INTERREG, HEALFISH , WP2, Genetic studies. Marja-Liisa Koljonen Finnish Game and Fisheries Research Institute

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INTERREG, HEALFISH 2010-2013, WP2, Genetic studies Marja-Liisa Koljonen Finnish Game and Fisheries Research Institute 16.10.2014 1

Genetic structure of Finnish and Russian brown trout populations in the Gulf of Finland area Marja-Liisa Koljonen, Aki Janatuinen, Jarmo Koskiniemi and Ari Saura 16.10.2014 2

DNA samples Samples from 34 river systems draining into the Gulf of Finland (+ 5 hatchery stock samples), over 70 individual samples and 3430 fish in Finnish and Russia river data 709 catch samples, three sea areas 11 estonian stocks added to the baseline from the Estonian data sets Analysis of 16 DNA microsatellites 16.10.2014 3

Sampled rivers and their state - this presentation 4

Diversity levels Number of alleles: 27, Purilanjoki 182, Danish hatchery Allelic richness: mean 4.7 /11 individuals, 6.2 /30 individuals 1.7 Purilanjoki, 2.4 Mustijoki 7.4 Denmark, 6.2 Urpalanjoki Mean diversity: 0.62 0.22 Purilanjoki, Punassuon Lohioja 0.39 0.77 Denmark, Urpalanjoki In Finnish populations: Over 6 alleles on average mixed populations: Fiskarsinjoki, middle and upper Vantaanjoki, Summanjoki upper reaches. Over 5 alleles: Aurajoki, Kiskonjoki-Latokartanonkoski, Kiskonjoki- Perniönjoki, Karjaanjoki-Nummenjoki, Mankinjoki, low Vantaanjoki, Kymijoki, Summanjoki main branch. 5

Diversity levels for areas Comparison between areas: Diversity tend to increase towards east. South west significantly lower. N all All Rich for 11 ind. All Rich for 30 ind. Mean DIV South west 81,6 4,1 5,7 0,55** Uusimaa 99,6 4,6 5,8 0,63 South east 104,2 4,9 5,7 0,64 Border rivers 90,3 4,4 6,3 0,61 Russian rivers 107,9 5,2 6,6 0,65 6

Relatedness Ne/N: mean 0.77 (range 0-2) (0.16 Upper Vantaanjoki 1.91 Kuokkalanpuro, RUS) Number of Families in samples; mean 40 (<10, Kilpeenjoki 5, Purilanjoki 6, Karjaanjoki-Mustionjoki 8, 258 Mustajoki) 50% siblings, 25% halfsibs, 12,5% first cousin, 3% second cousin. Mean relatedness; 6% (16,7% Purilanjoki 2.2% Kiskonjoki, Perniönjoki, 3,2% Kuokkalanpuro RUS) Over 10%: Purilanjoki 16.7, Kisko-Perniönjoki 12.4, Karjaa- Mustionjoki 10.5, Espoo-Ryssänoja 14.1, Mustijoki 10.7. Vilajoki 10.4. 7

Genetic differentiation among populations No differences at all (same origin) four river pairs Aurajoki Fiskarsinjoki (Fst 0.01), Ingarskilanjoki Koskenkylänjoki (Fst 0.01), Vantaanjoki_Palojoki Koskenkylänjoki (Fst 0.01) and Isojoki Kymijoki (Fst 0.01) Very high similarity also: Mankinjoki Low Vantaanjoki (Fst = 0.02) Low Vantaanjoki Middle Vantaanjoki (Fst = 0.02) Summanjoki-Kymijoki, Isojoki Summanjoki Main branch Kymijoki Summanjoki (Fst = 0.02), and Inojoki Kuokkalanpuro (Fst = 0.02) the only wild pops. 8

Within rivers system differentiation Riversystems: Kiskonjoki 5; Latokartanonkoski, Myllyjoki, Kisko-Koorla- Resident, Perniönjoki anadromous, Perniönjoki - Metsänoja Karjaanjoki 5; Mustionjoki, Nummenjoki, Nuijajoki, Vihtijoki, Saavajoki Siuntionjoki 2; Kirkkojoki and Passilankoski Espoonjoki 2 ; Main branch and Ryssänniitynoja Vantaanjoki 4; Low, Middle, Upper and Palojoki Summanjoki 2; Main branch and Upper reaches (very similar) Virojoki 2; Upper reaches and anadromous Saarasjärvenoja Only one: Uskelanjoki, Mankinjoki, 9

