Effects of ultraviolet radiation on pigmentation, photoenzymatic repair, behavior, and community ecology of zooplankton.

In this report, we provide a perspective on how zooplankton are able to respond to present and future levels of ultraviolet (UV) radiation, a threat that has been present throughout evolutionary time. To cope with this threat, zooplankton have evolved several adaptations including behavioral responses, repair systems, and accumulation of photoprotective compounds. Common photoprotective compounds include melanins and carotenoids, which are true pigments, but also mycosporine-like amino acids (MAAs) and several other substances, and different taxa use different blends of these compounds. It is not only the level of UV radiation, however, that determines the amount of photoprotective compounds incorporated by the zooplankton, but also other environmental factors, such as predation and supply rate of the compounds. Furthermore, compared to taxa that are less pigmented, those taxa with ample pigmentation are generally less likely to exhibit diel migration. The photoenzymatic repair of UV damages seems to be more efficient at intermediate temperature than at low and high temperatures, suggesting that it is less useful at high and low latitudes, where UV radiation is often extremely high. While predicted future increases in UV radiation are expected to substantially affect many processes, recent studies show that most zooplankton taxa are well adapted to cope with such increases, either by UV avoidance behavior or by incorporation of photoprotective compounds. Hence, we conclude that future increase in UV radiation will have only moderate direct effects on zooplankton biomass and community dynamics.

Proteolytic degradation of cold-water fish gelatin solutions and gels.

The stability of cold-water fish gelatin (FG), both in solution and in the gel phase, has been studied as function of both temperature and exposure towards novel proteases of marine origin. A 1% (w/v) FG solution was readily degraded by such proteases above 20 degrees C, which was expected since FG at this temperature is a random coil molecule lacking the protective triple helical structure found in collagen. The dynamic storage modulus for a 10% (w/v) FG gel increased monotonically at 4 degrees C. Ramping the temperature to 6, 8 or 10 degrees C led to a drastic reduction in G', but an apparent partial recovery of the network (increasing G') was observed with time at all temperatures. In the presence of proteases, a lower storage modulus was observed. At constant 4 degrees C, an apparent maximum value was reached after curing for 2h followed by a decrease in G' indicating protease activity. Ramping of temperature in the presence of proteases led to an even more drastic reduction in G' and no recovery of structure was observed with time. In this case, the overall rheological behaviour is a complex function of both thermal influence as well as proteolytic activity. In an endeavour to quantify the effect of the presence of proteolytic enzymes on the gelatin network, rheological investigation were undertaken where the dynamic storage moduli were recorded on different 10% (w/v) FG samples that had been acid hydrolysed to yield different average molecular weights. A significant reduction in storage modulus for average molecular weights below 50 kDa was found. This critical molecular weight most probably reflects the on-set of a regime where shorter chain lengths prevent percolation due to an increase in the loose end and sol fraction as well as a reduction in the average length of the pyrrolidine-rich regions reducing the number of possible junction zones.

Zooplankton chitobiase activity as an endpoint of pharmaceutical effect.

Numerous human and veterinary pharmaceuticals are constantly entering surface waters, despite little understanding of their potential impacts on aquatic ecosystems. To address this concern, an attempt to create a simple, reproducible, inexpensive, and sublethal toxicity bioassay for freshwater zooplankton was initiated. The approach was centered on characterizing the response of a zooplankton enzyme, chitobiase, to the presence of a toxicant. The aim of the present research was to develop a reproducible laboratory-based assay for Daphnia magna chitobiase activity and to screen four commonly prescribed pharmaceuticals using that assay. The four pharmaceuticals tested for potential effects on D. magna chitobiase activity were atorvastatin, lovastatin, fluoxetine, and sertraline. We were able to detect exposure-associated differences in chitobiase activity at concentrations of 0.1 mug/L fluoxetine after 24 and 72 hours of exposure. Differences were also detected for the other compounds. The response of chitobiase was found to be promising as an assay to measure sublethal effects in D. magna and perhaps other zooplankton species.

Proteolytic activity and protease classes in the zooplankton species Calanus finmarchicus.

The temperature optimum for the general proteolytic activity in the crude extract of Calanus finmarchicus was 50 degrees C and the pH optimum was found to be 7. The use of specific protease inhibitors resulted in the identification of at least three protease classes in the crude extract of C. finmarchicus. Those classes were serine, metallo and aspartic proteases. The serine and metallo proteases were found to be dominant under neutral to alkaline conditions and the aspartic proteases were dominant under acidic conditions. The cysteine proteases, if at all present, did not seem to be active or present in any substantial amount. The data presented points towards controlling and preserving the protein content in fresh C. finmarchicus.

