Multiple overwintering mechanisms inChymomyza amoena larvae (Diptera: Drosophilidae) and laboratory induction of freeze tolerance

Summary Chymomyza amoena larvae in apples in summer were found to be either potentially freeze tolerant or to supercool. By late September only potentially freeze tolerant larvae were recovered from apples. Larvae from walnut husks in winter supercooled to avoid freezing. However, freeze tolerance could be induced in laboratory grown larvae by placing them in apples around the seed area and maintaining them at chilling temperatures for several weeks. Overwintering mechanisms employed byC. amoena larvae in Michigan appear to depend upon larval feeding site.Prof. Davis at Michigan Agricultural College placed 9C. amoena emerging from apples in August and Sept. 1891 along with codling moth in the MAC (=MSU) Entomological Museum collection.     

Developmental patterns of supercooling capacity in a subantarctic wingless fly

Anatalanta aptera is a wingless fly which lives in subantarctic islands, particularly in sea bird colonies. Developmental patterns of supercooling capacity were studied in an experimental population reared at 5°C and fed ad libitum. Mean supercooling points of the eggs, second and third instar larvae, pupae, teneral and mature adults were –28.0°C, –10.8°C, –8.8°C, 17.6°C, –16.6°C and –8.2°C respectively. Low mean supercooling points were found for inactive stages (eggs, pupae) and, unexpectedly among Diptera, teneral adults. Mature adults had a combination of low supercooling ability, high lipid content and physogastry. The results are compared with those obtained in winged species from other families of Diptera.     

Overwintering ofChymomyza amoena larvae in apples in Michigan and preliminary studies on the mechanism of cold hardiness

Summary Chymomyza amoena, a member of the family Drosophilidae, is now breeding in fallen apples in the state of Michigan.C. amoena larvae can overwinter in fallen apples, completing development and emerging as adults in the summer. Cold resistance during the overwintering phase may not depend upon the accumulation of glycerol or other sugar alcohols.     

Temperature-dependent activity of mosquito larvicidal factor(s) present inBacillus sphaericus 1593–4 and 1691

Summary The larvicidal factor(s) ofBacillus sphaericus 1593–4 and 1691 was more active at high temperature (33–35 °C) as compared to low temperature (23–25 °C) for the larvae ofCulex fatigans andAnopheles subpictus.     

Amino acid and allozyme frequency changes in overwinteringChymomyza amoena (Diptera: Drosophilidae) larvae

Summary Proline is accumulated by overwinteringC. amonea larvae. Mortality averages over 50% by spring. PGM^F in the population shifts from 70% in summer to less than 50% during winter, increasing again in summer. Product synthesis from stored glycogen may be mandated in dipteran larvae also using larval proteins for cold hardiness when completion of development is delayed until spring.Thanks are gratefully extended to Derek Lamport and Patrick Muldoon of the MSU/DOE Plant Biology Laboratory for the amino acid analyses. Thanks are also extended to Jerry Wolfe, Director of Mt. Lake Biological Station in summer, 1985 and Blaine Cole, Director beginning summer, 1986 for research space, to the University of Virginia, Charlottesville for a fellowship for the work at MLBS in summer, 1986 and to all the property owners in Michigan and Virginia who allowed collection at their orchards, farms, gardens or residences. Apples in January and March, 1986 were from a university research orchard. — This paper is dedicated to Hampton L. Carson on his retirement.     

Aggregated oviposition byChymomyza amoena (Diptera: Drosophilidae)

Summary Aggregated oviposition byChymomyza amoena in Virginia and Michigan has been shown to be due to egg clustering, females attracted to fruits on whichC. amoena females have already oviposited, and to variation in fruit attractiveness. Negative binomialk values support behavioral similarities among females of this rarely seen species in both states. Discovery ofC. amoena in Europe may indicate that this larval overwintering drosophilid has a Holarctic distribution.The research in Virginia during summer 1986, conducted at Mt. Lake Biological Station, was supported by a fellowship from the University of Virginia. Thanks are gratefully extended to all property owners in Virginia and Michigan for allowing me to collect fruits and to Blaine Cole, Director and J.J. Murray, Chairman of the Biology Dept., University of Virginia for research space in 1986 and 1987. R. N. Band constructed the figures. Comments from reviewers have been helpful.     

The effect of proctolin on the adenylate and guanylate cyclases in theLocusta brain at various developmental stages

Summary Proctolin at concentrations 10^–8–10^–7 M elevated by 40% brain adenylate cyclase activity of adultLocusta migratoria migratorioides R.F. In moulting individuals, proctolin caused a decrease in brain adenylate cyclase activity, and it proved to be ineffective in the larvae. Proctolin caused only a slight decrease on guanylate cyclase activity of the brain at every developmental stage.     

