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Rediscovery of Knipowitschia goerneri and its molecular relationships with other European northern Mediterranean Knipowitschia species (Teleostei: Gobiidae)


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The Corfu dwarf goby, Knipozvitschia goerneri, known only from the Korission Lagoon catchment on Kerkyra Island (Greece), was described in 1991 on the only existing material, consisting of five specimens. It has not been observed since 1983, and was thought to be possibly extinct. In 2014, we collected nine specimens of K. goerneri from Korission Lagoon. We found this species to be morphologically and genetically well distinguishable from all other Knipozvitschia species. Analyses of the mitochondrial gene cytochrome b placed it as a sister species of K. milleri, which is another goby from the northern Ionian Sea basin.
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Ichthyol. Explor. Freshwaters, Vol. 26, No. 4
Ichthyol. Explor. Freshwaters, Vol. 26, No. 4, pp. 363-372, 3 figs., 2 tabs., March 2016
© 2016 by Verlag Dr. Friedrich Pfeil, München, Germany – ISSN 0936-9902
Rediscovery of Knipowitschia goerneri
and its molecular relationships with other European
northern Mediterranean Knipowitschia species
(Teleostei: Gobiidae)
Jasna Vukib*, Marcelo Kovacib**, Stamatis Zogaris*** and Radek Šanda****
The Corfu dwarf goby, Knipowitschia goerneri, known only from the Korission Lagoon catchment on Kerkyra Island
(Greece), was described in 1991 on the only existing material, consisting of five specimens. It has not been observed
since 1983, and was thought to be possibly extinct. In 2014, we collected nine specimens of K. goerneri from Koris-
sion Lagoon. We found this species to be morphologically and genetically well distinguishable from all other
Knipowitschia species. Analyses of the mitochondrial gene cytochrome b placed it as a sister species of K. milleri,
which is another goby from the northern Ionian Sea basin.
* Department of Ecology, Faculty of Science, Charles University in Prague, Vinicná 7, 128 44 Prague, Czech
Republic. E-mail: (corresponding author)
** Prirodoslovni muzej Rijeka, Lorenzov prolaz 1, HR-51000 Rijeka, Croatia.
*** Hellenic Centre for Marine Research, Institute of Marine Biological Resources and Inland Waters, Anavissos,
Attiki 19013, Greece. E-mail:
**** Department of Zoology, National Museum, Václavské nám. 68, 115 79 Prague, Czech Republic.
A renewed interest in the European fish fauna has
resulted in the recent discovery or recognition of
many new species, especially from the Mediter-
ranean region (e. g., Freyhof et al., 2005; Kovacib,
2005; Kottelat, 2007; Miller & Šanda 2008; Buj et
al., 2010). This is an area with an exceptionally rich
freshwater ichtyofauna (Freyhof & Brooks, 2011),
and the number of species continues to increase
(e. g., Buj et al., 2014; Freyhof et al., 2014). Many
freshwater fish species in the region are endemic
to a single or a few neighbouring rivers, and are
under threat from human activities (Smith &
Darwall, 2006; Freyhof & Brooks, 2011).
Several fish species from the Mediterranean
area are currently considered extinct (Smith &
Darwall, 2006; Geiger et al., 2014). However, it is
possible that some of these have been overlooked
due to their cryptic behaviour, small size, absence
of real search effort, lack of interest, political or
social situation, or administrative obstruction.
One of the endemic species with an unclear
conservation, as well as taxonomic status, is the
Corfu dwarf goby, Knipowitschia goerneri Ahnelt,
1991. Known only from the Korission Lagoon
Vukib et al.: Rediscovery of Knipowitschia goerneri
catchment on Kerkyra Island (Corfu), Greece
(Ahnelt, 1991), this goby was described from the
only existing material (five specimens) collected
in 1968 and 1983, and taken from a freshwater
spring and the brackish coastal lagoon, respec-
tively. It was not observed in nature after 1983
(Kottelat & Freyhof, 2007), although at least one
ichthyological survey was conducted at various
localities on Kerkyra Island since this time (Kalo-
gianni et al., 2006), and K. goerneri was suggested
to be extinct (Economidis & Chrysopolitou, 2009).
In the IUCN red list, this goby is listed as data
deficient (Crivelli, 2006), as it is one of the least
studied freshwater fish species with practically
nothing known about its biology or ecology.
During an ichthyological survey on Kerkyra
Island in July 2014, several specimens of Knipo-
witschia were collected in Korission Lagoon
(Fig. 1). The aim of this work is to investigate
the taxonomic status of this population and to
provide information on its molecular relationships
with other Knipowitschia species known from the
European northern Mediterranean region.
Materials and methods
Counts and measurements follow Miller (1988),
as other recent descriptions of Knipowitschia spe-
cies also use the same methodology (Ahnelt 1995,
2011; Kovacib, 2005; Kovacib & Šanda, 2007; Miller,
2009). The length of the specimens is presented
as standard length, and the terminology of the
cephalic sensory systems follows Sanzo (1911)
and Miller (1972). All specimens were stained in
a 2 % solution of Cyanine Blue in distilled water
using a reversible staining method (Saruwatari et
al., 1997) to more clearly visualise scales and the
cephalic sensory systems.
Abbreviations: SL, standard length; COI,
cytochrome c oxidase subunit 1. Collection codes:
NMP, National Museum, Prague; NMW, Natur-
10 11
Southern Kerkyra (Corfu)20°
22° 24°
0 50 100 km
Fig. 1. a, Location of Korission Lagoon, locality of Knipowitschia goerneri; b, sampling site at Korission Lagoon
(indicated by arrow); c, Localities from where populations of Knipowitschia have been recorded in Greek Ionian
Sea drainage (Economou et al., 2007; Barbieri et al., 2015): 1, Korission Lagoon on Kerkyra Island (K. goerneri);
2, Acheron River (K. milleri); 3-15, still unidentified populations: 3, Kalamas delta; 4, Louros River; 5, Arachthos
River; 6, Vlychos spring; 7, Voulkaria Lake; 8, Acheloos delta; 9, Trichonis Lake; 10, Evinos delta; 11, Mornos
delta; 12, Prokopos-Kotychi Lagoons; 13, Pinios River on Peloponnese; 14, Alfios delta; and 15, Zakynthos Island.
Ichthyol. Explor. Freshwaters, Vol. 26, No. 4
historisches Museum, Wien; PMR, Natural His-
tory Museum, Rijeka.
DNA extraction and PCR. DNA was extracted
from fin tissue preserved in 96 % ethanol using
following the manufacturers instructions. Mito-
chondrial gene cytochrome b was analysed. The
amplification primers used were GluF and ThrR
(Machordom & Doadrio, 2001). PCR reactions,
amplification protocol, and PCR product purifica-
tion follow Šanda et al. (2008). Sequencing was
carried out by the Macrogen Service Centre (Seoul,
South Korea) using the amplification primers.
Molecular data analysis. Sequences were aligned
manually and revised in BioEdit (v.7.0.9). The
final alignment included 1119 base pairs of cy-
tochrome b. Pomatoschistus bathi was used as the
outgroup. From the Adriatic basin, K. panizzae
and K. montenegrina were included in the analyses
as the other two freshwater endemics from this
area (K. radovici and K. mrakovcici) are genetically
almost identical to K. panizzae (Geiger et al., 2014).
The best fitting model of nucleotide substitu-
tion was assigned using JModeltest 0.1.1 (Posada,
2008). Using Akaike information criterion (AIC),
the GTR+I model was selected. Two different ap-
proaches were used: Maximum Likelihood and
Bayesian inference. Maximum Likelihood phylo-
genetic analyses were performed using the BEST
approach implemented in PhyML, version 3.0.1,
which combines nearest neighbour interchanges
(NNI) and subtree pruning and regrafting (SPR)
algorithms to maximize tree likelihood (Guindon
& Gascuel, 2003). Five random starting trees were
used. Branch support was quantified by bootstrap
analysis with 1000 random replicates.
A Bayesian analysis was conducted using
MrBayes v. 3.1.2 (Huelsenbeck & Ronquist, 2001).