Structure of Vantaanjoki brown trout populations Tributary 4 Luhtajoki 2 36 84 98 Tributary 3 Lepsämänjoki Upper reaches Käräjäkoski Upper reaches Toronmäenkoski 98 Tributary 5 Epranoja 4 99 35 87 32 36 Middle Nukarinkoski Low Middle Nukarinkoski Mid Lower Vantaankoski Lower Pitkäkoski Tributary 1 Longinoja Lower Ruutinkoski 3 0.1 43 Tributary 2 Palojoki Rannikonmäki 96 Tributary 2 Palojoki Juvankoski 10

3 Purilanjoki AO Genetic distances among Finnish and Russian anadromous. (border FIN-RUS). Watersheds are numbered from west to east. Bootstrap numbers for the branches are shown as percentage from 1000 repeats. A for anadromous and O for original. 2 large groups, structure follows geographic order originally. 4 5 5 19 45 22 3 4 14 17 2 Paimionjoki AO 45 5 Uskelanjoki AO 7 Kiskonjoki Perniönjoki AO 7 Kiskonjoki AO 12 Mankinjoki AO 57 10 Ingarskilanjoki AO 58 15 Sipoonjoki AO 13 Espoonjoki AO 52 20 Summanjoki AO 22 Urpalanjoki FIN-RUS AO 21 Virojoki AO 34 24 Vilajoki FIN-RUS AO 31 Vammeljoki RUS AO 30 Pikkuvammeljoki RUS AO 16 28 Notkopuro RUS AO 28 23 29 Inojoki RUS AO 25 32 Kuokkalanpuro RUS AO 33 Rajajoki RUS AO 49 34 Luga RUS AO Group A Group B 9 Karjaanjoki Mustionjoki AO 41 Group C 11 Siuntionjoki AO Group D 23 Santajoki FIN-RUS AO 50 25 Rakkolanjoki FIN-RUS AO Group E 26 Mustajoki FIN-RUS AO 60 27 Kilpeenjoki FIN-RUS AO Group F Group G 16.10.2014 11

Genetic distances among Finnish and Russian anadromous and resident brown trout stocks. Populations from the same riverssytem often in the same branch not always, however. 0 0 0 1 2 1 1 4 0 9 1 25 70 60 51 48 57 61 0 8 16 40 2 11 19 9 2 Paimionjoki AO 3 Purilanjoki AO 16 Mustijoki RO 7 Kiskonjoki Myllyjoki RO 7 Kiskonjoki Perniönjoki AO 7 Kiskonjoki Perniönjoki Metsänoja RO 7 Kiskonjoki Latokartanonkoski AO 1 Aurajoki AI 8 Fiskars Risslaån RI 8 Fiskarsinjoki AI 14 Vantaanjoki Middle reaches AI 14 Vantaanjoki Lower reaches AI 12 Mankinjoki AO 15 Sipoonjoki AO 14 Vantaanjoki Tributary 2 Palojoki RI 50 31 38 61 81 97 2 16 6 52 81 83 24 38 5 39 83 43 51 48 32 10 Ingarskilanjoki AO 18 Koskenkylanjoki AI 9 Karjaanjoki Nummenjoki RO 9 Karjaanjoki Nuijajoki RM 13 Espoonjoki AO 13 Espoonjoki Ryssänniitunoja RO 19 Kymijoki AM 20 Summanjoki Main stream AO 20 Summanjoki Upper branches RO 14 Vantaanjoki Upper reaches RM 11 Siuntionjoki Kirkkojoki AO 11 Siuntionjoki Passilankoski AO 21 Virojoki RO 9 Karjaanjoki Saavajoki RM 9 Karjaanjoki Vihtijoki RO 5 Uskelanjoki AO 7 Kiskonjoki RO 4 Halikonjoki ARO 24 Vilajoki FIN-RUSAO 6 Punassuon Lohioja RO 9 Karjaanjoki Mustionjoki AO 17 Porvoonjoki RO 22 Urpalanjoki FIN-RUS AO 21 Virojoki AO 23 Santajoki FIN-RUS AO 25 Rakkolanjoki FIN-RUS AO 26 Mustajoki FIN-RUS AO 26 Mustajoki Kananoja 2006 FIN-RUS AO 27 Kilpeenjoki FIN-RUS AO 31 Vammeljoki RUS AO 30 Pikkuvammeljoki RUS AO 28 Notkopuro RUS AO 29 Inojoki RUS AO 16.10.2014 12

