Zootaxa 5477 (3): 367–379 ISSN 1175-5326 (print edition)

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Copyright © 2024 Magnolia Press
Article ZOOTAXA
ISSN 1175-5334 (online edition)
https://doi.org/10.11646/zootaxa.5477.3.6
http://zoobank.org/urn:lsid:zoobank.org:pub:41523CF9-C99B-490D-BB77-3336A59D1E07

Microdous hanlini (Gobiiformes: Odontobutidae), a new species of the fine-

toothed sleepers from Guangxi, China
FANGXIN WANG1, 2, MULAN WEI3, YOU HE4 & CHENHONG LI1,2
1
Shanghai Universities Key Laboratory of Marine Animal Taxonomy and Evolution, Shanghai Ocean University, Shanghai 201306,
China
� [email protected]; https://orcid.org/0000-0002-5917-0552
2
Engineering Research Center of Environmental DNA and Ecological Water Health Assessment, Shanghai Ocean University, Shanghai
201306, China
3
Du’an Yao Autonomous County Aquatic Technology Station, Hechi, Guangxi 530700, China.
� [email protected]; https://orcid.org/0009-0000-0254-5096
4
Shanghai Synchrotron Radiation Facility, Shanghai Advanced Research Institute, Chinese Academy of Sciences, Shanghai 201204,
China.
Corresponding authors: � [email protected]; https://orcid.org/0009-0004-8027-0760; � [email protected]; https://orcid.
org/0000-0003-3075-1756

Abstracts

Microdous (Gobiiformes: Odontobutidae) is a genus of freshwater sleepers distributed in southern China and Vietnam.
There are two described species in this genus, M. chalmersi widely distributed in Hainan Island and Guangxi province
of China and M. amblyrhynchos with a very restricted distribution range in Baise, Guangxi. It has been reported that M.
chalmersi of Guangxi might be a cryptic species, which is different from M. chalmersi of Hainan. Here, we describe the
cryptic species from Guangxi as a new species, Microdous hanlini, and compare it with the other species of Microdous.
Microdous hanlini can be distinguished from M. chalmersi by its wider head (head width/head length = 0.51–0.58 vs.
0.48–0.53 in M. chalmersi) and wider interorbital width (interorbital width/head length = 0.15–0.19 vs. 0.09–0.13 in M.
chalmersi). Microdous hanlini can be distinguished from M. amblyrhynchos by its longer snout, (snout length/head length
= 0.29–0.35 vs. 0.26–0.28 in M. amblyrhynchos); slenderer head (head width/head length = 0.51–0.58 vs. 0.61–0.65); and
protruding eyes (vs. not protruding). Phylogenetics analyses based on partial sequence of cytochrome c oxidase subunit
I (COI) gene (~1500 bp) showed that M. hanlini, M. chalmersi and M. amblyrhynchos formed reciprocal monophyletic
clades and M. hanlini is more closely related to M. amblyrhynchos than to M. chalmersi. Genetic distance between the
three species is greater than the interspecific distance between some species of Odontobutis.

Key words：Odontobutidae, skin teeth, freshwater sleepers, taxonomy, cryptic species

Introduction

The family Odontobutidae is a small group of freshwater sleeper fishes distributed in East Asia from the Far East region
of Russia to northern Laos and Vietnam, and in the Japanese archipelago. It was first established by Hoese and Gill
(1993) based on three genera including Odontobutis, Perccottus, and Micropercops. Three other genera, Sineleotris
(Herre, 1940), Neodontobutis (Chen et al. 2002) and Terateleotris (Shibukawa et al. 2001) were successively added to
this family. Li et al. (2018) established the genus Microdous for Phylipnus chalmersi (Nichols and Pope, 1927), also
named as Sineleotris chalmersi during their phylogenomic study of the Odontobutidae. Microdous is distinguished
from other odontobutid genera by a combination of characters: lateral line absent (present in Terateleotris); barbel-
like projection on sensory papilla absent (present in Neodontobutis); cephalic sensory canals complete (moderate in
Micropercops, absent or reduced in Odontobutis); and absence of vertical bands on the sides and a dark band under
eye (present in Sineleotris).
Subsequently, a new species, Microdous amblyrhynchos Hu & Li was discovered in Baise, Guangxi, which has
a blunter snout and wider interorbital width than that of M. chalmersi (Hu, 2023). Furthermore, a population of M.

