Fifth-instar Caterpillars Research Articles

Developmental and reproductive effects of clothianidin exposure in monarch butterflies (Lepidoptera: Nymphalidae)

AbstractNeonicotinoid insecticides are used to reduce crop damage caused by insect pests, but sublethal levels could affect development and reproduction in nontarget insects, such as monarch butterflies (Danaus plexippus) (Lepidoptera: Nymphalidae). To investigate the impact of field-realistic concentrations of the neonicotinoid clothianidin on monarch butterflies, we grew swamp milkweed (Asclepias incarnata) (Apocynaceae) in either low (15 ng/g of soil) or high (25 ng/g of soil) levels of clothianidin, or in a control (0 ng/g), then raised monarchs on the milkweed. Morphological traits of monarch caterpillars were measured during development and, once they eclosed, were mated as adults to quantify egg size and mass and the number of eggs laid. Although the effects of the treatment had complex effects on caterpillar length, width and volume of late-instar caterpillars were negatively affected. Fifth-instar caterpillars from the high-dose insecticide treatment had lower mass than other groups. Adult monarch butterflies raised on treated milkweed were larger than controls, but clothianidin exposure did not affect the number of eggs laid or egg size. Although the magnitude of the effect depends on clothianidin concentration, our results suggest that exposure to clothianidin during early life can impact monarch caterpillar development but is unlikely to reduce female reproductive output.

Pyrethroid Exposure Reduces Growth and Development of Monarch Butterfly (Lepidoptera: Nymphalidae) Caterpillars.

Insecticide exposure has been identified as a contributing stressor to the decline in the North American monarch butterfly Danaus plexippus L. (Lepidoptera: Nymphalidae) population. Monarch toxicity data are currently limited and available data focuses on lethal endpoints. This study examined the 72-h toxicity of two pyrethroid insecticides, bifenthrin and β-cyfluthrin, and their effects on growth and diet consumption. The toxicity of bifenthrin to caterpillars was lower than β-cyfluthrin after 72 h. Survival was the most sensitive endpoint for bifenthrin, but diet consumption and caterpillar growth were significantly reduced at sublethal levels of β-cyfluthrin. Using AgDRIFT spray drift assessment, the aerial application of bifenthrin or β-cyfluthrin is predicted to pose the greatest risk to fifth-instar caterpillars, with lethal insecticide deposition up to 28 m for bifenthrin and up to 23 m for β-cyfluthrin from treated edges of fields. Low boom ground applications are predicted to reduce distances of lethal insecticide exposure to 2 m from the treated field edge for bifenthrin and β-cyfluthrin. Growth and survival of fifth-instar monarch caterpillars developing within the margins of a treated field may be significantly impacted following foliar applications of bifenthrin or β-cyfluthrin. These findings provide evidence that pyrethroid insecticides commonly used for soybean pest control are a potential risk to monarch caterpillars in agricultural landscapes.

Effect of Host Plants on the Viability of Black-veined White Aporia crataegi L. at Low Natural Population Density

Patterns of interaction of an oligophagous insect, black-veined white Aporia crataegi L., with host plants in years with a low density of its natural population are analyzed. In years with low A. crataegi population abundance in the south of Sverdlovsk region (Sysertsky district), the species composition of caterpillar host plants gradually decreases from several woody rosaceous species of the genera Padus, Sorbus, Malus, and Crataegus to one species (Padus avium Mill.). It is shown that the mortality rate is higher for fifth-instar caterpillars developing on mountain ash than for those developing on bird cherry. Adult A. crataegi developing on bird cherry are not only larger but also grow faster than those developing on mountain ash, which may be an additional factor responsible for the observed chronographic variation during the emergence of A. crataegi generation in nature.

Secondary Metabolites of Asclepias curassavica (Apocynaceae) and its Effects on Food Preference and Mortality of Spodoptera frugiperda (Lepidoptera: Noctuidae)

