A new species of Lepidocephalichthys (Teleostei: Cobitidae) with distinctive sexual dimorphism and comments on relationships in southern lineages of Cobitidae

Abstract

Lepidocephalichthys (Telostei:Cobitidae) is diagnosed as being unique among cobitids in having the 7–8th pectoral rays of mature males modified. Recently collected material from Thailand included a new species of Lepidocephalichthys in which mature males have a large (extending over ~75% of the fin-ray length) dorsally projecting and serrated flange and a ventrally projecting flange. The ventrally projecting structure is unique among cobitids. An expanded phylogenetic analysis of cobitids, including previously published sequences and new material including the new species, reinforces the monophyly of Lepidocephalichthys. Relationships within southern lineages of cobitids, and the unusual habitat of the new species are discussed.

Full text

Accepted by R. Pethiyagoda: 22 Jun. 2010; published: 3 Aug. 2010 1
ZOOTAXA
ISSN 1175-5326 (print edition)
ISSN 1175-5334 (online edition)
Copyright © 2010 · Magnolia Press
Zootaxa 2557: 1–18 (2010)
www.mapress.com/zootaxa/Article
A new species of Lepidocephalichthys (Teleostei: Cobitidae)
with distinctive sexual dimorphism and comments on
relationships in southern lineages of Cobitidae
JUSTIN C. HAVIRD1,2, LAWRENCE M. PAGE2,6, WEERAPONGSE TANGJITJAROEN3,
CHAVALIT VIDTHAYANON4, CHAIWUT GRUDPAN5, & SURIYA UDDUANG5
1Florida Museum of Natural History, University of Florida, Dickinson Hall, Gainesville, FL 32611, USA. E-mail: jhavird@ufl.edu
2Department of Zoology, University of Florida, 211 Bartram Hall, Gainesville, FL 32611, USA
3Department of Equine Science, Faculty of Veterinary Medicine, Chiang Mai University,Chonlapratan Rd, Mae Hea, Muang, Chiang
Mai, Thailand, 50100
4The Northeastern Research Institute for Petrified Wood and Mineral Resources, Nakorn Ratchasima Rajabhat University, 184 Moo 7
Tambon Suranaree, Mouang District, Nakorn Ratchasima, Thailand, 30000
5Department of Fisheries, Faculty of Agriculture, Ubonratchathani University 85, Sathollamark Rd., Warinchamrap, Ubon
Ratchathani, Thailand, 34190
6Corresponding author. E-mail: lpage1@ufl.edu
Abstract
Lepidocephalichthys (Telostei:Cobitidae) is diagnosed as being unique among cobitids in having the 7–8th pectoral rays
of mature males modified. Recently collected material from Thailand included a new species of Lepidocephalichthys in
which mature males have a large (extending over ~75% of the fin-ray length) dorsally projecting and serrated flange and
a ventrally projecting flange. The ventrally projecting structure is unique among cobitids. An expanded phylogenetic
analysis of cobitids, including previously published sequences and new material including the new species, reinforces the
monophyly of Lepidocephalichthys. Relationships within southern lineages of cobitids, and the unusual habitat of the
new species are discussed.
Key words: Cypriniformes, loaches, phylogenetics, dichromatism, Thailand, Vietnam, Mun River, Mekong
Introduction
Species of Lepidocephalichthys (Cobitidae) are small, spined loaches with sexually dimorphic pectoral fins.
They are widely distributed throughout South and Southeast Asia, including the islands of Java and Borneo.
Recent systematic studies on Lepidocephalichthys have included a taxonomic revision of the genus (Havird &
Page 2010) and an analysis of phylogenetic relationships within Cobitidae (Šlechtová et al. 2008). Šlechtová
et al. (2008) examined relationships among cobitids using the mitochondrial cytochrome b gene (cyt b) and
the nuclear recombination activation gene-1 (RAG-1). Both datasets supported a large monophyletic lineage
referred to as the ‘northern clade’ and including species of Cobitis, Iksookimia, Niwaella, Kichulchoia,
Koreocobitis, Misgurnus, Paramisgurnus and Sabanejewia. Most genera in this clade were not found to be
monophyletic. Genera that fell outside the northern clade (Acanthopsoides, Acantopsis, Canthophrys,
Kottelatlimia, Lepidocephalichthys, Lepidocephalus, Neoeurirrhichthys and Pangio) did not form a
monophyletic group and were referred to as the ‘southern lineages.’ These genera were recovered as
monophyletic groups based on mitochondrial (with the exception of Acanthopsoides) as well as nuclear DNA
data. Lepidocephalichthys was found to be sister to Pangio in the RAG1 analysis, a relationship not recovered
in earlier analyses of morphological data (Nalbant 1963, 1994; Sawada 1982).
Although several morphological characters distinguish genera in the southern lineages, sexually
dimorphic traits are among the most useful (Table 1). In particular, mature males have modifications of the

