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Intraspecies Variability of the Chub (Squalius Cephalus L.) in the Czech Republic and Possibilities of Its Morphological Determination

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Intraspecies Variability of the Chub (Squalius Cephalus L.) in the Czech Republic and Possibilities of Its Morphological Determination November 8–9, 2017, Brno, Czech Republic 24 years INTRASPECIES VARIABILITY OF THE CHUB (SQUALIUS CEPHALUS L.) IN THE CZECH REPUBLIC AND POSSIBILITIES OF ITS MORPHOLOGICAL DETERMINATION LUKAS JUREK Department of Zoology, Fisheries, Hydrobiology and Apiculture Mendel University in Brno Zemedelska 1, 613 00 Brno CZECH REPUBLIC [email protected] Abstract: The aim of the study was to find whether it might be possible to discover individual lines of selected fish species, in our case the chub (Squalius cephalus), using meristic and plastic traits. The lines were detected using genetic methods; in particular, using analysis of mitochondrial (COI) as well as nuclear (RP1S7) markers. The studied fish originated from all river basins within the Czech Republic. A total of 30 plastic and 20 meristic traits were determined in 18 samples. Only in three traits, the level of significance was found to be lower than P = 0.05. In practice, however, not even these three traits can categorize particular individuals into the given lines. Therefore, no trait was found that would clearly define the morphological differences between the studied lines of the chub. Key Words: chub, Squalius cephalus, morphometric characteristics, plastic traits, intraspecies variability INTRODUCTION With the advent of genetic methods and their increasing availability, molecular methods have collided with classical ichthyology based on morphological traits. For the needs of taxonomy, however, we need to find a common language for both these methods. We are interested in understanding the intraspecies diversity mainly for the sake of a more efficient protection of the biodiversity of our ichthyofauna. The genus Squalis is a relatively variable group of fish; its taxonomy and exact number of species in Europe have not yet been completely determined. This is supported by the fact that presently new species are being found, such as Squalius janae reported by Bogutskaya and Zupančič (2010) from Slovenia Bogutskaya and Zupančič (2010). A number of authors have studied the genus Squalis; for example, Sanjur et al. (2003), and Doadria and Carmona (2006) on the Iberian Peninsula, Zupančič et al. (2010) in Croatia, Seifertová and Šimková (2010) in Europe, Lerch (2010, 2012) in the Balkans, and Saleh et al. (2017) in Iran. The chub is an omnivorous fish species, commonly found in all types of running and sometimes even stagnant waters within the Czech Republic. Historically, no lines/subspecies of chub have been reported on the territory of the Czech Republic or formerly, Czechoslovakia. Hrabě et al. (1973) who otherwise mention subspecies in fish, do not describe any subspecies in this particular species. Baruš and Oliva (1995) note minute differences in the branched rays in the dorsal and anal fins in individual fish from the Danube, Odra, and Elbe rivers, however, without defining specific subspecies. Using genetic methods, the study Molecular biodiversity inventory of the ichthyofauna of the Czech Republic (Mendel et al. 2012) revealed that at least three chub lines inhabit the territory of the Czech Republic. My aim was to determine whether any differences can be found among these lines at the level of morphology, and thereby, whether the lines can be reliably distinguished without using DNA analysis. 