Unrooted tree from anadromous Finnish and Russian brown trout stocks. Six groups, two hatchery groups. 26 Mustajoki Kananoja 2006 FIN-RUS AO Uskelanjoki group Finnish-Russian border rivers, Bay of Viborg 7 Kiskonjoki Latokartanonkoski AO Aurajoki group 25 Rakkolanjoki FIN-RUS AO 23 Santajoki FIN-RUS AO 27 Kilpeenjoki FIN-RUS AO 26 Mustajoki FIN-RUS AO 22 Urpalanjoki FIN-RUS AO 11 Siuntionjoki Kirkkojoki AO 9 Karjaanjoki Mustionjoki AO 3 Purilanjoki AO 5 Uskelanjoki AO 2 Paimionjoki AO 7 Kiskonjoki Perniönjoki AO 8 Fiskarsinjoki AI 1 Aurajoki AI 13 Espoonjoki AO 21 Virojoki AO 24 Vilajoki FIN-RUSAO 10 Ingarskilanjoki AO 18 Koskenkylanjoki AI 15 Sipoonjoki AO 12 Mankinjoki AO 14 Vantaanjoki Lower reaches AI 14 Vantaanjoki Middle reaches AI 19 KymijokiAM 20 Summanjoki Main stream AO Russian rivers 32 Kuokkalanpuro RUS AO 29 Inojoki RUS AO The Karelian Isthmus 28 Notkopuro RUS AO 30 Pikkuvammeljoki RUS AO 31 Vammeljoki RUS AO 33 Rajajoki RUS AO 34 Luga RUS AO Ingarskila group Isojoki group 16.10.2014 13

Recommendations 1 Soutwest coast: 1. Still local genetic material left in the Uskelanjoki group. 2. Currently some too small populations, only few families left, new local genetic material needed somewhere, fish transfers possible. 3. Own local hatchery stock to be considered. 4. The distribution of Aurajoki only to agreed rivers and sea ranching, not into river systems with own local stocks, like Kiskonjoki 5. Improving of spawning and nursery areas opening of migration routes 6. Fishing regulation to sustainable level 14

Recommendations 2 Uusimaa coast: 1. Local genetic material left in several rivers of the Ingarskilanjoki group. 2. Ingarskila releases recommended only when needed. 3. Diversity levels high on average. 4. The distribution of Isojoki and Aurajoki only to agreed rivers and sea ranching, not into river systems with own local stocks 5. Differentiation among river systems should be supported by closing down releases when possible 6. Improving of spawning and nursery areas, opening of migration routes 7. Fishing regulation to sustainable level 15

Recommendations 3 Southeast coast: 1. Local genetic material left in the Bay of Viborg group 2. Virojoki and Urpalanjoki belong to that group 3. Mustajoki releases recommended when needed possibly for Kymijoki as well 4. Diversity levels high on average 5. Several native selfsustaining stocks left 6. The distribution of Isojoki and Luutajoki at most to Kymijoki 7. Improving of spawning and nursery areas, opening of migration routes 8. Fishing regulation to sustainable level 16

Stock identification of sea trout catches in the Gulf of Finland Marja-Liisa Koljonen 16.10.2014 17

