Early Instar Stages Research Articles

The Wnt pathway regulates wing morph determination in Acyrthosiphon pisum

Wing dimorphism occurs in insects as a survival strategy to adapt to environmental changes. In response to environmental cues, mother aphids transmit signals to their offspring, and the offspring either emerge as winged adults or develop as wingless adults with degeneration of the wing primordia in the early instar stage. However, how the wing morph is determined in the early instar stage is still unclear. Here, we established a surgical sampling method to obtain precise wing primordium tissues for transcriptome analysis. We identified Wnt as a regulator of wing determination in the early second instar stage in the pea aphid. Inhibiting Wnt signaling via knockdown of Wnt2, Wnt11b, the Wnt receptor-encoding gene fz2 or the downstream targets vg and omb resulted in a decreased proportion of winged aphids. Activation of Wnt signaling via knockdown of miR-8, an inhibitor of the Wnt/Wg pathway, led to an increased proportion of winged aphids. Furthermore, the wing primordia of wingless nymphs underwent apoptosis in the early second instar, and cell death was activated by knockdown of fz2 under the wing-inducing condition. These results indicate that the developmental plasticity of aphid wings is modulated by the intrinsic Wnt pathway in response to environmental challenges.

Uncovering the Microbiota of Bagworm Metisa plana (Lepidoptera: Psychidae) in Oil Palm Plantations in Malaysia.

Bagworm Metisa plana is one of the major pests in Malaysia's oil palm plantation, with infestation resulting in huge economical loss. Currently, the microbial profile of the bagworm has yet to be study. Understanding the biology of the pest such as the bacterial community is crucial as bacteria associated with insects often provide benefits to the insect, giving the insect host a better chance of survival. Here, 16S amplicon sequencing was used to identify the bacteria community of M. plana. Additionally, two comparisons were made, the bacterial communities between two larval stages (early instar stage and late instar stage) from outbreak area; the bacterial communities of late instar stage larvae from non-outbreak between outbreak areas. From this study, it was found that the bacterial community of M. plana consisted of Proteobacteria, Actinobacteria, Bacterioidetes, Firmicutes and other minor phyla, with Proteobacteria being the most dominant phylum. Furthermore, bacterial genera of M. plana consisted of Pantoea, Curtobacterium, Pseudomonas, Massilia and other minor genera, with Pantoea being the most dominant. It was also found that the alpha and beta diversity in both comparisons were not significantly different. We present our data as a first insight towards the bacterial community of M. plana, paving a way towards understanding the biology of the bagworm M. plana.

Fitness differences between parthenogenetic litters of the synanthropic scorpion Tityus stigmurus (Scorpiones: Buthidae)

Abstract Clonal lineages in similar environments may be influenced by non-Mendelian inheritance, such as maternal age effects and developmental instabilities. These mechanisms may affect the developmental fitness of parthenogenetic litters. In this study, the scorpion Tityus stigmurus (Thorell, 1876) was used to analyze the effects of non-Mendelian variation on parthenogenetic litters. A total of 75 juveniles from five females were reared under the same controlled conditions, while their development was observed and evaluated through differences in offspring fitness traits (litter size, prosoma size, developmental time, and mortality) between the litters. First and second litters had a similar litter size, although second litters exhibited longer developmental time in the early instar stages (second and third) than first litters. These results indicate that T. stigmurus females allocated nutrient resources to maximize litter size rather than developmental fitness in subsequent litters. Differences in developmental time found in early instars but not in late instars may have occurred as a result of stochastic developmental variations in young individuals. Such variation in duration of development has the potential to influence survival of juveniles in natural environments because individuals that grow faster may avoid large predators and consume larger prey, compared to those that do not.

Relative larvicidal property of common oxide nanostructures against Culex quinquefasciatus.

Common oxide nanostructures such as silicon-di-oxide, magnesium-oxide, zinc-oxide, and copper-oxide (CuO) having useful functional and bioactive properties have been synthesised and characterised. All these nanostructures have been found to be larvicidal towards Culex quinquefasciatus mosquito especially against lower instars in comparison with higher instars in 48 h. Only, CuO is larvicidal against late instar stages after 48 h. Moreover, CuO is larvicidal against first instar stages after 24 h (LC50 157 mg/l). However, none of these nanostructures are pupicidal. Post mortality larval morphology was found to be distorted under bright field microscopy and scanning electron microscopy images of affected larval surface appeared to be rough and uneven. Fluorescent images showed that nanostructures infiltrated inside visceral organs of larvae. Nanostructures also caused tissue oxidative stress in larvae. These results indicate that above stated oxide nanostructures are effective larvicidal agents against early instar stages of Culex larvae.

