Morphological study on the immature stages of the Banana fruit caterpillar,Tiracola plagiata (Walker, 1857) and its close relative,Tiracola aureata Holloway, 1989 (Noctuidae: Noctuinae: Leucaniini) with their host plant information

Imature stages of the Banana fruit caterpillar, Tiracola plagiata (Walker, 1857) and its close relative, Tiracola aureata Holloway, 1989 are described and illustrated. We revealed the following four facts. 1: The sixth instar is estimated to be the last instar larvae in both species based on the width of head capsules. 2: Morphologically they are distinguishable in the third to sixth instar larvae, but not distinguishable in the eggs, first to second instar larvae and pupae by our study. 3: Both species are polyphagous. 4: Some of the formerly known host plant records of T. plagiata should be excluded because they are revealed to be the hostplants of T. aureata.     

First record of the sedge feeder Bactra verutana Zeller (Lepidoptera: Tortricidae) in Chile based on morphology and DNA barcodes

The sedge-feeding moth Bactra verutana Zeller, 1875 (Lepidoptera: Tortricidae: Olethreutinae: Bactrini), described from Dallas, Texas, USA, is widespread, recorded throughout much North America, Central and South America, including the Caribbean, and Africa. The species is recorded for the first time from Chile based on specimens collected in the coastal valleys of the Atacama Desert, where its larvae feed on Cyperus corymbosus Rottb. var. subnodosus (Nees & Meyen) Kük. (Cyperaceae). A single DNA barcode haplotype, which is widespread in USA, was found in two Chilean specimens sequenced.     

External morphology of the leaf-tying larva and pupa of the Neotropical micromoth <Emphasis Type="Italic">Eccopsis razowskii</Emphasis> (Lepidoptera: Tortricidae)

Eccopsis razowskii Vargas (Lepidoptera: Tortricidae: Olethreutini) is a Neotropical micromoth native to the Atacama Desert with leaf-tying larvae that feed on Acacia macracantha Willd. (Fabaceae). The external morphology of its last instar and that of its pupa are described and illustrated for the first time with the aid of light and scanning electron microscopy. The morphology of each of these life stages is highly similar to that of the only sympatric congeneric, the pest species E. galapagana Razowski & Landry, the larvae of which feed on Prosopis spp. (Fabaceae). The pupae of the two species cannot be separated based on morphology; they can only be accurately separated based on the form of the mandible. The findings are discussed in terms of their significance for the taxonomy and systematics of Eccopsis. In addition, a comparison with the two other Tortricidae associated with A. macracantha in the Atacama Desert is provided.     

The hormonal regulation of the larval diapause in the codling moth, Laspeyresia pomonella (Lep. Tortricidae)

The hormonal control of the facultative diapause of the codling moth has been investigated. The diapause can be divided into 4 phases or periods: (1) diapause induction by short-day conditions (SD) in young larvae, (2) initiation of the diapause in the early last larval instar by a high titre of juvenile hormone, (3) onset and maintenance of diapause with inactivity of the neuroendocrine system, as evidenced by the results of neck-ligation experiments, (4)termination of diapause by the production of ecdysteroid.Diapause-induced larvae pupated after spinning the cocoon, if the state of induction was changed by injection with the anti-juvenile hormone precocene II at the beginning of the last larval instar and subsequent results of neck-ligation experiments, (4) termination of diapause by the production of ecdysteroid. treated with juvenile hormone during the first 1.5 days after the last larval moult and subsequently reared under SD. Under LD, continuous application of juvenile hormone during the last larval instar and after spinning did not prevent the insects from moulting to either a supernumerary larva, a pupa or a larval-pupal intermediate. Termination of diapause, i.e. pupation, was achieved by injecting diapausing larvae with 20-hydroxyecdysone. Although juvenile hormone was found to have a prothoractropic effect in diapausing larvae, no pupal moult could be induced by the application of the hormone. Contrary to the hormonal situation before pupation of nondiapausing larvae, no juvenile hormone could be detected before or during the pupation of larvae after diapause.     

