Rapid assessment of different oxygenic phototrophs and single-cell photosynthesis with multicolour variable chlorophyll fluorescence imaging

We present a new system for microscopic multicolour variable chlorophyll fluorescence imaging of aquatic phototrophs. The system is compact and portable and enables microscopic imaging of photosynthetic performance of individual cells and chloroplasts using different combinations of blue, green, red or white light. Automated sequential exposure of microscopic samples to the three excitation colours enables subsequent deconvolution of the resulting fluorescence signals and colour marking of cells with different photopigmentation, i.e., cyanobacteria, green algae, red algae and diatoms. The photosynthetic activity in complex mixtures of phototrophs and natural samples can thus be assigned to different types of phototrophs, which can be quantified simultaneously. Here, we describe the composition and performance of the new imaging system and present applications with both natural phytoplankton and microalgal culture samples.     

Diets of calanoid copepods on the West Florida continental shelf: Relationships between food concentration,food composition and feeding activity

Bottle incubations were conducted in March, July/August and October 1992. to measure the daily rations (R) and objectively characterize the diets of the calanoid copepodsEucalanus elongatus, Undinula vulgaris, Centropages velificatus andTemora stylifera from the west Florida continental shelf. Daily rations,R, were clustered around two, order-of-magnitude different means, 1.3 and 11.2% of body C d^–1, representative of quiescent and active feeding modes, respectively. The food concentration at which the transition from quiescent to active mode occurred was influenced by food particle size. In the quiescent mode, diets were dominated by nanoplankton, whereas no food type dominated the diet in the active mode. Selective feeding, defined as a statistically significant difference between the frequency distributions of foods in the diet and environment, occurred in both quiescent and active copepods. However, what appeared to be selective feeding in quiescent copepods could be explained by processes that passively modified the distribution of the diet relative to that of the food supply. Conversely, selective feeding in active copepods apparently resulted from foraging for particles >5 m in diameter in food environments dominated by nanoplankton (<5 m).     

Genotoxicity biomarkers in the amphipod Gammarus elvirae exposed in vivo to mercury and lead,and basal levels of DNA damage in two cell types

The present work deals with the application of genotoxicity biomarkers by means of the Comet assay in haemocytes and spermatozoa of the crustacean Gammarus elvirae exposed in vivo to heavy metals. Furthermore, a basal levels (BLs) study of DNA damage in the two cell types considered for two different gammarids species, G. elvirae and Echinogammarus veneris, was carried out. It is important to identify factors that influence the outcome of the assay in order to obtain reliable and reproducible results usable for risk assessment purposes. Our results highlight that the Italian legal limits for Hg and Pb, respectively, 0.5 and 50?µg/L, are inadequate for establishing safety thresholds in the aquatic environment. Furthermore, the freshwater invertebrate G. elvirae, used for the first time to measure the effect of genotoxicants, is a good candidate for evaluating the genotoxicity damage induced by heavy metals. Our results concerning spermatozoa show excessively variable responses and high BLs.     

Residues of PCDD,PCDF, and PCBs in some marine organisms in Lake Temsah,Ismailia, Egypt

Residues of PCDDs/F, non-ortho, mono-ortho PCBs, and other PCBs were monitored in the tissues of mullet fish, bolti fish, bivalves and crab taken from Lake Temsah, at Ismailia, Egypt.

Results showed that 2,3,7,8 Tetra CDD and 1,2,3,7,8 Penta CDD were the most frequently detected PCDD congeners. Similarly, 2,3,7,8 Tetra CDF, 1,2,3,7,8 Penta CDF and 2,3,4,7,8 Penta CDF were the most frequently detected PCDF congeners. No relationship was apparent between the concentrations of detected PCDDs congeners and the degree of chlorination, except with crab samples in which an increase in the chlorination coincided with a decrease in the concentrations of the congeners.

