Antarctic nemerteans: Heteronemertea– descriptions of new taxa, reappraisals of the systematic status of existing species and a key to the heteronemerteans recorded south of latitude 50oS

Twenty-three species representative of 15 heteronemertean genera are recorded from Antarctic and Subantarctic waters south oflatitude 50^oS. Five new genera are established and seven new species are fully described and illustrated. These are Cephalurichus antarcticus gen. et sp. nov., Dokonemertes magellanensis gen. et sp. nov., Dokonemertes macquariensis sp. nov., Micrellides multiperula gen. et sp. nov., Paramicrurinella falklandica gen. et sp. nov., Parborlasia landrumae sp. nov. and Zodionemertes chilensis gen. et sp. nov. Baseodiscus antarcticus Baylis, Antarctolineus scotti (Baylis), Lineus longifissus (Hubrecht) and Parborlasia fueguina Serna de Esteban & Moretto are redescribed and illustrated and the nature of rhynchodaeal pouches protruding into the cephalic blood lacunae of Parborlasia corrugatus (Mcintosh) is reexamined and discussed. The main anatomical characters of the remaining 11 species are recorded and it is concluded that of these only Parapolia grytvikenensis Wheeler, Cerebratulus malvini Wheeler, Chilineus glandulosa (Burger), Huilkia ushuaiensis Serna de Esteban & Moretto, Micrura pacifica Friedrich and Wiolkenia friedrichi Serna de Esteban & Moretto can at present be accepted as valid, although incompletely described, taxa. Cerebratulus larseni Wheeler, Cerebratulus validus Bürger, Lineus autrani Joubin and Lineus turqueli Joubin are so poorly and inadequately described that their taxonomic status cannot be verified, and the recorded occurrence of Lineus nigricans Burger in southern latitudes cannot be substantiated on the available evidence. Maps showing the recorded distribution of all the species are included and a key to the 18 validated species is provided.     

Stylet penetration by the bayberry whitefly, as affected by leaf age in lemon, Citrus limon

Bayberry whitefly (Parabemisia myricae [Kuwana]) crawlers were placed on young and mature lemon leaves and were allowed 7–9 days to settle. Afterwards, the nymphs were fixed and sectioned in situ on the leaves and the area of leaf under each whitefly was examined at 1 000 x for stylet penetration. Both stylets and stylet tracks were readily visible in the sections. The path of penetration was mostly intercellular and the objective appeared to be the phloem. Passage of the stylets through the plant tissue did not cause detectable damage to most cells; however, damaged plant cells occasionally were noticed. Nymphs that had moulted during the 7–9 day settling period reached the phloem significantly more often than those that were still in their first instar. In each of the three replicates, penetration in the mature leaf occurred significantly less often than penetration in the young leaf (4% vs. 72%, p<0.01, ²). Penetration appears to be inhibited in the mature leaf either by the leaf cuticle or by factors detected by the nymphs after very shallow penetration into the leaf. The cuticle of mature leaves was much thicker than the cuticle of young leaves and may have been a barrier to stylet penetration.

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Résumé Des larves de premier stade (avant la fixation) de l'aleurode, Parabemisia myricae (Kuwana), ont été placées, pour qu'elles se fixent, pendánt 2 à 9 jours, sur des feuilles jeunes ou mûres de citronnier. Des morceaux de feuilles avec des larves fixées ont été trempés dans 2% d'agar; ce procédé a permis de maintenir les larves à leurs sites de fixation finale. Des pièces d'agar contenant les morceaux de feuilles avec les larves ont ensuite été trempées dans de la paraffine, et sectionnées en séries de 10 . Il était nécessaire de maintenir une basse température pendant l'opération afin d'obtenir de bonnes sections des feuilles mûres durcies. Les stylets et leurs traces étaient faciles à voir dans les sections teintées aux safranins, et fast green. Presque toutes les traces des stylets étaient intercellulaires et leur destination semblaient être de phloème. En général, la plupart des cellules ne semblaient pas endommagées, les larves qui avaient mué pendant la période de 7 à 9 jours de fixation avaient atteint significantivement plus le phloeme que les larves du premier stade. Dans trois essais, la pénétration des feuilles mûres était significativement moins fréquente que celle des feuilles jeunes (p<0.01, ²).La pénétration dans des feuilles mûres semble être empêchée par la cuticule ou par des facteurs perçus par les nymphes après une pénétration superficielle. La cuticule des feuilles mûres était beaucoup plus épaisse que la cuticule des jeunes feuilles et pourrait donc représenter une barrière à la pénétration des stylets.

     

Biochemical and biophysical characterization of collagen in Tenebrio molitor L. (Coleoptera, Tenebrionidae)

The collagen from the mesenteric sheath of the tenebrionid insect Tenebrio molitor was extracted by limited pepsin digestion and purified. This collagen was characterized using CM-cellulose chromatography, sodium-dodecylsulfate disc-gel electrophoresis and aminoacid analysis. This molecule was found to be assembled from three identical alpha chains and could be represented by the formula (alpha) 3. The amino acid composition is characteristic of collagen (one-third glycine, high iminoacid content), with high content of hydroxylysine and low content of alanine. Cyanogen bromide digests of these chains indicated that they are not related to any of the known invertebrate or vertebrate chains of interstitial collagens. The molecular weight (M = 280000D) and length (290 nm) were typical, and the banding patterns of the segment-long-spacing crystallites (SLS) and of the reconstitued fibrils were very similar to type I collagen. The denaturation temperature (Td) was 30.7 degrees C and correlated with the total pyrrolidine content as observed in other collagens (von Hippel & Wong's relation). It was concluded that the collagen from this insect showed the classical biochemical and biophysical features of other invertebrate interstitial "primitive" collagens.     

