Development of a standardized protein immunomarking protocol for insect mark–capture dispersal research

A field study was conducted to test the marking efficiency of broadcast spray applications of protein marks on stationary (represented by cadavers) and free‐roaming lady beetles Hippodamia convergens Guérin‐Méneville that were strategically placed in blooming alfalfa plots. The marks tested included three different concentrations of egg albumin from chicken egg white, casein from bovine milk and trypsin inhibitor from soy milk. The cadaver and free‐roaming beetle treatments served to measure the acquisition and retention of each protein treatment regime by direct contact with the spray solution and by residual contact with protein‐marked residue on alfalfa, respectively. In addition, the vertical distribution of marking efficacy was determined by sampling alfalfa plant tissue and beetle cadavers that were located on the upper and lower portion of the plant canopy. The data indicated that the backpack spray apparatus was very effective at uniformly administering the various protein marks, regardless of the concentration, throughout the entire plant canopy. Also, the free‐roaming beetles readily self‐marked by contact exposure to protein‐treated plants. We also identified concentrations of each protein type that will mark about 90% of the resident beetle population. Moreover, if a mark–capture‐type study only requires two unique protein marks, we determined that concentrations of 25% for egg white and 100% for bovine milk could be used to mark 98% of the population. Our results provide a significant step towards standardizing protein immunomarking protocols for insect mark–capture dispersal research. In addition, we identify several areas of research that are needed to further standardize the protein mark–capture procedure.     

Susceptibility of last instar red flour beetles and confused flour beetles (Coleoptera: Tenebrionidae) to hydroprene.

Last instar larvae of the red flour beetle, Tribolium castaneum (Herbst), and the confused flour beetle, Tribolium confusum Jacquelin du Val, were either exposed for 8-144 h on concrete treated with 1.9 x 10(-3) mg(AI)/per cm2 hydroprene, or continually exposed on concrete treated with 9.8 x 10(-4) to 1.9 x 10(-3) mg[AI]/per cm2 hydroprene. In both tests, larvae were exposed and held at 27 or 32 degrees C and 40, 57, or 75% RH. When larvae were exposed with no food to hydroprene for different time intervals, then transferred to untreated concrete containing flour, consistent effects were produced only at 144 h. At this exposure interval, the percentage of beetles arrested in the larval stage after 3-4 wk was generally greater at 75% RH compared with 40 and 57% RH, but there were no differences between species or temperature. The percentages of dead adult red flour beetles and live adults with morphological deformities were also greatest at 75% RH, and defects were more prevalent in red flour beetles than in confused flour beetles. When larvae were continually exposed to different concentrations of hydroprene on concrete that contained flour, the percentage of arrested larvae, dead adults, and live adults of both species generally increased with concentration. There were more deleterious effects at 75% RH compared with either 40 or 57% RH, and effects were more pronounced in the red flour beetle compared with the confused flour beetle. In both experiments, temperature effects were variable and inconclusive. Results indicate that continual exposure of last instar red flour beetle and confused flour beetle to hydroprene can limit population development, but exposure intervals of >6 d may be required for maximum effectiveness.     

Feeding behaviour and reproductive biology of Colorado potato beetle adults fed transgenic potatoes expressing the Bacillus thuringiensis Cry3B endotoxin

Colorado potato beetle (Leptinotarsa decemlineata Say) adult longevity and fecundity were studied on transgenic potato clones expressing a Cry3B endotoxin of Bacillus thuringiensis (Bt). Adult longevity and fitness were studied for the first 3 weeks after emergence. Beetle reproductive biology on highly resistant clones, intermediary resistant clones and control potato plants was monitored by dissecting females after 7–15 days of feeding and also by analysing haemolymph protein content after 3 days of feeding. Feeding behaviour on transgenic plants expressing high toxin concentrations and on control plants was monitored individually for 36 newly emerged adult beetles feeding on leaf disks during the first two meals. Lethal Time₅₀ for adult beetles feeding on transgenic clones as the sole source of food was not significantly shorter than for beetles on control clones reared in a growth chamber. Differences tended to be larger when the experiment was conducted in a greenhouse with a less optimal temperature range (LT₅₀ = 9.52 and 10.45 days for two transgenic clones and 13.86 for control). In contrast, female egg production on transgenic plants was almost totally inhibited. Dissection studies indicated that adult males living on high-level Bt-expressing transgenic potatoes were still able to mate and produce mobile sperm, but the females were impaired in their reproductive ability since their ovaries were generally not fully developed. An examination of the haemolymph revealed the protein concentration in females living on transgenic plants to be dramatically reduced ( 50%), and electrophoresis showed a reduced content of vitellogenin in these samples.Feeding behaviour of adult Colorado potato beetles was not affected by the different food plants; this indicates that transgenic potato plants were readily accepted as host plants by beetles. The effects of these findings on the use of transgenic plants as a means of L. decemlineata control are discussed.     

