Determination of photosynthetic pigment composition in an individual phytoplankton cell in seas and lakes using fluorescence microscopy; properties of the fluorescence emitted from picophytoplankton cells

To develop a method for the determination of photosynthetic pigment species in individual phytoplankton cells, especially natural picophytoplankton cells, the fluorescence spectra of intact cells were studied with cultured phytoplankton. The study was made mainly with phycoerythrin-containing picophytoplankton collected off Japan in 1982 with reference to diatomal species and phycoerythrin-free cyanophycean species. The spectra were measured for cell suspensions with an ordinary spectrofluorometer, and for individual cells with a microscope spectrofluorometer, paying special attention to the effect of cell-fixation. Results indicated that: (1) the cell-fixation with the glutaraldehyde and paraformaldehyde mixture modified phycoerythrin emission from picophytoplankton markedly in its wavelength location and intensity, but (2) the emission from phycocyanin was affected far less, and (3) the emission from chlorophyll a was not altered. However, the phycoerythrin emission modified by the fixation was found to be easily distinguished from other emissions, and kept its intensity high enough for detection with a fluorescence microscope. The fluorescence properties after the fixation were kept unaltered for a long period of time. Based on the results, we propose a simple method for the determination of photosynthetic pigments in individual phytoplankton cells in seas and lakes using fluorescence microscopy. Results of our tests with natural samples of phytoplankton are presented, and problems for further improvement are also discussed.     

Molecular taxonomy of vestimentiferans of the western Pacific,and their phylogenetic relationship to species of the eastern Pacific III. Alaysia-like vestimentiferans and relationships among families

The phylogenetic status of Alaysia-like vestimentiferans, which were collected at eight sites in the western Pacific, was analyzed on the basis of the nucleotide sequence of part of a mitochondrial gene for cytochrome oxidase I. The 123 individuals analyzed were tentatively classified into four species, which inhabit, respectively, seep areas off the central part of the Japanese mainland, hydrothermal vent fields in the Mid-Okinawa Trough, hydrothermal vent fields in the southern Okinawa Trough, and both seep and vent sites in the Bismarck Sea. From six additional lamellibrachiid specimens from two sites in the Bismarck Sea, two new tentative species were recognized, namely, one from a seep area off Papua New Guinea and another from the hydrothermal vent field at the DESMOS site in the Manus Basin. Phylogenetic analysis using all available sequences of six vestimentiferan families and Alaysia-like vestimentiferans revealed that vestimentiferans could be divided into three groups, namely, lamellibrachiids, escarpiids, and others, even though the monophyly of the third group was supported by only low bootstrap probabilities. Within the first group, the earliest divergence of a tentative lamellibrachiid species from the DESMOS site was apparent. All Alaysia-like vestimentiferans formed a monophyletic group with Arcovestia ivanovi from the Manus Basin, and it is suggested that this group might be derived from vestimentiferans that are endemic to the hydrothermal sites in the Eastern Pacific Rise.     

Life-history patterns in two forms of Macrophthalmus japonicus (Crustacea: Brachyura)

Two behaviorally distinct forms — courtship signal — (Form L and V) of Macrophthalmus japonicus (De Haan) are known. Monthly samples of each form were collected from two areas in Japan: Form L from Amakusa, western Kyushu, between April 1985 and February 1986; Form V from Fukuoka, northern Kyushu, between November 1982 and December 1983. Life-history traits were compared between two populations and discussed. Individuals from the Amakusa population grew more slowly and matured at a smaller size than those from the Fukuoka population. Maturation period and longevity did not differ significantly. In the first year of reproduction, females in the Fukuoka population produced more, and larger broods than those in the Amakusa population and crab mortality was higher in every age group. Egg size did not differ significantly. The major environmental factors — air-temperature, nitrogen content of the surface mud and period of aerial exposure were similar in both areas, and it is suggested that the differences in life-history exhibited by these crabs are genotypic, and perhaps related to their different form.     

Molecular taxonomy of vestimentiferans of the western Pacific and their phylogenetic relationship to species of the eastern Pacific

The nucleotide sequence of part (624 bp) of a mitochondrial gene for cytochrome oxidase I was determined for 46 escarpiid vestimentiferans collected from seven sites in the western Pacific and 49 individual specimens of Arcovestia ivanovi from two sites in the Manus Basin. Phylogenetic analysis, based on the newly obtained and previously reported sequences, indicated that escarpiids in the western Pacific can be divided into two tentative species, as we proposed in a previous report. While members of the first tentative species have been collected exclusively from a seep area at a depth of 300 m off the coast of central Japan, the members of the second species inhabit some sites at depths greater than 1,100 m, namely, seep areas in Japanese and Papua-New Guinean waters as well as hydrothermal vent fields in the Okinawa Trough and the Manus Basin. We detected no genetic structure among populations of the second tentative species. The first tentative species was more closely related to a species in the eastern Pacific, Escarpia spicata, and to a species in the Gulf of Mexico, Escarpia laminata, than to the second tentative species in the western Pacific. Sequences obtained from all arcovestiids were identical with the exception of those from three individuals, each of which included a single synonymous nucleotide substitution relative to the dominant haplotype, and no genetic differences were detected between specimens from the two sites in the Manus Basin.     