Origin and characteristics of the zooplankton phosphatase activity in a coastal ecosystem of the Mediterranean sea (Toulon Bay).

In Toulon Bay (France), very high phosphatase activities have been found in the zooplankton fraction>90 microm. This work was intended to specify their origin. For that purpose, larvae, juvenile and adult Crustacea (Copepods: Calanoids, Cyclopoids, Branchiopods: Cladocera, and Cirripeds) were isolated. Their activities were measured using paranitrophenyl phosphate dissolved in sea water in order to calculate Km (the enzyme half saturation concentration) and Vmax (the reaction rate when the enzyme is saturated with substrate). Vmax were referred to protein contents of the isolated organisms to calculate specific activities. For all zooplankton groups high and low affinity phosphatase activities were found. The low affinity enzyme was responsible for at least 70% of the total phosphatase activity. Its specific activity was higher for larvae than for copepodites and adults. In Cirriped nauplii this activity was particularly high with values which were several hundred times higher than that in other Crustacea. These enzymes had optimum pH close to 8.4, magnesium requirement and were competitively inhibited by orthophosphate. Experiments with intact and lysed Cirriped nauplii confirmed that living organisms had only a weak external activity and showed that most of the activity of these larvae was primarily intracellular.

Seasonal changes in phosphatase activities in Toulon Bay (France).

We studied the characteristics of the phosphatase activity (Km and Vmax) in total seawater and in particulate material of the three main plankton classes (0.25-5, 5-90 and >90 microm) in a coastal marine ecosystem of Toulon Bay (French Mediterranean Sea). The measurement of the hydrolysis of sodium paranitrophenylphosphate (pNPP), a substrate of phosphatase, revealed low and high affinity components in unfiltered seawater and in particulate matter. In unfiltered seawater, the low affinity activity was predominant from October to March during phytoplankton development. The high affinity activity dominated from April to June and was significantly correlated with the bacterial abundances. The phosphatase behaviour in the particulate material differs from that in the unfiltered seawater. The activity of the three particulate classes was generally much lower than that of unfiltered seawater, particularly the low affinity activity. The >90 microm size fraction consisted in greater part of zooplankton. In this size class, the activity (nmol l(-1) h(-1)) of the low affinity component was predominant from May to August, when the abundance of the larvae of copepods (copepodites) was highest. Its high specific activity (Activity/Protein concentration as nmol l(-1) h(-1) microg(-1)) was particularly elevated during this period. The 5-90 microm fraction consisted of phytoplankton cells, especially Dinoflagellates. Between September and January, the activity (nmol l(-1) h(-1)) of this size class was mostly supported by the low affinity component. The specific activity (nmol l(-1) h(-1) microg(-1)) of the high affinity component was highest in June and August. No significant correlation was found between phosphatase activities and chlorophyll a or total cell abundance. In return temporary relationships with specific taxa exist in particular with Ceratium spp., Gymnodinium spp. and Protoperidinium spp. The contribution of the 0.25-5 microm size class exceeded rarely 20% of the total particulate activity. Between June and August, high specific activities (nmol l(-1) h(-1) microg(-1)) were observed for its high affinity component. In autumn, strong rainfall increased the phosphate and nitrate concentrations and led to a drop in salinity, which probably explains the low phosphatase activities (nmol l(-1) h(-1)) and cell densities observed during this period.

Preliminary results of biomarker responses in zooplankton of brackish environments.

Zooplankton is an essential component of the food chains of marine and brackish environments. Ecotoxicological risk to zooplankton, evaluated by the biomarker approach, can be used as an early warning signal of risk to the health of marine ecosystems. The aim of this project is to evaluate the application of certain biomarkers in zooplankton. In the present paper, laboratory and field studies were performed with the copepods Acartia margalefi, Acartia latisetosa and the mysid Siriella clausi. Acetylcholinesterase activity (AChE) was determined in homogenates of whole organisms. Mean AChE activity was 10.05 micromol min g(-1) for A. margalefi, 3.30 micromol min g(-1) for A. latisetosa and 79.70 micromol min g(-1) for S. clausi. A. margalefi and S. clausi also showed a linear increase in enzyme activity with increasing concentrations of samples. Preliminary laboratory work was also carried out with specimens of A. latisetosa, which were exposed to an organophosphorus insecticide (parathion). Moderate inhibition of AChE (19%) with respect to controls was observed. AChE activities were two orders of magnitude higher in the zooplankton than in hemolymph samples of the decapod Carcinus aestuarii (Fossi et al., 1996) indicating that these species have a high metabolic rate, which makes them suitable for biomarker studies.