Energy use by the mountain pine beetle (Coleoptera: Curculionidae: Scolytinae) for dispersal by flight

The mountain pine beetle Dendroctonus ponderosae Hopkins is a major native pest of Pinus Linnaeus (Pinaceae) in western North America. Host colonization by the mountain pine beetle is associated with an obligatory dispersal phase, during which beetles fly in search of a suitable host. Mountain pine beetles use stored energy from feeding in the natal habitat to power flight before host colonization and brood production. Lipids fuel mountain pine beetle flight, although it is not known whether other energy sources are also used during flight. In the present study, we compare the level of energy substrates, proteins, carbohydrates and lipids of individual mountain pine beetles flown on flight mills with unflown control beetles. We use a colorimetric method to measure the entire metabolite content of each individual beetle. The present study reveals that mountain pine beetles are composed of more protein and lipid than carbohydrate. Both female and male mountain pine beetles use lipids and carbohydrates as energy sources during flight. There is variation between sexes, however, in the energy substrates used for flight. Male mountain pine beetles use protein, in addition to lipids and carbohydrates, to fuel flight, whereas protein content is not different between flown and control females.     

Interspecific and intraspecific variation in diet preference in five Atlantic forest dung beetle species

1. Dung beetles are commonly assumed to be generalist feeders, but there has been limited work in identifying whether there is interspecific variation in feeding preference. Equally, there has been no work exploring whether generalist feeding behaviour in a species is a result of within‐species specialisation. 2. This study identified the individual and species‐level feeding preferences of five dung beetle species towards human, jaguar and pig dung using a choice experiment. 3. It was found that species varied in their preference for the dung types, but there was no evidence that within a species, individual beetles varied in their dung choice. These findings were similar to results from field experiments that are more typically used to assess feeding preferences in dung beetles. 4. The results suggest that individual specialism in feeding may not be common in dung beetles. However, there is variation in feeding preferences among species, which is often overlooked and can have implications for the ecosystem functions they provide. It is suggested that choice arenas can be used to assess feeding preferences in dung beetle species that are not abundant enough to reliably estimate dietary choice from field studies.     

Response of the elm leaf beetle to host plants induced by oviposition and feeding: the infestation rate matters

We investigated by olfactometry and feeding‐ and oviposition‐choice‐tests how the highly specialised elm leaf beetle, Xanthogaleruca luteola Müller (Coleoptera: Chrysomelidae), responds to conspecifically induced defences in the field elm Ulmus minor Miller (Ulmaceae). While egg deposition of the beetle induced elms to release volatiles attractive to the egg parasitoid Oomyzus gallerucae Fonscolombe (Hymenoptera: Eulophidae), feeding alone did not. In the present study, females of the elm leaf beetle showed preferences for the odours of twigs induced by low egg deposition and feeding over odours from uninfested twigs. In contrast, heavy infestation rendered elm odours less attractive to the beetles. Feeding and oviposition bioassays revealed an oviposition preference for leaves from uninfested twigs when compared to locally infested leaves. However, beetles preferred to feed upon systemically induced leaves compared to uninfested ones. The different preferences of the elm leaf beetle during host plant approach might be explained by a strategy that accounts for both gaining access to high quality nutrition and avoiding competition or parasitism.     

