The influence of storage on antibacterial activity of sea water. I. Experiments with sea water stored at 18°C

Three sea-water samples of about 25 l each, stored at 18°C in the dark, were examined for antibacterial activity and bacterial content. Serratia marinorubra mainly was used as test bacterium. During storage for 197 to 454 days, bactericidal capacity varied considerably. In raw sea water, extreme log (N ₃–N ₀) values (N: number of colony-forming test bacteria/ml after 0 and 3 days of any test) were higher than-1 and lower than-4. Antibacterial activity of filtersterilized and autoclaved, stored, sea water likewise varied, but to a lesser degree. The number of colony-forming marine bacteria fluctuated about 3 orders of magnitude. Variations in bacterial content and in bactericidal action of stored sea water were positively correlated during the first months of storage; later, this correlation became negative. From these results it is concluded that variations in nutrient concentration — due to successive periods of growth and autolysis of marine bacteria — were the primary reason for the changes observed in antibacterial activity of stored sea-water samples.     

Bactericidal properties of natural and synthetic sea water as influenced by addition of low amounts of organic matter

The influence of added organic nutrients on the bactericidal capacity of natural and synthetic sea water was investigated, with special reference to levels below the normal concentration of orgnnic matter in natural sea water. Resting cells of Escherichia coli, Staphylococcus aureus and Serratia marinorubra were used as test organisms. The inactivation of test bacteria in synthetic and filter-sterilized natural sea water could be increased by addition of ZoBell's 2216E-broth (containing peptone and yeast extract), peptone or glucose. This effect of the nutrients depended on chemical nature and concentration as well as on specific properties of the test strains. In regard to peptone, less than 10^-4 mg/ml must be added to induce bacterial kill exceeding that observed with unsupplemented controls. With raw sea water, no fundamentally different results were obtained; however, the influence of marine bacteria was clearly discernible. All findings support a previously offered hypothesis of nutrient-dependent antibacterial activity of sea water which is discussed in detail.     

Killing of marine phytoplankton by a gliding bacterium Cytophaga sp., isolated from the coastal sea of Japan

A marine gliding bacterium Cytophaga sp. (strain J18/M01) was isolated from Harima-Nada, eastern Seto Inland Sea, Japan in 1990. This bacterium preys upon various species of marine phytoplankton. All of the five raphidophycean flagellates, all of the four diatoms, and one of the two dinoflagellates examined were killed within a few days when cultured with the bacterium. The bacterium presumably achieves this by direct attack, because the culture filtrate in which host organisms were totally destroyed had no significant effects on the growth of the same host organism (Chattonella antiqua). If one or a few bacterial cells were inoculated into C. antiqua culture, all of the host organisms were killed. The bacterium proliferated in filter-sterilized seawater, suggesting its ubiquitous existence in the coastal sea. The killing of phytoplankton by bacteria such as Cytophaga sp. J18/M01 may be a significant factor influencing the population dynamics of phytoplankton in nature and may contribute to the sudden disappearance of red tides in the coastal sea. Bacterial destruction of phytoplankton may also be a factor that regulates primary productivity in marine ecosystems.     

Antibacterial activity of the marine blue-green alga Trichodesmium erythraeum in the gastro-intestinal contents of the sea gull Laurus brunicephalus

The gastro-intestinal contents of the sea gull Laurus brunicephalus Jerdon were found to possess antibacterial activity. This activity could be traced to heavy accumulations of the marine blue-green alga Trichodesmium erythraeum in the gut. During 1969, a bloom of T. erythraeum began in Porto Novo waters about the middle of February, and attained a peak during the second week of March. It has been reported previously by the author that antibacterial properties are exhibited by T. erythraeum maintained in laboratory cultures, as well as in water samples collected from a red tide area. It was also reported (Ramamurthy, 1970) that the gut contents in 2 pelagic fishes, Hilsa kanagurta and Rastrelliger kanagurta, collected during the same red tide bloom period, possessed antibacterial properties. During this period large numbers of these fishes were consumed by sea gulls L. brunicephalus. In view of this finding, experimental procedures were adopted to determine whether extracts of T. erythraeum occurring in the gut of the sea gulls might exhibit antibacterial activity. It was found that T. erythraeum collected from the gut of L. brunicephalus could inhibit both gram positive and gram negative bacteria. Gastro-intestinal extracts from L. brunicephalus collected during the non-bloom period of T. erythraeum showed heavy microbial growth of bacteria and fungi. Evidently, antibacterial or sterile conditions prevail in the gut of these tropical sea gulls in a manner similar to that observed in Polar penguins by Sieburth (1959, 1961).     

