Assessment of wastewater derived pollution using viral monitoring in two estuaries.

Human wastewater-derived pollution of the environment is an emerging health risk that increases the number of waterborne and foodborne illnesses globally. To better understand and mitigate such health risks, we investigated the prevalence of faecal indicator bacteria, Escherichia coli, and indicator virus (crAssphage) along with human and animal enteric viruses (adenoviruses, noroviruses, sapoviruses, hepatitis E virus) in shellfish and water samples collected from two shellfish harvesting areas in the UK. Human noroviruses were detected at higher detection rates in oyster and water samples compared to mussels with peaks during the autumn-winter seasons. Human enteric viruses were sporadically detected during the warmer months, suggesting potential introduction by tourists following the relaxation of COVID-19 lockdown measures. Our results suggest that viral indicators are more suitable for risk assessment and source tracking than E. coli. The detection of emerging hepatitis and sapoviruses, support the need for comprehensive viral monitoring in shellfish harvesting areas.

Geographical variation in the carbon, nitrogen, and phosphorus content of blue mussels, Mytilus edulis.

Shellfish farming contributes to nutrient removal in coastal and estuarine systems, as bivalves incorporate nutrients into their tissues and shells, which is removed from the marine system on harvest. Fourteen locations around the UK were surveyed to explore geographic variation in carbon, nitrogen and phosphorus content of tissue and shell in blue mussels. Phosphorus in tissue had a significant negative relationship with mean annual seawater temperature for both rope and bottom cultured sites. Per tonne of live mussel, rope culture removed significantly more nitrogen (8.50 ± 0.59 kg) and phosphorus (0.95 ± 0.07 kg) than bottom cultured (5.00 ± 0.013 kg nitrogen and 0.43 ± 0.01 kg phosphorus). Bottom culture, however, provides significantly more C removal in shell (60.15 ± 0.77 kg) than in rope cultured (46.12 ± 1.69 kg). Further studies are required to examine the effect of growth rate, on the nitrogen and phosphorus remediation, and carbon stored in shell, of rope culture and bottom cultured mussel aquaculture.

Individual and mixture effects of selected pharmaceuticals on larval development of the estuarine shrimp Palaemon longirostris.

Few ecotoxicological studies incorporate within the experimental design environmental variability and mixture effects when assessing the impact of pollutants on organisms. We have studied the combined effects of selected pharmaceutical compounds and environmental variability in terms of salinity and temperature on survival, development and body mass of larvae of the estuarine shrimp Palaemon longirostris. Drug residues found in coastal waters occur as mixture, and the evaluation of combined effects of simultaneously occurring compounds is indispensable for their environmental risk assessment. All larval stages of P. longirostris were exposed to the nonsteroidal anti-inflammatory drug (NSAID) diclofenac sodium (DS: 40 and 750 µg L(-1)), the lipid regulator clofibric acid (CA: 17 and 361 µg L(-1)) and the fungicide clotrimazole (CLZ: 0.14 and 4 µg L(-1)). We observed no effect on larval survival of P. longirostris with the tested pharmaceuticals. However, and in contrast to previous studies on larvae of the related marine species Palaemon serratus, CA affected development through an increase in intermoult duration and reduced growth without affecting larval body mass. These developmental effects in P. longirostris larvae were similar to those observed in the mixture of DS and CA confirming the toxic effects of CA. In the case of CLZ, its effects were similar to those observed previously in P. serratus: high doses affected development altering intermoult duration, tended to reduce the number of larval instars and decreased significantly the growth rate. This study suggests that an inter-specific life histories approach should be taken into account to assess the effect of emergent compounds in coastal waters.

Effects of food limitation and pharmaceutical compounds on the larval development and morphology of Palaemon serratus.

Few ecotoxicological studies consider the roles of maternal influences and suboptimal environmental conditions when assessing the impact of pollutants on organisms. We studied the combined effects of pharmaceutical compounds, food condition and maternal body size on growth, development, body mass and morphology of larvae of the marine shrimp Palaemon serratus. Limited food availability is considered a factor leading to reduced survival and growth in marine crustacean larvae. It is known that P. serratus responses to food limitation vary among larvae hatched from females of different body length. The pharmaceuticals tested were the anti-inflammatory and analgesic diclofenac sodium (DS: at 77 µg L-1 and 720 µg L-1) the lipid regulator clofibric acid (CA: at 42 µg L-1 and 394 µg L-1) and the fungicide clotrimazole (CLZ: at 0.07 µg L-1 and 3.16 µg L-1). We observed morphological abnormalities in larvae exposed to CLZ. In addition, effects of this compound were stronger under food limitation leading to (1) reduced survival by 30%, (2) reduced juvenile body mass (22%) and (3) reduction in the number of molt stages (from 13 to 9) during larval development. This latter effect may indicate that CLZ reduced the larval capacity to respond to food limitation because development through a longer route, with additional stages, is considered an adaptive response to prioritize maintenance over morphogenesis. CA and DS affected developmental rate under food limitation but not growth or body mass. The toxic effects of CLZ, at lower concentrations than CA and DS, were stronger in larvae with higher body mass, hatched from the largest females. This suggests that maternal influences and suboptimal environmental conditions should be further studied to inform modeling of the effects of emergent compounds on larvae of marine coastal species.

Effect of different diets on proteolytic enzyme activity, trypsinogen gene expression and dietary carbon assimilation in Senegalese sole (Solea senegalensis) larvae.

The effect of diet on larval growth, anionic trypsinogen gene expression (ssetryp1), and trypsin (EC 3.4.21.4) and chymotrypsin (EC 3.4.21.1) activities was assessed in Solea senegalensis. Changes in larval carbon stable isotope (δ(13)C) composition were used to estimate carbon assimilation. Diets were supplied for 20days to fish held in larval rearing tanks and consisted of live rotifers, Artemia sp. nauplii, rotifers followed by Artemia sp., rotifers co-fed with inert diet and inert diet alone. Growth was significantly faster in larvae fed only Artemia and those fed rotifers and Artemia (k=0.381-0.387day(-1)). Trypsin and chymotrypsin activities increased from 3 to 4days after hatching (DAH) in all dietary treatments, while ssetryp1 transcripts increased at 4-5 DAH only in larvae fed live prey. ssetryp1 gene expression was activated later in larvae fed only Artemia and this corresponded with Artemia δ(13)C values being reflected in larval tissue. Larval δ(13)C values also indicated greater selection and/or assimilation of rotifers in relation to the inert diet. Results demonstrate that during early larval development of sole, diet modulates ssetryp1 gene expression. The rapid and intense response to diets that promoted different growth and survival suggests the suitability of this biomarker as a nutritional status indicator in early sole larvae.