Bioluminescence in oceanology

For analytical purposes bioluminescence can be used in three main ways: 1. luminescence measurement of bioluminescent system components isolated in vitro; 2. determination of luminous organisms' reaction to the in vivo test-action; 3. measurement of bioluminescence in marine ecological systems. The majority of the reports of this Symposium are dealing with the first two topics. The aim of our presentation is to draw attention to the third one. The possibilities of bioluminescent analysis are wider than its traditional scheme of applications in the laboratory, when the emitting system is withdrawn from a native source and is placed in a cuvette of the light measuring device. The reverse scheme is also possible, i.e. the device can be introduced into light emitting system such as a marine biocenosis–the community of the sea inhabitants–where we obtain a highly sensitive and rapid means of gaining the information on the vital activity of marine ecosystems, i.e. their spatial structure, rhythms, man's influence upon them, etc. The present communication will consider the possibilities of this form of bioluminescent analysis.     

Sodiumd-glucose cotransport in the gill of marine mussels: Studies with intact tissue and brush-border membrane vesicles

Summary Glucose transport was studied in marine mussels of the genusMytilus. Initial observations, with intact animals and isolated gills, indicated that net uptake of glucose occurred in mussels by a carrier-mediated, Na⁺-sensitive process. Subsequent studies included use of brush-border membrane vesicles (BBMV) in order to characterize this transport in greater detail. The highest activity of Na⁺-dependent glucose transport was found in the brush-border membrane fractions used in this study, while basal-lateral membrane fractions contained the highest specific binding of ouabain. Glucose uptake into BBMV showed specificity for Na⁺, and concentrative glucose transport was observed in the presence of an inwardly directed Na⁺ gradient. There was a single saturable pathway for glucose uptake, with an apparentK _t of 3 m in BBMV and 9 m in intact gills. The kinetics of Na⁺ activation of glucose uptake were sigmoidal, with apparent Hill coefficients of 1.5 in BBMV and 1.2 in isolated gills, indicating that more than one Na⁺ may be involved in the transport of each glucose. Harmaline inhibited glucose transport in mussel BBMV with aK _i of 44 m. The uptake of glucose was electrogenic and stimulated by an inside-negative membrane potential. The substrate specificity in intact gills and BBMV resembled that of Na⁺-glucose cotransporters in other systems;d-glucose and -methyl glucopyranoside were the most effective inhibitors of Na⁺-glucose transport,d-galactose was intermediate in its inhibition, and there was little or no effect ofl-glucose,d-fructose, 2-deoxy-glucose, or 3-O-methyl glucose. Phlorizin was an effective inhibitor of Na⁺-glucose uptake, with an apparentK _i of 154nm in BBMV and 21nm in intact gills. While the qualitative characteristics of glucose transport in the mussel gill were similar to those in other epithelia, the quantitative characteristics of this process reflect adaptation to the seawater environment of this animal.     

Atmospheric input of inorganic nitrogen to the Western Mediterranean

Bulk inorganic nitrogen deposition was monitored over a period of 3 years at the Bavella Pass (Corsica, France). Annual fluxes range between 126 and 150mol.m^–2.d-^–1, increasing slightly with annual rainfall. Natural background average concentrations of rain water and associated fluxes were estimated from a classification of rain events into natural (Oceanic and Saharan), polluted and composite. Long range transport of incoming polluted air masses increases the atmospheric wet nitrogen input by at least a factor of 1.6 in this Mediterranean area. Extrapolation of atmospheric dissolved inorganic nitrogen input to the Western Mediterranean leads to fluxes of 80 to l00mol.m^–2.d-^–1. This atmospheric input is in the same order of magnitude as the inorganic nitrogen riverine input. As a consequence, the nitrogen budget for the Mediterranean has had to be reassessed. Atmospheric wet inorganic nitrogen input is of noticeable importance to marine Mediterranean ecosystems, representing on average 10 to 25% of new production in the Western Basin, with values of up to 60% in oligotrophic zones.     