A new Brazilian Passiflora leafminer: Spinivalva gaucha,gen. n., sp. n. (Lepidoptera,Gracillariidae, Gracillariinae), the first gracillariid without a sap-feeding instar

Male, female, pupa, larva and egg of a new genus and species of Gracillariidae (Gracillariinae), Spinivalva gaucha Moreira and Vargas from southern Brazil are described and illustrated with the aid of optical and scanning electron microscopy. A preliminary analysis of mitochondrial DNA sequences including members of related lineages is also provided. The immature stages are associated with Passiflora actinia, Passiflora misera and Passiflora suberosa (Passifloraceae), and build mines on the adaxial leaf surface. Initially the mines are serpentine in shape, but later in larval ontogeny become a blotch type. Although the larvae are hypermetamorphic as in other Gracillariidae, there is no sap-feeding instar in Spinivalva gaucha; the larva feeds on the palisade parenchyma, thus producing granular frass during all instars. Pupation occurs outside the mine; prior to pupating, the larva excretes numerous bubbles that are placed in rows on the lateral margins of the cocoon external surface. This is the second genus of gracillariid moth described for the Atlantic Rain Forest, and the second gracillariid species known to be associated with Passifloraceae.     

Interspecific competition between the braconid endoparasitoids Glyptapanteles porthetriae and Glyptapanteles liparidis in Lymantria dispar larvae

The effect of interspecific competition between the solitary endoparasitoid Glyptapanteles porthetriae Muesebeck (Hymenoptera: Braconidae) and the gregarious Glyptapanteles liparidis Bouché (Hymenoptera: Braconidae), was investigated in larvae of Lymantria dispar L. (Lepidoptera: Lymantriidae). Host larvae were parasitized by both wasp species simultaneously in premolt to the 2^nd or the 3^rd host instar or in an additional approach with a 4‐day delay in parasitization by the second wasp species. Host acceptance experiments revealed that both wasp species do not discriminate between unparasitized host larvae and larvae parasitized previously by the same or the other species. In more than 90% female wasps parasitized the larva they encountered first. During the period of endoparasitic development, larvae of the competing parasitoid species never attacked the egg stage of the other species. When host larvae were parasitized simultaneously by both wasp species, the rate of successful development of both species depended on the age of the host larva at the time of its parasitization; G. liparidis emerged successfully from 44% of host larvae parasitized during the premolt to 2^nd instar, G. porthetriae from 28%, and in 20% of the hosts both parasitoid species were able to develop in one gypsy moth larva. However, when host larvae were parasitized simultaneously during premolt to the 3^rd instar, G. liparidis was successful in 90% of the hosts, compared to 8% from which only G. porthetriae emerged. In the experiments with delayed oviposition, generally the species that oviposited first succeeded in completing its larval development. Larvae of the species ovipositing with four days delay were frequently attacked and killed by larvae of the first parasitizing species or suffered reduced growth. As the secondary parasitoid species, G. porthetriae‐larvae were never able to complete their development, whereas G. liparidis developed successfully in at least 12,5% of the multiparasitized host larvae. Thus, multiparasitism of gypsy moth larvae by both Glyptapanteles species corresponds to the contest type; however, G. porthetriae is only able to develop successfully as the primary parasitoid of young host larvae.     

Factors influencing the developmental capacity of Galleria larval epidermis

The nature of the cuticle secreted by integument from a day-1 penultimate instar larval Galleria when cultured in vivo in the abdomen of a last instar larva varied with the age of the host. When placed in a day-5 last instar larva, the implanted integument secreted a pupal cuticle at the time the host metamorphosed and became a pupa. However, when placed in a day-7 last instar larva the implant, from the same stage donor, secreted a larval cuticle at the time the host pupated. Experimental studies involving implantation of the integument for a 24 hr period, into various developmental stages of normal and ligated last instar larvae, pupae, and pharate adults, prior to placing it in a day-7 last instar larva suggest that a non-hormonal factor present in day-4 and -5 last instar larvae is important to initiate pupal syntheses.     