Purification and infectivity ofEntomophaga grylli (Fresenius) Batko Pathotype 2 againstMelanoplus differentialis (Thomas) [Ort.: Locustidae]

The life stages ofEntomophaga grylli (Fresenius) Batko Pathotype 2 were purified and separated by centrifugation in Percoll^R density-gradient medium. The ranges of buoyant densities for germinated resting spores, germ conidia, and resting spores respectively were: 1.040–1.050, 1.055–1.085, and 1.080–1.120 g/ml. Cuticular invasion by germinated germ conidia was the means by whichMelanoplus grasshoppers became infected. Scanning electron micrographs revealed germination of germ conidia on the visible host integument at 100% RH, but not at 90% RH. Significantly higher mortality (P<0.05) was obtained after 3 weeks with grasshoppers incubated in constant light than in constant dark for 24 h following treatment. The disease was not transmitted by ingestion of any life stage. Contribution N^o 85-153-J, Department of Entomology. Agricultural Experiment Station, Kansas State University, Manhattan, KS 66506.     

Neodiaptomus intermedius n. sp. (Calanoida: Copepoda) from South India

A new species of Neodiaptomus, N. intermedius n. sp. is described and illustrated. It is compared with related species of the strigilipes — physalipus species group.     

Comparison of peptidase activities in some fungi pathogenic to arthropods

Peptidases, highly specific toward several synthetic chromogenic peptides, were found in the mycelia of four arthropod pathogenic fungi: Aphanomyces astaci, Beauveria bassiana, Metarrhizium anisopliae, and Paecilomyces farinosus. A. astaci peptidases had high hydrolyzing activities toward most of the peptides, especially those with arginine in the P1 position, while those of B. bassiana and P. farinosus readily hydrolyzed peptides with valine and arginine, as well as proline and tyrosine in the P2 and P1 positions, respectively. The hydrolyzing capacities of M. anisopliae peptidases were similar to A. astaci, but showed lower specific activities. Casein or azocoll was only hydrolyzed by A. astaci peptidases. B. bassiana and M. anisopliae had a very low hydrolyzing capacity toward casein and could not degrade azocoll. P. farinosus had no hydrolyzing activity toward casein or azocoll. Only peptidases from the crayfish pathogen A. astaci could degrade the crayfish cuticle. The peptidase preparations of A. astaci and B. bassiana hydrolyzing MeO-Suc-Arg-Pro-Tyr-pNA or Bz-Phe-Val-Arg-pNA were of the serine type. The possible importance of peptidase activity of arthropod pathogenic fungi in the infection process is discussed.     

Factors to consider in performing survival studies with insect cells

Summary Insect cell lines are not well-suited to colony formation in liquid medium following low-density cell plating. The present studies demonstrate that the time of addition of fetal bovine serum to the culture medium and the number of γ-irradiated feeder cells added to each plate are important factors in developing a useful colony formation assay. TN-368 lepidopteran and WR69-DM-1 dipteran cell lines were used for these experiments. Both cell types display increased plating efficiencies if serum is added to the medium one or more days prior to plating as compared to adding serum immediately before plating. Growth curves obtained by seeding cells at higher densities also indicate that cell growth is slightly better if serum is added one or more days before seeding. These findings are especially important for survival and toxicity studies because the results demonstrate that even seemingly minor factors involved in cell survival assays may benefit treated cells to a greater degree than untreated control cells, thus providing an erroneous assessment of cell survival. This work was supported by USPHS grant R01-CA34158, awarded by the National Cancer Institute, DHHS, Bethesda, MD.     

Nemerteans of the Great Barrier Reef 6. Enopla Hoplonemertea (Polystilifera: Reptantia)

Two new genera and species of reptantic polystiliferous hoplonemerteans from the Great Barrier Reef province of Australia are described and illustrated. Urichonemertes pilorhynchus gen. et sp. nov. and Xenonemertes rhamphocephalus gen. et sp. nov. are both placed in the family Drepanophoridae (Polystilifera: Eureptantia: Aequifurcata).     

The establishment of cell lines from imaginal wing discs of Spodoptera frugiperda and Plodia interpunctella

Continuous cell cultures were established from imaginal wing discs of 2 Lepidoptera, Spodoptera frugiperda and Plodia interpunctella. The S. frugiperda line (IAL-SFD1) grows as multicellular vesicles and responds morphologically and biochemically to the insect hormone, 20-hydroxyecdysone. In contrast, the P. interpunctella cells (IAL-PID2) grow as attached monolayers of small spindle-shaped cells and do not appear to have specific responses to 20-hydroxyecdysone, although growth rates are slowed in these cells upon exposure to the hormone.