Rivers sampled for the baseline, only anadrmous stocks 16.10.2014 18

The genetic distances among anadromous brown trout populations from 45 rivers draining into the Gulf of Finland from Finland, Russia and Estonia. 8 similarity groups, all river stocks not possible 6 3 30 58 5 30 19 42 70 5 11 23 15 52 2 Paimionjoki AO 3 Purilanjoki AO 5 Uskelanjoki AO 6 Punassuon Lohioja A/R 49 14 9 Karjaanjoki Mustionjoki AO 11 Siuntionjoki Kirkkojoki AO 7 Kiskonjoki Perniönjoki AO 1 Aurajoki AI Aurajoki Group 76 75 7 Kiskonjoki Latokartanonkoski AO 8 Fiskarsinjoki AI 10 Ingarskilanjoki AO 99 18 Koskenkylänjoki AI 84 76 15 Sipoonjoki AO Ingarskilanjoki Group 12 Mankinjoki AO 62 14 Vantaanjoki AI 13 Espoonjoki AO 54 ISOJOKI ANA Released 58 Isojoki Group 19 Kymijoki AM 6020 Summanjoki Main stream AI 22 Urpalanjoki FIN-RUS AO 15 21 Virojoki AO 10 23 Santajoki FIN-RUS AO FIN-RUS Group 4025 Rakkolanjoki FIN-RUS AO 26 Mustajoki FIN-RUS AO 24 27 Kilpeenjoki FIN-RUS AO 24 Vilajoki FIN-RUS AO 31 Vammeljoki RUS AO 12 33 Rajajoki RUS AO 17 28 Notkopuro RUS AO Russian Group 30 Pikkuvammeljoki RUS AO 29 Inojoki RUS AO 14 24 14 48 19 47 31 57 4 5 32 Kuokkalanpuro RUS AO 34 Luga RUS AO 35 Torvajogi EST 42 Pirita Leiva EST 39 Selja EST 41 Valgejogi EST 38 Toolse EST 37 Kunda EST 40 Loobu EST 43 Vääna EST 36 Pyhäjogi EST 28 44 Keila EST 45 Vasalemma EST Uskelanjoki Group Siuntionjoki Group Estonian Group 16.10.2014 19

Brown trout population groups used for Gulf of Finland catch analysis. Group Populations 1 GAurajoki Aurajoki, KiskonjLatok, KiskonjPernion and Fiskarsinjoki 2 GUskelanjoki Paimionjoki, Purilanjoki, Uskelanjoki and Mustijoki 3 GSiuntionjoki PunasLohioja, Mustionjoki and Siuntionjoki 4 GIngarskilanjoki Ingarskilanjoki, Mankinjoki, Vantaanjoki, Sipoonjoki and Koskenkylanjoki 5 GIsojoki Espoonjoki, Kymijoki, Summajoki and Isojoki 6 FIN-RUS Virojoki, Urpalanjoki, Santajoki, Vilajoki, Rakkolanjoki, Mustajoki and Kilpeenjoki 7 RUS Notkopuro, Inojoki, Pikkuvammeljoki, Vammeljoki, Kuokkalanpuro, Rajajoki and Luga 8 EST Torvajogi, Pyhajogi, Kunda, Toolse, Selja, Loobu, Valgejogi, PiritaLeiva, Vaana, Keila and Vasalemma 16.10.2014 20

Finnish brown trout catch mixture samples from the Gulf of Finland. Location Main sites Year N Eastern Gulf of Finland Pernaja, Loviisa, Ruotsinpyhtää, Pyhtää, Kotka 2008, 2009, 2010, 2011, 2012 448 Middle Gulf of Finland Kirkkonummi, Espoo, Helsinki, Sipoo, Porvoo 2010, 2011, 2012 224 Western Gulf of Finland Raasepori, Inkoo 1999, 2003, 2011 10 Archipelago Sea Naantali, Turku, Parainen, Kemiönsaari 2010, 2011, 2012 21 River sample Kymijoki, Urpalanjoki 1997, 2010 6 In all 709 16.10.2014 21

