Über den einfluß des hydrostatischen druckes auf die ultrastrukturen von zellen und geweben. IV. Der effekt der temperatur auf die druckresistenz der membran-, filament- und mikrotubulisysteme verschiedener gewebe mariner muscheln und Branchiostoma lanceolatum

Temperature-pressure experiments have been carried out on gill epithelium of Modiolus modiolus and Mytilus edulis (Baltic and North Seas), as well as on the skin, pharynx, and trunk muscles of Branchiostoma lanceolatum (Pallas), employing the technique of pressure fixation. Temperatures of 0°, 5° and 15°C have been combined with pressures up to 800 atm. Low temperature (0°C) intensifies disintegration of ultrastructures by pressure in comparison to 5°C. The highest degree of structural disorganization can be observed at 600 atm and 0°C. Increased temperature (15°C) increases pressure resistance, but the structural difference is more quantitative than qualitative. In pressure experiments, temperature probably affects cell organelles at the molecular level.     

Method for indirectly defining optimum temperatures of inhabitancy for marine cold-blooded animals

Investigations on the influence of pressure on the activity of alkaline phosphatase of some marine invertebrates from the Baltic Sea were carried out at temperatures between 5° and 55°C. The results obtained indicate that the influence of pressure on alkaline phosphatase activity is modified by temperature. At low temperatures, pressure has a decreasing effect on enzyme activity, while it has an increasing effect at higher temperatures. The gill homogenates of the relatively pressure-resistent bivalves Cyprina islandica and Mytilus edulis show a lower decrease of activity under pressure at low temperatures than the gill-homogenate of Carcinus maenas and a purified enzyme preparation of a homoiothermal animal.     

Some aspects of the thermal tolerance of Trichomya hirsuta (Mollusca: Bivalvia) of Eastern Australia

The upper temperature tolerance range for Trichomya hirsuta L. is described for acclimation temperatures of 10° to 32°C at 30% S. Changes in thermal resistance with time are described along with the incipient lethal temperatures. The eurythermicity for the species is 240.7 C°-squared. The ultimate upper incipient lethal temperature is 35.1°C. Definition of the incipient sublethal temperature is given, and is equal to 32°C. Diet effects are shown not to be significant in short-term lethal experiments, but indicate that unfed mussels reduce their resistance after prolonged exposure to high temperatures. Size was also shown to have no effect on resistance and tolerance. A comparison of the tolerance triangle of T. hirsuta with those of the temperate species Mytilus edulis and Mya arenaria revealed that the 24-h LT₅₀ minus 2.2 C° approximates the incipient lethal temperature. The eurythermicities of the temperate species are Mytilus edulis, 363 C°-squared; and Mya arenaria, 415 C°-squared.     

Effect of environmental factors on byssal thread formation

The effects of various factors on byssal thread formation have been examined in the laboratory using Modiolus demissus and Mytilus edulis. With M. demissus thread formation (threads/mussel/h0 and proportion of mussels forming threads decreased with increasing size; prior exposure to air enhanced subsequent thread formation; mechanical agitation reduced thread formation. In addition, low salinity acclimated mussels adapted more rapidly to 32 ppt than high salinity acclimated mussels did to 16 ppt; threads were not formed in the absence of calcium and/or magnesium; and there was no reduction in thread formation at temperatures as high as 27° to 28°C. M. edulis died and thread formation approached zero at temperatures exceeding 26°C.     

