Amoebic gill disease (AGD) is a proliferative gill disease of marine cultured Atlantic salmon Salmo salar, with the free-living protozoan Neoparamoeba perurans being the primary aetiological agent. The increased incidence of AGD in recent years presents a significant challenge to the Atlantic salmon farming industry in Europe. In this study, a real-time TaqMan ® PCR assay was developed and validated to detect Neoparamoeba perurans on Atlantic salmon gills and further used to monitor disease progression on a marine Atlantic salmon farm in Ireland in conjunction with gross gill pathology and histopathology. The assay proved specific for N. perurans, with no cross-reactivity with the related species N. pemaquidensis, N. branchiphila or N. aestuarina, and was capable of detecting 2.68 copies of N. perurans DNA µl −1. Although the parasite was detected throughout the 18 mo period of this study, mortality peaks associated with clinical AGD were only recorded during the first 12 mo of the marine phase of the production cycle. The initial AGD outbreak resulted in peak mortality in Week 17, which was preceded by PCR detections from Week 13 onwards. Freshwater treatments were an effective method for controlling the disease, resulting in a reduction in the weekly mortality levels and also a reduction in the number of PCR-positive fish. In comparison to traditional diagnostic methods, our PCR assay proved to be highly sensitive and a valuable tool to monitor disease progression and, therefore, has the potential to provide information on the timing and effectiveness of treatments.

Gill diseases are a complex and multifactorial challenge for marine farmed Atlantic salmon. Co-infections with putative pathogens are common on farms; however, there is a lack of knowledge in relation to the potential effect co-infections may have on pathology. The objective of this study was to determine the prevalence and potential effects of Neoparamoeba perurans, Desmozoon lepeophtherii, Candidatus Branchiomonas cysticola, Tenacibaculum maritimum and salmon gill poxvirus (SGPV) during a longitudinal study on a marine Atlantic salmon farm. Real-time PCR was used to determine the presence and sequential infection patterns of these pathogens on gill samples collected from stocking until harvest. A number of multilevel models were used to determine the effect of these putative pathogens on gill health (measured as gill histopathology score), while adjusting for the effect of water temperature and time since the last freshwater treatment. Results indicate that between 12 and 16 weeks post-seawater transfer (wpst), colonization of the gills by all pathogens had commenced and by week 16 of marine production each of the pathogens had been detected. D. lepeophtherii and Candidatus B. cysticola were by far the most prevalent of the potential pathogens detected during this study. Detections of T. maritimum were found to be significantly correlated with temperature showing distinct seasonality. Salmon gill poxvirus was found to be highly sporadic and detected in the first sampling point, suggesting a carryover from the freshwater stage of production. Finally, the model results indicated no clear effect between any of the pathogens. Additionally, the models showed that the only variable which had a consistent effect on the histology score was N. perurans.

Gill health is fundamental to the success of finfish aquaculture. These delicate organs are responsible for gas exchange, acid-base balance, waste excretion and osmoregulation, yet are constantly exposed to a variety of microorganisms and environmental insults which can result in various gill diseases (Evans 2005, Mitchell & Rodger 2011). While the aetiologies of many disease pathologies are attributed to single pathological agents, the drivers of gill disease can be more complex (Gjessing et al. 2017, Herrero et al. 2018). A multifactorial approach to disease occurrence that considers the impact of environmental factors and microbial community structure, rather than focusing on a single agent, may therefore be a ra

Amoebic gill disease (AGD) caused by Neoparamoeba perurans, has emerged in Europe as a significant problem for the Atlantic salmon farming industry. Gross gill score is the most widely used and practical method for determining AGD severity on farms and informing management decisions on disease mitigation strategies. As molecular diagnosis of AGD remains a high priority for much of the international salmon farming industry, there is a need to evaluate the suitability of currently available molecular assays in conjunction with the most appropriate non-destructive sampling methodology. The aims of this study were to assess a non-destructive sampling methodology (gill swabs) and to compare a range of currently available real-time polymerase chain-reaction (PCR) assays for the detection of N. perurans. Furthermore a comparison of the nondestructive molecular diagnostics with traditional screening methods of gill scoring and histopathology was also undertaken. The study found that all molecular protocols assessed performed well in cases of clinical AGD with high gill scores. A TaqMan based assay (protocol 1) was the optimal assay based on a range of parameters including % positive samples from a field trial performed on fish with gill scores ranging from 0 to 5. A higher proportion of gill swab samples tested positive by all protocols than gill filament biopsies and there was a strong correlation between gill swabs tested by protocol 1 and gross gill score and histology scores. Screening for N. perurans using protocol 1 in conjunction with non-destructive gill swab samples was shown to give the best results.