New records of water mites from the Balkans revealed by DNA barcoding (Acari, Hydrachnidia) Author Pešić, Vladimir Department of Biology, University of Montenegro, Cetinjski put b. b., 81000 Podgorica, Montenegro. E-mails: anamanovic @ yahoo. com; milicajovanovic 93 @ yahoo. com vladopesic@gmail.com Author Jovanović, Milica Department of Biology, University of Montenegro, Cetinjski put b. b., 81000 Podgorica, Montenegro. E-mails: anamanovic @ yahoo. com; milicajovanovic 93 @ yahoo. com Author Manović, Ana Department of Biology, University of Montenegro, Cetinjski put b. b., 81000 Podgorica, Montenegro. E-mails: anamanovic @ yahoo. com; milicajovanovic 93 @ yahoo. com Author Karaouzas, Ioannis Institute of Marine Biological Resources and Inland Waters, Hellenic Centre for Marine Research, 46.7 km Athens- Sounio Av., Anavyssos, 19013, Greece. E-mail: ikarz @ hcmr. gr Author Smit, Harry Naturalis Biodiversity Center, P. O. Box 9517, 2300 RA Leiden, the Netherlands. E-mail: Harry. Smit @ naturalis. nl * Corresponding author. E-mail: vladopesic @ gmail. com text Ecologica Montenegrina 2021 2021-12-22 49 20 34 journal article 10.37828/em.2021.49.2 2336-9744 8044105 Torrenticola meridionalis Di Sabatino & Cicolani, 1990 Torrenticola meridionalis is a rhitrobiontic species, originally described from Italy ( Di Sabatino and Cicolani 1990 ) and subsequently reported from different parts of the Balkans (see Pešić et al . 2010 , 2018 ). During our 2019 survey in the Ohrid Lake region in North Macedonia we collected and successfully barcoded two specimens from the ancient Prespa Lake that matches the description of T. meridionalis . The finding of this species in the littoral zone of Prespa Lake is very interesting because this habitat is unusual for torrenticolid mites. The analysis involved 17 nucleotide sequences and the final data set comprised 669 nucleotide positions for 15 specimens of T. meridionalis , one specimen of T. elliptica (as most closely related species) and T. laskai (as a distant outgroup). The neighbour-joining (NJ) tree is presented in Fig. 2 . 84 DNCBD021-20| Torrenticola meridionalis |CCDB-3867-B09| Montenegro | BOLD:AED7519 DNCBD073-20| Torrenticola meridionalis |CCDB-3867-G01| Montenegro | BOLD:AED7519 DNCBD074-20| Torrenticola meridionalis |CCDB-3867-G02| Montenegro |BOLD:AED7519 DCDDJ038-21| Torrenticola meridionalis |CCDB 38361 D02| Montenegro | BOLD:AED7519 TorrM-I DNAEC022-20| Torrenticola meridionalis |15. CG2019 7 B12| Macedonia | BOLD:AED7519 DNCBD051-20| Torrenticola meridionalis |CCDB-3867-E03| Montenegro | BOLD:AED7519 DNAEC023-20| Torrenticola meridionalis |18. CG2019 7 C1| Macedonia |BOLD:AED7519 DNCBD049-20| Torrenticola meridionalis |CCDB-3867-E01| Montenegro |BOLD:AED7519 DCBDJ041-21| Torrenticola meridionalis |CCDB 38392 D05| Italy | BOLD:AEK9662 DCBDJ067-21| Torrenticola meridionalis | CCDB 38392 F07| Italy | BOLD:AEK9662 TorrM-II DCBDJ068-21| Torrenticola meridionalis |CCDB 38392 F08| Italy | DCCDB022-21| Torrenticola meridionalis |CCDB38233 B10| Montenegro | BOLD:AEI3402 DCDDJ020-21| Torrenticola meridionalis |CCDB 38361 B08| Montenegro |BOLD:AEI3402 TorrM-III DCCDB048-21| Torrenticola meridionalis | CCDB38233 D12| Montenegro |BOLD:AEI3402 DCDDJ037-21| Torrenticola meridionalis | CCDB 38361 D01| Montenegro |BOLD:AEI3402 DCCDB023-21| Torrenticola elliptica | CCDB38233 B11| Montenegro |BOLD:AEI9183 DNAEC073-20| Torrenticola brevirostris |42. M19 29A 5 G6| Montenegro |BOLD:AED9586 0.020 Figure 2. Neighbour-joining tree based on studied Torrenticola meridionalis COI sequences. We delineated three T. meridionalis haplogroups. The sequences from Italy forms a well-supported haplogroup (TorrM-II) recovered in the COI tree as a sister clade of haplogrup (TorrM-III) which includes specimens from Montenegro . Both haplogroups possesses a low intraspecific distance ( Table 4 ). The average K2P genetic distance between these two haplogrups was estimated to be 6.3±1.1% K 2P. Our sequences from Prespa Lake belong to a strongly-supported haplogroup (TorrM-III) which includes also sequences from Montenegro . In the COI tree, the later clade was recovered as a sister clade to other T. meridionalis haplogroups and a high interspecific distance which ranged from 6.7 to 7.2% K2P ( Table 5 ). Table 4. List of Torrenticola specimens used in this study. BINs are based on the barcode analysis from 10-12-2021.