Interspecific competition between the braconid endoparasitoids Glyptapanteles porthetriae and Glyptapanteles liparidis in Lymantria dispar larvae

The effect of interspecific competition between the solitary endoparasitoid Glyptapanteles porthetriae Muesebeck (Hymenoptera: Braconidae) and the gregarious Glyptapanteles liparidis Bouché (Hymenoptera: Braconidae), was investigated in larvae of Lymantria dispar L. (Lepidoptera: Lymantriidae). Host larvae were parasitized by both wasp species simultaneously in premolt to the 2^nd or the 3^rd host instar or in an additional approach with a 4-day delay in parasitization by the second wasp species. Host acceptance experiments revealed that both wasp species do not discriminate between unparasitized host larvae and larvae parasitized previously by the same or the other species. In more than 90% female wasps parasitized the larva they encountered first. During the period of endoparasitic development, larvae of the competing parasitoid species never attacked the egg stage of the other species. When host larvae were parasitized simultaneously by both wasp species, the rate of successful development of both species depended on the age of the host larva at the time of its parasitization; G. liparidis emerged successfully from 44% of host larvae parasitized during the premolt to 2^nd instar, G. porthetriae from 28%, and in 20% of the hosts both parasitoid species were able to develop in one gypsy moth larva. However, when host larvae were parasitized simultaneously during premolt to the 3^rd instar, G. liparidis was successful in 90% of the hosts, compared to 8% from which only G. porthetriae emerged. In the experiments with delayed oviposition, generally the species that oviposited first succeeded in completing its larval development. Larvae of the species ovipositing with four days delay were frequently attacked and killed by larvae of the first parasitizing species or suffered reduced growth. As the secondary parasitoid species, G. porthetriae-larvae were never able to complete their development, whereas G. liparidis developed successfully in at least 12,5% of the multiparasitized host larvae. Thus, multiparasitism of gypsy moth larvae by both Glyptapanteles species corresponds to the contest type; however, G. porthetriae is only able to develop successfully as the primary parasitoid of young host larvae.     

Storage Proteins of Diamondback Moth,Plutella xylostella: Purification and Expression Profile

Storage proteins (SPs) were significantly detected in the hemolymph during the late instar larvae of diamondback moth, Plutella xylostella. These SPs were resolved into three proteins (SP1, SP2, and SP3) at 7% SDS-PAGE. Their apparent molecular sizes were around 80 kDa. SP1 was synthesized later than SP2 and SP3 during the development of the last instar. Total soluble proteins of last instar larvae were extracted and fractionated sequentially with ammonium sulfate, size-exclusion chromatography, and ion-exchange chromatography. The SPs were purified and their developmental expression was discussed.     

The Immature Stages of Ptilocolepus extensus McLachlan, 1884 (Trichoptera: Hydroptilidae: Ptilocolepinae) with Notes on Biology

The last instar larva and the pupa of Ptilocolepus extensus McLachlan, 1884, an endemic of the Iberian Peninsula, are described for the first time. The final instar larva of Pt. extensus bears close resemblance to those of Ptilocolepus granulatus (Pictet, 1834), but the abdomen has a well-developed dorsal sclerotized plate on segment VIII; this is a morphological feature which has not been observed in any other Ptilocolepinae. Final instar larvae make dorsoventrally flattened, elongate-oval cases of leaf pieces from several moss and liverwort species; such cases are characteristic of the Ptilocolepinae. Some additional data on biology and geographical distribution are given.     

DNA barcoding‐based species delimitation increases species count of Eois (Geometridae) moths in a well‐studied tropical mountain forest by up to 50%

Abstract The genus Eois comprises an important part of megadiverse assemblages of geometrid moths in mountain rainforests of southern Ecuador. In this study we report: (i) on the construction of a DNA barcode library of Eois for identification purposes; and (ii) the exploration of species diversity through species delimitation by pair‐wise distance thresholds. COI barcode sequences were generated from 408 individuals (at least 105 species) collected on a narrow geographic scale (～40 km²) in the Reserva Biológica San Francisco. Analyses of barcode sequence divergence showed that species delimitations based solely on external morphology result in broad overlap of intra‐ and interspecific distances. Species delimitation at a 2% pair‐wise distance threshold reveals a clear barcoding gap. Fifty‐two previously unrecognized species were identified, 31 of which could only be distinguished by an integrative taxonomy approach. Twelve additional putative species could only be recognized by threshold‐based delimitation. Most splits resulted in two or three newly perceived cryptic taxa. The present study increased the number of Eois species recorded from that small area of Andean mountain forest from 102 to 154 (morphology‐ plus integrative taxonomy‐based) or even 166 (sequence‐based), leaving the species accumulation curve still far from reaching an asymptote. Notably, in no case did two or more previously distinguished morphospecies have to be lumped. This barcode inventory can be used to match larvae to known adult samples without rearing, and will therefore be of vital help to extend the currently limited knowledge about food plant relationships and host specialization.     