Accepted by Y.-H. Zhao: 7 Jun. 2024; published: 9 Jul. 2024 367

Licensed under Creative Commons Attribution-N.C. 4.0 International https://creativecommons.org/licenses/by-nc/4.0/
chalmersi of Guangxi was found divergent from a population of M. chalmersi of Hainan based on mitochondrial
genome data, but no new species was described (Jiang et al., 2023). Recently, we collected nine specimens of “M.
chalmersi” from Du’an Guangxi. We examined these specimens and compared them with M. chalmersi and M.
amblyrhynchos. We also collected molecular data from the specimens and compared them with related species of the
Odontobutidae, including representative species of Microdous, Sineleotris, Micropercops and Odontobutis. Here,
we report a new species of Microdous based on analyses of morphometric characters and DNA barcode sequences.

Material and methods

Sampling

Nine specimens were collected from Du’an, Guangxi, China (23°55.94′N, 108°6.07′E) on March 2020 (Fig.
1). The left pectoral fins of three of them were clipped and used for molecular analysis (SOU1801015–1,
SOU1801015–2, SOU1801015–3). Then all specimens were fixed in 10% formalin, and transferred to 75% ethanol
to observe morphological data. All specimens were deposited in the Fish Collection of Shanghai Ocean University
(SOU1801015–1~9; contact person: Dr Ya Zhang, email: [email protected]).

Figure 1. The sampling location of Microdous hanlini sp. nov., Du’an, indicated by the orange triangle. Additional distribution
sites previously known for M. hanlini (orange dots; Gan et al., 2017; Zheng et al., 1981; Xiao et al., 2023; Wu et al., 1991; Wu
et al., 1986. ), M. chalmersi (yellow square; Wu and Ni, 1986) and M. amblyrhynchos (green star; Hu et al., 2023).

Morphological examination

Thirty-two morphological characters were examined, including eight meristic counts and 24 morphometric
characters. Measurements were taken with an electronic caliper to 0.1 mm precision. Morphological measurements,
counts, and observation followed Wu et al. (2008). Meristic counts involved fins rays of dorsal fins (D1/D2), anal
fin (A), pectoral fins (P), pelvic fins (V), caudal fin (C), lateral scales (scale series located on the lateral line),
transverse scales (scale series from origin of first dorsal fin to the abdomen) and predorsal scales (scale series
from head to origin of first dorsal fin),. Morphometric characters include total length (TL), standard length (SL),
Preanal length (PL),, caudal peduncle length, caudal peduncle depth, length of the first dorsal fin base, length of the
second dorsal fin base, length of anal fin base, body depth at caudal fin base, pectoral fin length, pelvic fin length,
body depth at the first dorsal fin origin, body depth at the anal fin origin, body width at first dorsal fin origin, body

368 · Zootaxa 5477 (3) © 2024 Magnolia Press Wang et al.

 width at the anal fin origin, head length, maximum head width, maximum head depth, head depth on cheek (at the
middle of the preopercular), snout length, postorbital head length, eye diameter, interorbital width and lower jaw
length. Morphological data were also taken for specimens of M. chalmersi and M. amblyrhynchos of Ichthyological
Collections, Shanghai Ocean University, including type specimens (voucher numbers: SOU1801010–1~3,
SOU1801010–7~8, 70504, v8791, v9228, 832384, v8792).
A specimen from the newly collected samples (SOU1801015–9) and a specimen of M. chalmersi (SOU1801017)
were scanned using MicroCT Skyscan 1176 (Bruker, Belgium) with 45 kV tube voltage, 0.3-degree rotation step,
and 8.7 µm pixel resolution. The three-dimensional renderings were created, visualized, and manipulated in the VG
Studio Max v2.1 (Heidelberg, Germany), as the previously reported (He et al., 2016).

