European seabass respond more strongly to noise exposure at night and habituate over repeated trials of sound exposure.

Aquatic animals live in an acoustic world, prone to pollution by globally increasing noise levels. Noisy human activities at sea have become widespread and continue day and night. The potential effects of this anthropogenic noise may be context-dependent and vary with the time of the day, depending on diel cycles in animal physiology and behaviour. Most studies to date have investigated behavioural changes within a single sound exposure session while the effects of, and habituation to, repeated exposures remain largely unknown. Here, we exposed groups of European seabass (Dicentrarchus labrax) in an outdoor pen to a series of eight repeated impulsive sound exposures over the course of two days at variable times of day/night. The baseline behaviour before sound exposure was different between day and night; with slower swimming and looser group cohesion observed at night. In response to sound exposures, groups increased their swimming speed, depth, and cohesion; with a greater effect during the night. Furthermore, groups also showed inter-trial habituation with respect to swimming depth. Our findings suggest that the impact of impulsive anthropogenic noise may be stronger at night than during the day for some fishes. Moreover, our results also suggest that habituation should be taken into account for sound impact assessments and potential mitigating measures.

Influence of DNA isolation from historical otoliths on nuclear-mitochondrial marker amplification and age determination in an overexploited fish, the common sole (Solea solea L.).

Historical otolith collections are crucial in assessing the evolutionary consequences of natural and anthropogenic changes on the demography and connectivity of commercially important fish species. Hence, it is important to define optimal protocols for purifying DNA from such valuable information sources while avoiding any damage to the physical structure of the otolith. Before being able to conclude on the harmlessness of a method, it is important to validate protocols on different kinds of otoliths by testing purification methodologies under standardized conditions. Here we compare the effect of two DNA extraction methods on the success in identifying the age in an overexploited marine fish, the common sole (Solea solea L.). To ensure optimal future population genetic and demographic analyses, we assessed DNA quantity and tested the DNA quality by investigating the amplification success of a mitochondrial and nuclear marker. Our results show that the choice of the DNA extraction method had a significant effect on the success of using these otoliths in age and growth analyses. Standard commercial and published protocols resulted in a severe damaging of the otolith structure, hampering accurate preparation and analyses of the morphological structures of the otoliths. Shortening the lysis time and lowering the EDTA (ethylene diamine tetraacetic acid) and SDS (sodium dodecylsulphate) concentration turned out to be beneficial for the stability of otolith structure, while maintaining an overall high DNA quality measured through polymerase chain reaction amplification success. We therefore recommend that care should be taken when choosing the extraction method for a molecular study on archived samples, in order to enable the maximal use of information embedded in historical material.