Natural History and Comparative Morphology of Immatures of <Emphasis Type="Italic">Gamelia anableps</Emphasis> (C. Felder &amp; R. Felder) (Lepidoptera: Saturniidae,Hemileucinae)

Egg, larva, and pupa of Gamelia anableps (C. Felder & R. Felder, 1874) are described for the first time and the chaetotaxy of the first instar provided. Eggs and selected structures of larvae and pupae were also investigated by means of scanning electron microscopy. Eggs are laid in clusters, are ellipsoidal and laterally flat. Larvae passed through eight instars in about 78 days. Pupation occurs inside a poorly organized cocoon of yellowish silk. The dark brown pupa is cylindrical in shape with appendages firmly fused together and to the body wall. The chaeotaxy is compared with other Saturniidae species belongining to Hemileucinae, Ceratocampinae, and Saturniinae and the natural history of G. anableps compared with other Saturniidae.     

Description of a new and highly distinctive genus and species of Euptychiina (Lepidoptera: Nymphalidae: Satyrinae) from the Brazilian southern Amazon

A new genus and species of Euptychiina (Satyrinae), Cristalinaia vitoria Mota, Zacca & Freitas gen. et sp. nov., is described based on three specimens collected in the region of the Cristalino River, Alta Floresta, Mato Grosso, Brazil. This rare species is known only from this region, where it flies inside the dense bamboo patches typical of that area. The last instar larva and the pupa are described; the larva was observed feeding on mature leaves of the common bamboo Guadua aff. paniculata Munro.urn:lsid:zoobank.org:pub:D61EDE8D-CAE9-41C6-B24D-BB789873566E     

The parasitic wasp <Emphasis Type="Italic">Baryscapus crassicornis</Emphasis> (Erdös) (Hymenoptera,Eulophidae) developing in larvae of the weevil genus <Emphasis Type="Italic">Larinus</Emphasis> germar (Coleoptera,Curculionidae)

Baryscapus crassicornis (Erdös) was reared from the last instar larvae of three species of Curculionidae: Larinus idoneus Gyllenhal, developing in the inflorescences of Jurinea cyanoides (L.), L. turbinatus (Gyllenhal), developing in Cirsium arvense (L.) in Russia (Ul’yanovsk Province), and L. latus (Herbst), developing in Onopordum acanthium L. and O. leptolepis DC. (Asteraceae) in Iran. Data on the parasitoid and its hosts biology, variation of morphological characters, and diagnostic notes are provided. The 3rd instar larva and the pupa of B. crassicornis are described for the first time.     

Leaf age effects of elevated CO2-grown white oak leaves on spring-feeding lepidopterans

