Growth and fatty acid composition of pikeperch (Sander lucioperca L., 1758) larvae under altered feeding protocol including the copepod Apocyclops panamensis (Marsh, 1913).

Alternative live feeds for small and sensitive fish early life stages such as pikeperch (Sander lucioperca L., 1758) can improve the larval quantity, quality and performance in aquaculture. Therefore, this study evaluated the cyclopoid copepod Apocyclops panamensis (Marsh, 1913) as live feed for pikeperch larviculture from day 11 post hatch (dph) in two independent experiments. In both experiments, pikeperch larvae had the highest specific growth rate (SGR) when they fed on Brachionus plicatilis until dph 11 and A. panamensis until dph 16-18. SGR was related to a decrease in total fatty acids (FAs), saturated FAs and monounsaturated FAs in pikeperch larvae, indicating their use as energy for growth. Within the polyunsaturated FAs, docosahexaenoic acid (DHA) increased in larvae fed with A. panamensis and coincided with the highest SGR suggesting that DHA is accumulated in larvae as structural FA. Our study demonstrated a suitable pikeperch larval fatty acid composition for growth after feeding A. panamensis compared with Artemia sp. from dph 11 until dph 16 and previously fed with B. plicatilis. Moreover, it highlighted the importance of the dietary PUFAs in pikeperch rearing, specifically of linoleic acid (LA) from dph 4 until dph 11 and of DHA from dph 11 onwards.

The artificial humic substance HS1500 does not inhibit photosynthesis of the green alga Desmodesmus armatus in vivo but interacts with the photosynthetic apparatus of isolated spinach thylakoids in vitro.

Humic substances (HSs) can influence the growth and composition of freshwater phytoplankton assemblage. Since HSs contain many phenolic and quinonic moieties and cause growth reductions in eco-physiological field experiments, HSs are considered photosystem II herbicides. To test this specific mode of action in vivo and in vitro, respectively, we used intact cells of the green alga Desmodesmus armatus, as well as thylakoids isolated from spinach (Spinacia oleracea) as a model system for the green algal chloroplast. Photosynthetic electron transport was measured as oxygen evolution and variable chlorophyll fluorescence. The in vivo effect of the artificial humic substance HS1500 on algae consisted of no impact on photosynthesis-irradiance curves of intact green algae compared to untreated controls. In contrast, addition of HS1500 to isolated thylakoids resulted in light-induced oxygen consumption (Mehler reaction) as an in vitro effect. Fluorescence induction kinetics of HS-treated thylakoids revealed a large static quenching effect of HS1500, but no inhibitory effect on electron transport. For the case of intact algal cells, we conclude that the highly hydrophilic and rather large molecules of HS1500 are not taken up in effective quantities and, therefore, cannot interfere with photosynthesis. The in vitro tests show that HS1500 has no inhibitory effect on photosystem II but operates as a weak, oxygen-consuming Hill acceptor at photosystem I. Hence, the results indicate that eco-physiological field experiments should focus more strongly on effects of HSs on extracellular features, such as reducing and red-shifting the underwater light field or influencing nutrient availability by cation exchange within the plankton network.

Natural xenobiotics to prevent cyanobacterial and algal growth in freshwater: contrasting efficacy of tannic acid, gallic acid, and gramine.

Allelochemical action against planktonic phototrophs is one central issue in freshwater ecology and quality management. To determine some basic mechanisms of this toxic action, we exposed the coccal green alga, Desmodesmus armatus, and the coccal cyanobacterium, Microcystis aeruginosa, in a batch culture well-supplied with carbon dioxide to increasing concentrations of the polyphenols tannic acid and gallic acid and the alkaloid gramine. The phototrophs were checked after 2d and at the end of the culture for biomass-based growth rates, cell volume, maximum quantum yield of photosystem II (ΦPSIImax), chlorophyll a content (chla) after 2d and at the end of the culture, and lipid peroxidation only at the end of the culture. During the culture, the pH rose from 7.64 to 10.95, a pH characteristic of eutrophic freshwater bodies during nuisance algal blooms. All xenobiotics reduced the growth rate, ΦPSIImax, and chla during the first 2d with M. aeruginosa being more sensitive to the polyphenols than D. armatus. The efficacy of the polyphenols declined with increasing pH, indicating potential polymerization and corresponding reduced bioavailability of the polyphenols. In contrast to the polyphenols, gramine increased its toxic action over time, independent of the prevailing pH. All exposures caused slight to severe lipid peroxidation (LPO) in the phototrophs. Hence, one mechanism of growth inhibition may be oxidative stress-mediated reduction in photosynthesis. The presented results suggest that in successful field trials with leachate, the prevailing environmental conditions may inactivate polyphenols and xenobiotics other than polyphenols may be more effective.

