Final Instar Caterpillars Research Articles

Plant Quantity Affects Development and Survival of a Gregarious Insect Herbivore and Its Endoparasitoid Wasp.

Virtually all studies of plant-herbivore-natural enemy interactions focus on plant quality as the major constraint on development and survival. However, for many gregarious feeding insect herbivores that feed on small or ephemeral plants, the quantity of resources is much more limiting, yet this area has received virtually no attention. Here, in both lab and semi-field experiments using tents containing variably sized clusters of food plants, we studied the effects of periodic food deprivation in a tri-trophic system where quantitative constraints are profoundly important on insect performance. The large cabbage white Pieris brassicae, is a specialist herbivore of relatively small wild brassicaceous plants that grow in variable densities, with black mustard (Brassica nigra) being one of the most important. Larvae of P. brassicae are in turn attacked by a specialist endoparasitoid wasp, Cotesia glomerata. Increasing the length of food deprivation of newly molted final instar caterpillars significantly decreased herbivore and parasitoid survival and biomass, but shortened their development time. Moreover, the ability of caterpillars to recover when provided with food again was correlated with the length of the food deprivation period. In outdoor tents with natural vegetation, we created conditions similar to those faced by P. brassicae in nature by manipulating plant density. Low densities of B. nigra lead to potential starvation of P. brassicae broods and their parasitoids, replicating nutritional conditions of the lab experiments. The ability of both unparasitized and parasitized caterpillars to find corner plants was similar but decreased with central plant density. Survival of both the herbivore and parasitoid increased with plant density and was higher for unparasitized than for parasitized caterpillars. Our results, in comparison with previous studies, reveal that quantitative constraints are far more important that qualitative constraints on the performance of gregarious insect herbivores and their gregarious parasitoids in nature.

Revisiting macronutrient regulation in the polyphagous herbivore Helicoverpa zea (Lepidoptera: Noctuidae): New insights via nutritional geometry

Insect herbivores that ingest protein and carbohydrates in physiologically-optimal proportions and concentrations show superior performance and fitness. The first-ever study of protein–carbohydrate regulation in an insect herbivore was performed using the polyphagous agricultural pest Helicoverpa zea. In that study, experimental final instar caterpillars were presented two diets – one containing protein but no carbohydrates, the other containing carbohydrates but no protein – and allowed to self-select their protein–carbohydrate intake. The results showed that H. zea selected a diet with a protein-to-carbohydrate (p:c) ratio of 4:1. At about this same time, the geometric framework (GF) for the study of nutrition was introduced. The GF is now established as the most rigorous means to study nutrient regulation (in any animal). It has been used to study protein–carbohydrate regulation in several lepidopteran species, which exhibit a range of self-selected p:c ratios between 0.8 and 1.5. Given the economic importance of H. zea, and it is extremely protein-biased p:c ratio of 4:1 relative to those reported for other lepidopterans, we decided to revisit its protein–carbohydrate regulation. Our results, using the experimental approach of the GF, show that H. zea larvae self-select a p:c ratio of 1.6:1. This p:c ratio strongly matches that of its close relative, Heliothis virescens, and is more consistent with self-selected p:c ratios reported for other lepidopterans. Having accurate protein and carbohydrate regulation information for an insect herbivore pest such as H. zea is valuable for two reasons. First, it can be used to better understand feeding patterns in the field, which might lead to enhanced management. Second, it will allow researchers to develop rearing diets that more accurately reflect larval nutritional needs, which has important implications for resistance bioassays and other measures of physiological stress.

Macronutrient Balance Modulates the Temperature-Size Rule in an Ectotherm.

