Research Article |
Corresponding author: Branko Dragičević ( brankod@izor.hr ) Academic editor: Paraskevi Karachle
© 2022 Marcelo Kovačić, Branko Dragičević, Mišo Pavičić, Iva Žužul, Tanja Šegvić-Bubić.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Kovačić M, Dragičević B, Pavičić M, Žužul I, Šegvić-Bubić T (2022) New records of recently described Zebrus pallaoroi (Actinopterygii: Gobiiformes: Gobiidae) with notes on its morphology, ecology, and molecular identification. Acta Ichthyologica et Piscatoria 52(1): 13-19. https://doi.org/10.3897/aiep.52.79723
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The family Gobiidae represents the species-richest family of fishes in the Mediterranean Sea. However, many Mediterranean gobiid species are poorly known with just one or few species records in total. Among those species, the recently described Zebrus pallaoroi Kovačić, Šanda et Vukić, 2021 has been known only from the localities in the species description: the southern Adriatic, northern Ionian, and northern and western Aegean seas. The additional specimens of this species were collected in central and northern Adriatic Sea. The identification was confirmed by morphological and genetic methods. The morphological and preserved coloration characters were compared with the data from the limited type material and the alternative preserved coloration diagnostic character was suggested. The first data on alive and freshly dead coloration are reported. The phylogenetic analysis was performed on sequenced COI, cytochrome b, and rhodopsin genes. The analyzed molecular markers placed Z. pallaoroi in a distinct clade within Zebrus–Millerigobius group. The preference of this species for the very shallow habitat is confirmed and discussed.
Adriatic Sea, COI gene, habitat, morphology, new records, Zebrus pallaoroi
The family Gobiidae, with 48 species, represents the species richest family of fishes in the Adriatic Sea (
The aim of the presently reported study was to reveal the additional records of recently described Z. pallaoroi, expanding its known range, with notes on its morphology, ecology, and molecular identification.
Localities and sampling. The specimens were collected off the city of Split, the central Adriatic Sea, and off Cape Lipica, Oštro, the northern Adriatic Sea (Fig.
Material. Material collected (standard length + caudal fin length): 1 male, 26.66 mm, caudal fin damaged, PMR VP5043 (Fig.
Ecology and geographic distribution. Both individuals collected in Split were found below boulders of approximately 20 cm diameter, which were sparsely overgrown by photophilic algae at a small beach consisting of boulders and cobblestone on top of a layer of medium and coarse gravel, at depths 0.0–0.2 m at low tide. During the search, several other individuals presumably of the same species have been observed but not collected. The beach was sheltered, situated inside the port (Fig.
Morphological methods. The morphological data is a combination of characters that positively identify genus Zebrus and species Z. pallaoroi among family Gobiidae in the CLOFNAM area (
Genetic methods. Molecular analysis was conducted on specimens PMR VP5043 and PMR VP5093 from northern Adriatic and specimen IOR-Zpallaoroi12021 from Central Adriatic. Total genomic DNA was isolated from fin clips preserved in 96% ethanol by proteinase K digestion, followed by a simplified DNA isolation procedure (
Product purification and sequencing were performed by Macrogen Inc. (Amsterdam, the Netherlands) on an ABI 3730 automated sequencer. Sequencing was done from both directions. Chromas Pro 1.5 software (http://www.technelysium.com.au) was used to evaluate the quality of the chromatogram. BLASTn (NCBI, available online) was used for sequence identification. In addition, sequences of closely related species from GenBank and the Barcode of Life Database (BOLD; www.boldsystems.org) were used for phylogenetic analyses. Sequence alignment was run by the ClustalW tool in MEGA X software (
Morphological identification. Morphology was examined on specimens PMR VP5043 and PMR VP5093 from northern Adriatic and specimen PMR VP5094 from central Adriatic. The specimens were identified as Zebrus species by having (1) suborbital papillae of the lateral-line system without longitudinal row a; (2) all three head canals present; (3) predorsal area naked; (3) seven transverse suborbital rows; (4) two suborbital transverse rows below row b; (5) interorbital papillae absent.
The presently reported material matches Z. pallaoroi diagnosis and differs from Z. zebrus by all diagnostic morphological characters listed in
The diagnostic coloration character of preserved specimens in species description (
Coloration of alive under stress and freshly dead specimen. The coloration of the specimen from Split (IOR-Zpallaoroi12021) observed in situ was dark brown, almost blackish, with barely visible vertical bars on the flanks, slightly paler in comparison to the surrounding coloration and conspicuous curved pale band on the head. Very soon after capture, darker coloration subsided and the reticulated brownish and yellowish pattern became visible dorsally appearing as vertical bars laterally; conspicuous pale band visible on the head extended from pectoral fin bases and also partly from the base of the fin rays, over upper portions of operculum and connected at the coronal head section above the eyes.
Genetics. The nucleotide sequences of the cyt b gene and the rho were obtained to ensure species identification. Namely, cyt b (1141 bp) and rho (748 bp) fragments of the study specimens produced significant alignments with sequences of Z. pallaoroi from the GenBank with 99% and 98% similarity, respectively. Phylogenetic reconstruction inferred by cyt b gene and rho clearly aggregated our specimens with sequences of Z. pallaoroi in a separate clade, according to other sequences from the family Gobiidae (Fig.
Bayesian phylogenetic tree for the taxa of the gobiid genus Zebrus and Millerigobius, reconstructed from nuclear rhodopsin DNA sequences (A) and mitochondrial cytochrome b (B). Numbers by nodes refer to posterior probabilities. Sequences obtained in this study are labelled with A (IOR-Zpallaoroi12021), B (PMR VP5043), and C (PMR VP5093), while sequences from GenBank are labelled according to accession number. Distance scale provided.
Bayesian phylogenetic tree for the several gobiid genus estimated by partial mitochondrial cytochrome c oxidase subunit I (COI). Sequences obtained in this study are labelled A (IOR-Zpallaoroi12021), B (PMR VP5043), and C (PMR VP5093), while Sequences from Barcode of Life Database (BOLD; www.boldsystems.org) database are labelled according to accession numbers. Numbers by nodes refer to posterior probabilities with distance scale provided.
The morphology of collected individuals matches
It is well known that the phylogenetic reconstruction of gobiid genera is complex. In this study, the molecular analysis corroborated the results presented by
The findings reported herein extend the known distribution of recently described Z. pallaoroi to the central and northern sections of the Adriatic Sea. This extension was the result of an increased research effort driven by the recent recognition of this cryptic species. The two presently reported collection sites were quite sheltered, which was not emphasized in the original description while the then noted preference for the very shallow water is confirmed (