Molecular analyses

Genomic DNA was extracted from ethanol-preserved tissue using an Ezup Column Animal Genomic DNA Purification
Kit (Sangon, Shanghai, China). The extracted DNA was checked by agarose gel electrophoresis for fragment size, and their
concentration was quantified with a NanoDrop 3300 Fluorospectrometer (Thermo Fisher Scientific, Wilmington, DE, U.S.A).
Partial sequence of cytochrome c oxidase subunit I (COI) (~ 500 bp), covering the whole barcoding region was amplified
from three specimens (SOU1801015–1, SOU1801015–2 and SOU1801015–3). COI gene was amplified using a Vazyme 2x
Taq Plus Master Mix II kit (Sangon, Shanghai, China) with a forward primer, 5’-CCATTTTACCTGTGRCAATCACACG-
3’ and a reverse primer 5’-CAGAGCGGTTATGTRTCTGGCTTGAA-3’ according to Zhou et al. (2022). The PCR protocol
was the follows: denaturation at 94 °C for 5 min, 30 cycles of denaturation at 94 °C for 30 s, annealing at 55 °C for 30 s,
extension at 72 °C for 1 min, with a final extension of 7 min at 72°C (Zhou et al. 2022). The size of amplified products was
checked on 1 % agarose gels. Amplified product with one clear target band was sent to Azenta for sequencing (Suzhou,
Jiangsu, China).
COI sequences from one Sineleotris saccharae (SOU1801016_2), one Micropercops swinhonis (NC_021763), two
Microdous chalmersi (OQ319988, OQ319987), one “S. chalmersi” (NC_045932) and three M. amblyrhynchos (OP536373,
OP536374, OP536375; specimen number SOU1801010-1~3) were retrieved from NCBI (https://www.ncbi.nlm.nih.gov/,
accessed on March 2023) or amplified and sequenced following the protocol as above. Odontobutis yaluensis (NC_027160)
and O. haifengensis (NC_036056) were used as outgroup taxa. All sequences were aligned using MUSCLE algorithm
implemented in MEGA X (Kumar et al., 2018). The “MODELS” tool in the software MEGA X was used to search for
the evolutionary model that best explains the variation between these sequences. The best substitution model found was
HKY+I. A maximum likelihood (ML) tree was reconstructed with 100 bootstrap replications under the best model. The
resulting trees were viewed and edited using FigTree v. 1.4.4 (Rambaut, 2018). Pairwise genetic distance within and between
species was calculated as the p-distance between the three Microdous species, and six species of Odontobutis (NC_036056,
NC_022818, NC_027583, NC_022706, NC_010199, NC_027160) were used to examine their relationship.

Results

Microdous hanlini Wang, He & Li, sp. nov.

Holotype. SOU1801015–8, female, 112.0 mm standard length (SL). Holotype obtained from a river of Du’an
(23°55.94′N, 108°6.07′E), Guangxi, China; collected by J.-H. Lan, March 2020 (Fig.2).
Paratypes. SOU1801015–7, male, 84.3 mm SL; SOU1801015–1, male, 76.4 mm SL; SOU1801015–4, female,
82.3 mm SL; SOU1801015–3, male, 60.3 mm SL. Collection data same as for the holotype.
Etymology. The species name is named after Prof. Hanlin Wu in honor of his significant contributions to the
study of the gobiiform fishes of China.
Diagnosis. The species M. hanlini can be distinguished from M. chalmersi (Nichols & Pope, 1927) by the
following characteristics: wider head, head width/head length = 0.51–0.58 (vs. slender head, head width/ head length
= 0.48–0.53); wider interorbital width, interorbital width (bone)/head length = 0.15–0.19 (vs. slender head and narrow
interorbital width, interorbital width (bone)/head length = 0.09–0.13). The species can be distinguished from M.
amblyrhynchos by the following characteristics: snout pointed, snout length/head length ratio 0.29–0.35 (vs. blunt

A new species of the fine-toothed sleeper, Microdous hanlini Zootaxa 5477 (3) © 2024 Magnolia Press · 369
snout, snout length/head length ratio 0.26–0.28); slenderer head, head width/ head length = 0.51–0.58, (vs. head width/
head length =0.61–0.65); eye large and protruding outward (vs. eye not protruding) (Fig. 3, Table 1). The species M.
hanlini can be distinguished from Sineleotris saccharae by absence of dark band under eye. The species M. hanlini can
be distinguished from Sineleotris namxamensis by the following characteristics: lateral scale rows 40-43 (vs. 36-39);
transverse scale rows 14-16 (vs. 12-14). These distinguishing features showed no difference between male and female
individuals of M. hanlini (Table 2).