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A NEW SPECIES OF LEPIDOCEPHALICHTHYS
pectoral-fin rays, forming a structure termed the lamina circularis, which varies among genera.
Lepidocephalichthys is diagnosed as having the 7–8th pectoral-fin rays modified in mature males (Havird &
Page 2010). In cobitids, variation exists in the location of the lamina circularis, the number of rays modified,
and shape of the structure formed.
Here we describe a new species of Lepidocephalichthys from the Mekong River drainage with highly
distinctive sexually dimorphic pectoral fins. We also present an expanded phylogeny of the southern lineages
of cobitids to confirm the generic placement of the new species. A discussion of relationships and sexual
dimorphism within cobitids is provided.
Methods
Lengths were measured to the nearest 0.1 mm using digital calipers. Most measurements and counts follow
Hubbs and Lagler (1964), with modifications as described in Havird & Page (2010). Specimens were
classified as adults if they had a standard length (SL) equal to or greater than the SL of the smallest specimen
with modified pectoral fins (Table 2). Smaller specimens were classified as juveniles and were not included in
morphometric analyses. Adults were interpreted as male if they had modified pectoral fins and as female if
they lacked modifications. Since the sexes of most specimens could not be confirmed via dissection due to
small sample sizes, some specimens classified as females may be immature males. Sexually dimorphic
characteristics are those of adult males vs. those of large individuals without modified pectoral fins (most of
which are likely females).
TABLE 2. Adult standard length (SL, mm) and relative pectoral-fin lengths (% SL) for cobitids with large sample sizes.
Minimum adult SL is for the smallest specimen with a lamina circularis. Acantopsis spp. refers to pooled data for three
described (A. choirorhynchos, A. dialuzona, and A. octoactinotos) and several undescribed species. See Havird and Page
(2010) for data on previously described Lepidocephalichthys. P = p-value comparing male and female pectoral-fin
lengths calculated with a Student’s two-tailed, unpaired t-test assuming equal variance.
Institutional abbreviations are listed at http://www.asih.org/codons.pdf. In total, 490 specimens of cobitids
were examined, mostly within the southern lineages. Specimens examined in the recent review of
Lepidocephalichthys (Havird & Page 2010) were used in this study but are not listed in Material Examined
(unless novel GenBank numbers are given). Catalog numbers of specimens examined are followed by number
of specimens and range in SL in parentheses. A tilde (~) before geographic coordinates indicates that they
were estimated from general locality data. All statistics were calculated using Microsoft Excel 2003.
Species Min Adult SL
Mean Max Male pectoral fin
length Female pectoral fin
length P
Acanthopsoides molobrion 32.0 45.9 62.5 14.2 (n = 11) 11.6 (n = 15) < 0.001
Acantopsis spp. 39.0 98.6 153.4 16.6 (n = 14) 14.8 (n = 30) 0.001
Kottelatlimia pristes 23.6 31.1 42.9 18.9 (n = 49) 15.8 (n = 39) < 0.001
Lepidocephalichthys zeppelini 14.1 19.4 25.8 23.3 (n = 37) 20.2 (n = 91) < 0.001
Pangio cuneovirgata 27.1 32.9 38.9 8.8 (n = 14) 7.3 (n = 14) < 0.001
Pangio doriae 59.8 70.5 91.2 5.2 (n = 7) 3.2 (n = 5) 0.002
Pangio kuhlii 38.6 50.2 67.2 9.5 (n = 13) 7.3 (n = 15) < 0.001
Pangio malayana 36.7 40.7 47.1 8.9 (n = 13) 7.0 (n = 7) < 0.001
Pangio mauraniformis 36.3 43.9 48.9 10.8 (n = 12) 7.2 (n = 7) < 0.001
Pangio piperata 33.5 40.7 48.4 10.8 (n = 13) 7.6 (n = 15) < 0.001
Pangio semicincta 42.8 50.8 62.7 9.5 (n = 7) 7.5 (n = 5) 0.015
Pangio shelfordii 39.6 48.4 57.6 10.7 (n = 11) 7.5 (n = 8) < 0.001

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HAVIRD ET AL.6 · Zootaxa 2557 © 2010 Magnolia Press
Because the sexually dimorphic pectoral fin of the new species is unique among cobitids, we performed a
phylogenetic analysis with sequences from across Cobitidae to confirm that the new species grouped within
Lepidocephalichthys. Recently collected cobitid tissues from Thailand, Sumatra, and Malaysia (Table 3) were
analyzed along with published sequences (including outgroups) from Šlechtová et al. (2008). Whole genomic
DNA was extracted from fin clips preserved in ethanol using a 5% Chelex solution and 3 l of prteinase K
with overnight digestion. An approximately 1140 bp region including the complete mitochondrial cyt b gene
was amplified and sequenced using the primer pair Glu-L.Ca14337–14359 and Thr-H.Ca15568–15548
(Šlechtová et al. 2008). Approximately 910 bp of the RAG1 gene were amplified and sequenced using the
primers RAG-1F and RAG-RV1 (Šlechtová et al. 2008). PCR cycling parameters were those used by
Šlechtová et al. (2006) for cyt b and Šlechtová et al. (2007) for RAG1.
Bands were visualized with ethidium bromide staining on 1% agarose gels to verify that primers
amplified fragments of the appropriate sizes. PCR products were sequenced at the Interdisciplinary Center for
Biotechnology Research (ICBR), University of Florida, Gainesville. Chromatograms were viewed and
consensus sequences were assembled using CodonCode Aligner. Final alignments were generated using
Clustal X2 (Larkin et al. 2007) and corrected manually by eye.
A Bayesian phylogeny was generated using MrBayes, version 3.1.2 (Ronquist & Huelsenbeck 2003) with
five million generations and 45,000 trees sampled after a 10% burn-in for two runs. Posterior probabilities
were used as an indication of node support. The cyt b and RAG1 analyses were partitioned by codon position,
and each position was subjected to Modeltest version 3.7 (Posada & Crandall 1998) to determine the best-fit
model of sequence evolution using the Akaike Information Criterion. The GTR + I + G model was chosen for
each cyt b position. For the RAG1 dataset, the GTR + I + G, HKY + I, and K81uf + G models were chosen for
the first, second, and third positions, respectively.
A parsimony analysis was also performed using PAUP 4.0b10 (Swofford 2002) for the RAG1 and cyt b
datasets. Heuristic searches with random addition and tree bisection and reconnection parameters were
completed, and support values were generated based on 1000 bootstrap replicates. Tamura-Nei-corrected
sequence divergence values were calculated using PAUP 4.0b10.
Results
Lepidocephalichthys zeppelini Havird & Tangjitjaroen, new species
(Fig. 1; Tables 2–5)
Holotype. UF 174131, 21.1 mm-SL male, Thailand, Ubon Ratchathani, Mun River (tributary of Mekong
River), isolated pools in a rice field, Ubon Rajathanee University campus, 15°8’3.18”N, 104°55’27.78”E, 10
June 2008, L. M. Page, W. Tangjitjaroen, S. Udduang, and J. C. Havird.
Paratypes. CAS 219335, 8 males, 37 females, 17.0–25.8 mm SL, Thailand, Ubon Ratchathani market,
~15°9'23.73"N, ~105° 3'27.81"E, 11–13 Sept. 1990, T. R. Roberts. NIFI 3248, 1 male, 8 females, 15.5–23.1
mm SL, Thailand, Nong Khai, Beung Kan, Goot Ting marsh, ~18°13'34.15"N, ~103°39'0.33"E, 11 June 2003,
C. Vidthayanon. NIFI 3249, 1 male, 8 females, 15.9–20.0 mm SL, Thailand, Nong Khai, Beung Kan, Goot
Ting marsh, ~18°13'34.15"N, ~103°39'0.33"E, 23–27 March 1996, C. Vidthayanon. UF 171981, 3 juveniles,
11.3–12.7 mm SL, Thailand, Ubon Ratchathani, Mun River (tributary of Mekong River), rice field,
15°8’17.76”N, 104°55’30.54”E, 9 June 2008, L. M. Page, W. Tangjitjaroen, and J. C. Havird, GenBank
GQ174345, GQ174347, GQ174383, GQ174399. UF 171983, 1 juvenile, 12.7 mm SL, Thailand, Amnat
Charoen, Mekong drainage, small sandy stream, 15°54’13.56”N, 105°11’17.28”E, 10 June 2008, L. M. Page,
W. Tangjitjaroen, and J. C. Havird. UF 174130, 10 males, 18 females, 4 juveniles, 12.1–22.8 mm SL, data as
for holotype, GenBank GQ174346, GQ174348–51, GQ174384, 5. UF 174132, 1 female, 15.7 mm SL,
Thailand, Ubon Ratchathani, Mun River (tributary of Mekong River), marsh near Ubon Rajathanee
University campus, 15°10’45.84”N, 104°45’44.76”E, 10 June 2008, L. M. Page, W. Tangjitjaroen, and J. C.
Havird. UMMZ 227583, 11 males, 22 females, 15.0–23.3 mm SL, Vietnam, Long Xuyen, Mekong drainage,