How varying pest and trap densities affect Tribolium castaneum capture in pheromone traps

The red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), is an important insect pest in food processing facilities. Pheromone trapping is frequently used to monitor red flour beetle populations in structures; however, the optimal trap density and the relationship between trap captures and beetle density is not known. Two experiments were performed concurrently in environmentally controlled 30‐m² walk‐in chambers to determine the relationship between aggregation pheromone trap captures of red flour beetles and beetle and trap number. In one experiment, beetle density was kept constant at 200 individuals per chamber while trap number was varied from 1 to 8, and in the other experiment trap number remained constant at one per chamber while beetle density varied from 20 to 800 individuals. Results indicated that approximately one out of 23 red flour beetles were captured in a trap. Number of beetles captured in traps increased significantly as beetle density increased; however, the proportion of beetles captured remained consistent across beetle densities with a mean of 4.7 ± 0.6% of individuals captured. Trap captures varied significantly with trap placement within experimental chambers, indicating that subtle differences in the trapping environment can influence trap captures. Data suggested that trap densities of 0.07–0.10 m^?2 (2–3 traps per chamber) would maximize trap capture, whereas a trap density of 0.13 m^?2 (four traps per chamber) would maximize the predictive ability of a trapping equation estimating beetle density from trap captures. Results provide information needed to more thoroughly explore how environmental factors might influence red flour beetle trap capture in the absence of changes in beetle density. Further understanding of these relationships will allow for more accurate assessments of absolute beetle density from pheromone trap capture data.     

Do earthworms help to sustain the slug predator Pterostichus melanarius (Coleoptera: Carabidae) within crops? Investigations using monoclonal antibodies

Earthworms provide a major potential source of alternative food for polyphagous predators, such as carabid beetles, that are natural enemies of slugs, aphids and other agricultural pests. Non-pest prey may foster larger numbers of natural enemies, which then help to control pests, or alternatively may help to divert the predators away from pest control. An earthworm-specific monoclonal antibody was developed to study carabid-earthworm interactions in the field and assess the role of earthworms as alternative prey. The antibody could identify as little at 7 ng of earthworm protein in an ELISA, and could detect earthworm remains in the foregut of the carabid beetle Pterostichus melanarius for 64 h after consumption. Thirty-six per cent of field-collected beetles contained earthworm remains. Quantities of earthworm proteins in the beetle foreguts were negatively related to total foregut biomass, suggesting that earthworm consumption increased as total prey availability declined. There was also a negative relationship between foregut biomass and beetle numbers, but both quantities and concentrations of earthworm proteins in beetle foreguts were positively related to beetle numbers. This suggests that as beetle activity-density increased, total prey availability declined, or, as prey availability declined, beetles spent more time searching. In these circumstances, beetles fed to a greater extent on earthworms, an acceptable but nonpreferred food item. Earthworms may, therefore, provide an ideal alternative prey for P. melanarius, helping to sustain it when pest numbers are low but allowing it to perform a 'lying-in-wait' strategy, ready to switch back to feeding on pests when they become available.     

Influence of pea lectin expressed transgenically in oilseed rape (Brassica napus) on adult pollen beetle (Meligethes aeneus)

Adult pollen beetles were exposed to three transgenic lines of Brassica napus L. expressing pea lectin up to 0.7% of total soluble protein in anthers. Two lines not expressing the pea lectin were used as controls. Adult responses were recorded in two experiments where beetles were fed either racemes or anthers. There was no effect of plant line on feeding rate, weight change or oviposition rate. A slightly reduced egg size was recorded when beetles were feeding on one of the plant lines with high concentration of pea lectin. Previous studies on the effect of pea lectin on pollen beetles have only focused on larval performance. The results of this study suggest that pea lectin has no adverse effect on the pest in its most damaging life stage.     

Oviposition site selection by herbivorous beetles: a comparison of two thistle feeders, Cassida rubiginosa and Henosepilachna niponica

In insects that feed on plants in both adult and larval stages, it is often difficult to distinguish oviposition preference from adult feeding preference, because oviposition can occur at or in proximity to feeding sites. In the present study, characteristics of oviposition site selection of two beetle species, Cassida rubiginosa Müller (Coleoptera: Chrysomelidae) and Henosepilachna niponica (Lewis) (Coleoptera: Coccinellidae), were investigated in the field and laboratory, with particular attention to relationships with adult feeding sites. In the field, distances between adult feeding scars and egg masses differed for C . rubiginosa and H . niponica , with the former being very small and the latter averaging 24.6 cm. The same tendencies for the distances between adult feeding scars and egg masses of the two beetle species were confirmed in cages in which only female beetles were released. Cassida rubiginosa restricted egg laying to host plants in the field and to leaves in laboratory assays. On the other hand, H . niponica placed 8% of egg masses on plants adjacent to host plants in the field and often placed eggs on artificial substrates rather than leaf discs in laboratory assays. These results suggest that oviposition and female feeding sites are virtually inseparable in the case of C . rubiginosa , while H . niponica females do not necessarily keep to host plant leaves as oviposition substrates and they tend to oviposit at some distance from their feeding sites. Results are discussed in relation to proximate and ultimate causes of host selection behavior.     