Evidence for a symbiosis between bacteria of the genus Rhodobacter and the marine sponge Halichondria panicea : harbor also for putatively toxic bacteria?

Halichondria panicea (Pallas) is a marine sponge, abundantly occurring in the Adriatic Sea, North Sea, and Baltic Sea. It was the aim of the present study to investigate if this sponge species harbors bacteria. Cross sections through H. panicea were taken and inspected by electron microscopy. The micrographs showed that this sponge species is colonized by bacteria in its mesohyl compartment. To identify the bacteria, polymerase chain reaction (PCR) analysis of the 16S rRNA gene segment, typical for bacteria, was performed. DNA was isolated from sponge material that had been collected near Rovinj (Adriatic Sea), Helgoland (North Sea), and Kiel (Baltic Sea) and was amplified with bacterial primers by PCR. The data gathered indicate that in all samples bacteria belonging to the genus Rhodobacter (Proteobacteria, subdivision α) are dominant, suggesting that these bacteria live in symbiotic relationship with the sponge. In addition, the results show that the different samples taken contain further bacterial species, some of them belonging to the same genus even though found in sponges from different locations. The possibility of the presence of toxic bacteria was supported by the finding that organic extracts prepared from sponge samples displayed toxicity, when analyzed in vitro using leukemia cells. Received: 7 March 1997 / Accepted: 2 October 1997     

Antimicrobial activity of extracts of Caribbean gorgonian corals

Extracts of 39 species of Caribbean gorgonians were tested for antimicrobial activity against 15 strains of marine bacteria. The bacteria consisted of three opportunistic pathogens, Vibrio parahaemolyticus, Leucothrix mucor, and Aerococcus viridans, and 12 strains isolated from either healthy or decayed gorgonians. Overall, only 15% (79 out of 544) of the tests resulted in antibacterial activity with 33% (13 out of 39) of the gorgonians inhibiting only one bacterial strain and 23% (9 out of 39) showing no activity. The extracts of four Pseudopterogorgia species showed relatively high levels of activity, inhibiting 43 to 86% of the bacterial strains. The potency of the active Pseudopterogorgia species was variable, however, and three additional Pseudopterogorgia species were inactive against all bacterial strains. With the exception of one sensitive strain, Vibrio species were resistant to gorgonian metabolites. Our results indicate that organic extracts of most Caribbean gorgonians do not possess potent, broad-spectrum antibacterial activity inhibitory to the growth of opportunistic marine pathogens and bacteria associated with healthy and decayed gorgonian surfaces. These findings suggest that the inhibition of bacterial growth is not the primary ecological function of gorgonian secondary metabolites and that bacteria may not be important selective agents in the evolution of gorgonian secondary chemistry.     

Some observations on the influence of salinity on growth and photosynthesis inPorphyra umbilicalis

Notwithstanding the great importance of the salinity factor in the marine environment, the knowledge of influence of salinity on growth of marine benthic algae is very limited. Rate of growth (mg, cm²) and O₂ output of the intertidal red algaPorphyra umbilicalis from Helgoland, North Sea, were measured during a 3 week culture in 3 different salinities (1/2-, 1- and 2-concentrated artificial sea water; Table 1). Under hypertonic conditions (2-concentrated sea water) growth rate and photosynthesis rate were depressed, compared to values obtained in normal concentrated sea water. Under hypotonic conditions (1/2-concentrated sea water), growth expressed in mg was the same as in normal concentrated sea water, or higher when expressed in cm³. Rate of O₂ output was almost unaltered in one of the two experiments, lowered in the other. Cell size increased at higher salinity, while swelling of cell walls and intercellular substances as well as the intensity of colouring decreased with salinity. The discrepancies between growth and photosynthesis under hypotonic conditions cannot be completely explained by the observed influences of salinity on morphological structures (cell size, swelling of cell substances). Detailed studies on the time course of photosynthesis and respiration rates, and preparation of a metabolic balance for the algae are necessary.     