Refolding of trypsin-subtilisin inhibitor from marine turtle eggwhite

Trypsin-subtilisin inhibitor from marine turtle eggwhite refolded quantitatively from its fully reduced state atpH 8.5 in the presence of reduced and oxidized glutathione. The refolding process was studied by following the accompanying changes in inhibitory activity, fluorescence, sulfhydryl group titer, and hydrodynamic volume. The refolding process followed second-order kinetics with rate constants of 4.80×10² M^–1 sec^–1 for trypsin-inhibiting domain and 0.77× 10² M^–1 sec^–1 for subtilisin-inhibiting domain of the inhibitor at 30°C and their respective activation energies of the refolding process were 15.9 and 21.6 kcal/mol. Fluorescence intensity of the reduced inhibitor decreased with time of refolding until it corresponded to the intensity of the native inhibitor. The inhibitor contained 1–2%-helix, 40–42%-sheet, and 57–58% random coil structure. Refolded inhibitor gave a circular dichroic spectrum identical to that of the native inhibitor. A number of principal intermediates were detected as a function of the refolding time. Size-exclusion chromatography separated the intermediates differing in hydrodynamic volume (Stokes radius). The Stokes radius ranged from 23 Å (fully reduced inhibitor) to 18.8 Å (native inhibitor). Results indicated the independent refolding of two domains of the inhibitor and multiple pathways of folding were followed rather than an ordered sequential pathway.     

The Effect of Photoinhibition on Photosynthetic Oxygen Production in the Brown Alga Dictyota dichotoma

Photoinhibition of photosynthesis in the brown alga, Dictyota dichotoma, was studied with a PAM fluorometer (Walz, Effeltrich, Germany) and a homemade oxygen measuring device. As a measure of fluorescence, Fv/Fm, and for the photosynthetic yield, ΔF/Fm', were used. Oxygen measurements show clearly that the observed degree, as well as the time course, of photoinhibition depends on the fluence rate of the light used to measure changes of the production rate. After photoinhibition of photosynthesis the depression of oxygen production caused by non-saturating fluence rates was generally much more pronounced than that caused by saturating or nearly saturating fluence rates. At minimal photoinhibition the initial slope and the convexity of the fluence rate-response curve of oxygen evolution decrease, whereas the level of light saturation decreases only after strong photoinhibition. Nevertheless, at different degrees of photoinhibition, changes in the degree of the upper bending of the fluence rate-response curve of oxygen production are also linearly correlated to changes in the fluorescence ratios (Fv/Fm and ΔF/Fm'). The action spectrum of photoinhibition, calculated on the basis of changes of Fv/Fm, indicates that the reaction center of PS I is not involved in photoinhibition. The lower effectiveness of blue light in comparison to effects of green and red light may be due to chloroplast displacement, as in the so-called strong light position, the light absorbed by the thalli in vivo is decreased.     

Materials for the fungus flora of Japan (49)

Two interesting microfungi are described as new to Japan:Talaromyces galapagensis (anam.Penicillium galapagense), isolated from soil in Shizuoka; andPenicillium megasporum, isolated from marine sludge in Nagasaki. Some observations are recorded, particularly on ascomatal initials ofT. galapagensis, which are similar to those described forTalaromyces flavus.(48): Kaneko, S., Mycoscience 36: 359–360, 1995.     

Effect of vertebrate predation on the spatio-temporal distribution of cladocerans in a temperature eutrophic lake

We analysed the spatio-temporal distribution of zooplankton along a profile of 10 stations from the shore to the pelagic zone from April to September 1988, the period when the larvae and juveniles Rutilus rutilus, the most abundant species in the Lake, are in the littoral zone. The digestive tracts of the young roach were analysed. They fed essentially on rotifers and on cladocerans. For comparison, zooplankton was also analysed at one littoral area without fish fry. There was an increase of cladoceran density from the vegetated nearshore zone to the offshore zone. Considering the density of Bosmina longirostris, Daphnia longispina, Chydorus sphaericus and Ceriodaphnia quadrangula, we observed a different distribution pattern in the course of the year. In the nearshore zone, the relative abundance of small species, Bosmina and Chydorus, was much higher than that of the larger Daphnia. From April to September, predation pressure mainly affected the smallest species: in contrast to the inshore station without fish fry, the density of Bosmina decreased in May in the littoral with fish. Chydorus was concentrated in the littoral between February and April, then grew into the pelagic zone, where predation pressure obviously was low during the warm season. The number of Daphnia, which was eaten by the fish fry at any time, remained low in the nearshore zone, which suggests that the presence of fish may cause Daphnia to avoid this zone. Ceriodaphnia which was not affected by this predation, was scarce in the nearshore zone during mid-summer. The low density of the cladocerans in the nearshore zone is likely associated with vertebrate predation by roach fry and juveniles, the result of such a process being either a depletion in density of the prey, or an avoidance behaviour.     