Effects of the adenylate cyclase activator forskolin and its inactive derivative 1,9-dideoxyforskolin on insect cytochrome P-450 dependent steroid hydroxylase activity

The adenylate cyclase activator forskolin and its pharmacologically inactive derivative 1,9-dideoxyforskolin were found to inhibit in a dose-dependent fashion the ecdysone 20-monooxygenase activity associated with wandering stage larvae ofDrosophila melanogaster and fat body and midgut from last instar larvae of the tobacco hornworm,Manduca sexta. The concentrations of these labdane diterpenes required to elicit a 50% inhibition of the cytochrome P-450 dependent steroid hydroxylase activity in the insect tissues ranged from approximately 5×10^–6 to 5×10^–4 M.     

Enzymatic characterization of nine endoparasite species of small ermine moths (Yponomeutidae)

Summary Eight hymenopterous and 1 dipterous species, all endoparasitic in eggs, larvae, or pupae of small ermine moths (Yponomeuta) were investigated for their allozyme variation at 3–29 loci. The mean heterozygosity level of the hymenopterous species is one-third of that of the dipterous species. Zymogram patterns of the parasite larvae do not interfere with those of the host.     

Chemical mediators in the oviposition behaviour of the house longhorn beetleHylotrupes bajulus

Summary Oviposition behaviour in the timber pestHylotrupes bajulus is mediated by pheromones ((–)-verbenone and p-cymen-8-ol) produced in the frass of the wood-boring larvae of the species.Acknowledgment. We thank the S. R. C. for a studentship (MDH) and N. E. R. C. for research support and technical assistance.     

Octopamine and proctolin mimic spontaneous membrane depolarisation inLucilia larvae

Summary Octopamine and proctolin at concentrations below 10^–8 M reversibly induce a spontaneous rhythmic depolarization which occurs in body-wall muscles ofLucilia larvae. The effect appears to be postsynaptic and mediated by receptors specific for each substance.This work was supported by NIEHS grant No. ES00814 and EPA grant No. R804345.     

Cryopreservation of parasites

Summary In this review, advances in cryopreservation of helminth parasites are reported. Our own studies demonstrate that metacestodes ofEchinococcus multilocularis can be maintained in a viable state for at least 1–2 years by appropriate deep-freezing and storage in liquid nitrogen. Infective larvae of the nematodeToxocara canis cryopreserved for 1 week in liquid nitrogen were maintained after thawing in vitro in a chemically defined medium for 35 weeks. Although motility of previously deep-frozen larvae was reduced they produced secretory/excretory antigens of similar immunodiagnostic quality as those from unfrozen larvae. Whereas infective larvae of several species of trichostrongylids can be easily cryopreserved, the infective larvae of the cattle lungworm,Dictyocaulus viviparus, and muscle larvae ofTrichinella spiralis are more sensitive to damage by subzero temperatures. Therefore, survival rates after cryopreservation are low, but improvement of the cooling schedules appears to be feasible. It is concluded that cryopreservation of certain stages of helminth and protozoan parasites is a useful technique for long-term storage of defined isolates, which can contribute considerably to reducing the number of experimental animals usually required for serial passages.     

The phorbol ester TPA induces metamorphosis in Red Sea coral planulae (Cnidaria: Anthozoa)

Controlled experiments on the metamorphosis of marine invertebrate larvae require artificial inducers. These inducers can be used for studying the involvement of known signal transduction pathways in settlement and metamorphosis. The ability of the tumor-promoting phorbol ester TPA (12-O-tetradecanoylphorbol-13-acetate) to induce metamorphosis in planulae of the Red Sea soft coral speciesHeteroxenia fuscescens, Xenia umbellata, Dendronephthya hemprichii, Litophyton arboreum andParerythropodium fulvum fulvum, and the stony coralStylophora pistillata, was examined by using various concentrations of TPA. The chemical induced metamorphosis in all six species. The effect was unspecific and concentration-related. For all the corals except forX. umbellata the highest mean percentages of metamorphosis were obtained with 8.1×10^–7–10^–9 M TPA. ForX. umbellata, the percentage of metamorphosis was lower, and was obtained within a wider TPA concentration range. The present results, along with previous studies on Hydrozoa and Scyphozoa, demonstrate that TPA is the first common artificial inducer for these classes of Cnidaria. TPA is known to activate the enzyme protein kinase C (PKC) and therefore plays an important role in studying the phosphatidylinositol signal transduction system. Evidence for the involvement of this pathway in triggering metamorphosis has already been reported for Hydrozoa and Scyphozoa. Our results suggest that PKC is also involved in initiating metamorphosis in Anthozoa.     