Locality (country, name)	Lat/Long	BOLD Acc.	Voucher code	BIN
Nos.
Torrenticola meridionalis Di Sabatino & Cicolani, 1990
N Macedonia, Prespa Lake at Oteševo	40.979 N, 20.917 E	15. CG2019_7 B12	DNAEC022-20	BOLD:AED7519
N Macedonia, Prespa Lake at Oteševo	40.979 N, 20.917 E	18. CG2019_7 C1	DNAEC023-20	BOLD:AED7519
Montenegro, Drcka stream at Mateševo	42.7619 N, 19.5549 E	CCDB-3867-B09	DNCBD021-20	BOLD:AED7519
Montenegro, Rikavac stream	42.1001 N, 19.1432 E	CCDB-3867-E01	DNCBD049-20	BOLD:AED7519
Montenegro, Rikavac stream	42.1001 N, 19.1432 E	CCDB-3867-E03	DNCBD051-20	BOLD:AED7519
Montenegro, Orahovštica river	42.2476 N, 19.0798 E	CCDB-3867-G01	DNCBD073-20	BOLD:AED7519
Montenegro, Orahovštica river	42.2476 N, 19.0798 E	CCDB-3867-G02	DNCBD074-20	BOLD:AED7519
Montenegro, Bistrica stream at Crkvine	42.8054 N, 19.4456 E	CCDB 38361 D02	DCDDJ038-21	BOLD:AED7519
Montenegro, Kolašinska rijeka	42.8391 N, 19.5749 E	CCDB38233 B10	DCCDB022-21	BOLD:AEI3402
Montenegro, Bistrica stream at Crkvine	42.9871 N, 19.4338 E	CCDB38233 D12	DCCDB048-21	BOLD:AEI3402
Italy, Sila, Fossiata, stream	39.4011 N, 16.5884 E	CCDB 38392 D05	DCBDJ041-21	BOLD:AEK9662
Italy, Sila, Fossiata, stream	39.4011 N, 16.5884 E	CCDB 38392 F07	DCBDJ067-21	BOLD:AEK9662
Italy, Sila, Fossiata, stream	39.4011 N, 16.5884 E	CCDB 38392 F08	DCBDJ068-21
Montenegro, Bistrica stream at Crkvine	42.8054 N, 19.4456 E	CCDB 38361 B08	DCDDJ020-21	BOLD:AEI3402
Montenegro, Bistrica stream at Crkvine	42.8054 N, 19.4456 E	CCDB 38361 D01	DCDDJ037-21	BOLD:AEI3402
Torrenticola elliptica Maglio, 1909
Montenegro, Kolašinska rijeka	42.8391 N, 19.5749 E	CCDB38233 B11	DCCDB023-21	BOLD:AEI9183
Torrenticola brevirostris (Halbert, 1911)
Montenegro, Zeta river at Spuz	42.5113 N, 19.1982 E	42. M19_29A_5_G6	DNAEC073-20	BOLD:AED9586

The taxonomic status of these lineages across the whole distribution of this probable species complex needs to be investigated by morphometric data and additional molecular markers.