Response by macrozoobenthos biomass to water level regulation in some Finnish lake littoral zones

The relationship of the macrozoobenthos biomass in the littoral area to the yearly fluctuation in water level and the characteristics of the area or lake are studied using data collected from sheltered bays in regulated and natural waters. Most of the lakes were clear and oligotrophic. The benthos biomass at all depths in the littoral decreased with increased water level fluctuation, provided that the transparency of the water was uniform.The macrozoobenthos biomass in the 0–3 m depth zone could be predicted fromlog macrozoobenthos biomass (mg ODW) m^-2=4.25-1.33 (log Biomass Index) in which the Biomass Index is calculated as% MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGqiVu0Je9sqqrpepC0xbbL8F4rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGcbaGaaeOqaiaabM% gacaqGVbGaaeyBaiaabggacaqGZbGaae4CaiaabccacaqGjbGaaeOB% aiaabsgacaqGLbGaaeiEaiaab2dacaqGGaGaaeiiaiaabccacaqGGa% GaaeiiaiaabccacaqGGaGaaeiiaiaabccacaqGGaGaaeiiaiaabcca% caqGGaGaaeiiaiaabccacaqGGaGaaeiiaiaabccacaqGGaGaaeiiai% aabccadaWcaaabaeqabaGaae4DaiaabggacaqG0bGaaeyzaiaabkha% caqGGaGaaeiBaiaabwgacaqG2bGaaeyzaiaabYgacaqGGaGaaeOzai% aabYgacaqG1bGaae4yaiaabshacaqG1bGaaeyyaiaabshacaqGPbGa% ae4Baiaab6gacaqGGaGaaeyAaiaab6gacaqGGaGaaeiDaiaabIgaca% qGLbGaaeiiaiaabchacaqGYbGaaeyzaiaabAhacaqGPbGaae4Baiaa% bwhacaqGZbGaaeiiaiaabMhacaqGLbGaaeyyaiaabkhaaeaacaqGOa% GaaeyBaiaabUdacaqGGaGaae4yaiaabggacaqGSbGaae4yaiaabwha% caqGSbGaaeyyaiaabshacaqGLbGaaeizaiaabccacaqGMbGaaeOCai% aab+gacaqGTbGaaeiiaiaab2gacaqGVbGaaeOBaiaabshacaqGObGa% aeiBaiaabMhacaqGGaGaaeyBaiaabwgacaqGHbGaaeOBaiaabccaca% qG2bGaaeyyaiaabYgacaqG1bGaaeyzaiaabohacaqGPaaaaeaacaqG% tbGaaeyzaiaabogacaqGJbGaaeiAaiaabMgacaqGGaGaaeizaiaabM% gacaqGZbGaae4AaiaabccacaqG2bGaaeyyaiaabYgacaqG1bGaaeyz% aiaabccacaqGPbGaaeOBaiaabccacaqG0bGaaeiAaiaabwgacaqGGa% Gaae4CaiaabggacaqGTbGaaeyzaiaabccacaqGVbGaaeiCaiaabwga% caqGUbGaaeiiaiaabEhacaqGHbGaaeiDaiaabwgacaqGYbGaaeiiai% aabohacaqGLbGaaeyyaiaabohacaqGVbGaaeOBaiaabccacaqGOaGa% aeyBaiaabMcaaaaccaGae8hiaaIaaKiEaiab-bcaGiaaigdacaaIWa% GaaGimaiaac6caaaa!CBD8!\[{\text{Biomass Index = }}\frac{\begin{gathered} {\text{water level fluctuation in the previous year}} \hfill \\ {\text{(m; calculated from monthly mean values)}} \hfill \\ \end{gathered} }{{{\text{Secchi disk value in the same open water season (m)}}}} \user1{x} 100.\]The whole illuminated littoral shifts due to water level fluctuation, which disturbs the zonation of the benthos. Such an increase or decrease in benthic biomass has been observed after one year of disturbance due to water level fluctuation. It need, however, a study based on the carefully planned and collected data, in which it can be taken account by a multivariate statistical analysis also the interactions between the important factors affected the littoral benthos.