Most ectotherms mature at a larger body size in colder conditions, a phenomenon known as the temperature-size rule. While a number of hypotheses have been proposed to explain this rule, little work has been done to understand it from a nutritional perspective. We have used the final-instar caterpillars of Spodoptera litura to investigate how dietary protein∶carbohydrate (P∶C) balance influences the relationship between temperature and body size. The strength and direction of the thermal reaction norm for body size were significantly altered by dietary P∶C balance. The slope of the reaction norm was nearly flat for caterpillars raised on a balanced food ([Formula: see text]) but was significantly negative for those on nutritionally imbalanced foods (1∶5 or 5∶1), especially when carbohydrates were in considerable excess. These nutrient-dependent effects of temperature on body size were caused mainly by corresponding changes in body lipid storage. When allowed to choose between imbalanced diets, caterpillars increased their preference for carbohydrates to meet high energy demands at higher temperatures. The slope of the thermal reaction norm for body size was substantially reduced by such a temperature-driven shift in nutrient preference, indicating that the impact of high temperature on body size was buffered by altered food selection. This study highlights the importance of macronutrient balance as a key factor modulating the relationship between temperature and body size in ectotherms and provides a novel approach for understanding the temperature-size rule.

Eggs, final-instar caterpillars and metamorphosis of Cyclosia macularia Guérin Méneville (Lepidoptera: Zygaenidae) from its larval host plant Baccaurea motleyana

Information on the immature stages of Zyganidae moths is rather limited compared to that on the adults. An encounter with the caterpillars on the Baccaurea motleyana tree enabled a description of the eggs, final-instar caterpillar and characteristic cocoon of the Chalcosiine Day-flying Moth, Cyclosia macularia Guérin-Méneville, accompanied by illustrations. Sexual dimorphism of this species based on the adults is also described.http://zoobank.org/urn:lsid:zoobank.org:pub:411550D6-FC0F-45BA-A2D2-770757E3F009

The sub-lethal effects of repeated freezing in the woolly bear caterpillar Pyrrharctia isabella

Repeated freeze-thaw cycles are common and are increasing in frequency with climate change in many temperate locations, yet understanding of their impact on freeze-tolerant insects is extremely limited. We investigated the effects of repeated freezing and thawing on the freeze-tolerant final instar caterpillars of the moth Pyrrharctia isabella (Lepidoptera: Arctiidae) by subjecting individuals to either a single sustained 35 h freeze or five 7 h freezes. Sub-lethal effects were quantified with changes in three broad groups of measures: (1) cold hardiness, (2) metabolic rate and energy reserves and (3) survival after challenge with fungal spores. Repeated freeze-thaw cycles increased mortality to almost 30% and increased tissue damage in Malpighian tubules and hemocytes. Repeated freezing increased caterpillar glycerol concentration by 0.82 mol l(-1). There were no changes in metabolic rate or energy reserves with repeated freezing. For the first time, we report increased survival after immune challenge in caterpillars after freezing and suggest that this may be linked to wounding during freezing. We suggest that little repair of freezing damage is possible in P. isabella caterpillars and repeated freeze-thaw cycles may present significant challenges to survival in this species.

The interactive effects of protein quality and macronutrient imbalance on nutrient balancing in an insect herbivore

The present study evaluates the interactive effects of protein quality and the ratio of protein to digestible carbohydrate on herbivore performance, various aspects of nutritional homeostasis (feeding and post-ingestive regulation) and food choice. In the first experiment, final-instar caterpillars of generalist-feeding African cotton leafworm Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) were confined to one of 20 diets varying in protein:carbohydrate (P:C) ratio (35:7, 28:14, 21:21, 14:28 and 7:35) and in the quality of dietary protein (0%, 20%, 50% and 80% replacement of casein-based protein mix with the low-quality plant protein, zein). Results indicated that the negative impact of low-quality protein on survival, development and growth was amplified as the P:C ratio of the diet fell. Consumption differences were the main underlying cause of this response. The effect of low protein quality was to reduce the efficiency of nitrogen utilization and to increase the conversion rate of ingested nutrient to lipid growth, irrespective of P:C ratio. In the second experiment, caterpillars were allowed to self-compose their preferred diet, selecting between one of four high-protein diets (all 35:7, but containing 0%, 20%, 50% or 80% zein) and one of four equal-P:C-ratio diets (21:21, again containing 0%, 20%, 50% or 80% zein). Caterpillars showed a compensatory increase in their self-selected P:C intake in response to the moderate decline in protein quality of the 35:7 food. No such response was demonstrated for the insects presented with 35:7 food with the lowest protein quality. The significance of these findings is discussed within the context of herbivore food selection.