Growth and development of Calanus spp. (Copepoda) during spring phytoplankton succession in the North Sea

Experiments were carried out to determine growth and development rates of the herbivorous copepod Calanus finmarchicus (Gunnerus) under natural conditions during the phytoplankton spring bloom in the northern North Sea. From 28 April to 25 May 1983 copepodite stages I, IV and V were incubated for a 3-d period on board a ship in vessels with naturally occurring phytoplankton or cultured algae as food. Highest rates of growth and development were achieved while the diatom Chaetoceros sp. was the dominant phytoplankton organism. These rates decreased considerably when this chain-forming diatom was succeeded after one week by the small-celled diatom Thalassiosira conferta. Again one week later, during the bloom of the succeeding colonial microflagellate Corymbellus aureus, copepodite stage IV still managed to maintain moderate rates of growth and development, but these rates dropped to almost zero in CV, suggesting the start of a resting stage. Nevertheless, brood collected from this generation and from Calanus helgolandicus (Claus) was raised in the laboratory to the adult stage at high speed. Since temperature and the total phytoplankton concentration in the sea remained almost constant it seems that the retardation and arrestment of growth and development were an immediate response to a qualitative change of the food composition related to the successive blooms of different algal species.     

Succession and growth limitation of phytoplankton in the Gulf of Bothnia (Baltic Sea)

A one year field study of four stations in the Gulf of Bothnia during 1991 showed that the biomass was ca. two times, and primary productivity ca. four times, lower in the north (Bothnian Bay) than in the south (Bothnian Sea) during the summer. Nutrient addition experiments indicated phosphorus limitation of phytoplankton in the Bothanian Bay and the coastal areas in the northern Bothnian Sea, but nitrogen limitation in the open Bothanian Sea. A positive correlation between the phosphate concentration and the production/biomass ratio of phytoplankton was demonstrated, which partly explained the differences in the specific growth rate of the phytoplankton during the summer. Differences in photosynthetic active radiation between the stations also showed a covariation with the primary productivity. The relative importance of nutrient or light limitation for photosynthetic carbon fixation could not, however, the conclusively determined from this study. Marked differences in phytoplankton species composition from north to south were also observed. The number of dominating species was higher in the Bothnian Sea than in the Bothnian Bay. The distribution of some species could be explained as due to nutrient availability (e.g. Nodularia spumigena, Aphanizomenon sp.), while salinity probably limits the distribution of some limnic as well as marine species. The potentially toxic phytoplankton N. spumigena, Dinophysis acuminata and Chrysochromulina spp. were common in the Bothnian Sea but not in the Bothnian Bay. The pico- and nanoplankton biomass during late summer was higher than previously reported due to a revised carbon/volume ratio.     

Yeasts from the North Sea

Yeasts were isolated from twelve established sites in the North Sea from 1964 to 1966. A percentage frequency of 99% with populations varying from <10 to >3000 viable cells/L was observed. This mycota was characterized by considerable spatial and temporal fluctuation, with the dominant yeast present being the ascosporogenous species, Debaryomyces hansenii. This taxon, as well as other common North Sea yeasts, e.g., Rhodotorula rubra and Candida diddensii, have been reported frequently from other marine locales. Noteworthy concentrations of yeasts, especially D. hansenii, were observed during summer months, often in association with various stages of development of the dinoflagellate, Noctiluca miliaris. The population dynamics of the North Sea yeasts are discussed in relation to similar studies of other marine environments.     