Biological activity of Hyposidra talaca nucleopolyhedrovirus against all instar stages of a destructive tea pest Hyposidra talaca (Lepidoptera: Geometridae) in India

Hyposidra talaca (Walker) (Lepidoptera: Geometridae) is the most harmful pest of northeastern tea hub of India that devastates the tea production by feasting on the tea leaves. Hyposidra talaca nucleopolyhedrovirus (HytaNPV) is a natural enemy of the aforesaid pest, as it poses great obstruction in the multiplication of the pest by causing significant larval mortality. The study was undertaken to screen the virus activity against first to fifth instar H. talaca larvae. Early instar stages are found more susceptible than the late stages as they tend to reflect highest LC50 value for fifth instar as 4.3 × 107 POB/ml and lowest LC50 value for first instar as 7 × 104 POB/ml within seven days of inoculation. LT50 values vary between 2.47 and 8.45 days for neonates to fifth instar. The high record of virulence of HytaNPV indicates its bright prospect as a useful microbial biopesticide.

Purine biosynthesis-deficient Burkholderia mutants are incapable of symbiotic accommodation in the stinkbug

The Riptortus-Burkholderia symbiotic system represents a promising experimental model to study the molecular mechanisms involved in insect-bacterium symbiosis due to the availability of genetically manipulated Burkholderia symbiont. Using transposon mutagenesis screening, we found a symbiosis-deficient mutant that was able to colonize the host insect but failed to induce normal development of host's symbiotic organ. The disrupted gene was identified as purL involved in purine biosynthesis. In vitro growth impairment of the purL mutant and its growth dependency on adenine and adenosine confirmed the functional disruption of the purine synthesis gene. The purL mutant also showed defects in biofilm formation, and this defect was not rescued by supplementation of purine derivatives. When inoculated to host insects, the purL mutant was initially able to colonize the symbiotic organ but failed to attain a normal infection density. The low level of infection density of the purL mutant attenuated the development of the host's symbiotic organ at early instar stages and reduced the host's fitness throughout the nymphal stages. Another symbiont mutant-deficient in a purine biosynthesis gene, purM, showed phenotypes similar to those of the purL mutant both in vitro and in vivo, confirming that the purL phenotypes are due to disrupted purine biosynthesis. These results demonstrate that the purine biosynthesis genes of the Burkholderia symbiont are critical for the successful accommodation of symbiont within the host, thereby facilitating the development of the host's symbiotic organ and enhancing the host's fitness values.

Extraordinary morphological changes in valve morphology during the ontogeny of several species of the Australian ostracod genus Bennelongia (Crustacea, Ostracoda)

Ostracods belonging to the genus Bennelongia differ much in valve morphology between adults and juveniles. Adult valves are asymmetrical, characterised by a beak-like feature in the antero-ventral region of the left valve, and, with some notable exceptions, mostly have smooth or weakly-ornamented valves. Juvenile specimens, on the other hand, have valves that are almost symmetrical, with no beak-like feature and are often heavily ornamented.We have examined the last 3 - 4 juvenile stages of 6 Bennelongia species from 5 different lineages, in order to decipher the types of external valve ornamentation and their recurrences during ontogeny and across lineages. It is clear that ornamentation is more prevalent at the early instar stages compared to the last 2 pre-adult stages, and especially when compared to the adult stage itself.We also examined the surprising presence of a calcified inner lamella with a prominent inner list in the pre-adult stages of Bennelongia species, that is usually absent in juveniles of other ostracods, thus questioning if heterochronic processes have provided an intermediate valve morphology between the simple (normal) cypridinid juvenile state and the heavily derived and modified state of adult Bennelongia.We discuss the possible (speculative) functionality of the ornamentation in juveniles.

Effects of elevated CO2 on feeding preference and performance of the gypsy moth (Lymantria dispar) larvae

AbstractMultiple‐choice assays were conducted in the laboratory to examine the effect of CO2‐induced changes in leaf quality on growth, nutritional indices and preferences of the gypsy moth (Lymantria dispar) larvae. The tested tree species, one‐year‐old aspen (Populus pseudo‐simonii Kitag.), two‐year‐old birch (Betula platyphylla) and three‐year‐old oak (Quercus mongolica Fisch.) were transplanted to open‐top chambers at ambient or elevated CO2 (650 ppm) concentrations in May 2005. The present study was conducted in 2006. Leaves from the upper and lower crowns of each tree species were sampled and analysed, respectively. On both measurement dates (24 July 2006 and 16 August 2006), there were significant CO2 effects on levels of condensed tannin, starch, nitrogen and C : N ratio. But only on the latter date, were the concentrations of total phenolics, soluble sugar, carbon and total non‐structural carbohydrates significantly affected by elevated CO2. Leaf dry weight content and specific leaf weight were almost completely unaffected by CO2 enrichment. Gypsy moth larvae exhibited a clear selectivity for tested leaf types (leaves from the upper and lower crowns of each tree species) even in their early instar stage, with the upper leaves of P. pseudo‐simonii being the most preferable and the lower leaves of Q. mongolica being the least preferred. The changes in leaf quality significantly reduced larval growth and altered partial indices of insect performance (e.g., relative growth rate and relative consumption rate). However, at least in this short‐term choice feeding assay (13 days), the CO2‐induced changes in leaf quality had no significant effects on food preference behaviour of the gypsy moth larvae, neither within the limited range of host plants nor within the leaves at different canopy heights of the same tree species.

