Campbell C. Pert

RNA-interference methods for gene-knockdown in the sea louse, Lepeophtheirus salmonis: studies on a putative prostaglandin E synthase.

Harnessing the full utility of extensive gene sequences recently available for the economically important sea louse, Lepeophtheirus salmonis, requires the adaptation of modern molecular biology approaches to this non-model organism. Using a putative microsomal prostaglandin E synthase type-2 (PGES2) as a candidate gene, we investigated gene-knockdown by double-stranded RNA interference (dsRNAi) in the small free-living and the larger parasitic stages of L. salmonis. dsRNA was administered to nauplius and copepodid stages by immersion for 7 h. Pre-adult and adults received dsRNA by intra-haemocoelic injection. The extent, speed and persistence of the knockdown effects were determined by RT-PCR. LsPGES2 was abundantly expressed in all life stages, including the non-parasitic stages. Administration of dsRNA to nauplius and copepodids by immersion had no effect on mortality rates and moulting through to copepodids was observed. Dramatic knockdown of LsPGES2 was observed within 7 h and persisted for at least 48 h. Injection of dsRNA had no effect on mortality in pre-adults and adults, but knockdown of LsPGES2 was apparent within 24 h, reaching 95% over the 72 h and was persistent for at least 120 h. The methods developed resulted in rapid and persistent knockdown in L. salmonis suitable for studies in the different stadia.

A survey of wild marine fish identifies a potential origin of an outbreak of viral haemorrhagic septicaemia in wrasse, Labridae, used as cleaner fish on marine Atlantic salmon, Salmo salar L., farms

Viral haemorrhagic septicaemia virus (VHSV) was isolated from five species of wrasse (Labridae) used as biological controls for parasitic sea lice predominantly, Lepeophtheirus salmonis (Krøyer, 1837), on marine Atlantic salmon, Salmo salar L., farms in Shetland. As part of the epidemiological investigation, 1400 wild marine fish were caught and screened in pools of 10 for VHSV using virus isolation. Eleven pools (8%) were confirmed VHSV positive from: grey gurnard, Eutrigla gurnardus L.; Atlantic herring, Clupea harengus L.; Norway pout, Trisopterus esmarkii (Nilsson); plaice, Pleuronectes platessa L.; sprat, Sprattus sprattus L. and whiting, Merlangius merlangus L. The isolation of VHSV from grey gurnard is the first documented report in this species. Nucleic acid sequencing of the partial nucleocapsid (N) and glycoprotein (G) genes was carried out for viral characterization. Sequence analysis confirmed that all wild isolates were genotype III the same as the wrasse and there was a close genetic similarity between the isolates from wild fish and wrasse on the farms. Infection from these local wild marine fish is the most likely source of VHSV isolated from wrasse on the fish farms.

Factors influencing the long-term dynamics of larval sea lice density at east and west coast locations in Scotland

Sea lice (Copepoda: Caligidae) are marine copepods that parasitize finfish, and in cases of high infestation can result in severe epithelial damage and mortality. In Scotland, 2 species of sea louse, Lepeophtheirus salmonis and Caligus elongatus, pose a significant economic burden to the marine Atlantic salmon aquaculture industry and potentially impact wild salmonids. The purpose of this study was to determine how the density of pelagic sea lice is affected by external variables, in order to improve our understanding of sea lice dynamics. Long-term data from 2 sampling sites on the east and west coasts of Scotland were modelled independently in conjunction with environmental and anthropogenic variables. Statistical analysis identified that at the east coast site, the most influential factor affecting lice density was salinity. On the west coast, salinity, rainfall and farmed salmon production year were most influential. Molecular and morphological techniques also showed that the individuals recorded on the east coast were C. elongatus, a generalist copepod parasite, whereas only the salmonid-specific L. salmonis were found on the west. These results reiterate the role of environmental factors in influencing sea lice dynamics, and that salmonids are the primary hosts of sea lice on the west coast, but there could be non-salmonid host species as well as salmonid species influencing east coast sea lice densities.

Developing a method to investigate infestation pressure from sea lice on migratory salmonids utilising towed & static sentinel cages

A number of studies have linked the presence of fish farms to sea lice in the environment (e.g. Harte et al 2017) and infestations of wild salmonids (Middlemas et al. 2010 Vollset et al. 2016a) and subsequent population dynamics in marine aquaculture areas (V⊘llestad et al. 2009; Vollset et al. 2016a). Although there are studies looking at the effect of migration behaviour on mortality (e.g. Vollset et al. 2016b), no study has ever been carried out to design and deploy equipment to measure the specific infestation pressure salmon smolts may experience from sea lice as they enter the marine environment and migrate down a sea loch towards the open sea in a region containing Atlantic salmon aquaculture.