Double-labelling methods used to diagnose apoptotic and necrotic cell degradations in copepod nauplii

Two double-labelling methods, Tunel+propidium iodide and Annexin V-FITC+propidium iodide, have been tested to diagnose cell degradation processes in N1–N2 nauplius stages produced by spawning females of the copepod Calanus helgolandicus fed either non-toxic (PRO: control) or toxic (TR1) diets under laboratory conditions. Observation of labelled samples with a confocal laser-scanning microscope revealed that the maternal-food effect, following absorption of the toxic diet, induced cell apoptosis and necrosis in 80–100% of the offspring, which sooner or later died. Similar cell damages were also observed in 18–50% of the nauplii produced by females incubated in filtered seawater. Such cell degradation processes and high mortality were not observed in nauplii produced by females fed the non-toxic diet. Our protocols allowed detection of cell degradations before death of unhealthy larvae, preferentially with a confocal microscope or a conventional visible-fluorescent-light microscope.Communicated by O. Kinne, Oldendorf/Luhe     

Changes in otolith microstructure and microchemistry during larval development of the European conger eel (<Emphasis Type="Italic">Conger conger</Emphasis>)

Otolith microstructure and chemical composition (Sr:Ca ratios) of the European conger eel (Conger conger) were examined during the larval developmental stages by scanning electron microscopy and wavelength dispersive spectrometer. Back-calculated hatching dates from the otolith microstructure of the developing leptocephali indicate a protracted spawning season from December to June. The early age of our developing specimens captured south of the Azores Islands suggests that the conger eel has another spawning area closer to Azores than the Mediterranean. Otolith increment width, which was relatively constant and narrow in the developing leptocephalus stage, increased sharply at age 170-250 days. Sr:Ca ratios in the otolith, which increased during the developing leptocephalus stage, showed a rapid drop coinciding with the increase in increment width. These coincidental changes were regarded as the onset of metamorphosis for this species. A close linear relationship between the age at metamorphosis and otolith growth rate indicates that the faster-growing larvae metamorphose earlier, suggesting that somatic growth should play an important role in the timing of metamorphosis. As shown in earlier work, the existence of an otolith marginal zone with unclear rings during metamorphosis prevents an accurate estimate of the larval stage duration of this species.     

Whitefly wax as a cue for phoresy in the broad mite, <Emphasis Type="Italic">Polyphagotarsonemus latus</Emphasis>(Acari: Tarsonemidae)

Summary. Broad mite, Polyphagotarsonemus latus (Acari: Tarsonemidae) exhibits a specific phoretic relationship with whiteflies. Under field conditions most broad mites, caught in sticky traps, are attached to whiteflies. Under laboratory conditions, attachment occurs equally well in the dark and light. Mites do not differentiate between the sexes of their phoretic host Bemisia tabaci. However, mite attachment to B. tabaci is greatly diminished by washing the host with various organic solvents, chloroform in particular. The effect of whitefly waxy particles on broad mite behavior was studied using wax from the whitefly Aleyrodes singularis and from the mealybug Planococcus citri. Broad mites were not only attracted specifically to the A. singularis waxy particles-treated leaf areas but were also attached to leaf trichomes in this area. The results of this study suggests the importance of olfactory cues from the whitefly waxy particles in the recognition process of the phoretic host and/or the induction of the attachment behavior to whitefly legs or leaf trichomes.     

Reproductive cycle of<Emphasis Type="Italic"> Bispira volutacornis</Emphasis> (Polychaeta: Sabellidae) on the west coast of Ireland

The reproductive cycle of Bispira volutacornis, a dioecious polychaete, was followed over two spawning periods in Lettercallow Bay, on the west coast of Ireland. In the younger generation of sexually mature adults, gender can be determined on the basis of colour, the dorsal aspect of the males being cream at maturation, whilst the females are pink at the previtellogenic stage and an olive green at maturation. An annual reproductive cycle of 12 months shows that oocyte/sperm maturation is extraovarian. The male reproductive cycle proceeded at a more rapid pace than that of the female cycle, with sperm maturing earlier. Spawning in B. volutacornis occurred between late July and early September. B. volutacornis is a polytelic species, surviving spawning and releasing all gametes in a single batch, with a mean spawned oocyte diameter of 118 m. The reproductive cycle appears to be regulated by a seasonal change in water temperature, and spawning is timed to produce larvae during the autumnal planktonic bloom in Lettercallow Bay.Communicated by J.P. Thorpe, Port Erin     

Inter-sexual differences in T-cell-mediated immunity of black-headed gull chicks (<Emphasis Type="Italic">Larus ridibundus</Emphasis>) depend on the hatching order