Scanning electron microscopy of foreleg tarsal sense organs of the poultry red mite, Dermanyssus gallinae (DeGeer) (Acari:Dermanyssidae)

Scanning electron microscopy (SEM) studies on the poultry red mite, Dermanyssus gallinae (DeGeer), showed that the tip of the foreleg tarsus of both sexes and early instar stages (larvae, protonymph and deutonymph) bears a cluster of 10 blunt peg-type sensillae. Diversity in size, shape and surface features of the wall of these sensillae characterize the sensilla complex (sensory field). Two short setae (1 and 2) with smooth walls lack detectable pores, whereas three medium-sized setae (4, 5 and 8) bear pores in the wall. Four medium or long setae (3, 6, 7 and 9) have longitudinal grooves or ribs in the wall, whilst one seta (10) possesses both pores and grooves in the wall. Based on the morphology, it is suggested that the porous setae could be olfactory receptors, and the grooved setae could be mechanoreceptors.

Lepidopteran pest populations and crop yields in row intercropped broccoli

Abstract To determine how the presence of non‐host plants impact populations of lepidopteran pests and yield of broccoli, Brassica oleracea var. italica L., broccoli was intercropped with tomato, Lycopersicon esculentum Miller, or yellow sweetclover (YSC), Melilotus officinalis L., and compared with broccoli monoculture grown at two densities (high and low). During a 1997 study, numbers of early instar imported cabbageworm, Artogeia rapae L., were greater in broccoli monoculture compared with intercropped plantings. In contrast, late instar A. rapae numbers were significantly higher in intercropped broccoli during most of the season, indicating greater survival of early instar stages in intercropped habitats. Cabbage looper, Trichoplusia ni Hübner, eggs and larvae were more abundant in broccoli monoculture than intercropped broccoli in 1997. In addition, T. ni populations were found at significantly lower levels in broccoli‐YSC compared with broccoli‐tomato habitat throughout the season. During a 1998 study, numbers of early and late A. rapae instars were greater in intercropped broccoli on most sampling dates. The mean percentages of harvested broccoli heads infested with insects and associated frass were higher in monocultures than intercropped treatments in 1997. Additionally, a significantly lower number of contaminants were found in the crowns of broccoli harvested from broccoli‐YSC (YSC) compared with broccoli‐tomato (tomato) habitat. However, broccoli heads were significantly smaller in intercropped habitats. Variations in insect responses and challenges to using crop diversification as a management tool are discussed.

Life tables of the processionary caterpillar Ochrogaster lunifer Herrich‐Schäffer (Lepidoptera: Thaumetopoeidae) at local and regional scales

A major ongoing debate in population ecology has surrounded the causative factors underlying the abundance of phytophagous insects and whether or not these factors limit or regulate herbivore populations. However, it is often difficult to identify mortality agents in census data, and their distribution and relative importance across large spatial scales are rarely understood. Here, we present life tables for egg batches and larval cohorts of the processionary caterpillar Ochrogaster lunifer Herrich‐Schäffer, using intensive local sampling combined with extensive regional monitoring to ascertain the relative importance of different mortality factors at different localities. Extinction of entire cohorts (representing the entire reproductive output of one female) at natural localities was high, with 82% of the initial 492 cohorts going extinct. Mortality was highest in the egg and early instar stages due to predation from dermestid beetles, and while different mortality factors (e.g. hatching failure, egg parasitism and failure to establish on the host) were present at many localities, dermestid predation, either directly observed or inferred from indirect evidence, was the dominant mortality factor at 89% of localities surveyed. Predation was significantly higher in plantations than in natural habitats. The second most important mortality factor was resource depletion, with 14 cohorts defoliating their hosts. Egg and larval parasitism were not major mortality agents. A combination of predation and resource depletion consistently accounted for the majority of mortality across localities, suggesting that both factors are important in limiting population abundance. This evidence shows that O. lunifer is not regulated by natural enemies alone, but that resource patches (Acacia trees) ultimately, and frequently, act together to limit population growth.