Stock group composition of Finnish sea trout catch based on Bayesian estimation. Medians of probability distributions and their 2.5%-97.5% probability intervals are shown. Gulf of Finland All over Eastern Middle Western Archipelago Stock group MED 2.5% 97.5% MED 2.5% 97.5% MED 2.5% 97.5% MED 2.5% 97.5% MED 2.5% 97.5% 1 Aurajoki 5 3 7 4 2 6 2 1 5 0 0 11 26 10 48 2 Uskelanjoki 0 0 0 0 0 0 0 0 0 0 0 9 0 0 4 3 Siuntionjoki 0 0 0 0 0 0 0 0 0 0 0 6 0 0 3 4 Ingarskilanjoki 23 20 26 16 13 19 37 31 44 18 3 46 31 15 53 5 Isojoki 53 49 57 61 56 65 40 34 47 74 45 93 14 4 33 6 FIN-RUS 0 0 1 0 0 1 0 0 2 0 0 10 3 0 18 7 RUS 2 1 3 2 1 4 0 0 2 0 0 11 18 6 38 8 EST 17 14 20 17 13 20 19 14 25 0 0 16 0 0 7 16.10.2014 22

Stock group proportions in the sea trout catches. West 10 fish Middle 224 East 448 Useful method for future. 100 % 90 % 80 % 70 % 60 % 50 % 40 % 30 % 20 % 10 % EST RUS FIN-RUS Isojoki Ingarskilanjoki Siuntionjoki Uskelanjoki Aurajoki 0 % Arch. West Middle East All 23

Result from catch analysis in the Gulf of Finland: - The Finnish sea trout catch is mainly composed of Isojoki and Ingarskilanjoki trout, 86 % in together. -The proportion of Isojoki trout is higher in the east (61%), than in the middle (40%) of the Gulf, where Ingarskila is more common (37%). - Aurajoki trout occurred in the Archipelago Sea area 26%. - Estonian fish was also observed in the estern and middle parts of the Gulf of F. catches, 17-19%. - Less spots in the Estonian trouts. - Regularly fishes announced having less less spots than usually were of Estonian origin. - Fish were more salmon like trouts. 24

Individual fish samples from the Eastern Gulf of Finland No Prob.1 Stock 1 Prob.2 Stock 2 Site Time mm g HUOM. 284 0.84 Kuokkala 0.16 Inojoki Pitkäviiri SW 3.1.2012 403 537 Vähän pilkkuja, lohimainen. T-ZE 3019 285 0.52 Vääna 0.34 Vasalem Pitkäviiri W 3.1.2012 519 1393 Vähän pilkkuja, T-ankkuri ZE 3020 286 0.99 Kymijoki 0.01 ISOJOKI Pitkäviiri SW 3.1.2012 441 815 REL, T-ankkuri ZE 3021 287 1.00 Vasalem 0.00 Vaana Pitkäviiri SW 3.1.2012 410 610 Vähän pilkkuja, T-ankkuri ZE 3022 288 0.56 Kymijoki 0.44 ISOJOKI Mustaviiri W 9.1.2012 632 2510 289 0.69 Koskenk 0.31 Ingars Mustaviiri W 9.1.2012 586 1749 T-ankkuri ZE 3023 290 1.00 Kymijoki 0.00 Vasalem Mustaviiri W 9.1.2012 466 881 REL, T-ankkuri ZE 3027 291 0.56 Koskenk 0.44 Ingars Pitkäviiri W 9.1.2012 445 732 REL, T-ankkuri ZE 3026 292 0.50 Vääna 0.47 Vasalem Pitkäviiri W 9.1.2012 413 555 Vähän pilkkuja, T-ankkuri ZE 3024 293 0.84 Koskenk 0.16 Ingars Pitkäviiri W 9.1.2012 438 689 REL, T-ankkuri ZE 3025 294 0.99 Vasalem 0.01 Vaana Pitkäviiri W 9.1.2012 423 706 Vähän pilkkuja, T-ankkuri ZE 3029 295 0.71 Vääna 0.20 Selja Pitkäviiri W 9.1.2012 354 428 Vähän pilkkuja, T-ankkuri ZE 3030 296 0.98 Koskenk 0.02 Ingars Pitkäviiri E 10.1.2012 478 1062 REL, T-ankkuri ZE 3032 297 0.64 Koskenk 0.36 Ingars Pitkäviiri E 10.1.2012 397 550 REL kuvan perusteella. T-ankkuri ZE 3033 298 1.00 Kymijoki 0.00 ISOJOKI Pitkäviiri E 10.1.2012 456 837 REL 299 0.56 Kunda 0.32 Selja Mustaviiri E 10.1.2012 649 2789 25

Photo by Antti Koli Thank you 26