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A NEW SPECIES OF LEPIDOCEPHALICHTHYS
rice paddy NW Long Xuyen, ~10°23'22.49"N, ~105°24'56.46"E, 22 July 1974, Rainboth, Smith, and
Weidenbach. ZRC 51882, 3, 17.0–22.9 mm SL, data as for holotype.
Diagnosis. A species of Lepidocephalichthys sensu Havird & Page (2010). Lepidocephalichthys zeppelini
is distinguished from other Lepidocephalichthys by having the 7–8th pectoral-fin rays of the mature male
having a large (extending along > 75% of the length of the 7–8th pectoral rays) dorsally projecting, rounded
rectangular flange with about 25 fine serrations and a smaller ventrally projecting, rounded flange (Fig. 1C,
D); a forked caudal fin; small barbels (not reaching orbit); dark reticulations on caudal fin; and small size (to
25.8 mm SL).
FIGURE 1. Lepidocephalichthys zeppelini, new species, UF 174131 (holotype), 21.1 mm-SL male, Thailand, Ubon
Ratchathani, Mun River (tributary of Mekong River), isolated pools in a rice field, Ubon Rajathanee University campus:
(A) dorsal view, (B) lateral view, (C) lamina circularis on pectoral fin, (D) medial view of pectoral fin removed from UF
174130 (22.2 mm-SL male paratype) showing lamina circularis.
Comparisons. Lepidocephalichthys zeppelini is distinguished from all other Lepidocephalichthys by
having the 7–8th pectoral-fin rays forming a large, dorsally projecting, rounded rectangular flange and a
smaller (although large compared to the dorsal projections of other species), ventrally projecting rounded
flange (Fig. 1C, D). It is also distinguished from all other species of Lepidocephalichthys by its small adult
body size (to 25.8 vs. 30.8 mm SL).
The new species is further distinguished from all other congeners except L. manipurensis Arunkumar, L.
furcatus (de Beaufort), L. goalparensis Pillai & Yazdani, and L. micropogon (Blyth) by its forked (vs. rounded
or truncated) caudal fin. It is distinguished from L. micropogon, L. goalparensis, and L. manipurensis by its
darkly reticulated (vs. barred) caudal fin, and from L. furcatus by having fine serrations (vs. smooth edge) on
the dorsal flange on the 7–8th pectoral-fin rays of males (Fig. 1C, D), barbels not reaching orbit (vs. barbels
reaching orbit), fewer predorsal scales (about 40 vs. 50–60), and smaller size (to 25.8 vs. 30.8 mm SL).

HAVIRD ET AL.8 · Zootaxa 2557 © 2010 Magnolia Press
Description. Morphometric data in Table 4; small adult size (averaging 19.4 mm SL); body depth
increasing from snout to nape, then decreasing to caudal peduncle (Fig. 1B); dorsal fin with 2 unbranched, 6
branched rays, last branched ray split to base; anal fin with 2 unbranched, 5 branched rays, last branched ray
split to base; pectoral fin with 1 unbranched, 7 branched rays; pelvic fin with 1 unbranched, 6 branched rays;
16 caudal rays (7 branched, 1 unbranched in each lobe); caudal fin forked; dorsal-fin origin anterior to pelvic-
fin origin; in mature males 7–8th pectoral-fin rays with large (extending along > 75% of length of rays),
dorsally projecting rounded rectangular flange with about 25 fine serrations and smaller, ventrally rounded
flange (Fig. 1C, D); axial process in well-preserved specimens as small pocket of skin projecting from
pectoral-fin base, not fused to rays; 30–46 predorsal scales (average about 40); no scales on top and side of
head; anterior nostril on end of small outwardly projecting tube; suborbital bifid spine with anterior projection
smaller than posterior; barbels: 2 rostral pairs, 1 maxillary pair at corner of mouth, medially split mandibular
lobe forming 2 medially thickened mouth flaps; barbels small: first pair of rostral barbels not reaching anterior
nostril, other pairs not reaching orbit; ventral mouth flaps usually without fringes.
Color in alcohol. Lepidocephalichthys zeppelini has pale yellow to white belly, background overlain with
dark-brown marks on side, dorsum, head, and fins. Six–14 dark-brown blotches on side disconnected in
females, connected by narrow stripe in males. Dark-brown chevron (<- shaped mark) at base of caudal fin
points towards head, similar to that in L. furcatus (Kottelat & Lim 1992), but usually darker. Three to five
predorsal and 3–6 postdorsal irregular dark blotches, chevrons, or bars on dorsum, with one at dorsal-fin base
(Fig. 1A). Most specimens with small, inconspicuous dark spot on upper caudal-fin base spanning principal
rays 4–7. Caudal, anal, and dorsal fins darkly reticulated. Dark stripe extends from snout, through eye, onto
top of head, forming two dark spots on top of head where it terminates; otherwise, top of head with small
dark-brown spots forming no pattern.
TABLE 4. Morphometric data for holotype and paratypes of Lepidocephalichthys zeppelini. Min = minimum, Max =
maximum, SD = standard deviation.
L. zeppelini (n = 128)
Character Holotype Mean Min max SD
Standard length (mm) 21.1 19.4 14.1 25.8 2.45
Percents standard length
Total length 123.5 126.2 111.4 140.2 4.408
Predorsal length 47.8 50.8 45.9 57.2 2.20
Prepelvic length 51.5 54.5 42.7 63.7 2.96
Preanal length 77.2 80.8 73.6 94.2 3.57
Body depth 21.3 21.4 17.8 25.1 1.56
Body width 8.69 11.0 7.17 16.4 1.71
Pectoral-fin length (all) –21.1 13.1 28.4 2.89
Pectoral-fin length (male) 23.9 23.3 17.4 28.4 2.53
Pectoral-fin length (female) –20.2 13.1 26.6 2.50
Pelvic-fin length 16.4 16.7 5.34 22.5 2.26
Dorsal-fin height 22.5 22.0 6.37 29.1 3.23
Caudal penduncle depth 10.6 10.8 7.75 14.5 0.99
Head length 20.4 23.7 19.0 29.9 1.80
Percents head length
Snout length 22.6 26.2 16.3 35.5 3.64
Orbit diameter 25.2 24.9 17.3 32.2 3.18
Anterior rostral-barbel length 14.5 11.3 5.41 19.6 3.20
Maxillary-barbel length 39.2 23.5 11.7 39.2 4.70
Interorbital width 23.3 18.5 10.9 31.1 3.75