In PCDF congeners, detected residues have had a reversed relationship with chlorination increase. In PCDD congeners, Octa CDD had the highest detected concentrations in the two fish species, while in the bivalves and crab, 2,3,7,8 Tetra CDD had the highest concentrations. The mullet fish had the highest total PCDDs concentration, at 0.398?pg/g fresh weight, followed by crab at 0.395?pg/g fresh weight, then bivalves and bolti fish at 0.187 and 0.062?pg/g fresh weight, respectively. In all the examined organisms, the total concentrations of PCDFs were much higher than the total concentrations of the PCDD congeners. The WHO–TEQ values were 11.92, 39.12, 25, and 3.6?pg/g fresh weight, for mullet fish, bolti fish, bivlaves and crab, respectively. The concentration of the mono-ortho congeners CB 118 was the highest detected of all non-ortho and mono-ortho PCBs congeners, with values of 0.382, 0.022, 0.231 and 0.357?ng/g fresh weight, in mullet fish, bolti fish, bivalves and crab, respectively. The WHO–TEQ concentrations were 0.799, 0.003?pg/g fw, 0.05?pg/g fresh weight, 0.676?pg/g, and 0.799?pg/g fresh weight, for the same species, respectively. The total concentration of PCBs 28, 52, 95, 99, 101, 105, 110, 118, 138, 146, 149, 151, 153, 170, 177, 180, 187 were 6.86?ng/g fresh weight for mullet fish, 0.2?ng/g fresh weight, for bolti fish, 2.72?ng/g fresh weight for bivalves and 2.8?ng/g fresh weight for crab, respectively.     

Effects of symbiont elimination and reinfection on the life processes of the planktonic foraminifer Globigerinoides sacculifer

Laboratory experiments were carried out to determine the influence of symbiotic dinoflagellates (zooxanthellae) on the shell growth, longevity, and reproductive potential of Globigerinoides sacculifer (Brady). Its symbionts were eliminated by 72-h treatment with a photosynthetic inhibitor (DCMU). Symbiont elimination resulted in earlier gametogenesis (shortened survival time) and smaller shell sizes of G. sacculifer when compared to untreated foraminifera grown in sea water. Individuals kept in continuous darkness in untreated sea water also exhibited early gametogenesis, short survival times and small shell sizes. Aposymbiotic foraminifera formed on the average one or two chambers fewer per individual and their rate of shell size increase is slower than symbiont-bearing foraminifera. Symbionts were lysed within perialgal vacuoles of G. sacculifer when subjected to DCMU treatment or kept in continuous darkness. One DCMU-treated group was reinfected with symbionts from crushed G. sacculifer donors. Soon after reinfection, these foraminifera resumed a shell growth rate and exhibited developmental stages that were nearly equivalent to those of untreated individuals, as deduced from their shell size, frequency of sac-like chambers, rate of gametogenesis, and survival time. Our experiments indicate that the symbionts aid in calcification and that elimination of symbionts triggers gametogenesis, thus shortening the life span of the foraminiferal host. The results imply that shell growth in symbiont-bearing planktonic foraminifera occurs mainly in the euphotic zone and that they do not survive for long periods below it.     

Abundance,distribution and prey composition of scyphomedusae in the southern North Sea

The annual cycle of abundance and distribution of the scyphozoan medusae Aurelia aurita, Cyanea lamarckii, C. capillata and Chrysaora hysoscella were studied in the southern North Sea in 2004 and 2005. Three different patterns of seasonal occurrence of medusae were distinguished: (1) the early occurring C. lamarckii (February–August), (2) C. capillata and A. aurita (April–August) and (3) the late appearing C. hysoscella (July/August–September). Cyanea lamarckii was the most frequently encountered species in this study; its highest mean abundance was 1.8 ± 2.7 ind. 100 m⁻³. The prey spectra of C. lamarckii, C. capillata and C. hysoscella contained several copepod and other crustacean species and thus make them potential competitors with fish larvae. Medusae in this study also consumed fish eggs and larvae, including clupeids, in all months analysed. Although peak spawning of sprat (Sprattus sprattus) coincides with the maximum abundance of medusae (May–June) the relative low abundance of all medusae species in this study makes jellyfish predation unlikely to be a factor controlling sprat recruitment in the time frame investigated.     