Hatching asynchrony in avian species leads to age and size differences between nestlings within a brood, handicapping last-hatched chicks in the sibling rivalry. Starvation due to this competitive disadvantage has been regarded as the primary cause of an increase in mortality with hatching order. However, for gulls it has also been suggested that disease is the cause of mortality for last-hatched chicks, possibly through reduced immunocompetence and thereby an enhanced susceptibility to infection. In addition, the male-biased mortality reported for several gull species may be related to a higher vulnerability to diseases in males compared to females. To determine the potential influence of the immune system on these mortality patterns, we investigated the T-cell-mediated immunity (CMI) of black-headed gull chicks in relation to hatching order and sex. We found a significant decrease in the CMI with hatching order. This result may be causally related to systematic changes in maternal yolk steroids and carotenoids within the laying sequence. For second-laid eggs, male CMI was significantly lower than female CMI. This is possibly linked to higher plasma levels of testosterone in male embryos which might have an immunosuppressive effect. If so, this effect is masked in eggs of either high (first egg) or low (last egg) quality. Chicks with low CMI showed enhanced mortality rates. Thus the differences in immune response are likely to contribute to the observed mortality patterns. However, hatching order significantly affected mortality independently of CMI, suggesting that competitive disadvantage due to hatching asynchrony is also important.Communicated by M. Webster     

Change in the uptake and cellular Si:N ratio in diatoms responding to the ambient Si:N ratio and growth phase

A change in the Si:N ratio of diatom cells during growth was examined for Chaetoceros socialis and Thalassiosira sp., with different initial silicate to nitrate (Si:N) ratios in the media. During exponential growth, C. socialis assimilated silicate and nitrate with a molar ratio of 0.5, independent of the ratio in the media, but after the depletion of nitrate, silicate continued to be taken up, and the Si:N ratio in the stationary phase increased to 2 as a function of the Si:N ratios in the media. In contrast, the ratio of silicate to nitrate taken up by Thalassiosira sp. increased with an increase in the Si:N ratio in the media. The Si:N ratio in the cells during the stationary phase increased in response to an increase in this ratio in the media. The Si:chl  a ratio also increased with the increase in the initial Si:N ratio in the media, while the N:chl  a ratio did not change to a great extent, indicating the changes in the cellular Si:N ratio was derived from changes in the Si content of the cells. These results indicated that the cellular Si:N ratio changed with the Si:N ratio in the medium, and the Si:N uptake ratio during the growth phase was different depending on diatom species. Thus, the dominance of different diatom species may affect nutrient composition and dynamics in the ocean.     

Genetic structure of the scleractinian coral<Emphasis Type="Italic"> Platygyra sinensis</Emphasis> in Hong Kong

Genetic structure was studied in Platygyra sinensis from six sites in Hong Kong by allozyme electrophoresis using an isoelectric focusing gel (IEF). Seven variable loci were detected using six enzyme systems. The number of alleles per locus ranged from 3.3 to 3.6 in the six samples, and heterozygosity from 0.429 to 0.540. Genotype frequencies were generally close to Hardy–Weinberg expectations. The values of unique multilocus genotype to the number of individuals for each sample (N_G/N) and observed genotypic diversity to expected genotypic diversity (G_O/G_E) were high, indicating a high level of sexual reproduction. Variation in allele frequencies among sites was low (F_ST=0.017), as was Nei's unbiased genetic distance (D), suggesting genetic similarity among sites.Communicated by T. Ikeda, Hakodate     

Worker piping in honey bees (<Emphasis Type="Italic">Apis mellifera</Emphasis>): the behavior of piping nectar foragers

This study investigates the brief piping signals ("stop signals") of honey bee workers by exploring the context in which worker piping occurs and the identity and behavior of piping workers. Piping was stimulated reliably by promoting a colony's nectar foraging activity, demonstrating a causal connection between worker piping and nectar foraging. Comparison of the behavior of piping versus non-piping nectar foragers revealed many differences, e.g., piping nectar foragers spent more time in the hive, started to dance earlier, spent more time dancing, and spent less time on the dance floor. Most piping signals (approximately 99%) were produced by tremble dancers, yet not all (approximately 48%) tremble dancers piped, suggesting that the short piping signal and the tremble dance have related, but not identical, functions in the nectar foraging communication system. Our observations of the location and behavior of piping tremble dancers suggest that the brief piping signal may (1) retard recruitment to a low-quality food source, and (2) help to enhance the recruitment success of the tremble dance.     

Factors influencing length of maternal care in brown bears (<Emphasis Type="Italic">Ursus arctos</Emphasis>) and its effect on offspring

Length of maternal care, i.e. the interval between successfully raised litters, is the most important factor explaining the variation in reproductive rate among brown-bear (Ursus arctos) populations. In this paper, we examine the variation in length of maternal care in radio-marked brown bears and its effect on their offspring in northern Sweden. Young stayed with their mothers for 1.4–1.5 or 2.4–2.5 (in one case 3.5) years and were weaned with body masses varying from 17 to 69 kg. The probability of yearling litters staying with their mother for a 2nd year increased with decreasing yearling body mass, and was higher for litters with two offspring than for litters with one or three to four offspring. Staying with their mothers for a 2nd year had a positive effect on mass gain in yearlings and this effect was more pronounced in litters with two than three to four offspring. Body mass of 2-year-olds was not related to age of weaning, suggesting that keeping offspring for an additional year mainly compensated for low yearling body mass. If large offspring body mass positively affects later offspring survival and reproduction, mothers may be able to optimize the length of maternal care according to the litter size and the size of their yearlings.Communicated by F. Trillmich