Assessing indicators of arsenic toxicity using variable fluorescence in a commercially valuable microalgae: Physiological and toxicological aspects.

Indicators signaling Arsenic (As) stress through physiology of microalgae using non-destructive methods like variable fluorescence are rare but requisite. This study reports stress markers indicating arsenic (As) toxicity (in two concentrations 11.25 µg/L and 22.5 µg/L compared to a control) exposed to a microalga (Diacronema lutheri), using fast repetition rate fluorometry (FRRf). Growth and physiological parameters such as cell density, chl a and the maximum quantum yield Fv/Fm showed coherence and impeded after the exponential phase (day 9 - day 12) in As treatments compared to the control (p < 0.05). On contrary photo-physiological constants were elevated showing higher optical (aLHII) and functional [Sigma (σPSII)] absorption cross-section for the As treatments (p < 0.05) further implying the lack of biomass production yet an increase in light absorption. In addition, As exposure increased the energy dissipation by heat (NPQ-NSV) showing a strong relationship with the de-epoxidation ratio (DR) involving photoprotective pigments. Total As bioaccumulation by D. lutheri showed a strong affinity with Fe adsorption throughout the algal growth curve. This study suggests some prompt photo-physiological proxies signaling As contamination and endorsing its usefulness in risk assessments, given the high toxicity and ubiquitous presence of As in the ecosystem.

Effects of different routes of exposure to metals on bioaccumulation and population growth of the cyclopoid copepod Paracyclopina nana.

We examined effects of the three metals cadmium (Cd), copper (Cu), and nickel (Ni) on two subpopulations of the cyclopoid copepod Paracyclopina nana. We sought to investigate the effects of metal exposure on population growth and structure of P. nana and to understand the parameters affecting the metal bioaccumulation in copepods. A first experiment tested the hypothesis of competition between these metals in a mixture using a P. nana mass culture in 10 L beakers with the sublethal concentrations (1/3 of LC50) as determined for E. affinis. A second experiment pursued the same with a P. nana population which was adapted to a higher Cu concentration for several generations (226.9 ± 15.9 µg g-1 dw Cu in copepods) and using the proper sublethal concentrations for P. nana. After 96 h of exposure, results from the first experiment showed a decreasing population growth and instead of an increasing metal accumulation in copepods. Cd also appeared to be more accumulated when it was alone, confirming the hypothesis of metal competition in mixture. Results from the second experiment revealed less marked effects. When metal concentrations increased in the treatment it decreased in copepods, indicating depuration activity in the population already adapted to metal exposure. This paper is the first one investigating the parameters affecting the bioaccumulation capacity of P. nana in response to metals. It offers a better understanding of copepod responses to metal contamination in a complex aquatic environment.

Population response of the estuarine copepod Eurytemora affinis to its bioaccumulation of trace metals.

We evaluated the acute toxicities of metals cadmium (Cd), copper (Cu) and nickel (Ni) to a widely-distributed copepod Eurytemora affinis isolated from the Seine estuary. Both sexes of adult E. affinis were exposed separately to the three metals at concentration gradients to determine its 50% lethal concentration (LC50). After 4 days of exposure, both males and females showed a higher sensitivity to Cu (male LC50: 25.0 µg.L-1 and female LC50: 38.0 µg.L-1) than to Ni (male LC50: 90.0 µg.L-1 and female 161.0 µg.L-1) and Cd (male LC50: 127.8 µg.L-1 and female LC50: 90.0 µg.L-1). To assess for the first time, the extend of metal bioaccumulation and its effect at population scale, late stages (>200 µm) were collected and exposed to each metal at the concentration of 1/3 LC50, and to their mixture during 144 h without feeding. The Cd concentration consistently increased with time until the end of the experiment, whereas the Ni and Cu concentrations reached a plateau after 24 h and 72 h exposure, respectively. The results revealed that the copepods could accumulate Cu faster than Ni and Cd either in the treatment alone (0.58 L g-1.d-1) or in the three-metal mixture (0.72 L g-1.d-1) after 50% of exposure time (72 h). The number of individuals decreased in copepod populations except for the Cd treatment, where the number of nauplii increased. In addition, all treatments of metal exposure negatively affected bacterial densities in the copepod cultures, where the Cu treatment showed a negative remarkable effect compared with Cd and Ni treatment did.

Differences in lethal response between male and female calanoid copepods and life cycle traits to cadmium toxicity.

This study determined the effect of cadmium (Cd) toxicity comparatively on two copepods, Eurytemora affinis (Poppe 1880) from a temperate region (Seine Estuary, France) and Pseudodiaptomus annandalei (Sewell 1919) from a subtropical region (Danshuei Estuary, Taiwan), according to their sex and reproductive stages. In addition, the effect of Cd to their life cycle traits was quantified. In the first experiment, both copepod sexes were exposed to 40, 80, 150, 220, and 360 µg/L of Cd and a control cultured in salinity 15, except that the temperature was 18 °C for E. affinis and 26 °C for P. annandalei. This allowed calculating median lethal concentration (LC50) of Cd after 96 h. This was 120.6 µg/L Cd for P. annandalei males which were almost twice as sensitive as P. annandalei females (LC50 = 239.5 µg/L Cd). For E. affinis females, the LC50 was 90.04 µg/L Cd, reflecting a 1.4 times higher sensitivity of females than of males (LC50 = 127.75 µg/L Cd). The males of both species were similarly sensitive; however, the E. affinis females were 2.7 times more sensitive than the P. annandalei females. We also compared the sensitivity of ovigerous females (OVF) and non-ovigerous females (NOF) of both species to Cd. Mortality was higher in NOF than in OVF of both copepod species in both the control and the 40 µg/L Cd treatment. Finally, the total population, fecundity and female morphology of both copepod species were estimated after exposing one generation cycle (nauplius to adult) to 40 µg/L Cd (for E. affinis) and 160 µg/L Cd (for P. annandalei). A significant decrease in cohort production, survival and clutch size but no significant difference in the prosome length of both copepod species exposed to Cd were detected. The ratio of OVF:NOF was high in both copepod species exposed to Cd. Cd toxicity did not significantly affect the M:F sex ratio and % OVF of E. affinis. However, the effect of Cd toxicity in P. annandalei was significant in the M:F sex ratio and was in favor of females and their reproductive activities due to an increase in % OVF. Moreover, there was a significant decrease in total production of P. annandalei due to high mortality in their nauplii and copepodid developmental stages. Toxicity to Cd appears to be affected by multiple factors including sex, reproductive life stage and species. The ecological implication of Cd toxicity on E. affinis and P. annandalei copepod ecology is more related to a skewed sex ratio, low egg production, reduced hatchability and reduced survival that affects the recruitment potential of the copepod nauplii resulting in a decreasing copepod population. Mortality, reproduction and population growth of model species may provide important bio-indicators for environmental risk assessment.