Phoxinus

Palandacic, Anja, Diripasko, Oleg A., Kirchner, Sandra, Stefanov, Tihomir & Bogutskaya, Nina G., 2024, An integrative approach highlights the discrepancy in the genetic, phenotypic, and presumptive taxonomic structure of Phoxinus (Actinopterygii, Leuciscidae, Phoxininae) in Bulgaria, Journal of Fish Biology 105 (1), pp. 214-238 : 216-220

2.1 | Data on distribution of Phoxinus in Bulgaria

Data on the distribution of Phoxinus in Bulgaria were collected from the literature ( Apostolou et al., 2023; Apostolov, 2005; Chichkoff, 1939; Dikov et al., 1994; Drensky, 1926, 1948, 1951;

Hydrological region/ Sample sampling location number Drainage

Middle Danube Black Sea

Topotypical csikii Ibar 1 Danube

Nishava 2 Danube

Lower Danube Black Sea

Beli Vit 3 Danube

Palakaria 4 Danube

Non‐ Danube Black Sea

Kamchiya 1 5 Kamchiya

Kamchiya 2 6 Kamchiya

Fakiiska 7 Fakiiska (formerly Mandra Lake tributary)

Izvorska 8 Izvorska

Karaagach 1 9 Kitenska [Karaagach]

Karaagach 2 10 Kitenska [Karaagach]

Morphology Molecular

Country Species name and/or clade (n) analyses (n)

Montenegro P. csikii , Clade 5 YES (6) YES (CO1 1)

Bulgaria P. csikii , Clade 5 YES (30; 5 a) YES (CO1 10; RAG1 10)

Bulgaria P. csikii , Clade 5 YES (33; 5) YES (CO1 10; RAG1 8)

Bulgaria P. csikii , Clade 5 YES (55; 5) YES (CO1 10; RAG1 10)

Bulgaria P. strandjae , Clade 14 YES (12) NO

Bulgaria P. strandjae , Clade 14 YES (51; 8) YES (CO1 10; RAG1 10)

Bulgaria P. strandjae , Clade 14 YES (14) YES (CO1 3; RAG1 3)

Bulgaria P. strandjae , Clade 14 YES (40, 2) YES (CO1 6; RAG1 6)

Bulgaria P. strandjae , Clade 14 YES (42, 12) YES (CO1 10; RAG1 10)

Bulgaria P. strandjae , Clade 14 YES (24) NO

Sample data

NMW 51266 (neotype), 98673, Rožaje, Ibar (Zapadna Morava‐ Velika Morava system), ca. 42°50′39″N 20°09′59″E, 1917. SL (mm) 84.4 (neotype); 78.9– 91.1

NMW 99088, Nishava R. at Tuden, Velika Morava system, 43°0′13''N 23°0′26''E, 02.11.2017. SL (mm) 46.2–68.1

NMW 99089, Beli Vit R. at Ribaritsa, 42° 50′16''N 24°22′52'' E, 06.11.2017. SL (mm) 45.1–67.3

NMW 99090, Palakaria R. at Yarlovo,

Iskar system, 42°28′58''N 23°16′12''E, 20.10.2017. SL (mm) 50.9–78.4

NMW 100449, Luda Kamchiya R., downstream of Ichera, 42°46′43.5″N 26°29′24″E, 26.07.2010. SL (mm) 43.1–50.1

NMW 100450, Medvenska R., at Dabovitsa, 42°48′47″N 26°37′37″E, 13.08.2011. SL (mm) 25.0–63.3

NMW 100451, Fakiiska R., 1 km

upstream from Fakia, 42°02′51″N 26° 46′12″E, 13.10.2010. SL (mm) 35.0– 44.3

NMW 100452, Izvorska R., at Indzhe Voyvoda, 42°09′50″N 27°05′52″E, 20.07.2010. SL (mm) 35.1–55.7

NMW 100453, Orlyashka R. at Vizitsa, 42°08′19″N 27°36′53″E, 26.09.2014. SL (mm) 33.5–64.3

NMW 100454, Uzunchairska R. at Pismenovo, 42°13′45″N 27°42′24″E, 20.07.2009; NMW 100455, same data, 26.09.2014. SL (mm) 25.6–44.4

(Continues)

T A B L E 2 (Continued)

Hydrological region/ Sample sampling location number Drainage

Topotypical strandjae 11 Veleka [Kocadere] Veleka

Sapanca 12 Lake Sapanca basin, Sakarya

Non‐ Danube Marmara Sea Gönen 13 Gönen

East Aegean

Maritsa (Vacha) 14 Maritsa [MeriÇ, Evros]

Maritsa (Tundzha) 15 Maritsa

West Aegean

Eleshnitsa 16 Struma [Strymon]

Topotypical 17 Strymon strymonicus

Angitis 1

Topotypical 18 Strymon strymonicus

Angitis 2

Morphology Molecular

Country Species name and/or clade (n) analyses (n)

Bulgaria P. strandjae , Clade 14 YES (42; 6) YES (CO1 10; RAG1 10)

Turkey P. strandjae , Clade 14 YES (24) YES (CO1 2; RAG1 2)

Turkey P. strandjae , Clade 14 YES (16) NO, previously published b

Bulgaria P. strandjae , Clade 14 YES (7) YES (CO1 6)

Bulgaria P. strandjae , Clade 14 YES (136; YES (CO1 10; 15) RAG1 10)

Bulgaria P. csikii , Clade 5 and P. YES (69; 8) YES (CO1 11; strymonicus , Clade 15 RAG1 11)

Greece P. strymonicus , Clade 15 YES (8) YES (CO1 5; RAG1 5)