Figure 2. Microdous hanlini sp. nov., SOU1801015–8, holotype, 112.0 mm standard length, Du’an, Guangxi. A lateral B
ventral C dorsal.

Description. First dorsal fin VIII or VIIII; second dorsal fin I, 9–11; anal fin I, 7–9; pectoral fin rays 13–15; pelvic fin I,
4–5; caudal fin rays 14–16; longitudinal scale rows 40–43; transverse scale rows 14–16; predorsal scales 18–23 (Table 2).
Body stout, cylindrical front, slightly compressed posteriorly. Head medium large and blunt, height of the head
equal to the width of the head. Eye large and protruding. Anterior end of the eye behind the posterior one-third of the
upper jaw. Gill openings extending to under front part of eyes. Gill rakers thin and pointed. Cephalic sensory canals
complete. Nostrils separated, two on each side, anterior nostril with a short tube, posterior nostril rounded, both located
in front of the eye. Smooth or tiny teeth–like dermal projections in vomerine region (Fig. 4).
The origin of the first dorsal fin aligned with or slightly posterior to the origin of pelvic fin. The second dorsal
fin originates before the origin of the anal fin. Caudal and pectoral fins oval. Pelvic fins separated, posterior ends not
reaching anus. Lateral lines absent, body surface and opercula covered with weakly ctenoid scales. The body color of
formalin-fixed specimens is grayish brown, with light-colored ventral surface, multiple dark spots on the dorsal fins,
caudal fin and lateral surface of the body, and an irregular black spot on the upper base of the pectoral fins (Fig. 2).

370 · Zootaxa 5477 (3) © 2024 Magnolia Press Wang et al.

 Figure 3. Lateral head (A) and dorsal (B) view of Microdous chalmersi (top, SL 83.0 mm, SOU 76V8792, Holotype), M.
hanlini sp. nov. (middle, SL 84.3 mm, SOU1801015–7, Paratypes) and M. amblyrhynchos (bottom, SL 74.9 mm, SOU1801010–
7, Holotype)

Figure 4. Vomerine region of A Microdous hanlini sp. nov. SOU1801015_10; B M. hanlini SOU1801015–2; C M. chalmersi
SOU1801009–1. Abbreviation: vt, vomerine teeth–like dermal projections.

A new species of the fine-toothed sleeper, Microdous hanlini Zootaxa 5477 (3) © 2024 Magnolia Press · 371
 Table 1. Comparison of measurements of Microdous hanlini sp. nov. vs M. chalmersi and M. amblyrhynchos.
Species Microdous hanlini M. chalmersi M. amblyrhynchos

70504
v8791
v9228
v8792

832384
Specimen

SOU1801015-1
SOU1801015-2
SOU1801015-3
SOU1801015-4
SOU1801015-5
SOU1801015-6
SOU1801015-7
SOU1801015-8
SOU1801010-7
SOU1801010-8
SOU1801010-1
SOU1801010-2
SOU1801010-3

Percentage of head length (%)

Maximum
52.17 51.95 57.34 55.09 58.34 56.06 51.36 58.01 48.86 49.23 53.33 48.34 49.60 60.98 62.73 64.87 57.95 62.16
head width
Interorbital
14.75 18.74 17.47 16.81 16.77 18.58 14.87 15.39 8.67 11.92 13.33 12.31 9.73 26.78 26.2 27.23 26.92 26.13
width

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Snout length 30.97 33.15 35.27 28.8 34.09 31.40 30.75 34.24 31.45 34.17 39.59 31.07 33.30 26.39 27.33 27.32 27.78 27.03

Percentage of head width (%)

Interorbital
53.28 49.78 46.45 61.97 52.99 58.76 62.25 48.15 37.93 42.39 45.03 47.75 47.16 41.97 46.46 42.03 43.93 41.77
width

Wang et al.
 Table 2. Morphological characters of the eight specimens of Microdous hanlini sp. nov.