Acid and alkaline phosphatase activities in the clam Ruditapes philippinarum

Acid and alkaline phosphatase activities have been partially characterized in Ruditapes philippinarum (Adams and Reeve, 1850). Two activity peaks at pH=4.5 and pH 10.5 were detected in the gill, digestive gland, mantle, siphon and foot. Acid phosphatase activity was higher than that of alkaline phosphatase. The highest activity for both enzymes was observed in the digestive gland and, in decreasing order, the gill, foot, siphon and mantle. Alkaline phosphatase activity was similar in the mantle, siphon and foot. K _m values were determined for both enzymes in the gill and digestive gland. Hill coefficients were near 1, indicating no allosteric behaviour for either enzyme in the two organs. The optimum temperature was the same for acid phosphatase in both gill and digestive gland (50 °C), while for alkaline phosphatase it differed for these two organs (gill, 40 °C; digestive gland, 35 °C). The apparent activation energy was obtained from Arrhenius plots, and ranged from 8.61 kcal/mol for alkaline phosphatase in the gill, to 10.84 kcal/mol for acid phosphatase in the digestive gland. The effects of metals (1 mM conc) on both enzyme activities were assayed in vitro. Hg strongly inhibited the enzyme activities in the gill and digestive gland, probably because of its affinity for the sulphydryl group. Histochemically, acid phosphatase in the gill was located in a granular form throughout the gill cells, but was undetectable in the ciliate epithelium of the gill filaments. Alkaline phosphatase was located in the gill skeleton. Clam size and phosphatase activities were inversely related, probably reflecting a decrease in shell deposition with inereasing size. As a function of season, both enzymes were present in lowest amounts in winter, when undifferentiated sex cells were predominant in the germinative epithelium, and highest in summer, when ripe individuals of both sexes were more frequent.     

Filtration rate,using a new indirect technique,in thirteen species of suspension-feeding bivalves

A method for determining filtration rates in undisturbed suspension-feeding bivalves is described. Concentrations of particulate matter in the water collected in the inhalant (C _i) and exhalant (C _e) currents were estimated with an electronic particle counter. The clearance was calculated as , where Fl=flow rate through the tube collecting exhalant water. Only above critical levels of water flow (Fl) were clearances representative of filtration rates. At 10° to 13°C, the filtration rates (F, 1 h^-1) within one or two orders of magnitude of dry weight (w, g), in Cardium echinatum L., C. edule L., Mytilus edulis L., Modiolus modiolus (L.) and Arctica islandica (L.) followed the allometric equations: 4.22w ^0.62, 11.60w ^0.70, 7.45w ^0.66, 6.00w ^0.75 and 5.55w ^0.62, respectively. Five species of bivalves [Spisula subtruncata (da Costa), Hiatella striata (Fleuriau), Cultellus pellucidus (Pennant), Mya arenaria L. and Venerupis pullastra (Montagu)] filtered with the same rates as individuals of Cardium echinatum and A. islandica of equivalent soft weight. In Pecten furtivus and P. opercularis filtration rates were about twice the rates measured in individuals of Mytilus edulis of comparable body weight. The gill area in M. edulis increases with size at the same rate as the filtration rate.     

Fluid motion and particle retention in the gill of Mytilus edulis: Video recordings and numerical modelling

Particle trajectories of 6.4m Latex spheres were recorded by video, both near an isolated blue mussel, Mytilus edulis, gill filament and, in place of an intact interfilamentary canal, in a model canal of width 200, 100 or 70m, formed by a transparent plate positioned next to a gill filament. Each arrangement was placed in a 2 x 10 x 10 cm test vessel filled with seawater. Serotonin (nerve-transmitter) stimulation was used to activate lateral cilia and to either lock latero-frontal cirri at the end of an active stroke (10^-5 M), or to activate them (10^-6 M), yielding lateral cilia beat frequencies of 19 and 16 Hz, respectively. With latero-frontal cirri locked, image analysis of particle tracks gave maximum velocities of ca. 2.9±0.2mm s^-1 close to the tips of lateral cilia, for both isolated filament and model canal cases. Experimental velocity profiles along the 200-m wide model canal were recorded and used as good approximations to the fluid velocity because of the low Reynolds number. A two-dimensional steady model was proposed for the gill pump, assumed to only comprise lateral cilia. This model was solved numerically for the experimental model, canal in the vessel and the results showed satisfactory agreement with experimental volocity profiles from particle tracks. The numerical approach was also applied to a model of a single interfilamentary canal in the vessel. The resulting mean velocity in the canal was 1.70 mm s^-1, but the resistance to flow in the model was less than that in an intact mussel gill. Video graphs of particle tracks indicated that active latero-frontal cirri play a role in the transfer of particles from through current to frontal current, probably by means of a strong interaction through the motion of intervening fluid rather than through a direct physical contact. M. edulis specimens used in the present study were collected in 1990 at Helsingør and in 1991 at Kerteminde, Denmark.     