Folivorous insect responses to elevated CO₂-grown tree species may be complicated by phytochemical changes as leaves age. For example, young expanding leaves in tree species may be less affected by enriched CO₂-alterations in leaf phytochemistry than older mature leaves due to shorter exposure times to elevated CO₂ atmospheres. This, in turn, could result in different effects on early vs. late instar larvae of herbivorous insects. To address this, seedlings of white oak (Quercus alba L.), grown in open-top chambers under ambient and elevated CO₂, were fed to two important early spring feeding herbivores; gypsy moth (Lymantria dispar L.), and forest tent caterpillar (Malacosoma disstria Hübner). Young, expanding leaves were presented to early instar larvae, and older fully expanded or mature leaves to late instar larvae. Young leaves had significantly lower leaf nitrogen content and significantly higher total nonstructural carbohydrate:nitrogen ratio as plant CO₂ concentration rose, while nonstructural carbohydrates and total carbon-based phenolics were unaffected by plant CO₂ treatment. These phytochemical changes contributed to a significant reduction in the growth rate of early instar gypsy moth larvae, while growth rates of forest tent caterpillar were unaffected. The differences in insect responses were attributed to an increase in the nitrogen utilization efficiency (NUE) of early instar forest tent caterpillar larvae feeding on elevated CO₂-grown leaves, while early instar gypsy moth larval NUE remained unchanged among the treatments. Later instar larvae of both insect species experienced larger reductions in foliage quality on elevated CO₂-grown leaves than earlier instars, as the carbohydrate:nitrogen ratio of leaves substantially increased. Despite this, neither insect species exhibited changes in growth or consumption rates between CO₂ treatments in the later instar. An increase in NUE was apparently responsible for offsetting reduced foliar nitrogen for the late instar larvae of both species.     

DNA barcoding the geometrid fauna of Bavaria (Lepidoptera): successes, surprises, and questions

Background

The State of Bavaria is involved in a research program that will lead to the construction of a DNA barcode library for all animal species within its territorial boundaries. The present study provides a comprehensive DNA barcode library for the Geometridae, one of the most diverse of insect families.

Methodology/Principal Findings

This study reports DNA barcodes for 400 Bavarian geometrid species, 98 per cent of the known fauna, and approximately one per cent of all Bavarian animal species. Although 98.5% of these species possess diagnostic barcode sequences in Bavaria, records from neighbouring countries suggest that species-level resolution may be compromised in up to 3.5% of cases. All taxa which apparently share barcodes are discussed in detail. One case of modest divergence (1.4%) revealed a species overlooked by the current taxonomic system: Eupithecia goossensiata Mabille, 1869 stat.n. is raised from synonymy with Eupithecia absinthiata (Clerck, 1759) to species rank. Deep intraspecific sequence divergences (>2%) were detected in 20 traditionally recognized species.

Conclusions/Significance

The study emphasizes the effectiveness of DNA barcoding as a tool for monitoring biodiversity. Open access is provided to a data set that includes records for 1,395 geometrid specimens (331 species) from Bavaria, with 69 additional species from neighbouring regions. Taxa with deep intraspecific sequence divergences are undergoing more detailed analysis to ascertain if they represent cases of cryptic diversity.     

Towards a global barcode library for Lymantria (Lepidoptera: Lymantriinae) tussock moths of biosecurity concern

Background

Detecting and controlling the movements of invasive species, such as insect pests, relies upon rapid and accurate species identification in order to initiate containment procedures by the appropriate authorities. Many species in the tussock moth genus Lymantria are significant forestry pests, including the gypsy moth Lymantria dispar L., and consequently have been a focus for the development of molecular diagnostic tools to assist in identifying species and source populations. In this study we expand the taxonomic and geographic coverage of the DNA barcode reference library, and further test the utility of this diagnostic method, both for species/subspecies assignment and for determination of geographic provenance of populations.

Methodology/Principal Findings

Cytochrome oxidase I (COI) barcodes were obtained from 518 individuals and 36 species of Lymantria, including sequences assembled and generated from previous studies, vouchered material in public collections, and intercepted specimens obtained from surveillance programs in Canada. A maximum likelihood tree was constructed, revealing high bootstrap support for 90% of species clusters. Bayesian species assignment was also tested, and resulted in correct assignment to species and subspecies in all instances. The performance of barcoding was also compared against the commonly employed NB restriction digest system (also based on COI); while the latter is informative for discriminating gypsy moth subspecies, COI barcode sequences provide greater resolution and generality by encompassing a greater number of haplotypes across all Lymantria species, none shared between species.

Conclusions/Significance

This study demonstrates the efficacy of DNA barcodes for diagnosing species of Lymantria and reinforces the view that the approach is an under-utilized resource with substantial potential for biosecurity and surveillance. Biomonitoring agencies currently employing the NB restriction digest system would gather more information by transitioning to the use of DNA barcoding, a change which could be made relatively seamlessly as the same gene region underlies both protocols.