Two runs, each consisting of four Monte Carlo
Markov Chains, were run simultaneously for
1 000 000 generations with sampling trees every
100 generations. The first 20 % of trees were dis-
carded as burn-in and the remaining trees were
used to construct a 50 % majority-rule consensus
tree. The posterior probabilities were used to
indicate the branch supports in the final tree.
Genetic divergences between and within
species of Knipowitschia, based on cytochrome b
nucleotide sequences, were calculated in Mega
5.2 (Tamura et al., 2011) as uncorrected pairwise
genetic distance (p-distances).
Knipowitschia goerneri
(Fig. 2)
Material examined. NMP P6V142862-142869, 1 male,
17.0 mm SL, 7 females, 13.1-20.6 mm SL; PMR VP3359,
1 female, 18.1 mm SL; Greece: Kerkyra Island: Korission
Lagoon, 39°27'02" N 19°54'20" E; R. Šanda & J. Vukib, 26
July 2014 (GenBank accession numbers: KT809439-
KT809446). – NMW 90253:1, 1, paratype, male, 17.8 mm
SL; Greece: Corfu (Kerkyra) Island: NW beach of Koris-
sion Lagoon; P. Keymar, Aug 1983.
Generic identification. The following combi-
nation of characters identify the specimens as
belonging to a species of the genus Knipowitschia:
1) suborbital row a of sensory papillae present;
2) pelvic disc with anterior transverse membrane;
3) cheek with several suborbital transverse rows of
sensory papillae; 4) anterior oculoscapular canal
ending in interorbital area, with paired pores λ,
or at doubled pore κ (K. goerneri), or head canals
absent; 5) no perianal organ; and 6) oculoscapu-
lar row tra of sensory papillae not reaching close
to suborbital row b.
Diagnosis. The genus Knipowitschia includes
16 species. There are no characters unique to
K. goerneri. Knipowitschia goerneri is easily dis-
tinguished from K. byblisia, K. ephesi, K. monte-
negrina, K. mrakovcici and K. thessala by having
the body squamation continuous from the axilla
to the caudal part along the lateral midline (vs.
body squamation not continuous). Knipowitschia
goerneri differs from K. milleri, K. ephesi and
K. montenegrina by the presence of at least some
head canals (vs. head canals absent). Knipo-
witschia goerneri is most clearly distinguished from
K. radovici, K. cameliae, K. caucasica, K. caunosi,
K. longecaudata and K. panizzae by the absence of
the preopercular head canal (vs. preopercular
head canal present). Knipowitschia goerneri differs
from K. iljini, K. cameliae, K. caucasica, K. caunosi,
K. longecaudata, K. panizzae and K. bergi by having
reduced squamation along the second dorsal fin,
i. e. dorsal naked along second dorsal fin up to
the base of the penultimate or of last articulated
ray (vs. squamation along the second dorsal fin
not reduced up to the base of the penultimate
or of last articulated ray). Knipowitschia goerneri
differs from K. mermere by having the base of the
last ray of the second dorsal fin before the vertical
through the base of the last anal-fin ray (vs. last
ray of second dorsal fin at vertical of the base of
the last anal-fin ray). Knipowitschia goerneri further
differs from K. mermere by having a wider head
(head width 17.9-22.3 % SL vs. 11.9-14.9; 58-69 %
HL vs. 38-48) and a shorter snout (snout length
16-21 % HL vs. 22-31).
Description. Body moderately elongate, cross
section circular anteriorly, compressed towards
caudal peduncle. Head large and moderately
depressed. Snout oblique, relatively longer in
smaller specimens, shorter in large specimens.
Eye large, dorsolateral, eye contour just slightly
above dorsal profile, i. e. above snout and predor-
sal area. Interorbital space distinctly smaller than
eye diameter and moderately narrow. Anterior
nostril short, tubular, erect, without process from
rim; posterior nostril pore-like, near orbit. Mouth
oblique, with lower jaw slightly projecting for-
ward to upper jaw, posterior angle of jaws below
pupil. Branchiostegal membrane attached along
entire lateral margin of isthmus. Body proportions
given in Table 1.
Fins. First dorsal VI; second dorsal I/7-8; anal
I/7-8; caudal 13 branched rays, 15-16 segmented;
pectoral 15-16; pelvic I/5 + 5/I. First dorsal fin
arising behind pectoral-fin base. First dorsal-fin
spines III-V when depressed extending to second
dorsal fin I in adult male, not reaching second
dorsal in female. Interdorsal space distinct. Second
dorsal fin commencing over or immediately be-
hind vertical of anus, with last ray before vertical
of last anal-fin ray. Anal fin commencing below
second to third articulated ray of second dorsal
fin. Caudal fin rounded. Pectoral fin extending
Table 1. Standard length and proportional measurements of Knipowitschia goerneri in this study and of type
material (Ahnelt, 1991). Morphometric ratios from Ahnelt (1991) were recalculated as percentages.
present research type material
range mean SD range
Standard length (mm) 13.1-20.6 17.8-21.8
In percent of standard length
Head length 30.0-33.8 31.7 1.3 30.0-34.2
Head width 19.1-22.3 20.3 1.1 17.9-20.0
Distance from snout to origin of first dorsal fin 39.2-42.2 41.2 1.0 39.1-43.9
Distance from snout to origin of second dorsal fin 56.1-61.7 59.1 2.0 54.6-61.3
Distance from snout to vertical of anus 55.2-59.7 57.1 1.4 55.2-61.3
Distance from snout to vertical of anal-fin origin 59.1-65.5 61.5 1.9 61.7-66.2
Distance from snout to vertical of pelvic-fin origin 31.2-35.9 32.9 1.5 30.3-34.5
Caudal-peduncle length 24.6-27.5 26.1 0.9 23.6-27.9
First dorsal-fin base 9.2-14.1 10.9 1.7 10.7-13.8
Second dorsal-fin base 13.0-16.0 14.9 1.1 16.1-18.3
Anal-fin base 13.5-17.1 15.2 1.2 16.1-17.9
Caudal-fin length 21.5-25.2 23.1 1.3 24.3-27.9
Pectoral-fin length 18.3-22.8 20.3 1.5 22.6-25.3
Pelvic-fin length 18.3-24.8 21.1 2.1
Body depth at pelvic-fin origin 21.0-23.8 22.7 1.1 20.7-23.9
Body depth at anal-fin origin 15.2-19.6 16.4 1.4 15.6-17.9
Body width at anal-fin origin 8.4-11.7 10.0 1.0
Caudal-peduncle depth 10.1-11.6 10.8 0.6 9.2-11.4
Distance from pelvic-fin origin to anus 22.5-30.4 25.9 2.4
In percent of head length
Snout length 17-21 19.2 1.3 16-20
Eye diameter 21-25 23.4 1.5 17-27
Postorbital length 49-61 54.9 3.7 53-61
Head width 58-69 63.9 3.6 59-61
Caudal-peduncle depth in percent of its length 38-47 41.5 2.8 36-51
Interorbital width in percent of eye diameter 42-55 47.2 5.2 39-54
Pelvic-fin length in percent of distance from pelvic origin to anus 74-94 81.9 6.2 79-98
Vukib et al.: Rediscovery of Knipowitschia goerneri
Ichthyol. Explor. Freshwaters, Vol. 26, No. 4
back to below posterior end of first dorsal fin.
Pelvic disc rounded, nearly reaching anus in
male, shorter in female; anterior membrane in
midline 1
3 to 1
2 of pelvic spine length, free edge
of anterior membrane smooth. Lengths of fins and
fin bases in proportion to standard body length
given in Table 1.
Scales. Side of body and caudal peduncle covered
with ctenoid scales; predorsal and dorsal region,
back to base of penultimate or of last articulated
ray of second dorsal fin naked, breast and belly
naked; 30 to 33 in lateral series. Body squamation
continuous from axilla to caudal part along lateral
midline. Upper edge of scaled area extending from
behind upper part of pectoral axilla backwards
and up, reaching second dorsal fin at its posterior
end. Lower edge of scaled area extending from
behind pectoral-axilla bottom backwards and up
to vertical of anus and interdorsal space, and then
downwards to anal fin, variably reaching it. Nar-
rowest section of scaled area along lateral midline
between interdorsal space and anus.