female
SOU1801015-1

male
SOU1801015-2

female
SOU1801015-3

male
SOU1801015-4

female
SOU1801015-5

male
SOU1801015-6

female
SOU1801015-7

male
SOU1801015-8
Characters

Counts
Dorsal fins VIII, I–9 VIII, I–9 VIII, I–9 VIII, I–9 VIII, I–9 VIII, I–9 VIIII, I–9 VIII, I–11
Anal fin I–8 I–7 I–8 I–7 I–8 I–8 I–8 I–9
Pectoral fin rays 13 14 14 14 14 14 14 15
Pelvic fin I–5 I–5 I–5 I–5 I–4 I–5 I–5 I–5
Caudal fin rays 15 15 14 15 15 16 16 15
Lateral scale (rows) 41 40 42 40 41 40 40 43
Transverse scale (rows) 15 16 15 15 15 15 14 15
Predorsal scales 20 22 20 22 23 21 22 18
Measurements (mm )
Total length 76.4 115.1 112.3 82.3 82.5 80.3 84.3 111.8
Standard length 59.3 97.9 103.4 70.7 70.8 67.0 71.0 93.8
Preanal length 37.5 60.4 60.3 39.9 42.2 40.8 43.8 53.6
Caudal peduncle length 17.9 27.5 25.7 18.2 19.3 17.6 18.2 28.2
Caudal peduncle depth 5.1 9.5 10.2 6.5 6.7 6.8 6.4 9.3
Length of first dorsal fin base 7.37 12.3 12.13 8.25 9.1 9.53 9.57 13.34
Length of second dorsal fin base 9.5 18.1 17.7 11.6 10.8 11.6 12.1 18.5
Length of anal fin base 6.2 10.7 11.2 8.7 7.7 7.8 7.7 12.1
Body depth at caudal fin base 5.8 10.3 9.00 8.2 6.6 7.2 7.2 10.3
Pectoral fin length 11.7 23.3 22.2 15.7 14.0 14.3 16.8 21.9
Pelvic fin length 12.9 17.2 19.3 12.0 13.5 11.4 12.7 16.3
Body depth at first dorsal fin 11.4 18.0 17.8 12.6 13.6 11.9 13.9 16.4
origin
Body depth at anal fin origin 8.2 15.7 16.9 10.8 9.4 10.4 11.2 15.4
Body width at first dorsal fin 8.6 14.5 14.7 10.1 10.6 9.3 10.5 14.8
origin
Body width at anal fin origin 4.5 10.2 9.7 6.7 7.7 7.6 6.1 11.5
Head length 19.9 35.7 33.9 22.7 22.1 21.4 24.6 29.8
Postorbital head length 10.1 19.6 16.1 11.8 10.8 10.8 11.5 13.5
Maximum head width 10.4 18.5 19.5 12.5 12.9 12.0 12.6 17.3
Maximum head depth 10.2 17.5 18.9 13.5 11.6 11.6 12.6 17.5
Head depth at preopercle 7.9 14.8 15.3 10.9 8.8 8.9 10.2 14.5
Snout length 6.2 11.8 12.0 6.5 7.5 6.7 7.6 10.2
Eye diameter 5.5 6.7 6.6 5.0 5.5 4.6 5.1 6.6
Interorbital width 2.9 6.7 5.9 3.8 3.7 4.0 3.7 4.6
Lower jaw length 7.1 11.4 10.2 7.7 7.2 7.3 7.9 9.1

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 Coloration in life. Head brown with black dots on cheek. Body side dark brown with irregular black spots.
Abdomen dark. Fins dim and transparent, with banded black spots. An irregular black fleck on upper part of base
of the pectoral fins.
Coloration preserved. Body grayish brown, with light–colored ventral surface, multiple dark spots on the
dorsal fins, caudal fin and lateral surface of the body, and an irregular black spot above the base of the pectoral fin.
Ventral side of abdomen light brown. Head brown, cheek light brown, a horizontal black band crossing the eyes.
Urogenital papilla whitish, with sparse black pigment.
Sexual dimorphism. Urogenital papilla elongates with a wide base, tapering and with a narrow tip. No distinct
opening in male; clear ovipore in female (Fig. 5).

Figure 5. Urogenital papilla of Microdous hanlini sp. nov. SOU1801015–8 (A, male) and SOU1801015–7 (B, female).