Chronic effects of Cu on reproduction of Polypedilum nubifer (chironomidae) through water and food

The effects of Cu on the reproduction of Polypedilum nubifer (Chironomid) through water and food were examined in a flowthrough aquarium by Cu exposure from the egg stage. The emergence success decreased to 74, 38, 16, and 2% of the control at 10, 20, 30, and 40 μg Cu/liter, respectively. However, the oviposition success (No. of egg clusters/female) remained at 64% of the control value even at 30 μg Cu/liter. From these results and the acute toxicity test, it was suggested that a decrease in emergence success was caused mainly by mortality in the early instar stage, when Cu sensitivity is high. There were no significant effects of Cu on the emergence success, oviposition success, and hatchability of the oviposited eggs in the midge which had been given food contaminated with about 330 μg Cu/g (dry weight). However, emergence success and hatchability of the oviposited eggs decreased to 30 and 40% of the control in the midge which had been fed food accumulating 1770 μg Cu/g, although the oviposition success was not impaired compared to the control. Emergence did not occur from the aquaria fed food accumulating 5200 μg Cu/g.

Estimation of the mortality of the immature stages of Aedes (Stegomyia) africanus Theobald in a tropical forest in Uganda

Data on the populations of larval and pupal stages of Aedes africanus , an important sylvan vector of arboviruses, were collected from tree-holes which are its natural breeding sites in forests. Comparable data were also simultaneously collected from bamboo pots stationed in the Zika Forest to simulate the natural breeding sites. Survivorship curves and time-specific life-tables were constructed. It was observed that the immature stages of A. africanus in both tree-holes and bamboo pots suffered very high mortality rates (an average of 91% in tree-holes and 83.2% in bamboo pots). It was further observed that mortality rates varied monthly. The survivorship curves constructed for the immature stages collected on a daily basis tended to show that a large proportion of the mortality rate was confined to the early instar stages, but the survivorship curves based on the combined data for the 2 years indicated constant mortality rates for all the stages. The high mortality rates of the immature stages of A. africanus appear to be attributed to predation by larvae of Toxorhynchites brevipalpis , with which it shares the same breeding places.

A population study of Phaulacridium vittatum Sjost (Acrididae)

Phaulacridium vittatum is a very abundant grasshopper in pastures in the Southern Tablelands area of New South Wales. It has a single generation per year, the active stages of which are present from late spring until mid-autumn of the following year. In grazed pastures female grasshoppers lay their egg pods in bare spaces between plants and there is a close correlation between the density distribution of egg pods and that of adults of the parent generation. An egg diapause occurs and mortality of eggs is relatively low. The first-instar nymph of P. vittatum feeds on prostrate and rosette-forming plants. However, irrespective of their abundance it is unable to locate these plants where grasses form the dominant plant cover and so fails to survive. Heavy spring rains which produce an abundance of annual and perennial grasses limit the amount of favourable space where the young nymphs can locate suitable food plants and thus result in high mortality. Heavy grazing and the introduction of mat-forming Trifolium subterraneum (L.) maintain open low pastures and favour survival of first-instar nymphs. When hatching has been relatively late, the seasonal drying off of annuals, particularly T. subterraneum, which is a favoured food plant of P. vittatum, results in heavy mortality of the early-instar stages. In most instances, populations have reached the fourth-instar stage by the time that the annuals dry out and dispersal then occurs. Frequently, dispersal consists merely of movement from sites where survival was high after hatching into areas in which the cover was initially unfavourable for post-hatching survival. However, where conditions were uniformly favourable for hatchling survival, mass movement of the grasshoppers to trees may occur. Under average rainfall and evaporation rates in summer very little plant growth occurs, so that to develop from the fourth instar to the sexually immature adult stage, the grasshoppers depend on the foliage accumulated by broad-leafed plants during the spring. As the numbers of fourth-instar nymphs are usually excessive in relation to the amounts of food accumulated during the spring growing period, heavy depletion of food occurs and numbers fall simultaneously. However, development of the grasshoppers is not interrupted by food shortage and, on reaching the sexually immature adult stage, numbers become stable. The persistence of populations under conditions of limited food is attributed to poor discrimination between favourable plants and those unfavourable for development or survival, the restriction of the movements of individuals to ambits or "home ranges" of limited area, an apparent inability of individuals to locate food plants when they wander away from their ambits in search of food and cannibalism of weakened individuals. In contrast to nymphs, adults can survive for prolonged periods in the sexually immature condition on a diet consisting exclusively of the fresh growth of those shallow-rooted perennial grasses which respond to light falls of rain. They do not become sexually mature under these conditions, but reproduce when sufficient rain falls to induce renewed growth of broad-leafed plants.