Behavioral responses of brown shrimp (Crangon crangon) to reduced seawater pH following simulated leakages from sub-sea geological storage of CO2.

Large-scale storage of CO2 within sub-sea geological formations is a viable option for reducing the volume of this greenhouse gas released directly to the atmosphere from anthropogenic activities. Risks to benthic marine life following possible leakage of gas through the seabed from this carbon capture and storage (CCS) initiative are not yet well established. This study examined behavior (activity patterns) in brown shrimp (Crangon crangon), exposed to a range of reduced seawater pH conditions (7.6, 7, or 6.5) simulating leakage scenarios of varying scales. Brown shrimp have an endogenous rhythmicity associated with their activity, which dictates they are most active during hours of darkness, presumably as protection against vision-dependent predators. This endogenous rhythm in activity continues to be expressed when shrimp are held under constant low-light conditions in the lab and provides an ecologically relevant endpoint to measure when examining the influence of reduced pH on the behavior of these animals. No marked differences in activity pattern were observed between control shrimp maintained at pH 8.1 and those at pH 7.6. However, changes in activity were evident at pH 7 and pH 6.5, where significant shifts in timing and intensity of activity occurred. There was an unexpected increase in activity within periods of expected light, probably signaling efforts by shrimp to migrate away from reduced seawater pH conditions. The loss of this important member of the benthic community due to migration may have important consequences for many of the resilient species that remain.

Selenium reduces the retention of methyl mercury in the brown shrimp Crangon crangon.

Methyl mercury accumulated at the top of aquatic food chains constitutes a toxicological risk to humans and other top predators. Because the methyl mercury enters the aquatic food chains at the lower trophic levels, uptake and elimination processes at these levels affect the methyl mercury content at the higher levels. Selenium modulates the biokinetics of mercury in aquatic organisms in fairly complex ways, increasing mercury retention in some aquatic mammals, but decreasing methyl mercury retention in fish. However, it is not known if selenium modulates methyl mercury accumulation at lower trophic levels in aquatic food chains. Here, we show that selenium administered via the food augments the elimination of methyl mercury from marine shrimp and that the effect is dose-dependent, demonstrable down to natural selenium concentrations in aquatic food items. Selenite, seleno-cystine, and seleno-methionine exert this effect but selenate does not. Our results suggest that the selenium naturally present at the lower trophic levels in marine food chains may play an essential role as a modifier of methyl mercury accumulation at these levels, thereby potentially also affecting biomagnification of methyl mercury toward the higher trophic levels in the aquatic food chains.

The heterodimeric ecdysteroid receptor complex in the brown shrimp Crangon crangon: EcR and RXR isoform characteristics and sensitivity towards the marine pollutant tributyltin.

Decapod crustaceans are characterized by multiple ecdysteroid receptor (EcR) and retinoid-X-receptor (RXR) isoforms, which likely exhibit variant dimerization and transactivation interactions. In the brown shrimp C. crangon we cloned C-terminally truncated CrcEcR and CrcRXR isoforms and isoforms exhibiting deletions within the hinge region. For the former, in silico modeling of the CrcEcR indicated that, where the conserved helices H10 and H11 of the ligand-binding domain (LBD) are missing, an alternative C-terminal α-helix repairs the ligand-binding pocket (LBP). The truncated CrcRXR isoforms lack a major part of the LBD (H4-H12), thereby compromising ligand binding and dimerization. Through an in vitro ecdysteroid responsive reporter assay, we showed that these natural receptor variations do not impair receptor functioning but probably alter the receptor dimerization preferences. By the same in vitro assay, using full-length CrcEcR and CrcRXR, the effect of tributyltin (TBT) on ecdysteroid-induced transactivation was evaluated. The transactivation by 10nM PonA was reduced with 64% by 20 nM TBT. In silico modeling confirmed that TBT fits in the full-length CrcRXR-LBD. Furthermore, semi-quantitative PCR indicated altered expression of CrcEcR and CrcRXR isoforms after in vivo acute exposure to TBT, especially in the ovaries.

Coping with the "dirt": brown shrimp and the microplastic threat.