Changes in microflora of a puffer fish Fugu niphobles,with different water temperatures

Grass puffer Fugu niphobles, captured in November 1986 at Shimoda, Japan, and then reared at different temperatures ranging from 10 to 29°C, was examined for microflora changes in the skin, gill and intestines. At 10°C, the skin and gill were colonized mainly by Pseudomonas, Flavobacterium and Moraxella, while at 29°C Vibrio and Flavobacterium appeared abundantly. The intestinal microflora, consisting of Vibrio, Pseudomonas and Flavobacterium, revealed little temperature dependence, although the intrageneric composition of Vibrio changed conspicuously depending on the change of water temperature. Vibrio group 1, tentatively identified as V. alginolyticus, was detected in all tissues examined at 20 and 29°C. This, along with the fact that V. alginolyticus produces tetrodotoxin, suggests that the tetrodotoxin contained in puffer is, at least partly, accounted for by this bacterial species inhabiting intestines.     

Diel variation in chlorophyll a,carbohydrate and protein content of the marine diatom Skeletonema costatum

Skeletonema costatum (Greville) Cleve isolated from Narragansett Bay, USA, was incubated at 3 light intensities (ca. 0.008, 0.040 and 0.075 ly min^-1) under a 12 h light: 12 h dark (12L:12D) photoperiod at 2°, 10° and 20°C. Cellular chlorophyll a increased at intensities less than ca. 0.040 ly min^-1; increases occured within one photoperiod at temperatures above 10°C. Cellular carbohydrate increased with light intensity at all temperatures; increases during the photophase were due to net production of the dilute acid-soluble fraction. Cellular protein increased during the photoperiod at 10° and 20°C; there was little difference in cellular protein among all cultures after one photoperiod. The rate at which cellular chlorophyll a increased in response to a decrease in light suggests that diel variation in cellular chlorophyll a is temperature-dependent in S. costatum. Protein: carbohydrate ratios ranged from ca. 0.5 to 2.0 over a diel cycle; ratios increased at lower intensities and higher temperatures. The diel range in protein:carbohydrate ratios equals that in cultures developing nitrogen deficiency; thus, use of this ratio as an index to phytoplankton physiological state must account for diel light effects.     

Isolation of DNA and RNA from ascidians

Combined effects of temperature, salinity and nutrition on larval survival and growth of the European oyster Ostrea edulis L. were studied over a period of seven days in the laboratory. Larvae were obtained in August 1985 from oysters reared under field conditions on the Mediterranean coast. Four temperatures (15°, 20°, 25°, 30°C), four salinities (20, 25, 30, 35 S) and two levels of nutrition (fed or unfed) were used in the experimental design; the fed larvae received a mixed algal diet of Isochrysis galbana and Chaetoceros calcitrans forma pumilum at a concentration of 100 cells per microlitre. Larvae survived over a wide range of temperature and salinity; statistical analysis indicated that nutrition had the greatest effect on the development of O. edulis larvae, explaining 85 to 88% of the variance in growth. Compared with temperature, the effect of salinity was very slight, usually statistically insignificant. The combined effects of temperature and nutrition produced the only significant interaction. Growth of starved larvae seems to be independent of both temperature and salinity within the range of levels tested.     

