Effect of micro-plastic particles on coral reef foraminifera.

Foraminifera are single-celled protists which are important mediators of the marine carbon cycle. In our study, we explored the potential impact of polystyrene (PS) microplastic particles on two symbiont-bearing large benthic foraminifera species-Heterostegina depressa and Amphistegina lobifera-over a period of three weeks, employing three different approaches: investigating (1) stable isotope (SI) incorporation-via 13C- and 15N-labelled substrates-of the foraminifera to assess their metabolic activity, (2) photosynthetic efficiency of the symbiotic diatoms using imaging PAM fluorometry, and (3) microscopic enumeration of accumulation of PS microplastic particles inside the foraminiferal test. The active feeder A. lobifera incorporated significantly more PS particles inside the cytoplasm than the non-feeding H. depressa, the latter accumulating the beads on the test surface. Photosynthetic area of the symbionts tended to decrease in the presence of microplastic particles in both species, suggesting that the foraminiferal host cells started to digest their diatom symbionts. Compared to the control, the presence of microplastic particles lead to reduced SI uptake in A. lobifera, which indicates inhibition of inorganic carbon and nitrogen assimilation. Competition for particulate food uptake was demonstrated between algae and microplastic particles of similar size. Based on our results, both species seem to be sensitive to microplastic pollution, with non-feeding H. depressa being more strongly affected.

Impact of heavy metals (Cu, Fe, Pb, Zn) on carbon and nitrogen uptake of the diatom-bearing benthic foraminifera Heterosteginadepressa.

Foraminifera are protists primarily living in benthic marine and estuarine environments. We studied uptake of inorganic carbon (C) and nitrogen (N) of the photosymbiont-bearing benthic coral reef foraminifera Heterostegina depressa in the presence of heavy metals. Incubation experiments were accomplished with artificial seawater enriched with copper, iron, lead and zinc at two different concentration levels (10 and 100 fold enriched in contrast to the usual culture medium). Additionally, isotopically labelled 13C-sodium bicarbonate and 15N-ammonium chloride were added to trace their assimilation over time (1 d, 3 d, 5 d, 7 d). Pulse-amplified modulated fluorescence measurements were performed to measure the potential impacts of heavy metals on chlorophyll fluorescence of the photosymbiont. Increased levels of copper (430.5 µg Cu/l) exhibited the greatest toxicity, while for low levels no effect on the overall metabolism of the foraminifera and the fluorescence activity of the photosymbiont could be detected. Iron (III) increased the symbiont activity, independent of concentration applied (44.5 and 513.3 µg Fe/l), which indicates Fe-limitation of the algal symbiont. Lead enrichment showed no detectable effect even at high concentration. Low concentrations of zinc (35.1 µg Zn/l) promoted the metabolism of the foraminifera, while high concentrations (598.4 µg Zn/l) were toxic. At low levels, two metals (Fe and Zn) promoted symbiont activity, at high levels, iron still boosted photosynthesis, but Zn and Cu had a negative impact on the obligatory photosynthetic symbionts.

Impact of pesticides on marine coral reef foraminifera.

Our laboratory study looked into how pesticides affect the foraminifera species Heterostegina depressa and their obligatory algal endosymbionts. We incubated the foraminifera separately with different types of pesticides at varying concentrations (1 %, 0.01 % and 0.0001 %); we included the insecticide Confidor© (active substance: imidacloprid), the fungicide Pronto©Plus (tebuconazole), and the herbicide Roundup© (glyphosate). Our evaluation focused on the symbiont's photosynthetically active area (PA), and the uptake of dissolved inorganic carbon (DIC) and nitrogen (nitrate) to determine the vitality of the foraminifera. Our findings showed that even the lowest doses of the fungicide and herbicide caused irreparable damage to the foraminifera and their symbionts. While the insecticide only deactivated the symbionts (PA = 0) at the highest concentration (1 %), the fungicide, and herbicide caused complete deactivation even at the lowest levels provided (0.0001 %). The fungicide had the strongest toxic effect on the foraminiferal host regarding reduced isotope uptake. In conclusion, all pesticides had a negative impact on the holosymbiont, with the host showing varying degrees of sensitivity towards different types of pesticides.

The change in metabolic activity of a large benthic foraminifera as a function of light supply.