Microplastic pollution is an emerging threat to marine biota. Uptake of microplastics can impair nutrition and affect the performance of organisms. However, the vulnerability to microplastics seems to vary between species for yet widely unexplored reasons. We investigated the stomach content of the brown shrimp, Crangon crangon, from the southern North Sea and performed feeding experiments and anatomical studies of the digestive organs to comprehend the distribution of fluorescent microparticles within the shrimp. Shrimp collected in their natural environment contained between 51 and more than 3,000 sand grains and fragments of bivalve shells in their stomachs. Sand grains may have been ingested to exploit the associated biofilm or to support maceration of food. Bivalve shell fragments were particularly abundant in summer when shrimp fed on freshly settled mussels. Shrimps' stomach can be cleaned from ingested particles by regurgitation. In an experimental approach, we administered fluorescent microbeads of 0.1, 2.1, and 9.9 µm diameter. Only the smallest particles (0.1 µm) entered the midgut gland, which is the principal site of nutrient resorption in crustaceans. A fine-meshed chitinous filter system in the stomach of the shrimp prevents the passage of particles larger than about 1 µm. C. crangon appears well adapted to handle natural microscopic particles. This trait might also be advantageous in coping with microplastic pollution.

Occurrence of neonicotinoids and fipronil in estuaries and their potential risks to aquatic invertebrates.

This study aimed to evaluate and qualify field-based potential risks of seven neonicotinoid and phenylpyrazole (fipronil) insecticides on aquatic invertebrates, including estuary-resident marine crustaceans. One hundred and ninety-three estuarine water samples, with salinity ranging from 0.5 to 32.7, were collected from four estuarine sites in the Seto Inland Sea of Japan, in 2015-2018 and the insecticide levels were measured. Five neonicotinoid and fipronil insecticides were successfully identified, and their occurrence varied temporally. Marine crustaceans were simultaneously harvested every month from one of the estuarine water sampling sites in 2015-2017. Three predominant crustacean species, kuruma prawn (Penaeus japonicus), sand shrimp (Crangon uritai), and mysid (Neomysis awatschensis), were captured and their seasonal presence was species independent. A 96-h laboratory toxicity study with the insecticides using kuruma prawn, sand shrimp, and a surrogate mysid species (Americamysis bahia) indicated that fipronil exerted the highest toxicity to the three crustaceans. Using both toxicity data and insecticide occurrence in estuarine water (salinity ≥10, n = 169), the potential risks on the three marine crustaceans were quantified by calculating the proportion of mixture toxicity effects (Pmix). The Pmix of seven neonicotinoids on the crustaceans was less than 0.8%, which is likely to be too low to indicate adverse effects caused by the insecticides. However, short temporal detection of fipronil (exclusively in June and July) significantly affected the Pmix, which presented the maximal Pmix values of 21%, 3.4%, and 72% for kuruma prawn, sand shrimp, and mysid, respectively, indicating a significant effect on the organisms. As for estuarine water (salinity <10), some water samples contained imidacloprid and fipronil exceeding the freshwater benchmarks for aquatic invertebrates. The present study provides novel insights into the seasonally varying risks of insecticides to estuarine crustaceans and highlights the importance of considering whether ecological risk periods coincide with crustacean presence.

Environmental pollution and natural populations: a biomarkers case study from the Iberian Atlantic coast.

The degradation of estuaries is a result of human activities which overloads the environment with substances of both industrial and/or natural origins. Bioindicators have been consistently used to interpret effects of contaminants in the environment. In this study, the use of biomarkers (particular measurable characteristics of a bioindicator organism) was used to evaluate the contamination by xenobiotics of Crangon crangon natural populations. The central aim was to evaluate the capability of a battery of biomarkers to discriminate sites with different types of contamination. The activity of the enzymes cholinesterases (ChE), lactate dehydrogenase (LDH) and glutathione S-transferases (GST) were used as biomarkers. In addition, the ChE form(s) present in the cephalotorax of C. crangon were characterised. Organisms were seasonally sampled from winter 2001/2002 to autumn of 2002, at "reference" sites and at sites that receive agricultural, industrial and/or urban effluents. Results obtained in the characterisation of ChE with different substrates and selective inhibitors demonstrate that the form of ChE present in the cephalotorax of C. crangon shows proprieties of vertebrates' AChE and therefore it may be classified as true AChE-like ChE. The battery of biomarkers exhibited seasonal and local variations, apparently related to agricultural, industrial or urban effluent contamination. The tested biomarkers proved to be able to discriminate sources of environmental contamination, and confirms C. crangon as a sensitive species suitable to be used as a bioindicator.