Distribution of the cladoceran Podon polyphemoides in the Chesapeake Bay

The distribution of the cladoceran Podon polyphemoides (Leuckart) in the Chesapeake Bay (USA) estuarine system was determined by a quantitative pump sampling method, and the patterns of abundance were correlated with temperature and salinity distributions. The species was seasonally recurrent, with distinct population maxima in the central portion of the bay. Population densities in excess of 60,000 podonids/m³ have been recorded. The podonids first appeared in the spring in the shallow tributaries, when water temperatures near the bottom reached 6°C. The vernal populations disappeared when summer temperatures exceeded 27°C, but reappeared in the fall as the water cooled. The species was euryhaline and eurythermal in its distribution, but the greatest concentrations were attained within relatively narrow zones of temperatures between 11^o and 26°C, and salinities between 8 and 18. The production of males, sexual females and sexual eggs occurred both in the spring and the fall between the thermal limits of 11^o and 17°C.     

Clearance rates of food suspension and food passage rates as a function of temperature in two north-sea bryozoans

The filtration rate has been determined for Electra pilosa and Conopeum reticulum, acclimated at 4 constant temperatures (6°, 12°, 18°, 22°C). Cryptomonas sp. was used as the test food-organism. Temperature affects filtration rate in both bryozoans. Clearance rate, measured at hourly intervals during the course of the experiment, was never uniform. Possibly, temporary reductions in ciliary activities were induced by the artificial conditions under which the experiments were carried out. The results obtained compare well with those reported by Bullivant (1968) on Bowerbankia imbricata, a cosmopolitan ctenostome. Rate of food passage through the alimentary canal is affected by temperature. The present study is the first to demonstrate the effect of temperature on the rate of filtration in bryozoans.     

Molecular mechanisms of adaptation to low temperature in marine poikilotherms. Some regulatory properties of dehydrogenases from two arctic species

The properties of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase from gill tissue of the tanner crab Chionocetes bairdi, and lactate dehydrogenase (LDH) and glyceraldehyde dehydrogenase from skeletal muscle of C. bairdi and the yellowfin sole Limanda aspera were examined over the physiological temperature range of the animals. Both animals were obtained in the Bering Sea in winter, and their enzymes appear to be remarkably cold-adapted. Affinity of sole LDH for substrate appears to increase with decreasing temperature, thus keeping reaction rate essentially independent of temperature at physiological concentrations of the substrate. Calculated values of activation energy are low, in keeping with the argument that organisms from cold environments have enzymes with a reduced energy of activation. In addition, Hill plots of the substrate saturation curves for lactate dehydrogenase from muscle of sole indicate that there is a facilitation of allosteric behaviour at low temperatures. Maximum affinity of sole LDH for substrate in the absence of univalent cations occurs at 3°C, while in the presence of 150 mN K⁺, it occurs between 0° to-2°C. The effects of Mg²⁺ on enzyme activity appear to be determined by concentration of substrate and temperature. Thus, glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase are stimulated more effectively by Mg²⁺ at low temperature and at low substrate levels whereas, at high concentrations of substrate, they are relatively independent of the bivalent cation. All four dehydrogenases are affected by the univalent cations Na⁺, K⁺ and NH₄ ⁺ in a manner which appears to be determined, in part at least, by concentration of substrate and by temperature. These findings suggest mechanisms for the maintenance and regulation of enzyme activity in poikilothermic tissues at low and changing temperatures.     

A functional interpretation of cilia and mucocyte distributions on the abfrontal surface of bivalve gills

The lack of fundamental data on the abfrontal surface of bivalve gills has prompted a comparative study of cilia and mucocytes on this surface, using scanning electron microscopy and histology on eight species of bivalves, representing seven families and the four major gill types: Mytilus edulis, Modiolus modiolus, Arca zebra, Placopecten magellanicus, Crassostrea virginica, Spisula solidissima, Mercenaria mercenaria, and Mya arenaria. Abfrontal cilia and mucocytes were found in all species studied, with types and densities differing within and between gill types. The three species of homorhabdic filibranchs presented different densities of abfrontal cilia and mucocytes, from very dense in M. edulis to sparse in A. zebra. The heterorhabdic gills had intermediate cilia and mucocyte densities, with highest concentrations of both abfrontal cilia and mucocytes on the principal filaments. The eulamellibranchs showed low ciliary densities together with high mucocyte densities, especially in S. solidissima, where the abfrontal mucocytes were glandular. These results indicate that: (1) the abfrontal surface is a vestigial mucociliary surface; (2) the abfrontal surface cannot participate in water pumping in most species, due to low ciliary densities; and (3) species with high densities of abfrontal mucocytes could utilize abfrontal mucus to reduce drag, especially in the highly fused gills, such as those of the eulamellibranchs. The differing distributions of abfrontal cilia and mucocytes may reflect different selective pressures acting on the gills within the various taxa. Received: 12 February 2000 / Accepted: 10 September 2000     