Coloration. Preserved specimens. Body dark
brown above and whitish below; male darker
than female. Body, except most of underside in
female and belly in male, covered with numerous
melanophores, variably and irregularly mottled
with hardly recognizable patterns. Five pale
dorsal saddles: at nape opposite opercle, origin
of first dorsal fin, interdorsal space and origin
of second dorsal fin, posterior end of second
dorsal fin and posterior part of caudal peduncle.
In female several darker blotches recognizable
along lateral midline. Entire head pigmented in
male, in female underside mostly whitish and
poorly distinguished preorbital bars extending
to form a Y-shaped blotch on chin. First dorsal
fin in adult female with pigmentation anteriorly
at base, oblique transparent band in lower half
above pigmented area at base. Upper half of fin
with broad dark oblique band above transparent
band and transparent upper edge at fin-spine tips.
First dorsal fin in adult male entirely pigmented,
with more intensive black spot in mid-height of
fin between spine IV and V. Second dorsal fin
and caudal fin pigmented without recognizable
pattern. Pectoral fin with dark blotch on upper-ray
origin and rarely scattered melanophores on rest
of upper part of fin. Pelvic fin whitish in female,
pigmented and covered with melanophores in
Lateral-line system. Cephalic canals present.
Anterior oculoscapular canal present, but much
reduced, divided into two or three parts: one
behind eye from pore κ (always paired) to pore α;
second more posteriorly from pore α' to pore ρ;
third short canal with pores λ and λ' between eyes
of variable occurrence in specimens. Posterior
oculoscapular and preopercular canals always
absent. Rows of sensory papillae: Preorbital
with three median rows and two rows lateral to
Fig. 2. Knipowitschia goerneri, NMP P6V142865, 17.0 mm SL, male, and NMP P6V142867, 18.9 mm SL, female;
Greece: Kerkyra Island: Korission Lagoon.
nostrils. Suborbital rows with infraorbital row
a extending forward to below anterior part or
anterior edge of pupil, posteriorly proliferated in
transverse row atp and a few shorter transverse
rows of 2-3 papillae. Longitudinal rows b not
extending anteriorly before vertical of posterior
border of orbit. Six or seven transverse c rows.
Longitudinal row d with continuous supralabial
and cheek parts. Three preopercular mandibular
rows: external row e and internal row i, both di-
vided, and mental row f. Oculoscapular rows tra,
z, x1, trp, x2, y present, with x1 divided by row trp
in anterior and posterior parts, additional papillae
present at position of missing preopercular canal
(below level of oculoscapular row z and slightly
in front of its vertical), and single papilla present
below posterior part of x1. Axillary rows as1, la1,
as2, la2 and as3 visible. Opercle with transverse
row ot, superior longitudinal row os and inferior
longitudinal row oi. Anterior dorsal row n trans-
versal, rows g and h longitudinal, rows o and m
absent. Interorbital longitudinal row p present.
Molecular relationships. Both Maximum Likeli-
hood and Bayesian inference analyses resulted in
an identical tree topology with a high statistical
support of branching (Fig. 3). Our results based on
cytochrome b nucleotide sequence analyses show
that K. goerneri is a sister species of K. milleri, the
species of Knipowitschia from the geographically
closest locality. The mean genetic divergence
between cytochrome b nucleotide sequences of
these two species is 3.2 % (Table 2), and they
form reciprocally monophyletic lineages (Fig. 3).
Morphology. Our morphological examination
confirmed that the newly collected Knipowitschia
from Korission Lagoon is K. goerneri. This species
cannot be confused with any other known goby
species from the European coast of the Mediter-
ranean, including K. milleri (Ahnelt & Bianco,
1990), the only other Knipowitschia species from the
Fig. 3. Maximum Likelihood estimation of phylogenetic relationships of analysed Knipowitschia species based on
cytochrome b nucleotide sequences. Numbers on branches are posterior probabilities/bootstrap support of nodes.
NMP P6V141422
NMP P6V142869
NMP D1326
NMP P6V141242
NMP P6V141302
NMP P6V142644
NMP P6V142867
NMP P6V142868
NMP P6V142866
PMR VP1985
NMP P6V142863
NMP P6V88957
NMP P6V142642
NMP P6V141423
NMP P6V89874
NMP P6V141418
NMP D1325
NMP P6V142865
NMP P6V88956
PMR VP1905
NMP P6V142646
NMP P6V142864
NMP P6V142643
PMR VP3359
NMP P6V142645
NMP P6V141300
Pomatoschistus bathi
Vukib et al.: Rediscovery of Knipowitschia goerneri
Ichthyol. Explor. Freshwaters, Vol. 26, No. 4
north-eastern part of the Ionian Sea basin. Unlike
K. goerneri, K. milleri completely lacks head canals.
The degree of reduction of both body squamation
and head canals in K. goerneri are unique among
all 16 named Knipowitschia species (Ahnelt, 1991,
1995, 2011; Miller, 2004a, 2009; Kovacib, 2005:
table 2; Kottelat & Freyhof, 2007; Kovacib & Šanda,
2007; Geiger et al., 2014) with the only exception
being K. mermere Ahnelt, 1995, a species from
Anatolia (Ahnelt, 1995) located so far away that
the presence of K. mermere in Korission Lagoon
is highly improbable. In addition, the values of
some morphometric characters of K. goerneri and
K. mermere do not overlap (head width, snout
length; see diagnosis), or overlap only slightly
(caudal-fin length 21.5-27.9 % SL, vs. 18.1-22.0;
body depth at pelvic-fin origin 20.7-23.9 % SL, vs.
19.2-21.6) (see Table 1 and Ahnelt, 1991, 1995).
Furthermore, K. mermere is a freshwater species,
which is related to K. thessala (Vinciguerra, 1921)
and is considerably divergent from K. milleri,
based on COI gene nucleotide sequence analysis
(Geiger et al., 2014), while K. goerneri is closely
related to K. milleri, based on cytochrome b gene
nucleotide sequence analyses (this study).
Comparison with the type material of K. goer-
neri showed that the morphology of the newly
collected specimens matches well with the original
species description (Ahnelt, 1991). Two exten-
sions of the original meristic character ranges
were observed in the new material of K. goerneri,
which is probably a result of the smaller number
of individuals used for the original description.
First, we observed both the second dorsal fin
and anal fin to have I/7-8, vs. I/8 for both fins
in the original description. Second, one specimen
exhibited 30 scales in the lateral series, vs. a range
of 32-33 in the original description. It was also
observed in the recent specimens that the pelvic
disc of a single male nearly reached the anus,
while in the original description the pelvic disc
of both males reached the anus.
The reduction of the cephalic lateral line canals
in the newly collected specimens of K. goerneri
matches with two types of reduction observed by
Ahnelt, 1991 (the types shown in his figures 3a,d
and 3b,e), both of which are well-represented in
the present sample. Some morphological details
missing in the original description were provided
in the present data (e. g. caudal-fin segmented-ray
count, exact limits of scaled area on body surface).
Ahnelt (1991) described the species based on only
five specimens, of which two were males and
three females. Measurements conducted on the
slightly larger sample size and better preserved,
freshly collected specimens, showed somewhat
higher morphological variability than previously
thought (Table 1; Ahnelt, 1991: table 1). However,
some morphometric characters (fin bases and fin
lengths) in the present study largely extend one
side of the range of values compared with the
original description. This is probably a result of
growth allometry, as five of the nine specimens
used in this study were small (13.1-15.7 mm SL)
compared to the comparatively larger specimens
used in the original description (17.8-21.8 mm
SL) (Ahnelt, 1991). The five new small specimens
had, proportionally to the standard length, shorter
dorsal- and anal-fin bases, as well as shorter cau-
dal- and pectoral-fin lengths compared with the
larger specimens in our sample and in the type
Molecular relationships. Based on cytochrome b
nucleotide sequence analyses, K. goerneri is closely
related to K. milleri, another species from the
north-eastern part of the Ionian Sea basin (Fig. 3).
