Gill diseases in maricultured Atlantic salmon in Norway - results from ongoing projects

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Gill diseases in maricultured Atlantic salmon in Norway - results from ongoing projects Terje Steinum, Mona Gjessing, Duncan Colquhoun, Anne Berit Olsen, Kai- Inge Lie* and Anne-Gerd Gjevre *FishVetGroup

«PGI» = Proliferative gill inflammation Inflammation Epithelial hyperplasia Cell death (necrosis) Circulatory disturbances Chronic gill disease Kvellestad et al. 2004, Agnalt et al. 2004, Kvellestad et al. 2005

Chronic gill diseases A significant problem in Norway Problems occurre in late summer and autumn Stress triggers clinical disease High risk farms A multifactorial aetiology strongly suspected Our studies focus on four agents Epitheliocyst forming bacteia Ca. Branchiomonas cysticola Ca. Piscichlamydia salmonis Microsporidian Desmozoon lepethophterii Amoeba Paramoeba perurans Correlation agent:histological lesions Epidemiology: Defining on site risk factors

Outline Results the «Outbreak study» (suspected gill disease) a «Longitudinal study» of one farm «at risk» In situ hybridization Metagenomics Infection trial «B. cysticola» New project: «Studies of multiple factors in «challenge models»

Results from «Outbreak study» 2012-2013 Farm location and material size 2012 and 2013 Samples from 25 seawater sites 41 % in South 32 % in Mid 27 % in North North 600 real time PCR analyses Four agents detected 112 gill sections scored Mid South

Results from «Outbreak study» 2012-2013 Monthly distribution of collected samples 7 6 Number of sites 5 4 3 2 1 0 2013 2012 Month

Results from «Outbreak-study» 2012-213 Distribution of agents in 21 farms < 50% infected fish low load medium load high load Area Farm no. Branchiomonas cysticola % positive fish Desmozoon lepeophtherii % positive fish Piscichlamydia salmonis % postive fish Paramoeba perurans % positive fish 1 (1) 100 83 100 not detected 1 (2) 60 100 100 not detected 2 100 94 53 not detected Southern 3 (AGD) 100 100 not detected 100 Norway 4 (AGD) 60 100 80 70 5 (AGD) 100 100 50 80 6 100 100 17 not detected 7 93 97 90 not detected 8 not detected 100 not detected not detected 9 (AGD) 100 100 40 100 10 100 90 70 not detected 11 100 97 93 not detected 12 100 100 not detected not detected Mid 13 97 97 not detected not detected Norway 14 100 100 not detected 20 15 100 80 3 not detected 16 100 100 27 not detected 17 100 60 50 not detected 18 (1) 100 not detected not detected not detected Nothern 18 (2) 100 not detected 7 7 Norway 19 100 not detected 13 not detected 20 80 30 67 not detected 21 100 17 7 3

Results from a «longitudinal study» of Farm X at «high risk» Temporal distribution of agents Site X followed from June-December 2013 12 samplings of at least 20 fish = 240 fish examined No gill disease outbreak during the study period All 4 agents present, 3 already detected by the end of June < 45% infected fish low load medium load high load Farm X Sample date Branchiomonas cysticola % positive fish Desmozoon lepeopherii % positive fish Piscichlamydia salmonis % positive fish Paramoeba perurans % positive fish 05.06.2013 not detected 5 not detected not detected 21.06.2013 55 not detected not detected 5 10.07.2013 100 5 not detected not detected 01.08.2013 100 5 not detected not detected 21.08.2013 100 25 not detected not detected 27.08.2013 100 45 10 not detected 11.09.2013 100 80 25 not detected 01.10.2013 100 100 90 not detected 10.10.2013 95 100 100 5 24.10.2013 95 100 100 20 12.11.2013 95 100 100 10 03.12.2013 85 100 100 45

Metagenomics Small scale metagenomics on total DNA isolated from salmon with chronic gill disease 1/8 plate > 106 000 sequences Less than 1 % non-salmonid sequences Binning (via blast) reveal sequences most likely representing the epithelocyst forming bacteria and the microsporidian Other sequences suggest the presence of other respiratory microorganisms A stepping stone for a new gill study at the NVI?

Infection trial at VESO Vikan: Can «B. cysticola» be transferred between fish? «Farm X» Gills from 25 salmon Conclusion We could not confirm transfer of B. cysticola to or development of gill disease in experimental fish within the study period Homogenization 4 week trial Sampling Gill and kidney Bath infection - postsmolt sea water Intra-peritoneal infection parr fresh water

Funding «FHF 900800: Proliferative gill disease in sea farmed Atlantic salmon»

«Gill disease in Atlantic salmon studies of multiple factors in challenge models» April 2014 to March 2017 Partners NVI and NIVA Main goals is to..esablish laboatory cultures of selected microbes...establish a multifactorial «gill challenge model» systematize the pathophysiological, histopathological and immunological effects of selected factors

Funding NFR 233585: «Gill disease in Atlantic salmon studies of multiple factors in challenge models»

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