314 November 8–9, 2017, Brno, Czech Republic 24 years MATERIAL AND METHODS The material used came from samplings collected within the study Molecular biodiversity inventory of the ichthyofauna of the Czech Republic (Mendel et al. 2012). Captured individuals (n = 18) originated from all major river basins in the Czech Republic. The genetically-determined lines didn´t correlate with the origin of the fish. The sex of the fish wasn´t known. This material was fixed using formaldehyde and subsequently transferred into 70% alcohol. It was stored in the depository of the National Museum in Prague, from where it was loaned for the purpose of my study. The methods were based on the studies of Hrabě et al. (1973), Baruš and Oliva (1995), and Kottelat and Freyhof (2007). A set of a total of 15 meristic and 28 plastic traits was evaluated in the fixed fish. Subsequently, biometric coefficients were calculated from the plastic traits. Meristic values were evaluated as numbers of hard and soft rays on all fins, and as numbers of scales at several levels of the body. The number of gill rakers on the first branchial arch was not determined due to protection of the museum material from damage, and the number of vertebrae weren´t determined for technical reasons. The size of studied fish ranged from 118 to 241 mm total length, or from 95 to 192 mm standard length. For biometric measurements, a digital calliper (Digital Calliper Powerfix Profi) with the accuracy to a tenth of a millimetre was used. In larger fish, some longitudinal dimensions (> 150 mm) were determined on the measuring board. Smaller fish were examined under a magnifying apparatus (Mantis Stereo Microscopes, Vision Engineering) at a ×2 or ×4 magnification. Measurements were done manually, and all horizontal dimensions were measured horizontally with the craniocaudal axis of the body. The determined values were processed statistically with the program Statistica 12 using the method of one-factor ANOVA and Canoco 5.04. RESULTS AND DISCUSSION Meristic traits Of the total number of 15 studied traits, only one was found to be statistically significant. It was the number of soft rays in pectoral fins, where the P value = 0.036. Practically speaking, however, even this trait is non-significant, as the minimum number of 16 soft rays occurred across all three lines, and therefore it cannot be used to distinguish among individual fish (Table 1). Table 1 Meristic traits by three lines of chub Feature line 1 (n = 8) line 2 (n = 7) line 3 (n = 3) Min Max Median Min Max Median Min Max Median Scale counts above lat. line 7.0 8.5 8.0 6.5 8.0 8.0 7.5 8.0 8.0 Lateral line scales min 43.0 47.0 44.0 44.0 45.0 44.0 44.0 46.0 45.0 Lateral line scales max 44.0 48.0 45.5 44.0 45.0 45.0 45.0 46.0 46.0 Scale counts below lat. line 3.0 3.5 3.0 3.0 4.0 3.0 3.0 3.0 3.0 Circumpeduncular scales 14.0 15.0 14.0 14.0 15.0 14.0 14.0 15.0 14.0 Predorsal scales 18.0 24.0 20.0 19.0 22.0 21.0 19.0 20.0 19.0 Postdorsal scales 19.0 23.0 20.5 19.0 21.0 20.0 20.0 23.0 22.0 Preventral