Preferential Feeding and Occupation of Sunlit Leaves Favors Defense Response and Development in the Flea Beetle,Altica brevicollis coryletorum – A Pest of Corylus avellana

The monophagous beetle, Altica brevicollis coryletorum, is a major leaf pest of Corylus avellana (common hazel). In contrast to majority of the other studied species of shrubs, sunlit leaves are grazed to a much greater extent than shaded leaves. Since the observation of a link between leaf irradiance level and A. brevicollis feeding is unique, we hypothesized that feeding preference of this beetle species is related to the speed needed to escape threats i.e. faster jumping. We also hypothesized that sunlit leaves are more nutritious and easier to consume than the leaves of shaded shrubs. Results indicated that beetle mass was greater in beetles occupying sunlit leaves, which is consistent with our second hypothesis. The study also confirmed under laboratory conditions, that larvae, pupae and beetles that were fed full-light (100% of full light) leaves were significantly heavier than those fed with shaded leaves (15% of full light). In the high irradiance conditions (higher temperature) duration of larval development is also reduced. Further results indicated that neither the concentration of soluble phenols, leaf toughness, or the number of trichomes could explain the insect’s preference for sunlit leaves. Notably, measurements of jump length of beetles of this species, both in the field and under laboratory conditions, indicated that the defense pattern related to jumping was associated with light conditions. The jump length of beetles in the sun was significantly higher than in the shade. Additionally, in laboratory tests, beetle defense (jumping) was more strongly affected by temperature (15, 25, or 35°C for 24h) than by leaf type. The effect of sunlit, higher nutrient leaves (greater level of non-structural carbohydrates) on defense (jumping) appears to be indirect, having a positive effect on insect mass in all developmental stages.     

Effect of pyriproxyfen and photoperiod on free amino acid concentrations and proteins in the hemolymph of the Colorado potato beetle, Leptinotarsa decemlineata (Say)

A North Dakota strain of the Colorado potato beetle, Leptinotarsa decemlineata (Say), was reared under both short- (8L:16D) and long-day (17L:7D) conditions. Age-related and pyriproxyfen- (JHA-) induced changes in hemolymph free amino acids and proteins were examined. Under a short-day photoperiod, the total free amino acid concentration in the hemolymph increased gradually up to 20 days of adult life, but the long-day beetles showed marked increases during the first 10 days and then decreased afterwards. Proline, glutamine and valine were the most abundant free amino acids in both sexes of beetles held under either short- or long-day photoregims. JHA treatment of diapausing adults, held under either short- or long-day conditions after treatment, terminated diapause as indicated by re-emergence from the vermiculite, feeding, mating, changes in free amino acid levels, the disappearance of diapause protein 1 and appearance of vitellogenin in the hemolymph. Furthermore, most of the JHA-treated females held under long-day conditions also matured oocytes and oviposited, but those held under short-day conditions did not.     

Effects of leaf toughness on mandibular wear of the leaf beetle, Plagiodera versicolora

Abstract. 1. The effects of leaf toughness on mandibular wear of the leaf beetle Plagiodera versicolora Laich. (Coleoptera: Chrysomelidae) were evaluated by feeding adult beetles young (tender) and old (tough) leaves of Salix babylonica and S. alba 'Tristis'.
2. Tough leaves erode the cutting surface of beetle jaws more so than tender leaves.
3. Beetles with worn jaws consumed leaves at a slower rate than ones with less mandibular wear.
4. Because rates of leaf consumption and egg production are positively correlated, increased mandibular wear may reduce beetle fecundity.
5. These results support the belief that leaf toughness may act as a potent defence affecting morphology, feeding behaviour, and ultimately spatial and temporal patterns of herbivores.     

