The NCC-based pattern matching operation in the Cartesian coordinate system also offers a measured 13-ms response time, which is twice faster than in the polar coordinate system. The moderate measured total accuracy of 88% and fast silkworm gender identification shows high potential for the deployment in the sericulture industry.

Control of Mosquitoes in Catch Basins in Connecticut With Bacillus thuringiensis israelensis, Bacillus sphaericus, and Spinosad

Catch basins are a major source of Culex pipiens pipiens, Cx. restuans, and Aedes japonicus in northeastern USA. VectoBac CG (Bacillus thuringiensis israelensis [Bti]), VectoLex CG (Bacillus sphaericus [Bs]), and VectoBac 12AS (Bti), each applied at maximum label rate of 1.8 g, 1.8 g, and 0.193 ml per catch basin, respectively, significantly reduced the numbers of larvae for 1 wk. The dosages on the labels for treatment of mosquito larvae in catch basins, where mosquito breeding is continuous, are not adequate for providing long-term control in the northeastern USA without the need for frequent retreatment. When applied at 3 times the maximum label rate, VectoLex CG, VectoBac 12AS, and VectoBac CG significantly reduced the numbers of larvae for 5, 4, and 2 wk, respectively. A single application of VectoMax WSP (Bti + Bs) (1 pouch containing 10 g) per catch basin significantly reduced the numbers of 3rd and 4th instars and healthy pupae in catch basins in 2008, but numbers of 3rd and 4th instars in treated catch basins at 21 days after treatment had increased to 40% of the numbers in untreated catch basins. A 2nd treatment of 1 pouch per catch basin reduced the numbers of 3rd and 4th instars and healthy pupae to near zero for the next 4 wk, into the middle of September 2008. In 2009, VectoMax applied as 1 pouch per catch basin on July 1 and again on August 18 significantly reduced the numbers of healthy pupae throughout the summer until the end of September. A 2nd application of VectoMax to catch basins is likely needed during summer, when rainfall averages 13.7 in. (approximately 34.25 cm) during June through September, to keep the numbers of Culex and Ae. japonicus significantly reduced to lower risk of human exposure to West Nile virus. The application of 1 Natular XRT tablet, each weighing approximately 40.5 g (6.25% spinosad), to individual catch basins in 2009 significantly reduced the total numbers of larvae for 5 wk.

Notes on the defoliation of Coconut Palm (Cocos nucifera) by the butterfly Opsiphanes quiteria quirinus in northeastern Costa Rica (Lepidoptera: Brassolidae)

The defoliation of two adjacent, mature trees of the palm Cocos nucifera by caterpillars of the butterfly Opsiphanes quiteria quirinus (Lepidoptera: Brassolidae) was studied in northeastern Costa Rica. During the early dry season (February, 1976), these trees were heavily striped of leaves by the caterpillars, producing a large number of healthy pupae that survived to produce adults. The population is characterized by non-overlapping generations on both trees. A single, non-overlapping generation produces a large number of adults that subsequently place many eggs on trees used by the previous generation. Oviposition occurs at dusk; eggs are placed singly or clustered on leaves, trunks, and the loose sheaths of fronds. Pupation occurs primarily on fronds sheaths and leaves. The infestation of caterpillars and pupae are greatest on lower fronds, presumably the result of a strong preference for oviposition on these leaves. Survivorship of at least fifth instar caterpillars in addition to pupae is high. Although Cocos palms are scarce at this locality, most of those present did not have an infestation, suggesting that either the observed heavy outbreak was unusual or that colonizing episodes are just beginning to occur here. While such outbreaks of Opsiphanes and other brassolids on banana have been documented many times in Central America, the outbreak of O. quiteria on Cocos palm in northeastern Costa Rica is a recent event, perhaps related to the increased removal of the original forest cover in recent years. On wild palms O. quiteria and other brassolids are far less abundant.

Semi-captive rearing of the African wild silkmoth Gonometa postica (Lepidoptera: Lasiocampidae) on an indigenous and a non-indigenous host plant in Kenya

The life cycle of the African wild silkmoth Gonometa postica Walker was studied between September 2000 and September 2001 in the Uasin Gishu District of western Kenya with Acacia mearnsii de Wild and Acacia hockii de Wild as host plants. The initial population of G. postica was set up from healthy live pupae in cocoons collected from host plants in the study area. Oviposition was carried out in net sleeves and small plastic cages. Moth emergence and oviposition were bimodal, occurring between September and October 2000 and between March and April 2001, respectively. Larvae were reared in semi-captivity by using net sleeves on the branches of the two host plants. Larvae passed through six developmental instars. The pupal stage lasted from December to February during the short rains and from June to September during the long rains. Moth fecundity in the net sleeves was higher than that of those kept in plastic cages and fewer eggs hatched in the plastic cages compared with the net sleeves. Larvae reared on A. mearnsii had a shorter developmental period compared with those reared on A. hockii. Male larvae spun cocoons earlier than females but moths of both sexes emerged simultaneously. Sexual dimorphism was exhibited in both pupal and adult stages, females being larger and heavier than males. The importance of the semi-captive rearing technique for sustainable utilization of wild silkmoths for mass production of cocoons is discussed.