A comparison of nutrient regulation between solitarious and gregarious phases of the specialist caterpillar, Spodoptera exempta (Walker)

Nutritional regulatory responses were compared between solitarious and gregarious phases of the African armyworm, Spodoptera exempta. When allowed to mix between two nutritionally imbalanced but complementary foods, final-instar caterpillars in both phases selected a diet comprising more carbohydrate than protein. This contrasts with other larval lepidopterans studied to date. Only minor differences were found in the position of the intake target for the two phases, despite their different energetic requirements for migration as adults. When restricted to nutritionally imbalanced diets, caterpillars of both phases were less disposed to overeat protein on high-protein diets than carbohydrate on high-carbohydrate diets, relative to the self-composed intake target. However, in both cases gregarious larvae overingested the excess nutrient to a greater degree than did solitarious larvae. Furthermore, gregarious larvae showed higher nitrogen conversion efficiency on an extreme protein-limiting diet, and accumulated more lipid per amount of carbohydrate consumed on carbohydrate-deficient diets. These phase-associated nutritional differences are consistent with the life-history strategies of the two phases.

The capacity of a Myrmica ant nest to support a predacious species of Maculinea butterfly

Caterpillars of Maculinea arion are obligate predators of the brood of Myrmica sabuleti ants. In the aboratory, caterpillars eat the largest available ant larvae, although eggs, small larvae and prepupae are also palatable. This is an efficient way to predate. It ensures that newly-adopted caterpillars consume the final part of the first cohort of ant brood in a nest, before this pupates in early autumn and becomes unavailable as prey. At the same time, the fixed number of larvae in the second cohort is left to grow larger before being killed in late autumn and spring. Caterpillars also improve their feeding efficiency by hibernating for longer than ants in spring, losing just 6% of their weight while the biomass of ant larvae increases by 27%. Final instar caterpillars acquire more than 99% of their ultimate biomass in Myrmica nests, growing from 1.3 mg to an estimated 173 mg. A close correlation was found between the weights of caterpillars throughout autumn and the number of large ant larvae they had eaten. This was used to calculate the number of larvae eaten in spring, allowing both for the loss of caterpillar weight during winter and the increase in the size of their prey in spring. It is estimated that 230 of the largest available larvae, and a minimum nest size of 354 M. sabuleti workers, is needed to support one butterfly. Few wild M. sabuleti nests are this large: on one site, it was estimated that 85% of nests were too small to produce a butterfly, and only 5% could support two or more. This prediction was confirmed by the mortalities of 376 caterpillars in 151 wild M. sabuleti nests there. Mortalities were particularly high in nests that adopted more than two caterpillars, apparently due to scramble competition and starvation in autumn. Survival was higher than predicted in wild nests that adopted one caterpillar. These caterpillars seldom exhaust their food before spring, when there is intense competition among Myrmica for nest sites. Ants often desert their nests in the absence of brood, leaving the caterpillar behind. Vacant nests are frequently repopulated by a neighbouring colony, carrying in a fresh supply of brood. Maculinea arion caterpillars have an exceptional ability to withstand starvation, and sometimes survive to parasitize more than one Myrmica colony. Despite these adaptations, predation is an inefficient way to exploit the resources of a Myrmica nest. By contrast, Maculinea rebeli feeds mainly at a lower trophic level, on the regurgitations of worker ants. Published data show that Myrmica nests can support 6 times more caterpillars of Maculinea rebeli than of M. arion in the laboratory. This is confirmed by field data.

Camouflage and selective predation in caterpillars of the poplar and eyed hawkmoths (Laothoe populi and Smerinthus ocellata)

Final instar caterpillars of the poplar hawkmoth (Laothoe populi) are usually yellow-green when they have been feeding on Salix fragilis but white when on Populus alba. Similarly final instar caterpillars of the eyed hawkmoth (Smerinthus ocellata) vary in colour from yellow-green when fed on Salix cinerea or S. fragilis to grey-green when fed on Salix ripens or S. alba. Most caterpillars of these two moths are thus very cryptic to the human eye. It has already been shown that colour matching is brought about by young caterpillars changing colour depending on the colour of the substrate perceived by their eyes. Evidence is presented here for selective predation, probably by birds. Yellow-green poplar hawk caterpillars disappeared from P. alba more rapidly than did white caterpillars, but there was no significant difference in the survival of the two colour forms on S. fragilis. Yellow-green caterpillars survived better on S. fragilis than on P. alba, but there was no significant difference in the survival of white caterpillars on the two plants. Reasons why selective predation did not occur in all of the experiments are discussed.