Long-term change of the marine environment and plankton in the Xiamen Sea under the influence of climate change and human sewage

ABSTRACT

Long-term observations of the Xiamen Sea in Fujian Province were used to analyze variations in sea temperature, salinity, inorganic nitrogen (N), activated phosphate (P), and phytoplankton, as well as the features of red tides. Results showed that in recent decades, sea temperature and concentrations of N and P nutrients increased while salinity decreased attributed to climate warming, rainfall, and human sewage. In addition, reduction in the number of phytoplankton species and rising abundance of phytoplankton indicated that the structure of phytoplankton community presented a simplified and minimized trend which magnified the dominance of dominant phytoplankton species. Since 2000, red tides have occurred more frequently, and the eutrophic-type diatom species, Skeletonema costatum appears to be the predominant species. Data suggested that variations of N and P nutrients exert a potent and rapid influence on phytoplankton than sea temperature and ocean salinity.     

The incidence of antibacterial activity in marine demospongiae; systematic and geographic considerations

Thirty species of temperate-zone sponges representative of all orders of the Demospongiae have been tested for antibacterial activity. The results indicate the following: that a higher percentage of temperate than tropical species produced active extracts (87% as opposed to 58%); that sponge extracts more frequently inhibited the growth of marine bacteria than they did non-marine bacteria (76.5% compared to 56%); that 46.5% of the extracts tested inhibited growth of Gram-negative bacteria while only 6.5% inhibited growth of Gram-positive bacteria. There is no obvious correlation between incidence of antibacterial activity and latitude of occurrence, systematic group or growth form of the sponge. It is suggested that antibacterial agents produced by sponges may have a role in enhancing the efficiency with which sponges retain bacterial food.     

Breeding and larval distribution of the pteropod Clione limacina in the North Atlantic,Subarctic and North Pacific Oceans

The pteropod Clione limacina (Phipps, 1774) is an arcticboreal, circumpolar species, which is widely distributed in the North Atlantic and Subarctic Oceans; it also occurs in the North Pacific Ocean (in the Oyashio and neighbouring waters) and along the Atlantic coast of North America in the waters of the cold Labrador current to the Cape Hatteras region (35° N). The distribution of C. limacina larvae in the plankton of the Norwegian, Barents and White Seas, the Bear Island-Spitsbergen region of the Greenland Sea, the Newfoundland Grand Bank and the Flemish-Cap Bank region of the North-western Atlantic Ocean, and the Kurile-Kamchatka region of the North-western Pacific Ocean has been studied, and information from literature concerning the reproduction and larval occurrence of the species is summarized. Throughout its distributional are, spawning of C. limacina is characterized by the same general ecological pattern. This species breeds and spawns in all types of water masses occurring within the vertical range which it commonly inhabits — from surface layers to 500 m water depth. In all local populations of the species, the most intensive spawning is correlated with the spring/summer period of annual heating of the local waters, and the highest abundance parallels maximum growth of phytoplankton which serves as food for veligers and early polytrochous larvae. After the end of this period, spawning intensity in all local C. limacina populations declines sharply, but spawning continues at low intensity during the autumn/winter season, being practically continuous throughout the year. Distribution patterns of C. limacina larvae are determined by those of their parental forms (the parental forms spawn in the zones permanently inhabited). The earliest larval stages of C. limacina (veligers) are present predominantly in the upper 100 or 200 m water layer, i.e. in the zone of high phytoplankton abundance. Polytrochous larvae, after becoming predaceous feeders, are distributed throughout the whole water column from the surface to 500 m depth, similar to adult C. limacina. As with the adults, larvae are present (within the species' distribution area) in all types of water masses. Since the beginning of the twentieth century, in the course of the warming of the Arctic Ocean, the southern race of C. limacina (formerly a summer/autumn seasonal invader in the Norwegian Sea) has become a permanent component of the plankton fauna of the Norwegian and Barents Seas in regions influenced by the Norwegian-Northcape Current System.     