Ontogenic change of gill chloride cells in leptocephalus and glass eel stages of the Japanese eel, <Emphasis Type="Italic">Anguilla japonica</Emphasis>

The development of gill chloride cells was examined in premetamorphic larvae (leptocephali) and juveniles (glass eels) of the Japanese eel, Anguilla japonica. Branchial chloride cells were detected by immunocytochemistry using an antiserum specific for Na⁺,K⁺-ATPase. The specificity and availability of the antiserum for the detection of Japanese eel chloride cells were confirmed by Western blot analysis. The chloride cells first appeared on the developing gill filaments in a mid larval stage of leptocephalus (32.2 mm). Both immunoreactivity and the number of chloride cells gradually increased as the fish grew to a late stage of leptocephalus over 54 mm. In glass eels just after metamorphosis, gill lamellae developed from the gill filaments, and a rich population of chloride cells was observed in the gill filaments. In glass eels collected at a coastal area, chloride cells were extensively distributed in the gill filaments. The chloride cell size decreased progressively in glass eels transferred from seawater (SW) to freshwater (FW), whereas there was no difference in cell number. In contrast, some Na⁺,K⁺-ATPase immunoreaction distinct from typical chloride cells was observed in the gill lamellae throughout FW-transferred fish, but disappeared in control fish maintained in SW for 14 days. These findings indicate that the gill and gill chloride cells developed slowly during the extremely long larval stage, followed by rapid differentiation during a short period of metamorphosis. The excellent euryhalinity of glass eels may be due to the presence of the filament chloride cells and lamellar Na⁺,K⁺-ATPase-immunoreaction, presumably being responsible for SW and FW adaptation, respectively.     

Some effects of temperature on the growth and metabolic rate of juvenile blue crabs,Callinectes sapidus,in the laboratory

Juvenile blue crabs, Callinectes sapidus Rathbun, were grown in the laboratory at different temperatures, and metabolic-rate determinations were made. Growth is shown to be dependent upon temperature. Crabs kept at high temperatures (34° and 27°C) grow faster than those kept at lower temperatures (13°, 15°, and 20°C). Increase in size per molt is less at higher temperatures than at lower ones. Mortality is directly proportional to temperature between 13° and 34°C and is very high during ecdysis at elevated temperatures. Metabolic rate increases with temperature, but various degrees of acclimation are seen after 4 weeks exposure. No acclimation of general activity to temperature was found. The findings are applied theoretically to crabs living in the region of heated discharge canals of electrical generators: the motile blue crab could extend its growing season without decreasing size at maturity by active selection of thermal surroundings.In part based on a thesis submitted in partial fulfillment of the requirements for the Degree of Master of Science at the University of Florida, USA.     

Effect of temperature on the proliferation of Gymnodinium breve and Gomphosphaeria aponina

The effect of temperature on the growth and proliferation of two marine microorganisms, the toxigenic dinoflagellate Gymnodinium breve, and a potential bio-control organism, the blue-green alga Gomphosphaeria aponina, was determined by culturing the organisms in thermal gradients established by heating and cooling the opposite ends of an aluminum bar that had been adapted to hold culture tubes. Gradients were linear and stable for the duration of each trial. There was no relationship between variations in light and growth of the organisms. Gymnodinium breve showed optimum growth at 22°C, and proliferated over a range of temperatures (17° to 30°C). Below 17°C cultures of G. breve declined in growth, and at 4°C the organisms died within 5 h. Above 31°C there was rapid decline in viability of cells, and at 33.5°C the organism died within 24 h. Gomphosphaeria aponina showed optimum growth between 24° and 29°C, with a maximum at 27°C. Growth at temperatures greater than 31°C was minimal, but the organism survived. Limitation may be due to repression of the bio-synthesis of an iron-transport compound.     