Impact of metal pollution on shrimp Crangon affinis by NMR-based metabolomics.

Both cadmium and arsenic are the important metal/metalloid pollutants in the Bohai Sea. In this work, we sampled the dominant species, shrimp Crangon affinis, from three sites, the Middle of the Bohai Sea (MBS), the Yellow River Estuary (YRE) and the Laizhou Bay (LZB) along the Bohai Sea. The concentrations of metals/metalloids in shrimps C. affinis indicated that the YRE site was polluted by Cd and Pb, while the LZB site was contaminated by As. The metabolic differences between shrimps C. affinis from the reference site (MBS) and metal-pollution sites (YRE and LZB) were characterized using NMR-based metabolomics. Results indicated that the metal pollutions in YRE and LZB induced disturbances in osmotic regulation and energy metabolism via different metabolic pathways. In addition, a combination of alanine and arginine might be the biomarker of Cd contamination, while BCAAs and tyrosine could be the biomarkers of arsenic contamination in C. affinis.

The effects of power station entrainment passage on three species of marine planktonic crustacean, Acartia tonsa (Copepoda), Crangon crangon (Decapoda) and Homarus gammarus (Decapoda).

Experiments have been undertaken exposing larval common shrimp (Crangon crangon) and lobster (Homarus gammarus) and adult copepods (Acartia tonsa) to the key stresses of entrainment within power-station cooling-water systems. The apparatus has enabled the testing of mechanical, thermal, chlorine and realistic pressure effects both alone and in combination, the range of stressors spanning the standard conditions found within a temperate coastal direct-cooled power station. Mechanical stresses affected only lobster larvae, pressure changes affected only the Acartia adults. Residual chlorine caused significant mortality of Acartia and shrimp larvae, but had no effect on lobster larvae even at 1 ppm. The temperature increment significantly affected all three species, with a synergistic effect on chlorine sensitivity in the shrimp larvae, but only temperatures higher than would be experienced in a normally-operating power station affected the copepods. The majority of individuals of each species would survive passage through a power-station system under normal conditions. It is notable that, within the species tested, generalizations from the responses of one species to those of another are not valid.

Absence of Ca2+-induced mitochondrial permeability transition but presence of bongkrekate-sensitive nucleotide exchange in C. crangon and P. serratus.

Mitochondria from the embryos of brine shrimp (Artemia franciscana) do not undergo Ca(2+)-induced permeability transition in the presence of a profound Ca(2+) uptake capacity. Furthermore, this crustacean is the only organism known to exhibit bongkrekate-insensitive mitochondrial adenine nucleotide exchange, prompting the conjecture that refractoriness to bongkrekate and absence of Ca(2+)-induced permeability transition are somehow related phenomena. Here we report that mitochondria isolated from two other crustaceans, brown shrimp (Crangon crangon) and common prawn (Palaemon serratus) exhibited bongkrekate-sensitive mitochondrial adenine nucleotide transport, but lacked a Ca(2+)-induced permeability transition. Ca(2+) uptake capacity was robust in the absence of adenine nucleotides in both crustaceans, unaffected by either bongkrekate or cyclosporin A. Transmission electron microscopy images of Ca(2+)-loaded mitochondria showed needle-like formations of electron-dense material strikingly similar to those observed in mitochondria from the hepatopancreas of blue crab (Callinectes sapidus) and the embryos of Artemia franciscana. Alignment analysis of the partial coding sequences of the adenine nucleotide translocase (ANT) expressed in Crangon crangon and Palaemon serratus versus the complete sequence expressed in Artemia franciscana reappraised the possibility of the 208-214 amino acid region for conferring sensitivity to bongkrekate. However, our findings suggest that the ability to undergo Ca(2+)-induced mitochondrial permeability transition and the sensitivity of adenine nucleotide translocase to bongkrekate are not necessarily related phenomena.