Veronique Creach

Laboratory and field validation of a simple method for detecting four species of non-native freshwater fish using eDNA

This paper presents the first phase in the development and validation of a simple and reliable environmental (e)DNA method using conventional PCR to detect four species of non-native freshwater fish: pumpkinseed Lepomis gibbosus, sunbleak Leucaspius delineatus, fathead minnow Pimephales promelas and topmouth gudgeon Pseudorasbora parva. The efficacy of the approach was demonstrated in indoor tank (44 l) trials in which all four species were detected within 24 h. Validation was through two field trials, in which L. gibbosus was detected 6-12 h after its introduction into outdoor experimental ponds and P. parva was successfully detected in disused fish rearing ponds where the species was known to exist. Thus, the filtration of small (30 ml) volumes of pond water was sufficient to capture fish eDNA and the approach emphasised the importance of taking multiple water samples of sufficient spatial coverage for detecting species of random or patchy distribution.

Comparing flow cytometry and microscopy in the quantification of vital aquatic organisms in ballast water

ABSTRACT The ability to quantify vital aquatic organisms in the 2–50 µm size range was compared between five different flow cytometers and several different microscopes. Counts of calibration beads, algal monocultures of different sizes as well as organisms in a Wadden Sea sample were compared. Flow cytometers and microscopes delivered different bead concentrations. These differences between the instruments became larger for algal monocultures and were even higher for the Wadden Sea sample. It was observed that the concentration differences were significant between flow cytometer and microscope counts, and that this difference increased with the size of the objects counted. Microscope counts were more accurate for larger (50 µm) objects because cytometers struggled with bigger particles that clogged the instruments. Contrary to microscopy, the flow cytometers were capable of accurately enumerating cultured cells in the 2–10 µm size range and cells in the lower size range of the 10–50 µm size class. Flow cytometers were also well-suited to assess low abundance samples due to their ability to process larger volumes than microscopes. The results were used to indicate which tools are suitable for ballast water monitoring: flow cytometry is a suitable technology for an indicative and real time analysis of ballast water samples whilst only microscopy would be robust enough for detailed taxonomical analyses.

Harmful algal blooms and climate change: exploring future distribution changes

Harmful algae can cause death in fish, shellfish, marine mammals, and humans, via their toxins or from effects associated with their sheer quantity. There are many species, which cause a variety of problems around north-west Europe, and the frequency and distribution of algal blooms have altered in the recent past. Species distribution modelling was used to understand how harmful algal species may respond in the future to climate change, by considering environmental preferences and how these may shift. Most distribution studies to date use low resolution global model outputs. In this study, high resolution, downscaled shelf seas climate projections for the north-west European shelf were nested within lower resolution global projections, to understand how the distribution of harmful algae may change by the mid to end of century. Projections suggest that the habitat of most species (defined by temperature, salinity, depth, and stratification) will shift north this century, with suitability increasing in the central and northern North Sea. An increase in occurrence here might lead to more frequent detrimental blooms if wind, irradiance and nutrient levels are also suitable. Prioritizing monitoring of species in these susceptible areas could help in establishing early-warning systems for aquaculture and health protection schemes.

Chlorine toxicity to Navicula pelliculosa and Achnanthes spp. in a flow-through system: The use of immobilised microalgae and variable chlorophyll fluorescence

Chlorination is a widely used antifouling method for freshwater and marine applications. Chlorine added to seawater reacts to form oxidants that are toxic to biofouling organisms. Further, the oxidants that result are short-lived, but may nevertheless affect non-target species in waterbodies receiving the antifouling effluent. This study evaluated the toxicity of chlorinated seawater (e.g. following sodium hypochlorite addition) on two different species of marine benthic diatoms (Achnanthes spp., and Navicula pelliculosa), which are representative of microphytobenthos communities - an important component in coastal habitats that may be exposed to chlorinated seawater. To evaluate the growth inhibition over a 72 h period, algae were immobilised in alginate beads and exposed to different levels of chlorination in a flow through system. Growth rates and physiological condition of the microalgae were evaluated using a Fast Repetition Rate fluorometer (FRRf). To determine whether alginate influenced the sensitivity of algal response, studies were also conducted in a static test system (without renewal of test solutions) using both free cells and immobilised cells with initial chlorine added to achieve a similar range of concentrations as those used in the flow-through study. Within the first hour of the exposure period there was an indication that, for both species, the free algal cells in the static system were more sensitive to exposure to chlorinated seawater than were alginate-immobilised cells in the flow through system. Immobilised cells in a static system with a single addition of chlorine were also less sensitive to chlorination than free algal cells. However, for periods of 24 h or more due to decay of TRO in the static system the exposure of immobilised algae in the flow through system had a greater impact and hence lower effect concentrations. For the flow-through studies Achnanthes spp. was the most sensitive after 72 h exposure with a potential no effect concentration EC10 value of 0.02 mg l-1 as Cl2 equivalents expressed as total residual oxidants (TRO) compared 0.04 mg l-1 TRO for N. pelliculosa. Immobilisation of algal cells in alginate was found to be an effective means of determining the impact of chlorination and is likely to be effective for other non-persistent substances. Based on the data produced, the extent and significance of ecological effects of chlorination upon algal species typical of microphytobenthos are likely to be limited providing discharges comply with a maximum allowable concentration of 0.01 mg l-1 TRO at the edge of an agreed mixing zone.