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A NEW SPECIES OF LEPIDOCEPHALICHTHYS
Sexual dimorphism. In addition to the modified pectoral fins and sexual dichromatism described above,
L. zeppelini has other sexual dimorphisms characteristic of Lepidocephalichthys. Males have significantly (P
< 0.001, Student’s two-tailed, unpaired t-test assuming equal variance) larger pectoral fins (Table 2) and
pelvic fins (Table 5) than females. Males are also significantly (P < 0.001, Student’s two-tailed, unpaired t-test
assuming equal variance) smaller than females: males average 18.1 mm SL and reach 21.7 mm SL whereas
females average 19.9 mm SL and reach 25.8 mm SL.
Relationships. Clades containing seven individuals of L. zeppelini for cyt b (Fig. 2A), and four
individuals for RAG1 (Fig. 2B), were resolved as a distinct lineage within Lepidocephalichthys with 100%
support in all analyses. In the cyt b analysis (Fig. 2A), L. zeppelini grouped most closely with L. thermalis
(Valenciennes); RAG1 sequences suggested a closer relationship between L. zeppelini and L. kranos Havird &
Page (Fig. 2B). The average intraspecific divergence for L. zeppelini was 0.1% for cyt b and 0.2% for RAG1.
The average interspecific divergence between L. zeppelini and L. thermalis was 22.0% for cyt b, and between
L. zeppelini and L. kranos was 8.9% for RAG1.
FIGURE 2. Phylogeny of species of Lepidocephalichthys based on Bayesian analysis of cyt b (A) and RAG1 (B).
Clades of species are alternatively shaded. Since the parsimony analysis gave a very similar topology, support values for
both analyses are presented at each node (numbers on left are posterior probabilities and numbers on right are bootstrap
values). Asterisks indicate Bayesian support values of 0.95 or greater and parsimony support values of 0.90 or greater;
single asterisks meet both criteria. Dashes indicate relationships were not supported in the parsimony analysis.
Lepidocephalichthys zeppelini, n. sp. and L. kranos, recently described by Havird & Page (2010), are bolded and their
support values are circled.
TABLE 5. Relative pelvic-fin lengths of adults of eight cobitid species with large sample sizes. Lengths are mean ±
standard deviation. P = p-value comparing male and female pelvic-fin lengths calculated with a Student’s two-tailed,
unpaired t-test assuming equal variance.
Pelvic Fin Length (% SL)
Species Male Female P
Acanthopsoides molobrion 12.1 ± 0.7 (n = 20) 11.1 ± 0.7 (n = 19) < 0.001
Kottelatlimia pristes 15.6 ± 1.6 (n = 28) 14.6 ± 1.1 (n = 33) 0.008
Lepidocephalichthys berdmorei 15.1 ± 0.6 (n = 20) 13.2 ± 1.5 (n = 22) 0.001
L. guntea 16.0 ± 1.1 (n = 20) 14.0 ± 1.5 (n = 26) < 0.001
L. hasselti 16.5 ± 1.5 (n = 32) 14.9 ± 1.2 (n = 29) < 0.001
L. kranos 17.8 ± 2.8 (n = 9) 16.2 ± 1.9 (n = 20) 0.145
L. zeppelini 17.6 ± 2.1 (n = 25) 15.6 ± 2.2 (n = 27) 0.002
Pangio semicincta 8.0 ± 0.5 (n =9) 6.0 ± 0.7 (n = 14) < 0.001
Eight species combined 15.9 ± 2.8 (n = 130) 14.0 ± 3.0 (n = 165) < 0.001

HAVIRD ET AL.10 · Zootaxa 2557 © 2010 Magnolia Press
FIGURE 3. Distribution of Lepidocephalichthys zeppelini. Black dots represent localities for specimens examined;
circle represents type locality.
FIGURE 4. Type locality of Lepidocephalichthys zeppelini. Individuals were in small ephemeral pools in otherwise dry
agricultural fields. Most other specimens are from similar habitats.
Distribution and habitat. Lepidocephalichthys zeppelini is known from the Mekong drainage in
Thailand and Vietnam (Fig. 3) where it has been collected with L. hasselti and L. kranos. It has been found
mainly in agricultural fields in small shallow pools (Fig. 4). One juvenile was collected in a small sandy
stream.

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A NEW SPECIES OF LEPIDOCEPHALICHTHYS
FIGURE 5. Phylogeny of cobitids based on Bayesian analysis of cyt b. Shading, support values, asterisks, and dashes as
in Figure 2.

HAVIRD ET AL.12 · Zootaxa 2557 © 2010 Magnolia Press
FIGURE 6. Phylogeny of cobitids based on Bayesian analysis of RAG1. Shading, support values, asterisks, and dashes
as in Figure 2.