The cytochrome b haplotypes of both species
form unambiguous reciprocally monophyletic
lineages (Fig. 3), and together they form a well-
supported clade, which is in a sister position to the
cytochrome b haplotypes of K. thessala, a species
endemic to the Thessalian Pinios River drainage in
the north-western Aegean Sea basin (on the other
side of the Pindos Range, a major biogeographic
barrier). The genetic distance between K. goerneri
and K. milleri inferred from the cytochrome b gene
sequences is around 3 % (Table 2). The intraspe-
cific genetic variability of both species based on
cytochrome b is several times lower; in K. milleri
it does not exceed 0.9 %, whereas in K. goerneri it
is much lower, with a maximum of 0.2 %.
Several other populations of Knipowitschia,
whose taxonomic status is not clarified, are known
from the north-eastern Ionian Sea basin (from the
Kalamas River to the western Peloponnese Alfios
River; Economou et al., 2007, Barbieri et al., 2015;
Fig. 1). Only some of these have been genetically
studied thus far (Vanhove et al., 2012; Geiger et
al., 2014). These studies show that several Knipo-
witschia populations (from the Acheloos and
Evinos River basins and the Prokopos-Kotychi
drainage in north-western Peloponnese) are
closely related to K. milleri. Each study used dif-
ferent mitochondrial genetic markers (12S and
16S in Vanhove et al., 2012; COI in Geiger et al.,
2014; cytochrome b in our study), thus it is not
possible to compare the results directly due to
the different mutational rates in different genes
(Wan et al., 2004). As K. goerneri is also closely
related to K. milleri, it is necessary to compare
all populations of Knipowitschia from the north-
eastern Ionian Sea basin with these two species
in order to delimit their exact distribution range
and to clarify the taxonomy of the Knipowitschia
populations from this area. This should ideally be
done by combining the genetic and morphological
approaches, and could potentially result in the
discovery of additional species.
Ecology and distribution. Knipowitschia goerneri
was originally found in a freshwater spring, as
well as in the brackish lagoon of Korission (Ah-
nelt, 1991). We discovered that the species is still
present in the lagoon, though we sampled at two
sites and the species was only found at one. The
sampling point at the lagoon was very shallow
(maximum depth of 40 cm), with a muddy bottom
containing numerous shell remains and a dense
filamentous algal covering. From the locality
where it was found, we assume that K. goerneri
is a euryhaline and eurythermal species. It is
probable that the conditions in the lagoon vary
substantially during the course of the year. The
size and status of the population are unknown.
Further research of the Korission Lagoon catch-
ment and its surroundings on Kerkyra Island is
necessary in order to characterise the ecological
and biological requirements of K. goerneri, its habi-
tat preferences and its distribution. This species
may occur in other wetland sites on the island,
as well as on the mainland opposite Kerkyra in
Greece and in Albania.
Conservation and threats. Ahnelt (1991) de-
scribed K. goerneri based on specimens from a
freshwater spring south-west of Korission La-
goon (the type locality), as well as the north-west
beach of the lagoon. Since the work of Ahnelt
(1991), the lagoon has not been mentioned as the
locality for K. goerneri (Economidis, 1995; Miller,
2004b; Economidis & Chrysopolitou, 2009), and
there is no published evidence that it has been
investigated for its presence since 1983. The as-
sumption that this species is critically endangered
or possibly extinct (Miller, 2004b; Economidis &
Chrysopolitou, 2009) was based on the destruction
of Gardiki Spring, situated north of the lagoon
(Economidis, 1995), which was erroneously men-
tioned as the only locality of K. goerneri (Miller,
2004b; Economidis & Chrysopolitou, 2009), even
though the species has never been found there.
Although K. goerneri was listed in the Red Data
Book of Greece as critically endangered and pos-
sibly extinct (Economidis & Chrysopolitou, 2009),
no conservation actions have been proposed
for this species (Economidis & Chrysopolitou,
2009). Despite the Korission Lagoon catchment
being designated as a Natura 2000 site (European
Environment Agency, 2014), it is threatened by
development, pollution, fisheries, and over-
pumping groundwater.
Conclusions. The rediscovery of K. goerneri in
this study, as well as another freshwater fish spe-
cies, Telestes miloradi (Jelib & Jelib, 2015), shows
that even in Europe species can be overlooked
for decades. Hence, it is possible that some other
species currently thought to be extinct (Smith &
Darwal, 2006; Kottelat & Freyhof, 2007; Geiger et
al., 2014) could in fact still persist (e. g. K. cameliae,
Chondrostoma scodrense). Intensive field work is
Table 2. Mean uncorrected pairwise genetic distance (p-distance; %) between analysed species of Knipowitschia
based on cytochrome b nucleotide sequence analysis. Number of analysed specimens and range of intraspecific
genetic variability included next to species names in parentheses, and mean intraspecific genetic variability
included in bold font.
K. goerneri K. milleri K. thessala K. montengrina K. panizzae K. caucasica P. bathi
K. goerneri (8; 0-0.18) 0.045
K. milleri (5; 0-0.89) 3.2 0.55
K. thessala (4; 0-0.09) 7.8 7.6 0.06
K. montenegrina (3; 0.09-0.18) 8.2 8.8 8.6 0.12
K. panizzae (2; –) 8.5 8.9 8.1 3.9 1.43
K. caucasica (4; 0-1.43) 10.3 10.1 9.9 8.7 8.5 0.95
Pomatoschistus bathi (1; –) 16.4 17.1 16.8 16.3 16.2 15.2 –
Vukib et al.: Rediscovery of Knipowitschia goerneri
Ichthyol. Explor. Freshwaters, Vol. 26, No. 4
needed to better evaluate the possible presence
of such taxa in the Mediterranean region.
Comparative material. Knipowitschia milleri: PMR
VP3212, 1 female, 22.9 mm SL; PMR VP3213, 1 male,
20.2 mm SL; PMR VP3214, 1 juvenile, 17.5 mm SL;
Greece: Acheron River.
Material used in molecular genetic analysis: K. milleri:
NMP P6V142642-142646, 5; Greece: Acheron River,
39°14'34" N 20°28'40" E (GenBank accession numbers:
K. thessala: NMP P6V141242, 1; Greece: Pinios drain-
age: Enipeas River, 39°33'43" N 22°4'45" E (GenBank
accession number: KT809433). – NMP P6V141418, 1;
NMP 141422-141423, 2, Greece: Pinios drainage: Lithe-
os River, 39°33'14" N 21°46'09" E (GenBank accession
numbers: KF214247, KT809431-KT809432).
K. montenegrina: NMP P6V89874, 1; NMP D1326-
D1325, 2; Montenegro: Moraca River, 42°18'37" N
19°11'56" E (GenBank accession numbers: KT809450-
K. panizzae: PMR VP1905, 1; Croatia: Privlaka,
44°16'37" N 15°07'53" E (GenBank accession number:
KT809453). – PMR VP1985, 1; Croatia: Cetina River,
43°26'21" N 16°45'02" E (GenBank accession number:
K. caucasica: NMP P6V88956-88957, 2; Bulgaria:
Pomorijsko Lake, 42°33'56" N 27°37'56" E (GenBank
accession number: KF214250, KT809449). – NMP
P6V141300, 1; NMP 141302, 1; Greece: Volvi Lake,
40°39'14" N 23°30'04" E (GenBank accession number:
Pomatoschistus bathi: NMP CG089B, 1; Montenegro:
Becibi: Adriatic Sea, 42°16'26" N 18°53'16" E (GenBank
accession number: KT809430).
This work was supported by the Ministry of Culture of
the Czech Republic (DKRVO 2014/14, DKRVO 2015/15,
National Museum, 00023272) and by the institutional
resources of the Ministry of Education, Youth, and
Sports of the Czech Republic (JV). MK received support
from the SYNTHESYS Programme project CZ-TAF-3784
at the National Museum Prague, financed by European
Community Research Infrastructure Action under the
FP7 Capacities Programme. We thank Anja Palandacib
(NMW) for the loan of the type material of K. goerneri.