scales 26.0 31.0 28.0 23.0 29.0 28.0 26.5 27.0 27.0 Preanal scales 7.0 10.0 8.3 7.0 9.0 8.0 7.0 9.0 9.0 Scales A – C fin 9.0 13.0 11.0 11.0 11.5 11.0 10.0 11.0 11.0 hard 3.0 3.0 3.0 2.0 3.0 3.0 3.0 3.0 3.0 D soft 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 C { } 19.0 20.0 19.0 19.0 21.0 19.0 19.0 19.0 19.0 hard 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 A soft 8.0 9.0 8.0 7.0 8.0 8.0 8.0 9.0 8.0 hard 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 V soft 8.0 8.5 8.0 8.0 8.0 8.0 7.5 8.0 8.0 hard 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 P soft 16.0 18.0 17.0 16.0 17.0 16.0 16.0 16.0 16.0 Legend: D – dorsal fin, C – caudal fin, A – anal fin, V – ventral (pelvic) fin, P – pectoral fin, lat. line – lateral line 315 November 8–9, 2017, Brno, Czech Republic 24 years Plastic traits Were found to be similarly non-significant. Of the 36 relative indices (Table 2), only two were found to be significant; the snout length in percentage of head length P = 0.002. The smallest snout length relative to head length was found in line no. 1 (on average 26.95%), followed by line 2 (28.32%), and the longest snout length on average was found in line 3 (28.69%). The other trait was the depth of caudal peduncle in percentage of body length, where P = 0.020. On average, the first group had 11.00%, the second 11.71%, and the third group 11.42%. Table 2 Plastic trait indexes by three lines of chub Feature line 1 (n = 8) line 2 (n = 7) line 3 (n = 3) % Min Max Average Min Max Average Min Max Average BD/SL 24.1 27.3 25.6 25.2 28.6 27.0 25.8 28.0 26.9 BD/BW 53.5 64.5 59.0 51.4 63.2 58.7 58.0 62.5 59.8 BW/SL 14.0 16.9 15.1 14.7 16.8 15.8 15.5 16.5 16.0 PD/ TL 42.8 46.5 44.4 43.2 46.7 44.6 43.3 44.5 44.0 PD/ SL 53.2 57.0 54.9 54.1 58.3 55.8 54.0 55.9 55.1 PD/PT 50.8 65.2 61.9 57.4 68.4 60.9 62.1 64.0 63.2 PT/TL 23.3 29.2 27.5 24.8 29.8 27.2 27.4 28.2 27.8 PT/SL 28.9 35.4 34.0 31.2 37.0 34.0 34.4 35.5 34.8 PVL/SL 48.7 52.0 50.6 49.8 51.7 50.9 50.0 51.9 51.2 PAL/SL 70.1 75.9 71.8 71.6 74.5 72.6 72.1 75.3 73.4 V-A/SL 19.9 23.8 21.7 20.6 22.9 22.3 21.8 23.8 23.1 LCP/SL 17.1 19.2 17.8 15.5 19.4 17.7 16.5 19.3 18.2 Von/SL 10.3 11.6 11.0 11.3 12.3 11.7 11.1 11.7 11.4 DCP/BD 41.3 44.3 43.0 39.5 45.4 43.4 41.8 43.1 42.6 WCP/BW 13.9 19.8 16.5 11.2 19.6 16.1 15.2 20.6 18.4 HL / TL 19.6 21.3 20.5 18.6 21.1 20.0 19.4 21.5 20.7 HL / SL 24.0 26.6 25.3 23.5 26.2 25.0 24.2 27.1 26.0 HL/BD 91.9 107.3 98.4 82.2 100.2 92.6 86.6 104.9 97.1 HL/PD 43.8 48.9 46.1 42.2 47.2 44.7 44.9 48.4 47.2 HW/BD 64.3 73.6 69.0 62.6 70.7 67.2 62.6 74.9 67.6 HW/BW 86.5 105.5 94.8 87.6 100.0 92.7 88.6 102.1 95.1 RL/HL 25.8 28.2 27.0 27.6 29.4 28.3 28.3 29.3 28.7 PL/HL 48.7 52.8 51.5 46.8 54.7 50.6 46.3 52.5 50.1 IW/HL 41.1 44.6 42.1 42.3 46.9 43.8 41.5 45.8 43.9 IW/HW 69.0 78.4 74.6 71.5 77.1 74.5 72.9 75.9 74.7 ED/SL 5.0 5.7 5.3 5.0 5.9 5.4 5.0 6.2 5.6 ED/HL 19.8 21.9 20.9 19.8 23.9 21.8 20.4 22.9 21.4 ED/RL 74.3 80.2 77.5 71.3 86.5 76.9 69.8 80.2 74.5 ED/IW 47.8 52.1 49.6 46.6 55.2 49.8 45.4 51.7 48.7 dD/SL 17.1 19.4 18.0 16.6 19.8 18.3 17.8 18.1 17.9 dC/SL 22.3 26.4 24.8 23.6 26.8 25.0 24.5 26.4 25.4 dA/SL 12.8 15.9 14.9 13.6 16.3 15.1 14.3 15.4 14.8 dV/SL 13.6 16.3 15.4 13.9 16.4 15.5 14.4 16.1 15.1 dP/SL 16.4 19.5 18.0 16.3 18.9 17.5 17.3 19.1 17.9 dD/BD 66.7 77.8 70.4 63.0 72.6
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