Seasonal changes in the diving parameters of king penguins (Aptenodytes patagonicus)

Contrasting conditions at-sea are likely to affect the foraging behaviour of seabirds. However, the effect of season on the dive parameters of penguins is poorly known. We report here on an extensive study of the diving behaviour of king penguins (Aptenodytes patagonicus) over the bird's complete annual cycle at the Crozet Islands. Time-depth recorders were used to record dive duration, bottom duration, post-dive interval, ascent rate and descent rate in breeding adults during different seasons in 1995 and 1996. Seasons included summer (n=6, incubation; n=6, chick brooding), autumn and winter (n=5 and n=3, respectively, chick at the crèche stage), and spring (n=4, birds at the post-moult stage). In all seasons dive duration increased with dive depth, but, for a given depth, dives were longer in winter (6.8 min when averaged over the 100-210 m depth layer) than in spring (4.6 min) and summer (4.4 min). The time spent at the bottom of the dives, which probably represents a substantial part of the feeding time, was much longer in winter (2.5 min per dive for dives over the 100-210 m layer) than during other seasons (1.0-1.4 min), i.e. there was a 2.5-fold augmentation for similar diving depths. Ascent and descent rates increased with increasing dive depth, but no difference in the relationships between rates of ascent and descent and dive depth was found among seasons. Furthermore, for all dive depths, ascent and descent rates were independent of the bottom duration. In all seasons post-dive intervals increased with dive duration and with dive depth, but they were longer in spring (2.3 min for dives over the 100-210 m layer) and summer than in autumn and winter (1.6-1.8 min). The diving efficiency decreased with increasing dive depth and was higher in autumn and winter (0.22-0.29) than in summer and spring (0.15-0.18). The large increase in bottom and dive duration from spring to winter is in agreement with the seasonal drop in prey density, with penguins spending more time searching for prey. In contrast, the consistency of the vertical velocity during contrasting conditions at-sea suggests that the transit time to depth is an important component of the foraging behaviour (scanning of the water column) that is independent of the prey availability. The time budget of the penguins during diving in a fluctuating environment appears to vary primarily during the bottom phase of the dives, with bottom duration increasing with diminishing prey supplies, while post-dive intervals shorten in the same time.     

Brood defense and filial cannibalism in the spottail darter (<Emphasis Type="Italic">Etheostoma squamiceps)</Emphasis>: the effects of parental status and prior experience

Male spottail darters (Etheostoma squamiceps) defend nest sites in which females deposit eggs over the course of several weeks. In laboratory experiments, I tested three predictions of the hypothesis that the presence of eggs increases the value of a nest site to male spottail darters: (1) males should preferentially defend nests with eggs over empty nests, and males that sired the eggs should (2) spend more time defending them and (3) consume less of their broods than should alloparent males. Parent males were tested using eggs they were guarding in the field; alloparents included males that were not defending eggs immediately prior to the experiment (inexperienced), and males that were defending eggs in the field when captured but were given eggs sired by a different male (experienced). In choice experiments in which males were offered nest sites differing only in the presence of eggs, males preferred to defend nest sites with eggs regardless of parental status. Parent males spent significantly more time defending eggs and consumed less of their broods than did inexperienced alloparent males. Experienced alloparent males were similar to parent males in brood defense but had levels of filial cannibalism similar to inexperienced males, suggesting that prior experience may influence brood defense but not egg consumption.     

Nutrients and biomass distribution in the Gulf of Aqaba (Elat), Red Sea

The distribution of phosphate, nitrate and silicate values obtained from 300 samples and of biomass determined by displacement volume in about 1,900 vertical plankton hauls (65, 200 and 330 m) collected from 8 oceanographic station during 1974–1975 in the Gulf of Aqaba (Elat) are summarized and illustrated by crosssections. Generally, the Gulf is poor in nutrients and the data indicate that it is filled with upper (150 m) Red Sea waters, flowing in over the sill of Tiran. Nutrient content of the upper and deep waters immediately outside the sill are well within the known range of those in the northern Red Sea. Biomass values are relatively low in the Gulf are generally similar to those reported from the northern Red Sea. Both nutrients and biomass values display seasonal and bathymetric variations in the Gulf and outside the sill.     

Potential dietary effects on the fatty acid composition of the common jellyfish Aurelia aurita

Fatty acid composition of the natural and aquarium-reared common jellyfish Aurelia aurita was investigated. Fatty acid composition of the aquarium-reared A. aurita clearly reflected that of the diet, brine shrimp (Artemia). In the same way, fatty acid composition of the natural A. aurita was assumed to reflect those of natural diets. Samples of natural A. aurita were collected from April 1995 to September 1995 in the Seto Inland Sea, Japan, and their fatty acids were analyzed by gas chromatography and mass spectrometry. Variation of fatty acid compositions was seasonal rather than dependent on body size. Two major seasonal groups were divided by the cluster analysis of the A. aurita fatty acid composition: the April–June and the August–September clusters. The April–June cluster was characterized by high contents of the (n − 3)-fatty acids of diatom origin, accumulated via the grazing food chain. By contrast, the August–September cluster was characterized by an increase in (n − 6)-fatty acids of macroalgal origin, probably transferred via the detritus food chain. These results suggest that the diet of natural A. aurita may shift between the diatom-based food chain and the detritus-based food chain. Received: 12 April 2000 / Accepted: 1 December 2000     

Development of a new antifouling method for a marine cooling water system

Variations in meristic characters occur within and between samples of milkfish Chanos chanos (Forsskal) collected on June 1977 from five localities in Philippine waters. The unbranched anal and ventral fin rays are the most plastic, and vertebral number is the most stable of nine meristic characters examined. The number of ventral fin rays is the only meristic feature which differed consistently enough to suggest distinct population groups: South China Sea, comprising samples from Ilocos and Panay Island; Pacific Ocean from Bicol; and Celebes Sea, from Zamboanga and Davao. A discriminant analysis which incorporated generalized distance (Mahanalobis D²) and percent overlap of a reduced set of characters indicated several morphometric subgroups of milkfish in Philippine waters, although geographic subgroups were not clearly demonstrated.SEAFDEC Contribution No. 218