We acknowledge the assistance of the colleagues from
the Hellenic Centre for Marine Research and particu-
larly of Alcibiades N. Economou. Aris Vidalis provided
assistance with cartography. We thank the editor and
two anonymous referees for their valuable comments,
and T. Kohler for language correction.
Literature cited
Ahnelt, H. 1991. A new species of Knipowitschia (Tele-
ostei: Gobiidae) from Corfu, Western Greece. Ich-
thyological Exploration of Freshwaters, 2: 265-272.
1995. Two new species of Knipowitschia from West-
ern Anatolia (Turkey). Mitteilungen aus dem Ham-
burgischen Zoologischen Museum und Institut, 92:
2011. Two new sympatric Knipowitschia species
(Teleostei: Gobiidae) from an eastern Mediterra-
nean coastal lake – examples of different dispersal
patterns? Zootaxa, 3114: 22-30.
Ahnelt, H. & P. G. Bianco. 1990. Orsinigobius milleri
n. sp., a new species of freshwater goby from W-
Greece (Pisces, Gobiidae). Annalen des Naturhis-
torischen Museums in Wien, Serie B, Botanik und
Zoologie, 91: 1-6.
Barbieri, R., S. Zogaris, E. Kalogianni, M. Stoumboudi,
Y. Chatzinikolaou, S. Giakoumi, Y. Kapakos, D.
Kommatas, N. Koutsikos, V. Tachos, L. Vardakas
& A. N. Economou. 2015. Freshwater fishes and
lampreys of Greece: an annotated checklist. Mono-
graphs on Marine Sciences No. 8. Hellenic Centre
for Marine Research, Athens, 130 pp.
Buj, I., R. Šanda, Z. Marcib, M. Baleta & M. Mrakovcib.
2014. Combining morphology and genetics in re-
solving taxonomy – a systematic revision of spined
loaches (genus Cobitis; Cypriniformes, Actino-
pterygii) in the Adriatic watershed. PLoS ONE, 9:
Buj, I., J. Vukib, R. Šanda, S. Perea, M. Baleta, Z. Marcib,
I. Bogut, M. Povž & M. Mrakovcib. 2010. Morpho-
logical comparison of bleaks (Alburnus, Cyprinidae)
from the Adriatic Basin with the description of a
new species. Folia Zoologica, 59: 129-141.
Crivelli, A. J. 2006. Knipowitschia goerneri. The IUCN
Red List of Threatened Species. Version 2014.3.
<> (accessed 20 February 2015)
Economidis, P. S. 1995. Endangered freshwater fishes
of Greece. Biological Conservation, 72: 201-211.
Economidis, P. S. & V. Chrysopolitou. 2009. Kni-
powitschia goerneri Ahnelt, 1991. Pp. 103-104 in:
A. Legakis & P. Maragkou (eds.), The red book of
threatened animals of Greece. Hellenic Zoological
Society, Athens.
Economou, A. N., S. Giakoumi, L. Vardakas, R. Bar-
bieri, M. Stoumboudi & S. Zogaris. 2007. The
freshwater ichthyofauna of Greece: an update based
on a hydrographic basin survey. Mediterranean
Marine Science, 8: 91-168.
European Environment Agency. 2014. Sites: Limnotha-
lassa Korission (Kerkyra). http://eunis.eea.europa.
eu/sites/GR2230002 (accessed 20 February 2015)
Freyhof, J. & E. Brooks. 2011. European Red List of
Freshwater Fishes. Publications Office of the Euro-
pean Union, Luxembourg, 62 pp.
Freyhof, J., H. Karst & M. Geiger. 2014. Valencia robertae,
a new killifish from southern Greece (Cyprinodon-
tiformes: Valenciidae). Ichthyological Exploration
of Freshwaters, 24: 289-298.
Freyhof, J., M. Kottelat & A. Nolte. 2005. Taxonomic
diversity of European Cottus with description of
eight new species (Teleostei: Cottidae). Ichthyo-
logical Exploration of Freshwaters, 16: 107-172.
Geiger, M. F., F. Herder, M. T. Monaghan, V. Almada,
R. Barbieri, M. Bariche, P. Berrebi, J. Bohlen, M.
Casal-Lopez, G. B. Delmastro, G. P. J. Denys, A.
Dettai, I. Doadrio, E. Kalogianni, H. Kärst, M. Kot-
telat, M. Kovacib, M. Laporte, M. Lorenzoni, Z.
Marcib, M. Özulug, A. Perdices, S. Perea, H. Persat,
S. Porcelotti, C. Puzzi, J. Robalo, R. Šanda, M. Schnei-
der, V. Šlechtová, M. Stoumboudi, S. Walter & J.
Freyhof. 2014. Spatial heterogeneity in the Mediter-
ranean biodiversity hotspot affects barcoding ac-
curacy of its freshwater fishes. Molecular Ecology
Resources, 14: 1210-1221.
Guindon, S. & O. Gascuel. 2003. A simple, fast, and
accurate algorithm to estimate large phylogenies
by Maximum Likelihood. Systematic Biology, 52:
Huelsenbeck, J. P. & F. Ronquist. 2001. MRBAYES:
Bayesian inference of phylogenetic trees. Bioinfor-
matics, 17: 754-755.
Jelib D. & J. Jelib. 2015. Telestes miloradi Bogutskaya,
Zupancic, Bogut & Naseka, 2012 and Delminichthys
ghetaldii (Steindachner, 1882) re-discovered in Croa-
tia, requiring urgent protection. Journal of Applied
Ichthyology, 31: 1133-1136.
Kalogianni, E., S. Giakoumi, S. Zogaris, Y. Chatz-
inikolaou, M. T. Stoumboudi, R. Barbieri, A. N.
Economou & B. Zimmerman. 2006. Rapid assess-
ment of the status of Valencia letourneuxi, the Greek
killifish. Final report. Hellenic Centre for Marine
Research & Zoological Society of London, 179 pp.
Kottelat, M. 1997. European freshwater fishes. Biologia,
Bratislava, 52 (Suppl. 5): 1-271.
— 2007. Three new species of Phoxinus from Greece
and southern France (Teleostei: Cyprinidae). Ich-
thyological Exploration of Freshwaters, 18: 145-162.
Kottelat, M. & J. Freyhof. 2007. Handbook of European
freshwater fishes. Kottelat, Cornol, and Freyhof,
Berlin, 646 pp.
Kovacib, M. 2005. A new species of Knipowitschia (Go-
biidae) from Dalmatia, Croatia. Cybium, 29: 275-
Kovacib, M. & R. Šanda. 2007. A new species of Knipo-
witschia (Perciformes: Gobiidae) from southern
Montenegro. Journal of the National Museum
(Prague), Natural History Series, 176: 81-89.
Machordom, A. & I. Doadrio. 2001. Evidence of a ceno-
zoic Betic-Kabilian connection based on freshwater
fish phylogeography (Luciobarbus, Cyprinidae).
Molecular Phylogenetics and Evolution, 18: 252-263.
Miller, P. J. 1972. Gobiid fishes of the Caspian genus
Knipowitschia from the Adriatic Sea. Journal of the
Marine Biological Association of the United King-
dom, 52: 145-160.
1988. New species of Corcyrogobius, Thorogobius and
Wheelerigobius from West Africa (Teleostei: Gobi-
idae). Journal of Natural History, 22: 1245-1262.
— (ed.) 2004a. The freshwater fishes of Europe. Vol.
8/II. Gobiidae 2. Aula, Wiebelsheim, 478 pp.
— 2004b. Knipowitschia goerneri Ahnelt, 1991. Pp. 370-
374 in: Miller (ed.), The freshwater fishes of Europe.
Vol. 8/II. Gobiidae 2. Aula, Wiebelsheim.
2009. A west Balkanian freshwater gobiid fish,
Knipowitschia mrakovcici sp. nov. (Teleostei: Gobi-
idae). Journal of Fish Biology, 74: 1499-1507.