Asclepias curassavica L. plant is toxic for vertebrates, and little is known about its effects on invertebrates, as well as whether its secondary metabolites have an influence on food preference and survival of insects. Thus, a study was conducted to verify the action of A. curassavica on Spodoptera frugiperda J.E.Smith. The plants were collected, dried and ground to compose a crude ethanolic extract. Food tests were carried out with and without opportunity of choice, using corn leaf discs immersed in the extract at concentrations of 1%, 2% and 4%, for assessment of food consumption (g) and preference index. Later, the extract was topically applied on second- and fifth-instar caterpillars, at concentrations of 1%, 2%, 4% and 6%, in order to observe its effect on survival. In parallel, analysis was conducted to verify the presence and measure the amount of total polyphenols and flavonoids in the extract. There was contact action between the extract and second-instar caterpillars at all concentrations, with treatment 6% causing 100% of mortality 72 hours after application. None of the treatments promoted contact action on fifth-instar caterpillars. In food tests with and without opportunity of choice, all treatments caused reductions in consumption, which classified all as phagodeterrent, with treatment 4% standing out as the least favorite. The presence of total polyphenols and flavonoids was found in the extract, presenting 58.75 ug/mL and 150.1 ug/mL, respectively. The A. curassavica extract proved promising in S. frugiperda control.

Antimicrobial resistance and genetic relationships of enterococci from siblings and non-siblings Heliconius erato phyllis caterpillars.

BackgroundStudies evaluating bacteria in insects can provide information about host–microorganism–environment interactions. The gut microbial community has a profound effect on different physiological functions of insects. Enterococcus spp. are part of the gut community in humans and other animals, as well as in insects. The presence and antimicrobial resistance profile of enterococci are well studied in different animals; however, data for Heliconius erato phyllis (Lepidoptera: Nymphalidae) do not yet exist. Therefore, the aims of this study were to evaluate the distribution of enterococcal species, their antimicrobial resistance profile and virulence genes, and the genetic relationships between enterococci isolated from fecal samples from sibling and non-sibling H. erato phyllis caterpillars collected from different sites in South Brazil.MethodsThree H. erato phyllis females were captured (two from a forest fragment and one from an urban area), and kept individually in open-air insectaries. Eggs were collected and caterpillars (siblings and non-siblings) were fed daily with Passiflora suberosa leaves. Fecal samples (n = 12) were collected from fifth-instar caterpillars, inoculated in selective medium, and 15 bacterial colonies were randomly selected from each sample. Enterococci were identified by PCR and MALDI-TOF, analyzed by disk diffusion antimicrobial susceptibility tests, and screened for resistance and virulence genes by PCR. The genetic relationships between the strains were determined using pulsed-field gel electrophoresis (PFGE).ResultsA total of 178 enterococci strains were identified: E. casseliflavus (74.15%; n = 132), E. mundtii (21.34%; n = 38), E. faecalis (1.12%; n = 2) and Enterococcus sp. (3.37%; n = 6). High rates of resistance to rifampicin (56%) and erythromycin (31%) were observed; 120 (67.41%) of the isolates showed resistance to at least one antibiotic and six (3.37%) were multidrug-resistant.None of the erythromycin-resistant strains was positive for the erm(B) and msrC genes. The virulence genes esp, ace, and gelE were observed in 35%, 7%, and 1% of the strains, respectively. PFGE separated the enterococci into 22 patterns, four being composed of strains from sibling caterpillars.ConclusionEnterococcus casseliflavus was the dominant species in fecal samples of fifth-instar caterpillars. Resistant enterococci strains may be related to environmental pollution or the resistome. The PFGE analysis showed genetic relationships between some strains, suggesting that the enterococci isolated from fecal samples of the sibling caterpillars might have come from common sources, e.g., via diet (herbivory) and/or vertical transmission (through the egg surface). Further studies will be conducted to better understand the role of Enterococcus in the microbial community of the gastrointestinal tract of these insects, and the mechanisms involved in acquisition and maintenance of enterococci.

Diet factors responsible for the change of the glucose oxidase activity in labial salivary glands of Helicoverpa armigera

We investigated the change of the glucose oxidase (GOX) activity in labial salivary glands of Helicoverpa armigera larvae fed with the artificial diet or host plant tobacco and the major factors responsible for such a change. Throughout larval development, the labial salivary GOX activities in caterpillars reared on the artificial diet were remarkably higher than those fed with the plant. After fifth-instar plant-fed caterpillars were transferred to the artificial diet, their labial salivary GOX activity increased quickly, which was closely correlated with the time spent feeding on the artificial diet. The total sugar content of the artificial diet was 68 times higher than that of the tobacco leaves. We hypothesized that sugars and secondary metabolites are the possible causes of induction of GOX activity. When fifth-instar caterpillars were fed with tobacco leaves coated with glucose or sucrose, their labial salivary GOX activity was significantly higher than those fed with leaves without sugar coating. Following native PAGE, 1 single band of the labial salivary GOX was observed in all the caterpillars fed with different diets, implying that only the activity of the isoenzyme was changed in response to different diets. Furthermore, the labial salivary GOX activity was determined after caterpillars were fed with artificial diets containing chlorogenic acid, rutin, and quercetin. The results showed that all these phenolic compounds had no effect on the GOX activity. We conclude that sugar in diets was a major factor influencing the labial salivary GOX activity of the larvae. Arch. Insect Biochem. Physiol. 2008.