Influence of nutrient levels in Tamarix on Diorhabda sublineata (Coleoptera: Chrysomelidae) survival and fitness with implications for biological control

Establishment of the saltcedar leaf beetle (Diorhabda spp.) has been unpredictable when caged or released in the field for saltcedar (Tamarix spp.) biocontrol. It has been observed that one caged tree might be voraciously fed upon by beetles while an adjacent tree in the cage is left untouched. We hypothesized that differences in the nutrient content of individual trees may explain this behavior. We evaluated survival, development rate, and egg production of beetles fed in the laboratory on saltcedar foliage from trees that had been grown under a range of fertilizer treatments. Tissue samples from the experimental trees and from the field were analyzed for percent nitrogen, phosphorus, and potassium. There was essentially no survival of beetle larvae fed foliage from saltcedar trees at nitrogen levels below 2.0%. At levels above 2.0% N, beetle larvae had corresponding increased survival rates and shorter development times. Multiple regression analyses indicated that nitrogen and phosphorus are important for larval survival and faster development rates. Higher levels of potassium were important for increased egg cluster production. The plant tissue analysis showed that the percentage of nitrogen in the experimental trees reflected the range of trees in the field and also that there is high variability within trees in the field. Our research indicates that if beetles are released on trees with poor nutrient quality, the larvae will not survive.     

Evaluating light attraction to increase trap efficiency for Tribolium castaneum (Coleoptera: Tenebrionidae)

The red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), is a major coleopteran pest in flour mills and storage facilities. An aggregation pheromone has been identified for this pest; however, the pheromone is of limited value for population monitoring. To develop more efficient methods to monitor this pest, experiments were conducted to determine whether light functioned as an attractant for the red flour beetle. Light-emitting diodes (LEDs) of various wavelengths were examined as light sources because they produce bright, narrow light spectra. A comparison of responses to light spectra across the visible and UV regions of the electromagnetic spectrum indicated that the beetle was most attracted to near UV LED at a 390 nm dominant wavelength. The use of LEDs in competitive laboratory experiments resulted in a 20% capture of released beetles, compared with a 1% capture with the aggregation pheromone alone. Even more beetles were captured with a combination of LEDs and commercially available chemical lures in traps. LEDs can easily be added onto existing trap designs or new traps can be designed to take full advantage of positive phototaxis.     

Effect of d-limonene on three stored-product beetles

d-Limonene was investigated for contact and fumigant toxicity, ovicidal effects, oviposition-deterrent, development inhibition, and feeding-deterrent activities against three stored-product beetles (Coleoptera): lesser grain borer, Rhyzopertha dominica (F.); rice weevil, Sitophilus oryzae (L.); and red flour beetle, Tribolium castaneum (Herbst). Contact and fumigant toxicity decreased as larvae aged. Contact toxicity was similar for adults of the three species tested, but R. dominica was most susceptible to fumigant activity. T. castaneum oviposition decreased as concentration of d-limonene increased and d-limonene reduced oviposition up to 92.3% at the concentration of 2.14 mg/cm2. Hatching of d-limonene-treated eggs of T. castaneum was reduced by 94.5% with no subsequent larval and adult survival at 2.14 mg/cm2 concentration. A flour disc bioassay indicated 87.7 to 96.8% feeding-deterrency by d-limonene toward all three insect species tested at the highest concentration of 60.0 mg/g food. These results suggest that d-limonene can be effectively used to suppress populations of stored-product beetles.     

The effects of the presence of stored product pests on the microfauna of a flour community

AIMS: To determine the effect that the presence of some beetles have on the species of bacteria found in their flour. METHODS AND RESULTS: Bacteria were isolated from flour that either did not contain beetles, contained Tribolium beetles in different environments, or contained either Stegobium paniceum or Lasioderma serricorne. These bacteria were tentatively identified by both the gas chromatography-fatty acid methyl esters (GC-FAME) method and partial sequencing of the 16S rRNA gene. All samples contained Bacillus species including the controls, but the non-Tribolium beetles and a Tribolium beetle line known to have low benzoquinones also contained Enterococcus and Enterobacter species. Additionally an unidentified bacteria isolate in the Enterobacteriaceae was also found in the L. serricorne sample. Our results also suggest incongruent identifications when using the GC-FAME method vs sequencing. CONCLUSIONS: Certain species of bacteria can be introduced by the presence of insect pests, but the diversity of species is far less in stocks of Tribolium beetles. SIGNIFICANCE AND IMPACT OF THE STUDY: Stored product pests can alter the bacterial community. Isolated species from this study show a strong genetic relationship to each other, suggesting an isolated evolving system. A unique bacteria was also isolated. GC and sequencing methods of identification are compared.     