We studied metabolic activity of the symbiont-bearing large benthic foraminifer Heterostegina depressa under different light conditions. Besides the overall photosynthetic performance of the photosymbionts estimated by means of variable fluorescence, the isotope uptake (13C and 15N) of the specimens (= holobionts) was measured. Heterostegina depressa was either incubated in darkness over a period of 15 days or exposed to an 16:8 h light:dark cycle mimicking natural light conditions. We found photosynthetic performance to be highly related to light supply. The photosymbionts, however, survived prolonged darkness and could be reactivated after 15 days of darkness. The same pattern was found in the isotope uptake of the holobionts. Based on these results, we propose that 13C-carbonate and 15N-nitrate assimilation is mainly controlled by the photosymbionts, whereas 15N-ammonium and 13C-glucose utilization is regulated by both, the symbiont and the host cells.

Spectroscopic analysis of sequestered chloroplasts in Elphidium williamsoni (Foraminifera).

Foraminifera are unicellular, marine organisms that occur worldwide. A very common species in the German Wadden Sea is Elphidium williamsoni. Some foraminifera (such as elphidia) are able to use kleptoplastidy, which allows them to incorporate chloroplasts from their algal food source into their own cell body. The experiments reported here are based on the fact that chlorophyll (a and c) can be detected in the intact cells with spectroscopic methods in the visible spectral range, which allows an indirect investigation of the presence of sequestered chloroplasts. Starving experiments of E. williamsoni in the light (24 h continuous) showed that the greatest decrease in chlorophyll content was recorded within the first 20-30 days. From day 60 on, chlorophyll was hardly detectable. Through subsequent feeding on a renewed algal food source a significant increase in the chlorophyll content in foraminifera was noticed. The degradation of chlorophyll in the dark (24 h continuous darkness) during the starving period was much more complex. Chlorophyll was still detected in the cells after 113 days of starving time. Therefore, we hypotheses that the effect of photoinhibition applies to chloroplasts in foraminifera under continuous illumination.

Photosynthetic performance of symbiont-bearing foraminifera Heterostegina depressa affected by sunscreens.

Foraminifera are abundant unicellular organisms that play an important role in marine element cycles. A large benthic foraminifer obligatory bearing photosymbionts is Heterostegina depressa. We studied potential impacts of sunscreens available on the market on the activity of photosymbionts on H. depressa by means of pulse-amplitude modulated (PAM) fluorescence microscopy. We included four different sunscreens, with two of them sold as "conventional" and two more stated as "eco-friendly". Further, the impact of pure Ensulizole (phenylbenzimidazole sulfonic acid) was tested, which is a common agent of sunscreens. Foraminifera were incubated at varying concentrations (10, 50 and 200 mgL-1) of different sunscreens and the pure Ensulizole for 14 days. The photosynthetic performance was measured after 1,3, 7 and 14 days. Pure Ensulizole had a strong negative impact on the photobionts, which was reflected by a significant reduction of the areal fluorescence signal. "Eco-friendly" sunscreens affected the health of foraminifera more severely compared to "conventional" ones. We assume that metal nanoparticles like titanium dioxide or zinc oxide of "eco-friendly" sunscreens are causing this impact, because these substances were already classified as toxic for several microorganisms.

Effects of heavy elements (Pb, Cu, Zn) on algal food uptake by Elphidium excavatum (Foraminifera).

Foraminifera are unicellular organisms and play a pivotal role in the marine material cycles. Past observations have shown that the species Elphidium excavatum is the most common foraminifera in the Baltic Sea. Feeding experiments showed that the food uptake and thus the turnover of organic matter are influenced by changes of physical parameters (e.g., temperature, salinity). Since many areas of the Baltic Sea are strongly affected by anthropogenic activity and are strongly contaminated by heavy elements from shipping in the past, this study examined the effect of heavy elements pollution on the food uptake of the most common foraminiferal species of the Baltic Sea, E. excavatum which was a subject of several previous studies. Therefore, Baltic Sea seawater was enriched with metals at various levels above normal seawater levels and the uptake of 13C- and 15N-labelled phytodetritus was measured by isotope ratio mass spectrometry. For each combination of metal type, concentration and time point 20 individuals of E. excavatum (three replicates) were fed with the green algae Dunaliella tertiolecta. The effect of dose parameters was measured in a two-way analysis of variance. Significant differences of food uptake were observable at different types and levels of heavy elements in sea water. Even a 557-fold increase in the Pb concentration did not affect food uptake, whereas strong negative effects were found for higher levels of Zn (144 and 1044-fold) and especially for Cu (5.6 and 24.3-fold). In summary it can be stated, that an increase in the heavy elements pollution in the Kiel Fjord will lead to a significant reduction in the turnover of organic matter by foraminifera such as E. excavatum.