Phylogeography and regional endemism of a passively dispersing zooplankter: mitochondrial DNA variation in rotifer resting egg banks.

We investigated the phylogeography of the salt water rotifer Brachionus plicatilis, a cyclical parthenogen with passive dispersal mechanisms, using resting eggs recovered from saline lake sediments. Individual resting eggs were obtained from a large selection of lakes which were representative of five endorheic basins and the chain of coastal ponds in the Iberian Peninsula. The novel use of resting eggs allows the integration of seasonal and annual variations as well as the impact of stochastic effects such as drift and local extinction. A 653 bp fragment of the mitochondrial cytochrome oxidase subunit I (COI) gene was sequenced from 98 eggs. Our results revealed a deep phylogeographical structure in this species, with a division into two main lineages with distinct geographical distributions, which probably diverged at the beginning of the Pleistocene period. Most of the mitochondrial DNA haplotypes were restricted to single lakes. Nested clade analysis supported Early Pleistocene fragmentation of populations, low gene flow and some long-distance colonization. These conclusions contrast strongly with previous ideas on rotifer biogeography and this pattern is consistent with a recolonization of the Iberian Peninsula from two glacial refugia. The results provide new insights into the processes responsible for the genetic diversification of passive dispersers, a life-history trait typical of zooplanktonic biotas.

Solar UVB-induced DNA damage and photoenzymatic DNA repair in antarctic zooplankton.

The detrimental effects of elevated intensities of mid-UV radiation (UVB), a result of stratospheric ozone depletion during the austral spring, on the primary producers of the Antarctic marine ecosystem have been well documented. Here we report that natural populations of Antarctic zooplankton also sustain significant DNA damage [measured as cyclobutane pyrimidine dimers (CPDs)] during periods of increased UVB flux. This is the first direct evidence that increased solar UVB may result in damage to marine organisms other than primary producers in Antarctica. The extent of DNA damage in pelagic icefish eggs correlated with daily incident UVB irradiance, reflecting the difference between acquisition and repair of CPDs. Patterns of DNA damage in fish larvae did not correlate with daily UVB flux, possibly due to different depth distributions and/or different capacities for DNA repair. Clearance of CPDs by Antarctic fish and krill was mediated primarily by the photoenzymatic repair system. Although repair rates were large for all species evaluated, they were apparently inadequate to prevent the transient accumulation of substantial CPD burdens. The capacity for DNA repair in Antarctic organisms was highest in those species whose early life history stages occupy the water column during periods of ozone depletion (austral spring) and lowest in fish species whose eggs and larvae are abundant during winter. Although the potential reduction in fitness of Antarctic zooplankton resulting from DNA damage is unknown, we suggest that increased solar UV may reduce recruitment and adversely affect trophic transfer of productivity by affecting heterotrophic species as well as primary producers.

[The lysozyme-antilysozyme functional system in hydrobionts and its role in forming aqueous biocenoses]. / Funktsional'naia sistema "lizotsim-antilizotsim" u gidrobiontov i ee rol' v formirovanii vodnykh biotsenozov.

Lysozyme activity has been evaluated in hydrobionts of different trophic levels: algae, zooplankton and germs. The existence of the functional system "Lysozyme of hydrobionts--antilysozyme of germs", functioning as a regulating element in the formation of aqueous microbiocenosis, has been substantiated on the basis of data provided by laboratory experiments and investigations carried out in natural water pools.

Phosphatases in lake water: characterization of enzymes from phytoplankton and zooplankton by gel filtration.

Sephadex gel filtration was used to characterize phosphomonoesterases in two small takes in northern Sweden. Two fractions, here termed phosphatase A and phosphatase B, were found both as free enzymes and associated with seston. The activity of phosphatase A was correlated with the presence of algal biomass. Phosphatase B, on the other hand, was derived from zooplankton. Phosphate served as an effective inhibitor of phosphatase A but had no such effect on phosphatase B. Both fractions had pH optima between 6.5 and 7.0.