Cephalic sensory canal system. Oculoscapular canal with three preorbita (lanterior interorbital) pores A’, B
and C, single interorbital pore D, single postorbital ( posterior interorbital) pore E, and seven pores F to L extending
posteriorly. Preopercular canal with five pores M to Q (Fig. 6).
Cephalic sensory papillae. Neuromasts numerous, small and densely set in mostly longitudinally arranged
rows (Fig. 6).

Head skeleton. The skeletons of M. hanlini and M. chalmersi were investigated using X-ray microtomography
(Fig. 7). A pair of the frontal bones constitutes the majority of the skull roof. The frontal includes a narrow anterior
supraorbital section and a widened postorbital section (or posterior section). The ratio of the width of anterior
section to postorbital section is much smaller in M. hanlini than that of M. chalmersi.

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 Figure 6. Patterns of main cephalic sensory pores (black spots) and papilla lines (rows of dots) of Microdous hanlini sp. nov. in
lateral (A); ventral (B) and dorsal(C) view. Preorbital pores A’, B, C; interorbital pore D; postorbital pore E; pores of posterior section of
oculoscapular canal F, G, H, I, J, K, L; preopercular pores Q, P, O, N, M.

Figure 7. Head skeleton of Microdous hanlini sp. nov. (SOU1801015–9) (A) and M. chalmersi (SOU1801017) (B) based
on microCT data.

A new species of the fine-toothed sleeper, Microdous hanlini Zootaxa 5477 (3) © 2024 Magnolia Press · 375
 COI analyses and genetic distance. The COI sequence of three paratype specimens were lodged in GenBank
with accession number PP481989-PP481991, and the Sineleotris saccharae as OQ382855. The results showed that
the three specimens of M. hanlini (SOU1801015–1~3) and the “Sineleotris chalmersi” sequence from Guangxi
formed a monophyletic clade with a bootstrap value of 100 (Fig. 8), suggesting that the “Sineleotris chalmersi” was
erroneously named for M. chalmersi, but is actually a sample of M. hanlini (Wang et al., 2019). Microdous hanlini
then formed a closer sister relationship to M. amblyrhynchos than to M. chalmersi (Fig. 8).

Figure 8. Maximum-likelihood (ML) tree of Microdous hanlini sp. nov. and other species of the Odontobutidae, O. yaluensis
and O. haifengensis as outgroups. The numbers by the nodes represent the bootstrap support values.

The intraspecific genetic distance of M. hanlini was 0.00065~0.03361, which is much smaller than that between
M. hanlini and M. chalmersi from Hainan (0.10759~0.11125) (Table 3). The average genetic distance between M.
hanlini and M. amblyrhynchos is 0.07283 (0.06878~0.08361) (Table 3). The average genetic distance between M.
hanlini and M. chalmersi is similar or greater than the genetic distance between some species of Odontobutis (Table
4).
Distribution. Specimens of M. hanlini examined in this study were collected from Du’an, Guangxi province
(Fig. 1). Other documented collection records of “M. chalmersi”, “Sineleotris chalmersi” or “Perccottus chalmersi”
in Guangxi include nearby regions of the Liu River, the Gui River, the Zuojiang River, the Youjiang River, the
Hongshui River and the Beilun River (Gan et al., 2017). The detailed distribution range of M. hanlini requires
further exploration.
Biology. According to the measurements of all specimens obtained in this research, the maximum size of M.
hanlini collected was 115.1 mm in standard length. A smaller M. hanlini was found in the stomach after dissecting
one specimen of M. hanlini, suggesting that M. hanlini is carnivorous.
Habitat. Microdous hanlini was collected from a rapid–flowing spring, which has rubble bottom substrate with
a vegetation-covered bank. Water depth 20–40 cm (Fig. 9).

376 · Zootaxa 5477 (3) © 2024 Magnolia Press Wang et al.

 Figure 9. Habitat of Microdous hanlini sp. nov.. An unnamed stream of Du’an (23°55.94′N, 108°6.07′E), Guangxi, China.