THE ENVIRONMENTAL AND GENETIC CONTROL OF SEASONAL POLYPHENISM IN LARVAL COLOR AND ITS ADAPTIVE SIGNIFICANCE IN A SWALLOWTAIL BUTTERFLY

Seasonal polyphenism, in which different forms of a species are produced at different times of the year, is a common form of phenotypic plasticity among insects. Here I show that the production of dark fifth-instar caterpillars of the eastern black swallowtail butterfly, Papilio polyxenes, is a seasonal polyphenism, with larvae reared on autumnal conditions being significantly darker than larvae reared on midsummer conditions. Both rearing photoperiod and temperature were found to have individual and synergistic effects on larval darkness. Genetic analysis of variation among full-sibling families reared on combinations of two different temperatures and photoperiods is consistent with the hypothesis that variation in darkness is heritable. In addition, the genetic correlation in larval darkness across midsummer and autumnal environments is not different from zero, suggesting that differential gene expression is responsible for the increase in larval darkness in the autumn. The relatively dark autumnal form was found to have a higher body temperature in sunlight than did the lighter midsummer form, and small differences in temperature were found to increase larval growth rate. These results suggest that this genetically based seasonal polyphenism in larval color has evolved in part to increase larval growth rates in the autumn.Corresponding Editor: T. Kawecki

Feeding, growth, and the thermal environment of cabbage white caterpillars, Pieris rapae L.

Laboratory studies of temperature effects on short-term feeding and growth rates were combined with field data on thermal environments to explore the consequences of temperature variation for growth of caterpillars of the cabbage white butterfly, Pieris rapae. Mean short-term (24-h) consumption and growth rates of fourth-instar P. rapae feeding on collard leaves increased continuously with increasing temperatures between 10 degrees and 35 degrees C, peaked at 35 degrees C, and declined rapidly with temperatures above 35 degrees C. Physical models can mimic temperatures of real fifth-instar caterpillars under collard leaves within 1 degrees -2 degrees C in sunny summer conditions in Seattle, Washington. Continuous recordings of operative temperatures of model caterpillars in a collard garden suggest that, at the timescale of the duration of the fifth instar (5-8 d in the field), P. rapae caterpillars frequently experience temperatures spanning a 25 degrees C range, they spend most of their time at temperatures well below those that maximize growth, and they encounter substantial variation in the frequency distribution of operative temperatures between time periods. Combining these data on growth rate as a function of temperature and the distribution of operative temperatures in the field, I illustrate how growth rates at higher temperatures can make disproportionate contributions to the overall mean growth rates even when higher temperatures are relatively infrequent. Fluctuating thermal conditions may generate variable patterns of selection on reaction norms for growth rate in the field.

Patterns and mechanisms of growth of fifth-instar Manduca sexta caterpillars following exposure to low- or high-protein food during early instars.

For many insect herbivores, variation in protein availability is a pervasive part of the environment. I explore how variable protein availability affects growth rates of fifth-instar Manduca sexta caterpillars and how growth is related to behavior and physiology. Groups of larvae were reared on low- or high-protein artificial diets (5.9% and 17.7% casein by dry weight, respectively) and then transferred in the fifth instar to the same or opposite diet. During or after the 24-h period following transfer, I measured growth rate, consumption rate, growth efficiency, midgut proteolytic activity, and masses of midgut contents and tissues. Fifth-instar caterpillars reared in earlier instars on high-protein diet grew about 20% more rapidly over 24 h than did caterpillars reared on low-protein diet. This growth pattern appears to be caused by differences in consumption and growth efficiency: caterpillars reared on high protein consumed more food, and used it more efficiently, than did caterpillars reared on low-protein diet. Over the short term (24 h), in contrast, fifth instars that received low-protein diet grew as rapidly as caterpillars that received high-protein diet. Increased (compensatory) consumption appears to be the primary mechanism by which caterpillars consuming low-protein food maintained growth rates.