Miller, P. J. & R. Šanda. 2008. A new West Balkanian
sand-goby (Teleostei: Gobiidae). Journal of Fish
Biology, 72: 259-270.
Posada, D. 2008. jModelTest: phylogenetic model aver-
aging. Molecular Biology and Evolution, 25: 1253-
Šanda, R., J. Vukib, L. Choleva, J. Krížek, A. Šedivá, S.
Shumka, & I. F. Wilson. 2008. Distribution of loach
fishes (Cobitidae, Nemacheilidae) in Albania, with
genetic analysis of populations of Cobitis ohridana.
Folia Zoologica, 57: 42-50.
Sanzo, L. 1911. Distribuzione delle papille cutanee
(organi ciatiforme) e suo valore sistematico nei Gobi.
Mitteilungen aus der Zoologischen Station zu Ne-
apel, 20: 249-328.
Saruwatari, T., J. Andrés López & T. W. Pietsch. 1997.
Cyanine blue: a versatile and harmless stain for
specimen observation. Copeia, 1997: 840-841.
Smith, K. G. & W. T. R. Darwall. 2006. The status and
distribution of freshwater fish endemic to the
Mediterranean basin. IUCN, Gland and Cambridge,
34 pp.
Tamura, K., D. Peterson, N. Peterson, G. Stecher, M.
Nei & S. Kumar. 2011. MEGA5: molecular evolution-
ary genetics analysis using Maximum Likelihood,
evolutionary distance, and Maximum Parsimony
methods. Molecular Biology and Evolution, 28:
Vanhove, M. P. M., A. N. Economou, S. Zogaris, M. H.
D. Larmuseau, S. Giakoumi, E. Kalogianni, F. A. M.
Volckaert & T. Huyse. 2012. Phylogenetics and bio-
geography of the Balkan sand gobies (Teleostei:
Gobiidae): vulnerable species in need of taxo-
nomic revision. Biological Journal of the Linnean
Society, 105: 73-91.
Vinciguerra, D. 1921. Descrizione di tre nuove specie
di pesci delle acque dolci di Grecia. Annali del
Museo Civico di Storia Naturale Giacomo Doria, 3:
Wan, Q. H., H. Wu, T. Fujihara & S. G. Fang. 2004.
Which genetic marker for which conservation genet-
ics issue? Electrophoresis, 25: 2165-2176.
Received 26 February 2015
Revised 10 November 2015
Accepted 6 January 2016
Vukib et al.: Rediscovery of Knipowitschia goerneri
... Kod Gobiidae raspored pora na glavi (Sl. 19, 71 i 75) predstavlja jedan od detrminirajućih karaktera za ovu grupu riba (Kovačić, 2005;Miler & Šanda, 2008;Vukić, et al. 2016). U poslednje vrijeme se u determinaciji vrsta, pogotovo u opisivanju novih, ovaj karakter često koristi. ...
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Drage kolege Zamolio bih kolege,naravno koji su zainteresovani, a koji razumiju jezik kojim je ovaj tekst (rukopis) napisan da daju svoj komentar ili mišljenje o istom. Mišljenje, preporuke, sugestije koje bi ste uradili, a bili u skladu s pravilima recenziranja bila bi uvrštena u proceduru a vaše ime u RECENZENTE. Mišljenje možete poslati preko RESEARCH GATE ili preko meila: Unaprijed zahvaljujem
... Similarly, small gobiids such as Knipowitschia spp. require taxonomic skills and molecular assessment in the laboratory, thus their taxonomy in several lentic ecosystems is still far from complete in several lakes (see Vukić et al., 2016Vukić et al., , 2017. The provenance and taxonomy of isolated populations of Knipowitschia on certain wetlands are still pending (Vanhove et al., 2011(Vanhove et al., , 2016Koutsikos et al., 2019a) as well as for some isolated lake populations (e.g., lakes Ozeros and Voulkaria (Barbieri et al., 2015). ...
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This study provides an annotated checklist of the freshwater fish species recorded in lentic ecosystems of Greece. Species distributional data were derived from an extensive review of published and unpublished sources and were evaluated for their consistency, adequacy and reliability. Overall, 650 freshwater lentic ecosystems (149 natural and 501 artificial) were investigated from which, 480 were fishless or fish data were unavailable. In the remaining 170 ecosystems, 122 taxa were recorded (belonging to 22 families and 57 genera). Some of the records concerned species with extirpated populations, species of doubtful occurrences and taxa of uncertain taxonomic status. The highest species richness (52 species) was recorded in the Ionian ecoregion while endemicity was highest in the Macedonia-Thessaly ecoregion (43.75%). The Eastern Aegean ecoregion presented the highest level of introduced species (75%). Only eight ecosystems hosted 20 or more species, while 92 ecosystems had over 50% of introduced species. The most widespread native species (Anguilla anguilla and Luciobarbus albanicus) were two highly migratory species, highlighting the importance of connectivity of aquatic ecosystems. The most widespread translocated species was Cyprinus carpio due to its high commercial and recreational value, whereas Gambusia holbrooki confirmed its high dispersal ability being the most widespread alien species. More than one-third of the species recorded (37.23 %) are threatened based on the IUCN Red List. The ultimate contribution of this study is to support the coordination and dissemination of baseline information on the freshwater fishes of Greek lentic ecosystems, required by a range of users. This knowledge will enable further understanding of the regional assemblage structure and other biogeographical patterns of the ichthyofauna of Greece and will contribute to species conservation by prioritising areas hosting species in need of protection. Finally, this study reveals the high knowledge gap regarding species composition in many Greek lentic ecosystems and highlights the need for relevant primary research including more systematic and standardised samplings.
... Malavasi et al., 2012;Thacker et al., 2019;Vanhove et al., 2012;Vukić et al., 2016Vukić et al., , 2017. However, the concatenation of cytb and rh1 provided here a highly resolved phylogeny at the interspecific level.Several molecular genetic studies have pointed to monophyly of the sand gobies sensu lato(Agorreta et al., 2013;Huyse et al., 2004;Thacker et al., 2019;Vanhove et al., 2012), with Gobiusculus Duncker, 1928 as a junior synonym of Pomatoschistus(Huyse et al., 2004;Knebelsberger & Thiel, 2014;Larmuseau et al., 2010;Thacker et al., 2019;Vanhove et al., 2012). ...
The Adriatic Sea and Adriatic Sea drainage system (or Adriatic region) is characterized by high levels of species richness. This especially applies to endemic fishes, as exemplified by the sand gobies. However, the diversity of this group is probably still underestimated. Several keys for gobiid identification, notably of the Adriatic region, have been proposed on the basis of morphological characters, but difficulties persist in species identification. Molecular phylogenies support the monophyly of the sand gobies, but relationships between genera and species remain poorly resolved. In this work, the diversity and evolutionary history of the Adriatic sand gobies (Pomatoschistus and Knipowitschia) were investigated by applying phylogenetic reconstructions, species delimitation tests, and molecular dating based on mitochondrial cytochrome b and nuclear rhodopsin 1 genes. The concatenation of both genes allowed, for the first time, the creation of a highly resolved phylogeny at the interspecific level. The generic diversity within the sand gobies is probably considerably higher than formerly assumed. Our results highlight cryptic diversity and suggest that new species remain to be described within Knipowitschia and Pomatoschistus. Most of all, our results emphasized the urgent need of a taxonomic revision of the sand gobies. Several past events were at the origin of sand goby diversification in the Adriatic region. It seems that glacial periods promoted the intraspecific diversification of the endemic species, whereas interglacial periods promoted the (re)colonization of the Adriatic Sea by more widespread species.
... A slightly higher intraspecific variability (1.5%) has been found in G. cruentatus by Čekovská et al. (2020). High genetic divergences were observed also between the species of other European gobioid genera (Knebelsberger and Thiel 2014;Vukić et al. 2016;Thacker et al. 2019). The divergence between G. xoriguer sp. ...