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Previous collectors of L. zeppelini described its habitat as marshes or rice paddies. While collecting in
Thailand in June at the beginning of the wet season, we originally targeted small streams, marshes, and
flooded rice fields with little success, yielding only a few juveniles. However, when small ephemeral pools in
otherwise dry agricultural fields were targeted, many adults were collected. At the type locality (Fig. 4), pools
less than 9 m2 contained the majority of L. zeppelini collected. Lepidocephalichthys hasselti was also
abundant in these pools, along with other common Southeast Asian freshwater fishes including species of
Anabas, Puntius, Rasbora, Channa, and Hampala.
Etymology. The species name zeppelini is a reference to the 1968–1980 band Led Zeppelin. Use of
the Gibson EDS-1275 double-neck guitar by Jimmy Page reminded us of the diagnostic double lamina circu-
laris of this species. It is Latinized as a noun in the genitive singular.
Discussion
In addition to supporting individuals of L. zeppelini as a distinct monophyletic clade within
Lepidocephalichthys, our expanded phylogenetic analysis of cobitids—50 new sequences and five additional
species from the southern lineages (Table 3) were added to the phylogenetic analysis of Šlechtová et al.
(2008)—presents an updated analysis of cobitid relationships. Analyses of cyt b and RAG1 sequences
included 1150 and 894 bps, respectively. Both gene trees had similar topologies and support values, and both
strongly supported the northern clade and other relationships found in Šlechtová et al. (2008).
As in Šlechtová et al. (2008), the phylogeny based on cyt b (Fig. 5) gave poor resolution of intergeneric
relationships among the southern lineages, but revealed within-species (geographical) structure in several
species with large samples. For example, Lepidocephalichthys hasselti (Valenciennes) from east Thailand
formed a clade distinct from that for samples from central Thailand. The RAG1 phylogeny (Fig. 6) gave
strongly supported relationships among genera but failed to provide much intraspecific geographic resolution.
With new sequences from Acanthopsoides molobrion Siebert included, the specimen of Acanthopsoides
robertsi Siebert from peninsular Malaysia (GenBank EF508482) included in the analysis by Šlechtová et al. is
recognized as a likely misidentification (A. robertsi is not known from peninsular Malaysia). Similarly,
including new sequences from Pangio oblonga (Valenciennes) resulted in the sample of Pangio pangia
(Hamilton) (GenBank EF508583) from the analysis by Šlechtová et al. falling within the P. oblon ga clade.
This also may be due to misidentification, or it may indicate a lack of reciprocal monophyly for the two taxa.
Although discussions of sexual dimorphism in Cobitidae are often limited to modifications of the pectoral
fins in males (Kim et al. 1997; Chen & Chen 2007; Kottelat & Tan 2008), other forms of dimorphism are
common in cobitids. As reported for other Lepidocephalichthys (Havird & Page 2010), L. zeppelini has dark
spots on the side of the body in the female and a narrow dark stripe in the adult male (Fig. 7G–I). Although
most pronounced in L. guntea (Hamilton) (Fig. 7D–F), this dichromatism was present in several cobitids
examined. Canthophrys gongota (Hamilton), Cobitis choii (Kim & Son), Cobitis sinensis Sauvage & Dabry
de Thiersant (Fig. 7A–C), and Acantopsis sp. have conspicuous spots on the female and a stripe on the male.
Males with the largest structure (lamina circularis) on their pectoral fins also have the most conspicuous
stripes, suggesting that circulating levels of sex hormones control the expression of both traits. This
hypothesis is supported by a study showing species of Misgurnus altering this structure in relation to sex
hormone treatments (Kim et al. 1997). Small males sometimes have a large lamina circularis and a
conspicuous stripe, and expression of the dimorphic traits does not seem to be strongly correlated with body
size. In addition to the lamina circularis and distinguishing color patterns, males also have significantly longer
pectoral fins (Table 2). Most males also have longer pelvic fins (Table 5) than do females.

HAVIRD ET AL.14 · Zootaxa 2557 © 2010 Magnolia Press
FIGURE 7. Relationship between color pattern (especially stripe intensity) and development of lamina circularis in
three species of cobitids: (A–C) Cobitis sinensis, UMMZ 240028, (A) 66.0 mm-SL female, (B) 71.4 mm-SL male, (C)
55.0 mm-SL male; (D–F) Lepidocephalichthys guntea, KU 29353, (D) 46.7 mm-SL female, (E) 45.9 mm-SL male, (F)
53.4 mm-SL male; (G–I) L. zeppelini, UF 174130, (G) 21.9 mm-SL female, (H) 16.2 mm-SL male, (I) 21.9 mm-SL
male.
The relatively dry agricultural fields where most specimens of L. zeppelini were found represent an
interesting habitat for loaches in Southeast Asia. Only juveniles were collected from larger ponds, small
streams, and marshes, suggesting either that juveniles were easier to catch or that adults were absent from
these habitats. Adults may only be found in the pools of otherwise dry agricultural fields—which likely vanish
completely during the dry season—begging an explanation for where the fishes go during the dry season.
Some may survive by burrowing into the substrate (M. Kottelat, pers. comm.); others may migrate to more