A peripheral input of thoracic origin inhibits chewing movements in the larvae of Manduca sexta

Fifth-instar caterpillars chew more or less continuously after their neck connectives have been cut, implying that the neural circuitry responsible for chewing is tonically inhibited by input arriving via the neck connectives. We have investigated the source of this inhibitory signal by lesion experiments. It appears to originate in peripheral structures innervated by the paired nerves of the thoracic ganglia. The responsible structures are not on the thoracic legs. Both the frequency of spontaneous chewing and the probability of obtaining long bouts of chewing in response to mechanical stimulation of either the labium or the labrum are raised in parallel by disinhibitory lesions to the thoracic nervous system. Disinhibition is as complete and symmetrical after cutting a single neck connective as after cutting both of them.

Insect cardioactive peptides: regulation of hindgut activity by cardioacceleratory peptide 2 (CAP2) during wandering behaviour in Manduca sexta larvae.

The functional relationship between cardioacceleratory peptide 2 (CAP2) and hindgut activity during wandering behaviour was investigated in fifth-instar larvae of the tobacco hawkmoth Manduca sexta. Inspection of the alimentary canal on the day prior to wandering showed that the gut, in preparation for metamorphosis, was voided of all contents by 18:00 h. Associated with this event, which we refer to as 'gut emptying', was an increase in the frequency of hindgut contractions measured in vivo. No change in heart activity was seen during this developmental period. Measurements of the amount of CAP2 in the central nervous system (CNS) of fifth-instar caterpillars revealed that CAP2 storage levels declined sharply on the day of gut emptying. The drop in CNS levels of CAP2 at gut emptying was temporally correlated with the appearance of CAP2 in the haemolymph. CAP2, when applied at physiological concentrations to an in vitro larval hindgut bioassay, caused changes in several parameters, including contraction frequency and amplitude, and basal tension. In vivo administration of CAP2 elicited hindgut responses that were qualitatively and quantitatively similar to those seen in vitro. Developmental studies on changes in CAP2 responsiveness during the last larval instar demonstrated that the hindgut is maximally sensitive to CAP2 on the day of gut emptying. Direct evidence in support of a role for CAP2 in fifth-instar larvae was provided by experiments in which the increase in gut activity in vivo seen at gut emptying was significantly reduced by injections of an anti-CAP antibody. Based on data from cobalt backfills and anti-CAP immunohistochemical staining, we propose that CAP2 exerts its effect on the larval hindgut at wandering via a local release from CAP-containing neurones in the terminal ganglion that project directly to the hindgut.

Effects of temperature on growth and efficiency of food utilization in fifth-instar caterpillars of the tobacco hornworm, Manduca sexta

Fifth-instar larvae of Manduca sexta were reared from hatching on artificial diet at 15, 20, 25, 30 and 35°C. Total development time decreased with increasing temperature. Very few larvae (12%) survived at 15°C, so this temperature was not considered further. There was some mortality at 30°C (11%), and at 35°C (50%). The absolute rate of growth in the fifth instar was faster at 25 than at 20°C, but was similar at 25, 30 and 35°C. This was true both for caterpillars that were chronically exposed to experimental temperatures (i.e. since hatching) and for those acutely exposed (i.e. reared up to fifth instar at 25°C). There was a progressive decrease with higher rearing temperatures in both the initial and final sizes of chronically exposed fifth-instar larvae. Acutely exposed caterpillars matched for initial size showed smaller temperature related differences in final size. Because of these size differences there were differences in relative growth rate which did not reflect true differences in absolute growth rate. Total food consumed by chronically exposed caterpillars was greatest at the lowest temperature (20°C), and decreased progressively with increasing temperature. The absolute rate of food consumption increased from 20 to 25°C, but did not vary significantly between 25 and 35°C. Differences in the sizes of the insects at the different temperatures meant that there were differences among relative measures of consumption that did not reflect absolute food consumption. For chronically exposed caterpillars, none of the three usual indices of food conversion efficiency (AD, ECI and ECD) varied significantly with temperature between 20 and 35°C. This implies that the effects of temperature on metabolic costs are closely matched to food consumption. Oxygen consumption increased with temperature between 20 and 25°C but was temperature compensated between 25 and 35°C. These findings are discussed in terms of their implications for the optimal temperature for growth in Manduca.

Head flicking by tent caterpillars: a defensive response to parasite sounds

Western tent caterpillars respond to sounds of 300–2100 Hz by head flicking. The sound frequencies to which they are most sensitive match the flight sound of a fly parasite which lays eggs on the heads of caterpillars. Caterpillars on tents are more likely to head flick when stimulated by sound and have lower parasitization levels than fifth-instar caterpillars which have left the tent.