Gobius xoriguer sp. nov., a new offshore species of goby (Teleostei: Gobiidae: Gobiinae) is described based on three specimens collected in 2010, 2012 and 2018 in the western Mediterranean, off Menorca (Spain), in the Gulf of Lion and off Corsica (France), at 51–104 m depth on coralline algae sea bed. It is easily distinguishable from its Atlantic-Mediterranean congeners by the combination of the following characters: large eyes, ~27-28% of head length; anterior nostril with a small triangular process; 14 soft rays on D2, 13 soft rays on A; enlarged first dorsal fin rays (adult males), with third D1 spine the longest, 23-27 % SL; uppermost P fin rays not free from membrane; long V-shape pelvic fins with vestigial frenum; 50-51 scales on LL; head oculoscapular canal with pores σ, λ, κ, ω, α, β, ρ, ρ1, ρ2, and preopercular canal with pores γ, δ, ε present; row x1 ending anteriorly behind pore β; a groove section between pore ρ and ρ1; suborbital row d discontinuous with large gap below suborbital rows 3 and 4; rows o separated; seven enlarged orange blotches on body side; white dotes on cheek and opercle on an orange background.With a known maximum size of 64 mm TL, it is among the smaller species of Gobius. Bayesian inference and Maximum likelihood phylogenetic tree topologies based on mitochondrial DNA COI sequences (barcoding region), including most Atlantic-Mediterranean Gobius species, support Gobius gasteveni Miller 1974 as the closest relative to Gobius xoriguer sp. nov. These sister species exhibit a high genetic divergence of 9.5% (uncorrected p-distance).
... Similarly, a study on Telestes pleurobipunctatus (Stephanidis 1939) revealed a high genetic differentiation between populations from different basins, suggesting the existence of a complex of species in the Ionian Sea slope (Buj et al., 2019). On the other hand, Knipowitschia goerneri Ahnelt 1991, which was believed to be restricted to a single lagoon system on Kerkira Island, is most probably much more widespread in the Ionian biogeographic region (Vanhove et al., 2016;Vukić et al., 2016Vukić et al., , 2017. Finally, molecular analyses of the species of Squalius Bonaparte 1837 from the south of the Peloponnese uncovered a past hybridization between evolutionary very distant lineages, which led to a complete mitochondrial DNA introgression (Perea et al., 2016). ...
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Pelasgus Kottelat & Freyhof 2007 (Leuciscidae) is a freshwater fish genus endemic to the Southern Balkans. The distribution of most of its species is insufficiently known. Pelasgus was molecularly studied only marginally, and the genetic diversity of individual species or populations, crucial for their conservation, is completely unknown. We studied distribution and genetic diversity of Pelasgus stymphalicus (Valenciennes 1844) and Pelasgus marathonicus (Vinciguerra 1921), two widespread species from southern Greece. Our data, based on cytochrome b sequences, confirmed that a number of populations whose taxonomic status had been uncertain, belong to one of these species. The distribution range of P. stymphalicus includes the Ionian mainland, from the Acheloos to Mornos rivers, and most of the Peloponnese, excluding the Evrotas and the headwaters of Alfios. The native range of Pelasgus marathonicus is the western Aegean mainland, from rivers near Athens to the Xerias river in the Pagasitikos Gulf. AMOVA showed that the genetic variance is slightly higher among than between populations. High F ST values indicated a pronounced genetic differentiation of the populations in both species. There was a high proportion of private haplotypes and a very small number of shared haplotypes between populations in both species, which indicates uniqueness of each population and their susceptibility to human-induced changes.
The high diversity of extant gobiids (Gobiidae: Teleostei) makes taxonomic and phylogenetic interpretation of fossil members of the clade a difficult task. To facilitate future taxonomic and systematic work on the group, we have assembled a morphological reference database encompassing skeletal characters, an otolith atlas and otolith morphometric data of 25 present-day species from the European Gobius lineage (s.l.) that represent 18 different genera and include all nine sublineages. We show that: (1) skeletal traits and morphometric otolith variables can be diagnostic for a sublineage; (2) otolith morphology allows identification at the genus and species levels; and (3) the number of anal-fin rays and details of the otolith margins can be used to discriminate closely related dwarf gobies. The skeletal and otolith characters are largely stable in the marine gobies analysed here, whereas freshwater gobies (Padogobius, Ponto-Caspian gobies) are far more variable. This might be related to the conquest by Padogobius and Ponto-Caspian gobies of freshwater and low-salinity habitats, in which environmental conditions can fluctuate widely. We anticipate that the database presented here can be used as a valuable reference tool to assess the relationships of fossil gobiids and increase our knowledge of the evolutionary history of the group as a whole.
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The gobies (Gobiidae) are the most diverse fish family in the Mediterranean Sea. Nevertheless, knowledge on their diversity, taxonomy, and phylogenetic relationships is still inadequate. The phylogenetic analyses reveal two genetically highly distinct clades among specimens identified as Zebrus zebrus . A new species, Zebrus pallaoroi sp. nov., is described based on an integrative approach. The neotype of Zebrus zebrus is designated. Genetic data confirm a pronounced level of divergence between Z. pallaoroi and Z. zebrus , with the mean genetic distance on cytochrome b being 18.1% and 1.07% on rhodopsin. Phylogenetic relationships within the Gobius -lineage were estimated on both markers. Morphologically, Z. pallaoroi is distinguished from the only congener Z. zebrus by having a snout longer than its eye, posterior nostril about 4/5–9/10 of the anterior nostril, eye diameter 4.3−4.7 in head length, ventrolateral head ridges transversally connected on the anterior side by a short transversal ridge, anterior membrane midline depth about 2/3 of the spinous ray, head canal pore α diameter about half of the distance between pore ρ and ρ 1 , suborbital sensory papillae row 5i going downwards to or near the level of row d , the distance between row 5i and row d absent or much smaller than the length of row 5i , and the body with ten to eleven vertical dark brown bands. Zebrus pallaoroi was recorded from the southern Adriatic, northern Ionian, and northern and western Aegean Seas, and is a cryptobenthic fish from very shallow waters.
Coastal lagoons are resilient and productive ecosystems that support high biological and habitat diversity, but are increasingly affected by several threats due to human exploitation. Many resident species of these areas show a restricted geographical range and a strict association with specific habitat, thus they could represent ecological indicators of these ecosystems. Data on the genetic variability distribution in populations of these species are crucial to identify the presence of different evolutionary units across their geographical distribution range and to plan actions for their management and conservation. In this context, the assessment of the genetic variability and structure of Italian specimens of the Adriatic dwarf goby Knipowitschia panizzae, a brackish species endemic in the Adriatic Sea, and included among the species of Community interest in Annex II of Habitats Directive, was carried out. To this purpose, goby samples were collected both from Italian sites where the species is native (Adriatic) and non-native (Tyrrhenian), probably introduced due to the practice of stocking lakes and coastal lagoons with juvenile of euryhaline species to sustain local fisheries. (CR)(COI) Results show the presence of high values of haplotype diversity, and no shared haplotypes between fish from sites where the species is native or introduced. Moreover, in Tyrrhenian Italian lagoons we identified an allochthonous species of Knipowitschia, previously undetected in Italian territories: the Corfu dwarf goby K. goerneri. This species was originally described as endemic to the Korission Lagoon catchment on Corfu Island but recently it has been reported also in Butrinti lagoon in Albania. The presence of this species, that almost totally replaced Adriatic dwarf goby in the Tyrrhenian lagoons under study, can be ascribed to the stockings of wild fry of commercially important species originating from Balkan countries.