Interactions between predatory ground beetles, the winter moth and an introduced parasitoid on the Lower Mainland of British Columbia

This study describes assemblages of predatory ground beetles in native birch forest and derived blueberry farms on the Lower Mainland of British Columbia and investigates their role in the successful biological control of winter moth ( Operopthera brumata L.) by the parasitoid Cyzenis albicans Fallén. Pitfall traps at four sites in 1991 and 1992 indicated that natural birch forest was dominated by native carabid species, while open blueberry was dominated by introduced species and some native xerophilous species. Predation by generalist invertebrates was an important mortality factor for both winter moth and C. albicans at all sites and was greatest in the first 2 weeks after pupation. Ground beetles including Harpalus affinus (Schrank) and Pterostichus spp. consumed winter moth pupae in arenas but adult beetles rarely consumed C. albicans pupae. Much of the overall predation was due to smaller invertebrates including beetle larvae. Predators actively searched for buried pupae and selected among pupal types based on condition (dead, parasitised or healthy) and size. Prey-selection differed among sites but C. albicans pupae were preferred at three of the four sites. At the remaining site, an organic blueberry farm, over 90% of both parasitised and healthy pupae were consumed. High levels of predation did not affect the levels of parasitism by C. albicans at the sites. This herbivore–parasitoid–predator system demonstrates the importance of naturally occurring predatory invertebrates for successful integrated pest management, even where they prey upon a biological control agent.

Potential mechanism for detection by Apis mellifera of the parasitic mite Varroa destructor inside sealed brood cells

Abstract The parasitic mite Varroa destructor Anderson &amp; Trueman is a major pest of the honeybee Apis mellifera L. throughout the world. Chemical agents currently used for mite control leave contaminating residues and promote pesticide resistance. As an alternative means of control, it would be useful to identify natural substances enabling bees to detect Varroa inside brood cells. These substances could then be used to trigger mite hygienic behaviour by bees.In this study several techniques were used to screen substances that might allow detection of infested brood cells by bees. Gas chromatography‐mass spectrometry analysis was performed on substances extracted in dichloromethane from the contents of brood cells. Solid phase microextraction and solid injection were performed on substances obtained from living and dead Varroa, respectively. Electroantennography was performed to assess the sensitivity of olfactory receptors in bee antennae to some of these substances.Principal component analysis based on proportions of cuticular substances allowed discrimination between bees and other cell contents. Foundress Varroa exhibited the greatest dissimilarity to healthy pupae that were used as controls. Immature Varroa and faecal material were intermediate. High molecular weight compounds, mainly dimethylalkanes, were proportionally the most characteristic components of foundress Varroa. This finding suggests that these compounds would be the most apt to induce uncapping of cells infested by Varroa. Solid‐phase microextraction and solid injection demonstrated the presence of aliphatic acids, esters, and one alcohol, eicosenol, in Varroa. Electroantennographic recordings showed that mite‐resistant bees were more responsive to some acids and one ester. We speculate that these compounds may be involved in recognition of living Varroa by honeybees.

Rearing Techniques for Creophilus maxillosus (Coleoptera: Staphylinidae), a Predator of Fly Larvae in Cattle Feedlots

Laboratory rearing methods were developed for the hairy rove beetle, Creophilus maxillosus L. Adults produced more eggs and lived longer when fed blowfly, Calliphora sp., larvae than when fed house fly, Musca domestica L., larvae. Well-fed adults produced 8.1 eggs per day compared with starved females, who produced 2.2 eggs per day. Wet sand was used as a rearing substrate; 4–6% moisture by weight was preferred by larvae, prepupae, and adults over that with 2% moisture. However, eggs hatched equally at all 3 moisture levels. Egg production decreased when sand became contaminated. Fresh sand resulted in increased egg laying. Survival was low (40%) from egg to 1st instar and during the prepupal stage (52%). Adults resulted from 87% of the healthy pupae. With 30% survival rate from egg to adult and a rate of lay of 500 eggs per adult female, an estimated 421,865 females could be produced in 3 generations.

Mulching with grass‐clippings in cauliflower: Effects on yield and brassica root flies(Deliaspp.)

Abstract The effects of mulching with organic materials on yield and damage by brassica root maggots (Delia floralis and D. radicum) in cauliflower, were studied in field experiments for 3 years. Mulching with grass‐clippings consistently resulted in increased yield and reduced damage by root maggots, as measured by wilting symptoms and root damage. During one of these years, the effects of mulching with grass‐clippings on root fly population dynamics was also studied. Mulching did not reduce egg‐laying but resulted in increased egg‐predation. The number of D. floralis pupae per plant was not reduced by mulching, probably because of higher larval mortality owing to competition in unmulched plots. Mulching decreased the rate of parasitization by Aleochara bilineata (Staphylinidae), resulting in a higher number of healthy pupae per plant in grass‐mulched plots, and these healthy pupae were heavier. All these effects were most pronounced when the mulch material completely covered the ground, even close to th...