Behavioral aspects of sperm competition in cuttlefish, Sepia officinalis (Sepioidea: Cephalopoda)

Sexual selection studies in cephalopods indicate that sperm competition is a central feature of their mating systems, yet this has not been studied experimentally in any detail. In 1998 we staged 20 matings of the cuttlefish, Sepia officinalis L., in the laboratory. Males rapidly initiated mating in the “head-to-head” position, with no apparent courtship. Mating lasted an average of 10 min (range 7 to 14 min). For the first 6 min (on average 63% of the mating duration), the male flushed strong jets of water directly at the female's buccal membrane, which sometimes resulted in the expulsion of parts of spermatangia placed there in recent matings. Then, in a single discrete movement that lasted an average of only 14 s, the male's modified fourth left arm – the hectocotylus – wrapped around a single large bundle of spermatophores (ca. 150 to 300) and transferred them to the female's buccal membrane. For the remainder of the mating (average 3 min, range 1.5 to 5.0 min), the hectocotylus repeatedly broke the spermatophores open, and manipulated them, so that sperm were released and many spermatangia were attached along the ventral buccal membrane, near the paired seminal receptacles. Approximately 140 spermatangia were attached in rows 3 to 5 deep around the ventral buccal membrane in a single mating; the rest were usually discarded during mating. Histology revealed that each of the seminal receptacles consists of a series of sperm storage bulbs connected by a central duct, which leads to a single pore at the surface of the buccal membrane. Baseline data on sperm motility were obtained, but the mechanism by which sperm enter the seminal receptacle remains unknown. Females seemed to initiate termination of mating, then males guarded their mates temporarily. These results, combined with other recent laboratory experiments, provide evidence that sperm competition may be a major feature of the mating system of S. officinalis. Received: 4 January 1999 / Accepted: 18 May 1999     

Assessing the extinction risk of insular,understudied marine species

Hydrothermal vents are rare deep-sea oases that house faunal assemblages with a similar density of life as coral reefs. Only approximately 600 of these hotspots are known worldwide, most only one-third of a football field in size. With advancing development of the deep-sea mining industry, there is an urgent need to protect these unique, insular ecosystems and their specialist endemic faunas. We applied the IUCN (International Union for the Conservation of Nature) Red List criteria to assess the extinction risk of vent-endemic molluscs with varying exposure to potential deep-sea mining. We assessed 31 species from three key areas under different regulatory frameworks in the Indian, West Pacific, and Southern Oceans. Three vent mollusc species were also examined as case studies of different threat contexts (protected or not from potential mining) to explore the interaction of local regulatory frameworks and IUCN Red List category assignment. We found that these assessments were robust even when there was some uncertainty in the total range of individual species, allowing assessment of species that have only recently been named and described. For vent-endemic species, regulatory changes to area-based management can have a greater impact on IUCN Red List assessment outcomes than incorporating additional data about species distributions. Our approach revealed the most useful IUCN Red List criteria for vent-endemic species: criteria B and D2. This approach, combining regulatory framework and distribution, has the potential to rapidly gauge assessment outcomes for species in insular systems worldwide.     

Ammonia emission from grassland and livestock production systems in the UK

Emissions of ammonia were measured from livestock excreta and fertilisers applied to grass swards, from grazed paddocks, from decomposing grass herbage and from an animal house containing dairy cows. Emissions from urine, dung, slurry and fertilisers were determined using a system of wind tunnels with each tunnel covering an area of 1 m(2). Emissions from grazed swards were determined using a micrometeorological mass balance method. From the results of these measurements, together with other published information, an inventory for ammonia emissions has been calculated for grassland and livestock production systems over the UK as a whole. It is estimated that emissions from grassland and cattle and sheep production amount to about 230 kt NH(3)-N annually, while emissions from pig and poultry production amount to about 40 kt and 80 kt NH(3)-N, respectively.     

Effect of ozone on net photosynthesis in oat (Avena sativa) and duckweed (Lemna gibba)

Short exposure to ozone depressed photosynthesis in both oat and duckweed at concentrations above 140 microg m(-3) and 300 microg m(-3), respectively. The effect on exposed oat flag leaves was age-dependent, with maximum susceptibility to ozone 10-20 days after emergence of the panicle. In duckweed, photosynthesis was more sensitive to differences in ozone concentration than to differences in duration of exposure.