Zootaxa 2557 © 2010 Magnolia Press · 15
A NEW SPECIES OF LEPIDOCEPHALICHTHYS
suitable habitat during the dry season. Long-term studies monitoring L. zeppelini and other species in these
fields could determine seasonal changes in habitat use or behavior.
Material examined
Acanthopsoides hapalias Siebert: Cambodia–UMMZ 234697 (26, 32.6–35.0).
Acanthopsoides molobrion Siebert: Borneo–ZRC 38854 (1, 26.9). ZRC 39980 (2, 42.9–50.6). Malaysia–
UF 173515 (12, 32.5–38.9). UF 173524 (17, 26.8–45.6), GenBank GQ174326, CQ174328, GQ174414. UF
173529 (2, 33.7–42.9), GenBank GQ174331, GQ174411. UF 173552 (27, 34.0–46.0). UF 173553 (27, 32.5–
44.4), GenBank GQ174328–30, GQ174406, GQ174408, GQ174413. ZRC 1471 (2, 40–46.9). ZRC 9309 (1,
54.9). ZRC 11107 (1, 39.8). ZRC 13764 (1, 36.6). ZRC 25577 (2, 32.0–45.7). ZRC 27566 (2, 33.7–38.7).
ZRC 28288 (1, 48.1). ZRC 29409 (2, 41.8–45.7). ZRC 29412 (2, 51.6–55.8). ZRC 39550 (4, 42.8–62.5). ZRC
40218 (2, 35.9–49.7). ZRC 42793 (1, 48.8). ZRC 42899 (1, 58.4). ZRC 42950 (1, 46.7). ZRC 44155 (1, 53.7).
Sumatra–UF 161609 (3, 38.2–43.8), GenBank GQ174313, GQ174410. UF 161622 (21, 31.1–35.5). UF
166885 (6, 45.0–51.3). UF 166898 (2, 28.2–33.8), GenBank GQ174325, GQ174409.
Acanthopsoides robertsi Siebert: Borneo–ZRC 37872 (1, 42.0). ZRC 40054 (4, 29.4–36.4). Sumatra–UF
166897 (1, 41.9).
Acantopsis choirorhynchos (Bleeker): Malaysia–ZRC 1449 (1, 113.0). ZRC 1461 (1, 75.1). ZRC 1463 (2,
115.1–135.4). ZRC 1490 (2, 104.4–111.9). ZRC 5898 (1, 93.4). ZRC 5901 (1, 98.6). ZRC 5915 (1, 107.6).
Myanmar: USNM 37844 (12, 36.3–90.4).
Acantopsis dialuzona van Hasselt: Malaysia–UF 173514 (2, 52.0–102.8), GenBank GQ174316,
GQ174318, GQ174403. UF 173516 (4, 84.8–101.2), GenBank GQ174314, GQ174317, GQ174381,
GQ174386. UF 173552 (8, 63.8–125.8), GenBank GQ174304–6, GQ174315, GQ174382, GQ174389,
GQ174391, GQ174412. ZRC 1491 (1, 113.4). ZRC 7412 (1, 153.4). ZRC 11775 (1, 63.8). ZRC 21488 (2,
48.6–118.1). ZRC 27576 (2, 61.6–62.9). ZRC 44190 (1, 49.8). Sumatra–UF 161621 (5, 55.5–60.2), GenBank
GQ174307, GQ174311, GQ174387, GQ174401. UF 161717 (9, 47.3–60.7), GenBank GQ174308–9,
GQ174390, GQ174402. UF 166890 (11, 48.8–58.9), GenBank GQ174310, GQ174388. ZRC 41665 (1, 43.1).
ZRC 42566 (1, 77.0). Thailand–ZRC 39328 (2, 82.1–112.4).
Acantopsis octoactinotos Siebert: Borneo–ZRC 37613 (1, 30.3). ZRC 37685 (1, 30.6). ZRC 40415 (2,
39.0–45.8).
Acantopsis spp. (several undescribed species): Aquarium–ZRC 50979 (4, 95.9–127.0). Malaysia–ZRC
1489 (2, 81.2–85.3). Myanmar–ZRC 43458 (1, 76.5). ZRC 43519 (1, 123.6). Thailand–UF 169951 (5, 59.6–
122.8). UF 169952 (6, 81.1–95.1). UF 169953 (3, 70.9–75.4). UF 169954 (14, 65.1–129.2). UF 170195 (1,
54.1). UF 172925 (1, 116.8). UF 172928 (1, 92.4), GenBank GQ174319, GQ1744407. ZRC 40898 (1, 73.6).
Sumatra–ZRC 38630 (2, 50.4–51.2).
Canthophrys gongota (Hamilton): Bangladesh–UF 172552 (2, 79.5–82.4). India–ZRC 38897 (4, 79.1–
97.9). Nepal–KU 28604 (2, 65.3–72.1). KU 29113 (4, 49.2–115.4). KU 29361 (1, 96.4). KU 29561 (2, 82.8–
84.2).
Cobitis bilineata Canestrini: Italy–UF 81156 (4, 42.9–58.0). UF 110731 (5, 45.6–65.8).
Cobitis calderoni Bacescu: Spain–UMMZ 212518 (10, 32.3–44.6).
Cobitis choii Kim & Son: Mongolia–ANSP 185225 (35, 21.0–55.0). ANSP 185388 (1, 42.9).
Cobitis elongata Heckel & Kner: Romania–UMMZ 185061 (2, 93.2–134.7). UMMZ 185062 (3, 108.5–
199.5).
Cobitis lutheri Rendahl: South Korea–UMMZ 240027 (6, 51.3–71.7).
Cobitis macrostigma Dabry de Thiersant: China–FMNH 14807 (2, 75.3–92.9).
Iksookimia longicorpa (Kim, Choi & Nalbant): South Korea–UMMZ 240026 (6, 88.7–111.9).
Iksookimia koreensis (Kim): South Korea–FMNH 95966 (3, 80.8–103.8). UMMZ 240025 (6, 61.1–78.2).
Iksookimia pumila (Kim & Lee): South Korea–UMMZ 240024 (6, 48.6–67.4).
Kottelatlimia katik (Kottelat & Lim): Malaysia–ZRC 9344 (holotype) (1, 12.9). ZRC 9345 (paratypes) (3,
11.2–12.5).