Delayed growth of mussel (Mytilus edulis) and scallop (Pecten maximus) veligers at low temperatures

Veliger larvae of Mytilus edulis (L.) from Menai Straits, North Wales, were maintained for up to 2 mo during 1981 at 5°C and then grown on to metamorphosis at 17°C. Larvae so treated showed similar low mortality and equivalent spat production to control larvae. Growth rate at 17°C was less in treated larvae than in controls, but treated larvae grew a little during the period at low temperature. Larvae of Pecten maximus (L.) from the Irish Sea suffered high mortality at low temperature but larvae surviving 2 wk at 8°C could be grown on to spat at 17°C. The longevity of M. edulis larvae is discussed in relation to the genetic homogeneity of adult populations around the UK.     

Grazing of Acartia hudsonica (A. clausi) on Skeletonema costatum in Narragansett Bay (USA): Influence of food concentration and temperature

Grazing experiments were performed with temperatureacclimated Acartia hudsonica fed the diatom Skeletonema costatum in concentrations ranging from 50 to 3×10⁴ cell ml^-1 at 5°, 10° and 15°C. The ingestion data were best fit by an Ivlev equation. Feeding threshold values of 39 and 59 cells ml^-1 were not significantly different from zero; however, filtration rates were depressed at low food concentrations. Maximum filtration rates increased exponentially with temperature, reaching a maximum with copepods collected at 14°–15°C, and then declining. Both the increase in ingestion rate with increasing food concentration and the maximum ingestion rate were significantly greater as experimental temperature was increased. Maximum ingestion rates were reached at concentrations greater than 6×10³ cells ml^-1. Percent of body carbon ingested per day at 5 g C L^-1 increased from 1.5% at 5°C to 6.7% at 15°C. At 500 g C L^-1, the ingestion increased from 84% (5°C) to 660% (15°C). Percent of body nitrogen at 0.5 g N L^-1 increased from 0.6% per day at 5°C to 2.5% per day at 15°C. At 50 g N L^-1, the ingestion was 42% body nitrogen at 5°C and 250% at 15°C. The influence of grazing by A. hudsonica on phytoplankton in Narragansett Bay, USA was estimated for 1972–1977. The percent of standing stock removed by grazing rarely exceeded 5% per day except during the late spring when S. costatum growth becomes nutrient limited and higher temperatures favor the rapid population growth of A. hudsonica.     

Blooms of the dinoflagellate Gyrodinium aureolum in a Long Island estuary:

A dense dinoflagellate bloom of Gyrodinium aureolum Hulburt in a shallow temperate zone estuary was monitored during the summers of 1982 and 1983. The bloom was typically extremely localized, its densest part exceeding 1000g chlorophyll a liter^-1 (2x10⁴ cells ml^-1). The bloom began at temperatures between 24.5° and 27°C, existed at as high as 30°C and terminated when water temperature dropped to between 19° and 22°C. The highest specific growth rate measured was 0.90d^-1 (1.3 divisions d^-1) and near the termination of the bloom decreased to 0.28d^-1 (0.4 divisions d^-1). A diel vertical migration of the bloom was observed. A box model analysis, based on division rates, vertical migration and water circulation patterns, indicated that the bloom must move downward at the estuary mouth to maintain itself in the estuary, either by means of a convergence system or by downward swimming. High growth rate, low grazing pressure, and a stratified water column are proposed to stimulate bloom formation. Decreasing growth rate appeared to reduce the intensity of the bloom and finally allowed its disappearance by estuarine flushing and mixing.Communicated by J. M. Shick, Orono