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This critical catalogue from the “The Age of Genes” exhibition presents readers with the comprehensive development of molecular phylogenetic and taxonomic research conducted at the National Museum. The reader will thus become acquainted with the Natural History Museum, which is part of the National Museum, and within which modern molecular genetic laboratories were established in 2003 at the Zoological Department and in 2016 at the Entomological Department. To facilitate orientation in this type of research, a general introduction is provided to present in brief a global-scale general advances in the field of genetics as well as a research into the DNA molecule itself, in the form of a historical survey. From Mendel‘s Laws of 1869 to the first real photo of the DNA helix taken by a transmission electron microscope in 2012. In addition, the actual structure of the DNA molecule, its placement, function and inheritance, which are among the basic attributes of this compound, are described in detail. This publication is written in Czech language. Certainly major milestones, and not just for zoology, were Sanger‘s DNA sequencing, i.e. determining the order of the individual components (nucleotides) in the DNA obtained from an organism, and then the Mullis polymerase chain reaction, which serves to simply amplify a selected DNA segment taken from along its entire length. These two molecular genetic methods are now quite routinely used in zoology, and these approaches have opened up a whole new level of knowledge. In general, these methods are used in a major way in research into the evolution of organisms, of their phylogenetic relationships, systematics and taxonomy in general and, of course, in the study of organism populations and their interrelationships. In addition to new, hitherto unresolved questions, these methods have an impact on many already existing results, which have often been transformed into quite different forms. The catalogue presents in essence the whole of the molecular taxonomic and phylogenetic research of the Natural History Museum. It is an up-to-date list of works making use of these methods and for selected studies such approaches are described in more detail for the lay public. The reader will thus learn more about the work of the Natural History Museum, which includes describing dozens of new species and subspecies from the world’s fauna every year. This type of research not only progressively reveals more fragments of the overall diversity of organisms on our planet, but can also be used very successfully in conservation practice. At the same time, research is pointed out that was directly funded by the NAKI II project, which also gave rise to The Age of Genes exhibition and this critical catalogue that accompanies it. The final part of the catalogue consists of individual exhibit photographs with details.
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A checklist of the freshwater fish fauna of Croatia is presented for the first time. It is based on 1360 publications of historical and recent data in the literature. According to the literature review, there were 137 fish species in 30 families and 75 genera recorded in Croatia. The checklist is systematically arranged and provides distributional data of the freshwater fish fauna as well as whether the species is endemic, introduced or translocated.
Technical Report
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This report presents the results of the first complete baseline investigation of V. letourneuxi aiming to provide appropriate information on its distribution, abundance, ecology, habitat conditions and local population status, upon which inferences about threats to the species, vulnerability to natural or human disturbances of its habitats and future viability prospects can be based. This information will also be used as a reference point for future surveys and can factor into the management plan for this species.
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Between 2003 and 2006, almost 80 localities in all main hydrological systems in Albania were sampled and data on the distribution of loach fishes gathered. The spined loach Cobitis ohridana Karaman, 1928 was found to be a common species in Albania, occurring in most of its river systems, from the Ohrid-Drin-Shkodra system in the east and north to the River Vjosë basin in the south. Cobitis meridionalis Karaman, 1924 occurs in Lake Prespa, while a spined loach with mtDNA of Cobitis sensu stricto origin is present at least in the Ohrid-Drin- Shkodra system. The most common stone loach in Albania was found to be Oxynoemacheilus pindus (Economidis, 2005). It was caught in the basins of the rivers Vjosë, Seman, Shkumbin and Erzen. Barbatula sturanyi (Steindachner, 1892) was recorded in the River Black Drin.
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Distribution records (historical, contemporary) for native and non-native freshwater fish species from 105 hydrographic basin areas were compiled and analysed in order to develop a nation-wide inventory (including transboundary river basins). Overall, 162 species, including diadromous and euryhaline, with documented occurrence records in freshwaters, and taxa of unclarified taxonomic status, are accommodated in the distributional compilation. An annotated checklist summarises the confirmed ichthyofauna of Greek freshwaters (161 species); a provisional supplementary list contains species recorded in brackish waters (55 species). In comparison to the last published (1991) checklist of freshwater fish of Greece, the present checklist shows an increase in species number of 53% (56 species). This increase has resulted mainly from taxonomic re-evaluations of existing taxa on the basis of new information and adoption of a new systematic concept. The current trend, as reflected in recent ichthyological publications, is towards abandonment of the biological species concept (BSC) and adoption of the phylogenetic species concept (PSC) for the delineation of species boundaries. The practical implications of the change in species concept on biodiversity conservation and watershed management are discussed. An overview of the composition and characteristics of the freshwater fish fauna of Greece is provided, especially with regard to the native and introduced status of species, and the spatial patterns of species richness and endemism. This systematic inventory may assist in efforts to develop nation-wide surface water bioassessment tools within the demands of the Water Framework Directive (WFD); it may further promote biodiversity conservation and biologically-orientated fishery management approaches.
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The peri-Mediterranean populations of Phoxinus usually identified as the wide-ranging palaearctic P. phoxinus represent a number of distinct species. Three new species are described: P. bigerri from Adour drainage, France and the upper Ebro drainage, Spain, P. septimaniae from Languedoc, France, and P. strymonicus from Strymon drainage, Greece. Species of Phoxinus differ in body proportions and shape (especially shape of caudal peduncle, anal fin, snout), shape of the patches of breast scales between and in front of pectoral fins, and live colouration. A neotype is designated for Cyprinus phoxinus and C. aphya, and C. phoxinus is made a nomen protectum.
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Taxonomic investigation of spined loaches from Dalmatia and Herzegovina was conducted on specimens from 14 localities. The results of the detailed morphological investigations were combined with genetic data (based on one mitochondrial and two nuclear genes) in order to resolve the taxonomic status of each Cobitis population. Among the investigated features of external morphology, the appearance of spots on the caudal fin base turned out to have the greatest diagnostic value. Furthermore, the number of branched fin rays enabled the discrimination of several species. No morphometric character alone could ensure determination of any Cobitis species. Nevertheless, groups of populations that are more similar in their body shapes correspond to mitochondrial phylogenetic lineages. Based on molecular genetic markers, Dalmatian and Herzegovinian spined loaches form independent lineages inside the Adriatic phylogenetic group. Mitochondrial DNA phylogenetic reconstruction revealed six monophyletic lineages, corresponding to six species distributed in the investigated area. The population distributed in Mostarsko blato karstic field in Bosnia and Herzegovina is described as a new species based on a unique combination of morphological characters: a single triangular Canestrini scale; usually 51/2 branched anal fin rays, 61/2 branched dorsal fin rays, 14 branched caudal fin rays; no spots in the surface pigmentation layer on the caudal fin base; scales on the body very small.
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Valencia robertae, new species, from the lower Pinios in northern Peloponnese and Mornos Rivers in southern mainland of Greece is distinguished from V. letoumeuxi and V. hispanica by having short lateral bars or vertically elongated small blotches along the midlateral body and an almost triangular anal fin in females, prominent lateral bars between the axial blotch and the caudal-fin base and a long anal fin reaching almost or to the first caudal-fin rays in males larger than 27 mm SL. It is also distinguished by 32 fixed, diagnostic nucleotide substitutions in the mtDNA COI barcode region.
The taxonomy of European species of Cottus (Cottidae) is revised. Results of molecular studies are summarised and the variability of morphological characters is reviewed. Molecular and morphological data support the rec-ognition of 15 diagnosable species in Europe. A neotype is designated for C. gobio; the type locality is in the lower Elbe drainage. Cottus gobio, C. hispaniolensis, C. koshewnikowi, C. microstomus, C. petiti, and C. poecilopus are re-diagnosed. Eight new species are described. Three of them are restricted to France: C. aturi to the Adour drain-age, C. duranii to the upper Dordogne, upper Lot and upper Loire drainages, and C. rondeleti to the Hérault drainage. Two new species are described from the Atlantic and North Sea basins: C. perifretum from Great Britain, and the Scheldt, Rhine, Seine, lower Loire and lower Garonne drainages, and C. rhenanus from the Meuse and lower and middle Rhine drainages. Cottus scaturigo is described from a single spring in northeastern Italy. In the Danube drainage, C. metae from the upper Save and C. transsilvaniae from the upper Arges are distinguished from the widespread C. gobio. Lectotypes are designated for C. ferrugineus and C. pellegrini. Cottus koshewnikowi Grat-zianow, 1907 is declared nomen protectum and C. gobio microcephalus Kessler, 1868 is declared nomen oblitum. The original spelling of C. milvensis is discussed.