HAVIRD ET AL.16 · Zootaxa 2557 © 2010 Magnolia Press
Kottelatlimia pristes (Roberts): Borneo– CAS 49352 (paratype) (1, 36.0). MCZ 56064 (paratypes) (2,
30.5–31.1). USNM 230265 (paratype) (1, 31.4). BMNH 2000.10.18.70 (1, 25.6). BMNH 2000.10.18.95 (1,
22.0). CAS 219322 (2, 23.2–30.7). ZRC 22836 (2, 32.2–36.0). ZRC 22855 (1, 35.8). ZRC 27850 (2, 26.1–
36.3). ZRC 29444 (3, 25.4–25.9). ZRC 29451 (2, 21.5–25.0). ZRC 37888 (1, 31.8). ZRC 38788 (3, 27.3–
32.0). ZRC 39512 (3, 27.9–35.3). ZRC 39856 (3, 28.1–30.0). ZRC 39870 (4, 29.4–33.0). ZRC 39893 (3,
28.7–31.0). ZRC uncataloged (1, 28.0), GenBank GQ174332. Malaysia–ZRC 14916 (1, 35.0). ZRC 14921 (3,
27.4–30.2). ZRC 14926 (3, 24.4–25.7). ZRC 15141 (2, 25.1–28.2). ZRC 17844 (2, 32.9–36.8). ZRC 20763 (2,
28.1–37.1). ZRC 20836 (3, 29.1–35.1). ZRC 27723 (2, 27.7–35.3). ZRC 38281 (3, 30.7–36.9). ZRC 38437 (2,
21.5–22.6). ZRC 43658 (2, 21.5–27.9). Sumatra–MZB 15302 (26, 27.1–31.5). MZB 15303 (21, 27.9–36.5).
UF 166980 (22, 29.7–35.6). UF 166981 (28, 23.6–31.2). UF 166984 (2, 29.2–34.5). ZRC 38514 (3, 30.2–
34.6). ZRC 38522 (3, 26.7–33.4). ZRC 38567 (1, 40.8). ZRC 38585 (2, 29.3–30.0). ZRC 38600 (2, 23.4–
32.5). ZRC 39084 (3, 25.3–33.5). ZRC 39153 (5, 30.3–31.4). ZRC 39177 (2, 33.9–34.3). ZRC 42259 (3,
30.0–40.0). ZRC 42318 (3, 36.6–39.0). ZRC 42400 (3, 28.1–38.4). ZRC 42437 (2, 20.7–29.7). ZRC 43029 (3,
30.9–42.9). ZRC 43089 (4, 31.1–40.1). ZRC 43128 (2, 24.6–31.0). ZRC 43140 (3, 28.4–35.1).
Lepdiocephalichthys berdmorei (Blyth): Thailand–UF 172831 (5, 56.3–74.1), GenBank GQ174337–8,
GQ174379, GQ174396.
Lepdiocephalichthys hasselti (Valenciennes): Thailand–UF 170276 (1, 29.2), GenBank GQ174333,
GQ174378. UF 171982 (51, 17.4–40.7), GenBank GQ174334–6, GQ174394–5, GQ174400. Sumatra–UF
161478 (1, 32.0), GenBank GQ174392. UF 161482 (2, 26.0–31.0), GenBank GQ174393.
Lepdiocephalichthys kranos Havird & Page: Thailand–UF 171980 (holotype) (1, 33.1), GenBank
GQ174342. UF 170286 (paratypes) (2, 26.6–33.6), GenBank GQ174341, GQ174380. UF 170287 (paratypes)
(4, 31.2–37.4), GenBank GQ174343. UF 170288 (paratypes) (10, 18.2–35.9), GenBank GQ174339–40. UF
173041 (paratypes) (1, 32.6), GenBank GQ174344.
Lepidocephalus macrochir (Bleeker): Sumatra–BMNH 1866.5.2.55 (syntype) (1, 77.7). Borneo–BMNH
2001.1.15.8066–8070 (4, 63.0–69.3). BMNH 2001.1.5.8071–8075 (1, 55.9).
Lepidocephalus spectrum Roberts: Borneo–USNM 230267 (paratype) (1, 50.3).
Misgurnus anguillicaudatus (Cantor): Florida–UF 143225 (1, 64.5). UF 148188 (10, 37.9–55.0). UF
163719 (4, 65.9–82.9).
Misgurnus fossilis (Linnaeus): Poland–UMMZ 185341 (8, 50.5–63.1).
Misgurnus mizolepis Günther: Taiwan–UMMZ 194439 (1, 71.0).
Misgurnus sp.: China–USNM 89204 (1, 94.0).
Neoeucirrhichthys maydelli Banarescu & Nalbant: Nepal–KU 29366 (1, 29.2).
Pangio agma (Burridge): Borneo–ZRC 8408 (1, 43.3). ZRC 31934 (2, 53.0–57.8). ZRC 40273 (2, 43.0–
44.3). ZRC 42720 (2, 40.7–44.6).
Pangio alcoides Kottelat & Lim: Malaysia–ZRC 41923 (1, 39.0). ZRC 40200 (3, 37.7–46.4).
Pangio alternans Kottelat & Lim: Borneo–ZRC 35037 (2, 31.0–31.8).
Pangio anguillaris (Vaillant): Laos–UMMZ 241968 (7, 50.1–62.5). Malaysia–ZRC 34884 (2, 64.1–65.6).
Thailand–ZRC 35647 (2, 42.4–44.3). Sumatra–ZRC 38629 (65.2–65.4).
Pangio cuneovirgata (Raut): Malaysia–ZRC 2060 (2, 33.2–36.5). ZRC 8414 (1, 28.6). ZRC 18461 (1,
37.2). ZRC 28178 (2, 31.7–34.6). ZRC 39553 (1, 36.9). ZRC 40179 (4, 27.1–38.9). ZRC 42774 (1, 33.2).
ZRC 42795 (1, 31.2). ZRC 42820 (1, 32.4). ZRC 42945 (2, 33.0–33.2). ZRC 44189 (2, 32.5–35.1). Sumatra–
ZRC 385642 (4, 27.9–33.6). ZRC 42299 (2, 29.6–31.5). Thailand–ZRC 42094 (4, 30.3–33.4).
Pangio doriae (Perugia): Malaysia–FMNH 62013 (1, 72.2). ZRC 1487 (2, 67.4–70.4). ZRC 14293 (1,
61.4). ZRC 42821 (2, 62.2–71.9). ZRC 42900 (2, 59.8–68.6). ZRC 42949 (1, 63.3). Sumatra–ZRC 41667 (4,
64.6–91.2).
Pangio filinaris Kottelat & Lim: Malaysia–ZRC 38742 (2, 33.3–35.7). ZRC 41872 (4, 34.4–49.0).
Pangio incognito Kottelat & Lim: Borneo–ZRC 39372 (1, 21.0).
Pangio kuhlii (Valenciennes): Borneo–ZRC 39431 (3, 42.0–56.0). Malaysia–ZRC 27946 (4, 43.4–54.3).
ZRC 28029 (3, 38.6–51.3). ZRC 28217 (2, 57.1–61.9). ZRC 39552 (1, 67.2). ZRC 40199 (2, 35.3–42.9). ZRC
40222 (2, 47.2–50.0). ZRC 42772 (2, 47.8–50.9). ZRC 42792 (2, 40.5–43.4). ZRC 42817 (1, 63.1). ZRC