Toxicity of hydroquinone to different freshwater phototrophs is influenced by time of exposure and pH.

The interaction of natural organic matter with phytoplankton communities in freshwater ecosystems is an intensively studied subject matter. Previous studies showed that apparently plant-derived phenols were able to inhibit algal and cyanobacterial growth. Furthermore, it was also assumed that humic substances (HS), which comprise the major part of dissolved organic carbon in freshwater ecosystems, directly interact with freshwater phototrophs. For example, quinoid building blocks of HS were thought to be algicidal. To identify key environmental variable for the toxic action of potential quinone algicides, we tested the toxicity of hydroquinone (HQ) to different eukaryotic and prokaryotic freshwater phototrophs in terms of growth performance and investigated also the effect of HQ oxidation at different pH values on its algicidal potential. It was shown that cyanobacterial species were much more susceptible to hydroquinone than coccal green algal species were, with Microcystis aeruginosa being the most sensitive species by far. In addition, it was obvious that the aging of hydroquinone-stock solution at pH 11 led to polymerization and, by this process, to a total loss of toxicity; whereas the algicidal potential sustained if the polyphenol was kept at pH 7. Since most lakes with heavy blooms of phototrophs possess pH values clearly above 7.0, it is questionable, if polyphenols in general and quinones in particular are the effective chemicals and if litter and straw leachates are applied as means to combat algal and cyanobacterial blooms.

Does quinone or phenol enrichment of humic substances alter the primary compound from a non-algicidal to an algicidal preparation?

Dissolved organic matter (DOM) has been shown to affect phytoplankton species directly. These interactions largely depend on the origin and molecular size of DOM and are different in prokaryotes and eukaryotes. In a preceding study, however, two humic substance preparations did not adversely affect coccal green algae or cyanobacterial growth even at high concentrations of dissolved organic carbon (DOC). These results contradicted previous findings, showing a clear, negative response of different phototrophs to much lower DOC concentrations. To test whether or not at least defined building blocks of humic substances (HSs) are effective algicidal structures, we enriched two humic preparations with hydroquinone and p-benzoquinone, respectively, and exposed two different green algae, Pseudokirchneriella subcapitata and Monoraphidium braunii, and two cyanobacterial species, Synechocystis sp. and Microcystis aeruginosa, to the unmodified and enriched HSs. As response variables, growth rates in terms of biomass increase, chlorophyll-a content, and photosynthetic yield were measured. The highest concentration (4.17 mM DOC) of the modified HSs clearly inhibited growth; the cyanobacterial species were much more sensitive than the green algal species. However, realistic ecological concentrations did not adversely affect growth. Aerating the exposure solution for 24 h strongly reduced the inhibitory effect of the modified HSs. The algicidal effect was obviously caused by monomers and not by polymerised high molecular weight HSs themselves. Furthermore, the maximum quantum yield (Φ PSII max) was stimulated in the green algal species by low and medium DOC concentrations, but reduced in the cyanobacterial species upon exposure to higher HS concentrations. The quinone- and phenol-enriched HSs only showed algicidal activity at high concentrations of 4.17 mM DOC and lost their effects over time, presumably by oxidation and subsequent polymerisation. This study confirms that the applied humic substances themselves are not effective algicides even if enriched in effective structures.

Removal of bisphenol A by the freshwater green alga Monoraphidium braunii and the role of natural organic matter.

Phytoremediation of waters by aquatic organisms such as algae has been recently explored for the removal of organic pollutants possessing endocrine disrupting capacity. Monoraphidium braunii, a green alga known for rapid growth and good tolerance to different natural organic matter (NOM) qualities, was tested in this study for the ability to tolerate and remove the endocrine disruptor bisphenol A at concentrations of 2, 4 and 10mgL(-1), either in NOM-free or NOM-containing media. NOM at concentrations of 2, 5 and 20mgL(-1) of DOC, was added because it may interfere with xenobiotics and modify their effects, modulate algal growth performances or produce a trade-off of both effects. After 2 and 4 days of algal growth, the cell number and size, the maximum quantum yield of photosystem II in the dark or light adapted state, and the chlorophyll a content were recorded in order to evaluate the algal response to bisphenol A. Moreover, the residual bisphenol A was measured in the algal cultures by chromatographic technique. Results indicated that after 2 and 4 days bisphenol A at the lower concentrations was not toxic for alga, whereas at the highest concentration it reduced algal growth and photosynthetic efficiency. The sole NOM and its combinations with bisphenol A at the lower concentrations increased the cell number and the chlorophyll a content of algae. After 4-day growth, good removal efficiency was exerted by M. braunii at concentrations of 2, 4 and 10mgL(-1) removing, respectively, 39%, 48% and 35% of the initial bisphenol A. Lower removal percentages were found after 2-day growth in the different treatments. NOM at any concentration scarcely influenced the bisphenol A removal. On the basis of data obtained, the use of M. braunii could be reasonably recommended for the phytoremediation of aquatic environments from bisphenol A.