Greece P. strymonicus , Clade 15 YES (14) NO

Sample data

NMW 99092, 100636 (neotype), Mladezhka R., 42°05′31″N 27°23′47″ E, 18.10.2017. SL (mm) 41.4–59.3

ZFMK 93593‐624, Kurtköy stream at Kurtköy, 40°42′46″N 30°10′30″E, 08.06.2006. SL (mm) 40.0–58.4

ZFMK 94196–215, Çanakkale, Gönen R. west of AkÇakoyun, Biga Peninsular, 39°49′36″N 27°08′36″E, 30.10.2007. SL (mm) 42.6–56.3

NMW 51267, Elitere [Vacha] R.at Sarambej [Septemvri], ca. 42°13′01″N 24°07′48″E, 1884. SL (mm) 50.8–61.9

15a: NMW 100456, Arapliiska R. (tributary to Popovska, Tundzha system), between Sitovo and Iglika, 42° 02′51″N 26°46′12″E, 12.10.2010; NMW 100458, same locality, 26.10.2018; 15b: NMW 100457, Arapliiska R. (tributary to Popovska, Tundzha system), at Lalkovo, 42°04′ 28″N 26°43′26″E, 12.10.2010; 15c: NMW 100459, Popovska R., 1 km above Voden, 42°03′44.5″N 26°43′ 15″E, 12.10.2010. SL (mm) 35.6–68.2

NMW 99091, Eleshnitsa R. at Vaksevo, 42°09′43″N 22°50′05″E, 12.10.2017; NMNHS uncat., locality data as NMW 99091, 23.04.2018. SL (mm) 29.9–69.1

ZFMK 55949‐53, 57256‐58, Kefalari spring west of Ano Kefalari, Angitis R. system, 41°03′41″N 24°15′57″E. SL (mm) 24.7–36.7

NMP P6V 141470–83, Kefalari spring at Drams, Angitis R. system, 41°04′24″N 24°15′54″E, 20.09.2011. SL (mm) 31.0–68.3

T A B L E 2 (Continued)

Hydrological region/ Sample sampling location number Drainage Morphology Molecular

Country Species name and/or clade (n) analyses (n) Sample data

Reference localities Adriatic Po 19 Po

Po 20 Po

Italy

Italy

P. lumaireul , Clade 1 YES (41) NO, previously published b

P. lumaireul , Clade 1 YES (43) NO, previously published b

ZFMK 45282–99, 46270–89, 50441–43, Piemont, Sangone stream W of Trana, 45°01′42″N; 7°23′43.5″E, 15.06.2011. SL (mm) 36.8–67.8

NMW 98662, Pinerolo, Chisone torrent at Contina Farm , 45°51´42″N; 7°29´43″E, 13.06.1014. SL (mm) 41.4–54.1

Skadar 21 Skadar Lake basin

Ohrid 22 Ohrid Lake basin

Reference localities Atlantic

P. bigerri 23 Estuary of Bilbao

Montenegro P. karsticus , clade 7 YES (44) NO, previously

YES (41) published b

Northern Phoxinus sp. , Clade 8 YES (9) NO, previously Macedonia published b

Spain P. bigerri , Clade 13 YES (154; 5) NO, previously YES (14) published b YES (3)

21a: ZFMK 93661‐93704, Črnojevica

Rijeka about 500 m upstream of village Črnojevica, 42°21′23''N 19°00′53''E, 13.05.2003. SL (mm) 41.7–56.2

21b: ZFMK 94100‐140, River MoraČa at village BioČe, 42°30′41″N 19°20′20″E, 20.07.2002. SL (mm) 38.2–57.2

22a: NMP P6 V 141381–89 , at Hydrobiological Institute , Ohrid, 41°06′ 08″N 20°48′50″E, 14.09.2011 .

22b: NMP P6 V 89747 –72, Sateska R., 41°19′02″N 20°47′43″E, 25.07.2006. SL (mm) 36.3–56.5

23a: NMW 51270, Arrigoriaga at Bilbao, August 1864. SL (mm) 29.7–60.9

23b: NMW 51276, Pau, 1869. SL (mm) 47.0–73.5

23c: NMW 51235, Bilbao. SL (mm) 46.7– 49.4

Abbreviations: CO1, the mt cytochrome oxidase 1 gene; n, sample number; NMNHS, National Museum of Natural History , Sofia, Bulgaria ; NMP P6 V, Natural History Museum , Prague, Czech Republic , Vertebrate collection; NMW, Fish Collection, Natural History Museum , Vienna, Austria ; nuclear RAG1 , recombination activating gene 1; SL, standard length; ZFMK, Zoological Research Museum Alexander Koenig , Bonn, Germany .

a Cleared & Alizarin Red S stained (if any) after semicolon. Species names based on localities (closeness to type localities) and taxonomic or molecular clade assignments as in PalandaČiĆ et al. (2017a, 2020) and Bogutskaya et al. (2019).

b Some samples were a part of previous studies (Geigher et al., 2014; PalandaČiĆ et al., 2017a, 2020); therefore, genetic information is available from the GenBank. In case of material from ZFMK and NMP, it is not clear which voucher specimens were genetically analysed.

Karapetkova & Zivkov, 1995; Marinov, 1989; Michajlova, 1965, 1970; Pehlivanov, 2000; Raikova-Petrova, 2000; Russev, 1964; Stefanov, 2007; Stefanov & Trichkova, 2004, 2006; Trichkova et al., 2009; Vassilev & Pehlivanov, 2005; Zivkov & Dobrev, 2001) and combined with unpublished field records from the National Museum of Natural History in Sofia (NMNHS) (Figure 1).