Zootaxa 2557 © 2010 Magnolia Press · 17
A NEW SPECIES OF LEPIDOCEPHALICHTHYS
42931 (1, 57.0). ZRC 42944 (1, 50.5). Sumatra–UF 166987 (6, 29.7–43.9). ZRC 38568 (2, 50.3–51.5). ZRC
42038 (1, 46.4). ZRC 42422 (2, 43.2–46.7).
Pangio malayana (Tweedie): Malaysia–UMMZ 238933 (3, 34.3–41.9). ZRC 17744 (1, 45.7). ZRC 28053
(1, 34.7). ZRC 28177 (1, 32.4). ZRC 34899 (2, 45.5–47.1). ZRC 35033 (1, 37.5). ZRC 38236 (1, 40.0). ZRC
42775 (2, 37.0–37.9). ZRC 42796 (2, 36.7–38.5). ZRC 42823 (4, 35.6–41.6). ZRC 42924 (2, 43.1–46.3). ZRC
42946 (2, 39.1–39.7). ZRC 42961 (2, 40.1–43.0). ZRC 44202 (2, 37.7–41.3).
Pangio mariarum: Borneo–ZRC 37644 (2, 31.0–33.9).
Pangio muraeniformis: Malaysia–FMNH 68663 (7, 33.7–35.7). ZRC 44200 (1, 40.8). Singapore–ZRC
1186 (4, 36.3–48.9). Sumatra–ZRC 30799 (2, 40.8–47.7). ZRC 30941 (2, 34.2–35.4). ZRC 32914 (2, 36.4–
48.7). ZRC 33040 (2, 46.5–48.1). ZRC 33389 (2, 40.2–43.4). ZRC 33622 (1, 46.4). ZRC 34296 (2, 39.7–
46.8). ZRC 42455 (3, 40.5–45.6).
Pangio myersi: Thailand–UMMZ 209439 (6, 54.6–64.8). ZRC 35807 (1, 38.7). ZRC 47141 (2, 64.8–
68.1).
Pangio oblonga: Java–ZRC 29150 (2, 54.8–56.1). Malaysia–ZRC 14322 (1, 36.6). ZRC 41038 (1, 45.3).
Thailand–UF 172938 (3, 37.3–39.3), GenBank GQ174323–4, GQ174397–8. ZRC 41972 (1, 48.6). Sumatra–
UF 161607 (2, 41.3–43.5), GenBank GQ174321. UF 166989 (4, 45.2–47.6), GenBank GQ174322,
GQ174405.
Pangio pangia: Malaysia–ZRC 496 (1, 41.0). ZRC 497 (2, 37.1–41.4). Myanmar–ZRC 43504 (2, 34.3–
35.7).
Pangio piperata: Malaysia–ZRC 34566 (1, 33.7). ZRC 38234 (2, 42.6–48.4). ZRC 38744 (4, 40.0–45.4).
ZRC 39554 (2, 33.5–38.9). ZRC 40221 (2, 40.3–47.9). ZRC 41871 (1, 44.0). ZRC 41934 (2, 43.9–46.5). ZRC
42699 (2, 38.3–38.8). ZRC 42770 (2, 42.8–43.1). ZRC 42787 (2, 35.9–37.0). ZRC 42824 (2, 38.4–38.6). ZRC
42864 (2, 43.7–46.7). ZRC 42922 (2, 34.3–35.7). ZRC 42948 (2, 33.7–39.8).
Pangio pulla: Borneo–ZRC 35022 (1, 59.7).
Pangio sandakanensis: Borneo–FMNH 44800 (paratypes of Acanthopthalmus sandakanensis) (2, 17.4–
23.5). FMNH 68159 (paratypes of Acanthopthalmus sandakanensis) (8, 24.8–35.5). FMNH 44798 (1, 27.3).
ZRC 37645 (17, 26.1–35.6). ZRC 45473 (2, 21.2–22.1).
Pangio semicincta: Malaysia–UF 173513 (2, 36.7–44.0), GenBank GQ174312, GQ174404. ZRC 17703
(1, 51.1). ZRC 17708 (2, 42.8–62.7). ZRC 43659 (2, 44.3–51.2). ZRC 4585 (3, 51.0–55.5). Sumatra–UF
161608 (2, 45.3–48.1), GenBank GQ174320. ZRC 43095 (46.5–51.0). Thailand–ZRC 42095 (2, 47.9–50.9).
Pangio shelfordii: Borneo–ZRC 37870 (1, 45.0). ZRC 37884 (2, 46.3–52.9). ZRC 39448 (3, 47.6–52.5).
Malaysia–ZRC 40233 (4, 45.4–53.7). ZRC 40255 (1, 47.4). ZRC 42771 (1, 46.2). ZRC 42825 (1, 43.0). ZRC
42836 (1, 57.6). ZRC 42850 (3, 47.1–55.6). Singapore–FMNH 60263 (5, 35.5–43.7). Sumatra–ZRC 37531
(2, 39.6–42.6).
Pangio sp.: Malaysia–ZRC 8916 (2, 34.7–42.0). Thailand–ZRC 39298 (1, 50.0). ZRC 41319 (1, 61.0).
Pangio superba: Borneo–ZRC 38787 (3, 30.9–36.4).
Sabanejewia larvata: Italy–UF 81157 (3, 28.7–43.6).
Sabanejewia romanica: Romania–UMMZ 185064 (10, 49.1–74.5).
Acknowledgements
This new species was independently discovered and under study by ichthyologists in three laboratories: Page,
Havird, and Tangjitjaroen; Vidthayanon; and Grudpan and Uddang. We combined our information and efforts
to produce this publication. Fieldwork was conducted and morphological data were assembled by all authors;
genetic analyses were done by Havird and Page. We thank curators, collection managers, and especially
collectors, of specimens from the following institutions: ANSP, BMNH, CAS, NIFI, FMNH, KU, MCZ,
MZB, UF, UMMZ, USNM, and ZRC. We thank B. Beamish, C. Beamish, J. Grudpan, B. Buasiyod for
assistance with collecting in Thailand; J. K. Parker and J. Yew for assistance with collecting in Malaysia; K.
Ng, T. Hui, K. Lim, T. Tan, I. Tan, M. Kottlelat, J. C. Mendosa, J. Lai, and M. Low for hospitality, assistance,

HAVIRD ET AL.18 · Zootaxa 2557 © 2010 Magnolia Press
and comments on this study during JCH’s visit to ZRC in Singapore and tissue samples of K. pristes; R.
Mayden and K. Conway for specimens from Bangladesh and Thailand as part of the Cypriniformes Tree of
Life Initiative (NSF DEB #0431326); G. Sheehy, R. Robins, A. Thomson, R. Johansen, A. Lopez, J. Sipiorski,
and J. Bloch for comments, technical assistance, and use of equipment. We are grateful to the All Catfish
Species Inventory award (NSF-DEB #0315963) to LMP and an East Asia and Pacific Summer Institute
fellowship from NSF and the National Research Foundation (Singapore) to JCH for funding.
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