Impact of two different humic substances on selected coccal green algae and cyanobacteria--changes in growth and photosynthetic performance.

BACKGROUND, AIM AND SCOPE: There is growing evidence to show that dissolved humic substances, HSs, can directly interact with freshwater organisms, such as phototrophic organisms, cladocerans, amphipods and fish. The responses are-at least in part-transcriptionally controlled. These interactions can lead to stress symptoms in the exposed organisms. In phototrophs, stress symptoms include a reduction in photosynthetic oxygen release and antioxidative stress. Besides the direct effects, HSs also cause indirect effects that provoke different physiological adaptations in the phototrophs. MATERIALS AND METHODS: The HS-influenced photosynthetic performance and stress response of two different green algae, Pseudokirchneriella subcapitata (Korsikov) Hindák and Monoraphidium braunii (Nägeli in Kützing) Komárková-Legnerová, and two cyanobacterial species, Synechocystis sp. (PCC 6803, Institut Pasteur) and Microcystis aeruginosa (PCC 7806, Institut Pasteur), were tested. Two humic preparations were applied, the synthetic HS1500 and HuminFeed, HF, which had previously been proven effective in bioassays with invertebrates and a water mould. RESULTS AND DISCUSSION: When the algae were grown near light saturation, most of the tested species were positively affected by HSs in growth rate or chlorophyll content. Cell sizes decreased with increasing HS concentrations for all eukaryotic phototrophs, except for the cyanobacteria. After 4 to 5 days of cultivation at the highest HS exposure, there was a decrease in total dry weight due to reduced cell sizes in contrast to an increase in cell numbers. With the exception of Synechocystis, the dry weight per cell ratio decreased with increasing HS concentration. The efficiency of utilizing absorbed light quanta increased with increasing HS concentrations; the maximum quantum yield of photosystem II (ΦPSIImax) was higher in all of the tested species, with the exception of M. aeruginosa, after exposure to HS. CONCLUSION: The applied humic preparations did not interact directly with PSII, but changed the physiological state of the algae, especially the photosynthetic performance. Neither the green algae nor the cyanobacteria were inhibited in growth or negatively affected in their photosynthetic performance. The exposure to lower concentrations of HS stimulated better growth of the phototrophs. The tested humic preparations obviously did not have the potency to act as xenobiotic stressors; furthermore, there was no sign of herbicide potency.

Enrichment of humic material with hydroxybenzene moieties intensifies its physiological effects on the nematode Caenorhabditis elegans.

Dissolved humic substances are taken up by organisms and interact on various molecular and biochemical levels. In the nematode Caenorhabditis elegans, such material can promote longevity and increase its reproductive capacity; moreover, the worms tend to stay for longer in humic-enriched environments. Here, we tested the hypothesis that the chemical enrichment of humic substances with hydroxybenzene moieties intensifies these physiological effects. Based on the leonardite humic acid HuminFeed (HF), we followed a polycondensation reaction in which this natural humic substance and a dihydroxybenzene (hydroquinone or benzoquinone) served as reaction partners. Several analytical methods showed the formation of the corresponding copolymers. The chemical modification boosted the antioxidant properties of HF both in vitro and in vivo. Humic substances enriched with hydroxybenzene moieties caused a significantly increased tolerance to thermal stress in C. elegans and extended its lifespan. Exposed nematodes showed delayed linear growth and onset of reproduction and a stronger pumping activity of the pharynx. Thus, treated nematodes act younger than they really are. In this feature the modified HF replicated the biological impact of hydroquinone-homopolymers and various plant polyphenol monomers, thereby supporting the hydroxybenzene moieties of humic substances as major effective structures for the physiological effects observed in C. elegans.