The effect of chronic heat stress on cortisol levels in the antarctic fishPagothenia borchgrevinki

Radioimmunoassay was used to determine levels of the stress-inducible glucocorticoid, cortisol, circulating in the plasma of the extremely stenothermal Antarctic fishPagothenia borchgrevinki at rest and after heat stress. Fish sampled immediately after capture (–1.9°C) had low cortisol levels (10.4±1.4 ng ml^–1, mean±SEM) as did fish which were laboratory rested for 3 days. Sudden exposure to 5°C (48h) resulted in a peak cortisol value after 3 h (69.9±6.8 ng ml^–1) whereas exposure to 8°C (6h) resulted in a peak value after 1 h (73.5±8.0 ng ml^–1). At both temperatures levels remained significantly elevated (p<0.05) for the entire period of exposure. Increased temperature also resulted in a significant change in haemoglobin, haematocrit and mean cell haemoglobin concentration (MCHC) (p<0.05). Plasma lactate was significantly elevated only after exposure to 8°C (p<0.05). Plasma cortisol levels fromP. borchgrevinki are reported here for the first time and show this cryopelagic Antarctic species to have an unusual hormonal stress profile.     

Influence of cyclic nucleotides on protein synthesis in vascular smooth muscle

Summary The incorporation of leucice-¹⁴C into protein in bovine mesenteric arteries was augmented by cyclic GMP (10^–3 M) and decreased by cyclic AMP (10^–3 M). There was no effect of 5 AMP (10^–3 M). The phosphodiesterase inhibiting drugs theophylline (10^–3 M) and papaverine (5×10^–5 g/ml) both decreased the leucine-¹⁴C incorporation.We are indebted to Mrs.Lena Burlin for hear assistance. Finacial support has been provided by the Swedish State Medical Research Council (No. 04X-101X-4498).     

Nematicidal activity of secondary and tertiary alkyl amides and amines

Summary Several C₁₁ to C₁₅ amides and amines that disrupt growth in certain insects showed high nematicidal activity in direct contact tests. Two amides and 9 amines killedPanagrellus at 5–10 ppm. Of these, 1 amide and 3 amines killedMeloidogyne larvae at 20 ppm.Nematology Laboratory.Insect Physiology Laboratory.     

Hemolymph proteins in an ascitic condition,induced by lethal1(3)gl tumorous tissue inDrosophila hydei

Summary Drosophila hydei larvae homozygous for the gene1(3)gl may survive to the age of 3 weeks, become bloated and be incapable of metamorphosis. Wild-type flies after the injection of a fragment of the1(3)gl disc, which has been previously culture in vivo for 40–500 days, also become bloated (ascitic reaction). In the hemolymph of both the advanced1(3)gl larvae and ascitic adult hosts the accumulation of a specific protein is observed.Acknowledgments. This work was supported by the Swiss National Science Foundation under Grant No.3.792-0.76 and the Georges und Antoine Claraz-Schenkung. We thank Prof. H. Gloor, University of Geneva, and Prof. P.S. Chen, University of Zürich, for advice and criticism.     

Bicarbonate and chloride transport across rat ileal basolateral membrane

The mechanisms of HCO ₃ ^– and Cl^– transport across basolateral membranes from rat ileum were investigated in isolated vesicles by means of uptake experiments. Neither Cl^–/HCO ₃ ^– exchanger nor Na⁺–(HCO ₃ ^– )_n cotransport seem to be present in ileal basolateral membranes. Moreover Cl^– uptake is unaffected bycis Na⁺ and/or K⁺ gradients, indicating the absence of Na⁺–Cl^–, K⁺–Cl^– and Na⁺–K⁺–2Cl^– symport activity. An electrically conductive pathway seems to be responsible for both HCO ₃ ^– and Cl^– fluxes. Evidence is also given for the presence of a Na⁺/H⁺ exchanger at the basolateral pole of ileal enterocytes.     

Zuchtmethode und semisynthetische Nährmedien für Apfelwickler

Summary A method for the individual rearing of the codling mothLaspeyresia pomonella is described. The method allows the collection of exactly dated eggs and the rearing of a large number of larvae in a small space. By using the second medium described, it is possible to watch all developmental stages at any time without disturbing them. The average larval survival rate is 70–80%.     

Transfer of isolated nuclei into protoplasts ofAspergillus nidulans

Nuclei were isolated from protoplasts of a haploid auxotrophicAspergillus nidulans strain. Transformation of protoplasts prepared from a complementary haploid auxotrophic strain with these purified nuclei resulted in both heterokaryotic and diploid colonies. The nutritionally-complementing colonies appeared at a frequency of 5×10^–7 to 10^–8.