Carotenoids and colouration of poplar hawkmoth caterpillars (Laothoe populi)

Carotcnoids and chlorophylls a and b were extracted from final instar caterpillars of the poplar hawkmoth (Laothoe populi) and the eyed hawkmoth (Smerinthus ocellata), as well as from their food plants. Both species of caterpillar absorb the two chlorophylls and the carotenoids lutein, cis-lutein and β-carotene in the gut and deposit lutein and cis-lutein in the integument. It is the lutein, together with pterobilin, that is largely responsible for the colour of the insect: yellow-green poplar hawkmoth caterpillars have more lutein in the integument than dull green ones which in turn have more than white ones. Yellow-green and dull green caterpillars both sequester lutein and cis-lutein in the gut wall, but the yellow-greens translocate more of these pigments to the integument than the dull greens. The white caterpillars absorb very little lutein and cis-lutein into the gut, and so they have much less also in the integument. The mechanism by which the reflected light perceived by the caterpillar is translated into differential absorption of pigment by the gut and deposition in the integument is not known.

Larvae of Maculinea rebeli, a large‐blue butterfly, and their Myrmica host ants: wild adoption and behaviour in ant‐nests

The behavioural interactions between caterpillars of Maculinea rebeli Hir. and their Myrmica ant hosts were studied, both in the wild at the time of adoption, and inside captive nests of six Myrmica species.In the wild, freshly moulted, final instar caterpillars left their food‐plants at a time of day that coincided with the peak foraging activity of Myrmica (18:00‐20:00 h). Once on the ground, caterpillars made no attempt to search for Myrmica but settled and waited for foraging ants to find them, which took up to 1.5 h. There was no adoption ritual: foragers of any Myrmica species picked up the caterpillars within 1–4 sec of discovery, and carried them directly to their nests.Caterpillars grew from < 2 mg to 110 mg in laboratory ant‐nests. About 60 mg was gained in autumn but 40% of this was lost during the winter, while the temperature was < 14 °C. Although caterpillars survived best with their normal host, Myrmica schencki, they could also survive in the nests of other Myrmica species. The presence of queen ants had no effect upon survival. The behaviour of the caterpillars was described and illustrated: this included the production of secretions that were drunk by the ants, begging for food and direct feeding by ants. The preferred solid food was ant eggs.The results are discussed in terms of the social biology of Myrmica ants. It is hypothesized that Maculinea rebeli caterpillars mimic the touch pheromones of ant worker‐larvae. This would explain the inability of ants to recognize caterpillars before touching them, their immediate adoption by any Myrmica species after discovery, host specificity inside wild ant‐nests, the absence of queen‐effect and the intimate attention of host workers.

The causes of colour and colour change in caterpillars of the poplar and eyed hawkmoths (Laothoe populi and Smerinthus ocellata)

Caterpillars of the poplar and eyed hawkmoths (Laothoe populi and Smerinthus ocellata respectively) were reared under different conditions in order to determine why final instar caterpillars vary in colour. Poplar hawkmoth caterpillars normally rest on the undersides of leaves. Dull green and redspotted caterpillars are genetically determined polymorphisms. Caterpillars that are not dull green, however, can become white when fed on Populus alba or yellow-green when fed on Salix fragilis. Experiments showed that it is the reflective qualities of the leaves that determines which colour the caterpillar develops: if the young larva sees white then it becomes white, but if it sees green, grey or black then it becomes yellow-green. Young eyed hawkmoth larvae always developed into grey-green final instar caterpillars under our rather poorly-illuminated rearing conditions, but when reared on wild plants in white muslin sleeves they became whitish-green. In this species also it appears that colour of the final instar is determined by the reflectance of the substrate perceived by the young caterpillar.