Laboratory rearing of the legume pod borer, Maruca testulalis Geyer (Lepidoptera: Pyralidae) on a semi-synthetic diet

The legume pod-borer, Maruca testulalis was reared in the laboratory for 10 successive generations using a semi-synthetic diet. The diet was composed of locally available soybean flour and cowpea flower powder as basal ingredients. The artificial medium was as suitable as the cowpea host plant for M, testulalis in supporting larval growth and development. Fecundity increased with advancing generations on the artificial medium. Field collected pupae were heavier than diet-reared pupae. However, the fecundity, fertility, adult life span, and sex ratio did not differ between the two rearing regimes. The laboratory-reared insects were of acceptable uniform quality and predictable performance even after 10 successive generations of laboratory rearing. Adult emergence ranged between 70–90%. One litre of diet produced on average 400 pupae or adults with a mean fecundity of 200 eggs per female. One person working 8 hr/day can produce a minimum of 1000 healthy pupae or adults daily. The laboratory rearing management protocol is described.

Mortality of Overwintering Pupae of the Cabbage Root Fly (Delia brassicae)

(1) During the winter of 1976-77, few pupae were removed by predators from either natural or introduced populations of pupae. (2) An increase in pupal density from one to thirty-two pupae/plant did not increase mortality. (3) Over three winters, about 15% of the pupae were killed by physical damage and pathogens. The very cold winter of 1978-79 did not increase mortality. (4) Healthy pupae and pupae parasitized by Idiomorpha rapae were killed by chlorfenvinphos at >5 ,g g-1 of soil at the start of winter. There was a linear relationship between the concentration of chlorfenvinphos and the logit of the proportion of healthy pupae killed. In contrast, the relationship for parasitized pupae, which were extremely sensitive to chlorfenvinphos at concentrations > 10 ,ug g91, was not linear. (5) The percentage of adults emerging from pupae was not affected by the depth of the pupae in the soil. Nevertheless, about 30% of flies that emerged at 3-75 cm to 15 cm and 80% at 30 cm deep failed to reach the soil surface. (6) Cultural practices most affected pupal mortality. When Wellesbourne soil was uncultivated, flies successfully reached the soil surface from nearly half of the total pupae whereas after autumn and spring ploughing only 12 and 31% respectively did so. Ploughing in early winter rather than in spring may therefore aid control of D. brassicae populations.

PREDATION BY LONCHAEA CORTICIS (DIPTERA: LONCHAEIDAE) ON THE WHITE PINE WEEVIL, PISSODES STROBI (COLEOPTERA: CURCULIONIDAE)

AbstractThe predatory behavior of Lonchaea corticis Taylor on the white pine weevil, Pissodes strobi Peck, in Sitka spruce, Picea sitchensis (Bong.) Carr., was studied by temporal sampling and dissection of terminal leaders, and by laboratory experiments. L. corticis oviposition occurred when mining P. strobi larvae were consolidating the feeding ring, an event that segregates the weevil larvae into healthy front-feeders and weak, starving "followers." The number of L. corticis within a Sitka spruce terminal was highly correlated with the number of weak and dying P. strobi larvae, but not with healthy larvae.L. corticis larvae experimentally deprived of dead P. strobi larvae, behaved as an effective predator, consuming both weak P. strobi larvae and healthy pupae, but apparently not healthy larvae. The transition of L. corticis from second to third instar appeared to occur only after sufficient weevils had been consumed. When an excess of prey was present, L. corticis larvae consumed a mean of 2.9 P. strobi pupae over their entire life cycle. In choice experiments, L. corticis larvae searched for and located mining P. strobi larvae, and fed preferentially on P. strobi pupae rather than granary weevil pupae, Sitophilus granarius L. Under certain circumstances, L. corticis could be an important regulatory agent of P. strobi populations.

Fatty acid and glycogen requirement of Heliothis virescens infected with cytoplasmic polyhedrosis virus

1. 1. Five-day-old larvae of Heliothis virescens infected with cytoplasmic polyhedrosis virus (CPV) had 66% more fatty acid calories than healthy larvae of the same age. 2. 2. Fatty acid caloric values of healthy and CPV-infected pupae were not significantly different. 3. 3. Infected pupae had six times the fatty acid calories of 10-day-old larvae; healthy pupae had a three-fold increase. 4. 4. Infected pupae had an average loss of 50% weight during metamorphosis to adults, 10% due to loss of fatty acids. 5. 5. Calculated on fresh weight basis, infected adult males had 31% less fatty acid calories than healthy adult males; infected females had 34% less than the healthy female. 6. 6. Infected males and females had 31 and 49% less glycogen, respectively, than healthy adults. 7. 7. During emergence of pupae to adults, approx 60 and 30 glycogen cal/insect were used by healthy and infected insects, respectively.