Iron-mediated effects on nitrate reductase in marine phytoplankton

The potential activity of nitrate reductase was determined in uni-algal cultures in the laboratory and in natural marine phytoplankton assemblages. In the laboratory bioassays, distinct differences in nitrate reductase activity were observed in iron replete versus depleted cultures for Emiliania huxleyi, Isochrysis galbana and Tetraselmis sp. Cells from iron-depleted cultures had 15 to 50 percent lower enzyme activity than those from iron-replete cultures. Upon addition of iron, nitrate reductase activity was enhanced in depleted cells up to levels comparable to those of the replete cells. Bioassays in the northern North Sea conducted in 1993, under low iron conditions, demonstrated similar results. Upon addition of 2.5 nM iron, a distinct enhancement, to a maximum of three times, of nitrate reductase activity was observed within 32 h after addition. Therefore, iron can stimulate nitrate reductase activity. In spite of the clean techniques used, some nitrate reductase activity was always observed. Iron deficiency was shown to impair nitrate reductase activity, but it is unlikely that nitrate reduction would cease completely.     

Seasonal and long-term variations in the body size of planktonic copepods in the northern Baltic Sea

Seasonal and long-term variations in the body size of planktonic copepods were studied in the northern Baltic Sea. The influence of temperature, salinity and phytoplankton concentration to the body size of Eurytemora affinis, Acartia bifilosa and Limnocalanus macrurus (Calanoida) was examined at three sea areas, differing in their hydrographical and trophic conditions (an archipelago area and an enclosed bay on the SW coast of Finland in 1992 and at an open sea station in the Gulf of Bothnia in 1991). There was an inverse relationship between copepod body length and temperature, while there was a direct effect of phytoplankton concentration. According to multiple regression analysis, the relative importance of these factors varied according to species, developmental stage and study area. In the archipelago area, copepod body size was mainly determined by temperature, while in the open Gulf of Bothnia, phytoplankton concentration was usually more important. Interannual variation in the summer body length of E. affinis and A. bifilosa was examined using samples collected over 18 years (1967 to 1984) in the archipelago area. In this analysis, the inverse relationship between water temperature and copepod body size disappeared; in E. affinis the average female length correlated positively with summer temperature. We suggest that the length and timing of the study period greatly influence the investigator's view of the factors regulating copepod body size.     

Continuous plankton records: Changes in the composition and abundance of the phytoplankton of the north-eastern Atlantic ocean and North Sea, 1958–1974

In most areas of the north-eastern Atlantic Ocean, diatoms have declined drastically in abundance in the last decade. Additionally, in areas to the north of 59°N Ceratium species and an index of total phytoplankton have also declined. South of 59°N the phytoplankton index has increased, diatoms have declined and Ceratium species have remained at a constant level of abundance. A possible explanation of the increase in the phytoplankton index at a time when the diatoms were declinig south of 59°N is the development of unidentified phytoplankton organisms such as microflagellates. As many of the variables influencing phytoplankton standing crop are governed in turn by the prevailing weather, the phytoplankton changes may well be a consequence of the general deterioration, since 1940, of North Atlantic weather. Changes in phytoplankton which may be attributed to an amelioration of climate since 1971 are evident as yet only in the southern North Sea.     

Bioactivity as an Options Value of Sea Cucumbers in the Egyptian Red Sea

Abstract:  The utility of a species can be divided into its direct, indirect, and options values. In the marine environment, direct consumptive values predominate and often lead to overexploitation at the expense of significant options values derived through bioprospecting for natural products. We surveyed the waters of the Egyptian Red Sea coast (Gulf of Aqaba [north] and the Red Sea [south]) for species of sea cucumbers and analyzed extracts from species for a range of bioactivities with potential biomedical applications. All habitat types were surveyed within these regions. We found 22 species of sea cucumber of which two, Holothuria fuscogilva and Holothuria flavomaculata , were recorded in Egypt for the first time. Although none of the species identified were unique to the Gulf of Aqaba, 10 species were only found in the Red Sea sector. Bioassay results showed that although no species had antibacterial activity, most extracts exhibited activity against Candida and Leishmania but were most active against a LoVo mammalian carcinoma cell line. Our most significant finding was the intraspecific variation in bioactivity in individuals collected from different habitat types and sectors of the coast. This variation may reflect the effect of environment on secondary metabolite production or may indicate significant genetic diversity between populations within a species. Our results indicate a potentially significant options value to sea cucumbers through bioprospecting. Given the importance of economic development in countries such as Egypt and the perceived low conservation value of invertebrates such as sea cucumbers, the linking of these factors to conservation is vital for the maintenance and sustainable exploitation of these animals .     