Field trial of diatomaceous earth in cotton gin trash against the larger black flour beetle, Cynaeus angustus (Coleoptera: Tenebrionidae)

The larger black flour beetle, Cynaeus angustus (LeConte) (Coleoptera: Tenebrionidae), is an agricultural and home nuisance pest in North America. In the Southern High Plains of Texas, the larger black flour beetle is associated with cotton gin trash, by-products of cotton ginning that are field stored in large piles for economic reasons. Larger black flour beetle overwinter in gin trash piles but may disperse by the millions in summer and autumn, entering houses as far as 2 km away where they cause distress to homeowners. Because > 1.2 billion kg of gin trash is produced annually in Texas alone, the potential consequences of the larger black flour beetle are enormous. We conducted a field experiment that evaluated the efficacy of diatomaceous earth (DE) on the abundance of the larger black flour beetle in gin trash. There were no significant differences in numbers of larger black flour beetle among treatments and controls (mean number of adults summed over time: controls = 115.41, layered treatment = 87.60, top and bottom treatment = 96.50, bottom treatment = 115.16). There were sufficient numbers of beetles in treated piles to still pose a potential home nuisance problem, likely because the moisture content of field-stored gin trash is too high for DE to work effectively. Therefore, treating cotton gin trash with diatomaceous earth will probably be unable to prevent home infestations of larger black flour beetle. Location within a gin trash pile and season influenced pest numbers, which has implications for long-term field storage of cotton gin trash.     

Impact of food source on survival of red flour beetles and confused flour beetles (Coleoptera: Tenebrionidae) exposed to diatomaceous earth

A series of experiments was conducted to determine the effect of a flour food source on survival of red flour beetle, Tribolium castaneum (Herbst), and confused flour beetle, Tribolium confusum (DuVal), exposed to the labeled rate (0.5 mg/cm2) of Protect-It, a marine formulation of diatomaceous earth. Beetles were exposed at 27 degrees C, and 40, 57, and 75% RH in 62-cm2 petri dishes. When beetles were exposed for 1 or 2 d in dishes with the labeled rate (0.5 mg/cm2, or 31 mg per dish) of diatomaceous earth or in dishes containing flour at varying levels from 0 to 200 mg mixed with the labeled rate of diatomaceous earth, survival of both species increased as the amount of flour increased, and quickly plateaued at levels approaching 100%. In a second set of experiments, beetles were transferred to dishes containing flour at varying levels from 0 to 200 mg after they were exposed for 1 or 2 d in dishes with the labeled rate of diatomaceous earth alone. There were no significant differences in beetle survival among the levels of flour, however, survival in dishes with flour was usually greater than survival in dishes with diatomaceous earth alone. In a third test, beetles were exposed for 1, 2, and 3 d in dishes with either the labeled rate of diatomaceous earth alone (clean dishes), dishes with diatomaceous earth and empty straws, or dishes with diatomaceous earth and approximately 300 mg of flour packed in the straws. Survival was not significantly different between clean dishes or dishes with straws, but survival in dishes containing the straws with flour was usually 100%, regardless of exposure interval. In all experiments, confused flour beetles were less susceptible to diatomaceous earth than red flour beetles. In addition, survival was negatively related to exposure interval and positively related to relative humidity.     

Impact of Bacillus thuringiensis – insecticides on population dynamics and egg predation of the Colorado potato beetle in North Carolina potato plantings

Field studies to assess the impact of Bacillus thuringiensis var. tenebrionis (Btt)-insecticides on Colorado potato beetle populations, egg survivorship and levels of predation on egg masses were conducted in replicated field research plots during two years. Stage-specific abundance of the Colorado potato beetle and predation on egg masses were monitored in Btt-treated and untreated potato plots in both years. The Btt-treatments significantly reduced densities of large (third and fourth instar) Colorado potato beetle larvae. The densities of large larvae remained below 0.5 and 3 per plant in the Btt-treatment while peak densities of 4.5 and 21 large larvae per plant occurred in the untreated control in 1992 and 1993, respectively. Regular sampling of egg masses indicated that predation rates in Btt-treated and untreated plots did not differ significantly although, in 1993, predation rates of up to 100% were recorded, only in Btt-treated plots. In a predator exclusion study carried out in 1992, survivorship of protected eggs was consistently higher than of eggs exposed to predation. Seasonal survivorship of exposed eggs was significantly lower in the Btt-treated than in untreated plots. Btt insecticides for control of Colorado potato beetles provided direct protection of the crop and were compatible with naturally-occurring biological control of Colorado potato beetle eggs due to predation.