Interspecific differences in the photosynthetic carbon metabolism of marine phytoplankton

Six species of marine phytoplankton of different sizes and taxonomic categories were grown in microcosms under identical experimental conditions; the species cultured were: Pavlova lutheri (Prymnesiophyceae), Dunaliella tertiolecta (Chlorophyceae), Phaeodactylum tricornutum (Baciollariophyceae), Eutreptiella sp. (Euglenophyceae), Alexandrium tamarense (Dinophyceae), and Phaeocystis pouchetii (Prymnesiophyceae). The photosynthetic carbon metabolism of these phytoplankton was studied throughout the exponential and lag phases of growth after nutrient depletion. The relative incorporation of carbon into protein was positively correlated with phytoplankton growth, while carbon assimilation into low molecular weight metabolites (LMWM) and storage products, i.e., lipid and polysaccharides, generally increased under nutrient-limiting conditions. Clear taxonspecific differences were observed in the proportions of carbon incorporated into cell constituents. A significant linear relationship was consistently found between the relative synthesis of protein to LMWM, and both the production normalised to chlorophyll (P:B) and the phytoplankton growth rate. However, ANCOVA revealed significant, interspecific differences in these relationships.     

Comparison of osmoregulation between Adriatic and North Sea Carcinus

In two spatially isolated species — North Sea Carcinus maenas L. and Adriatic Carcinus mediterraneus Czerniavsky —osmoconcentration in hemolymph and Na-transport were investigated. The experiments were performed on crabs which had been long-term acclimated to sea water (1190 mOsmol/l) and to 50% sea water, and on individuals immediately after transfer from sea water to 50% sea water, until new steadystate conditions had been reached. About 10 to 12 h after transfer, a new steady state of total osmoconcentration as well as of the concentrations of the most representative cations Na, K, Ca, and Mg was attained. The hemolymph contents of Na and total osmotically active substances of the North and Adriatic Sea populations are significantly different when they are long-term acclimated to sea water and to 50% sea water. Total sodium fluxes, as calculated from influx and outflux experiments, are in the range 12.1 to 15.2 M Na/g/h in both species when acclimated to sea water. In 50% sea water, sodium fluxes are retarded, especially in C. maenas (6.6 M Na/g/h). This value is significantly lower than the corresponding value in C. mediterraneus (11.4 M Na/g/h). Differences are discussed on the basis of adaptation of the populations to their different environments.     

Dissolved organic substances during a Phaeocystis pouchetii bloom in the German Bight (North Sea)

Dissolved organic substances (carbohydrates, organic nitrogen, free amino acids) were measured in the German Bight (North Sea) in June, 1981. During and before this survey, sea foam was observed in the east Frisian coastal water and it accumulated on the nearby beaches to an unusually high extent. In this coastal water area, a large Phaeocystis pouchetii Lagerheim bloom and very high concentrations of dissolved organic matter were found. The above dissolved organic substances were all positively correlated to a highly significant degree with P. pouchetii cell numbers in the bloom area. An influence of salinity (or river water) on this correlation could be excluded. Thus, exudation or decomposition products of P. pouchetii were most likely the cause of the unusually high concentrations of dissolved organic matter in the bloom area off the east Frisian coast, where P. pouchetii blooms have been reported for many years. Ammonia concentrations were very low in the P. pouchetii bloom area; this and the accumulation of dissolved organic substances might lead to speculation that decomposition of dissolved organic matter by bacteria could have been reduced due to antibiotic activity of P. pouchetii.