Table 3. Genetic distance between Microdous hanlini sp. nov. (SOU1801015–1~3, NC_045932.1), M. chalmersi
(OQ319988, OQ319987), and M. amblyrhynchos (SOU1801010-1~3, GenBank number OP536373.1~ 0P536375.1).
Species M. hanlini M. chalmersi M. amblyrhynchos
SOU1801015–1

SOU1801015–2

SOU1801015–3

NC_045932.1

OQ319987

OQ319988

OP536373.1

OP536374.1

OP536375.1
SOU1801015–1
SOU1801015–2 0.00130
SOU1801015–3 0.00196 0.00065
NC_045932.1 0.03500 0.03355 0.03362
OQ319987 0.10927 0.10759 0.10773 0.11125
OQ319988 0.10927 0.10759 0.10773 0.11125 0.00000
OP536373.1 0.07013 0.06878 0.06878 0.08361 0.10722 0.10722
OP536374.1 0.07013 0.06878 0.06878 0.08361 0.10722 0.10722 0.00000
OP536375.1 0.07013 0.06878 0.06878 0.08361 0.10722 0.10722 0.00000 0.00000

Table 4. Genetic distance between six species of Odontobutis.

O. haifengensis O. sinensis O. interrupta O. potamophilus O. platycephala O. yaluensis
O. haifengensis
O. sinensis 0.1442
O. interrupta 0.1554 0.1370
O. potamophilus 0.1527 0.1286 0.0393
O. platycephala 0.1501 0.1331 0.1288 0.1351
O. yaluensis 0.1521 0.1292 0.0747 0.0759 0.1281

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 Comparative material. All M. chalmersi and M. amblyrhynchos specimens examined for morphological
comparison were loaned from the Fish Collection of the Shanghai Ocean University, China with the registration
tags 76V8791, 76V8792, 76V9228 from Qiongzhong, Hainan, May 1976; HN832384 from Changjiang, Hainan,
May 1983, and 070504 from Wuzhishan, Hainan, May 1983; SOU1801009–1, from Wuzhishan, Hainan, August
2018; SOU1801010 (1~3, 7~8) from Baise City, Guangxi.

Discussion

Microdous chalmersi was described as Philypnus chalmersi (Nichols and Pope, 1927) originally from Hainan Island
of southern China. However, Philypnus is an obsolete genus name established for the eleotrid Gobiomorus dormitor
(Jordan and Gilbert, 1882) in the Western Hemisphere. Philypnus chalmersi has been shown genetically far apart from
G. dormitor and nested within the Odontobutidae, so the name of P. chalmersi has been corrected to M. chalmersi
(Li et al., 2018). Traditionally, Microdous chalmersi was thought to be distributed in Hainan Island and the Pearl
River basin of China and northern Vietnam (Wu and Ni, 1986). Recent analysis of mitochondrial genome sequences
suggested a potential cryptic species (Jiang et al., 2023), which has been confirmed by thorough examination of the
newly collected specimens in this study. Wang et al. (2019) reported a complete mitochondrial genome for Sineleotris
chalmersi. The specimen was collected from Dahua Yao Autonomous County (23°33′35.36″N, 107°20′36.06″E) in
the upper Pearl River (Guangxi, China). Our phylogenetic studies including this specimen indicated that it is a
specimen of M. hanlini.
Microdous hanlini can be distinguished from other species in the genus by: head width and interorbital width;
longer snout; eye protruding outward. Phylogenetics analyses showed that M. hanlini and M. chalmersi formed
reciprocal monophyletic clades and M. hanlini had a closer sister relationship to M. amblyrhynchos than to M.
chalmersi (Fig. 8). Microdous hanlini and M. amblyrhynchos occur closely in geological distance and probably have
sympatric distribution. The mechanism of their differentiation requires further study.
Sineleotris namxamensis has no dark band under eye but a black fleck at the base of pectoral fin (Chen &
Kottelat 2004), which is consistent with the species of Microdous but not Sineleotris. Therefore S. namxamensis
may be a species of Microdous. More samples should be collected from Lao PRD and Vietnam to compare using
both morphological characters and molecular data.

Ethical approval

All animal procedures performed in this research were done in accordance with the “Ethical Standards of the
Shanghai Ocean University (2020)”.

Competing interests

The authors declare no competing interests.

Author contributions

F. Wang and C. Li conceived and designed the study. M. Wei collected the specimens and analyzed morphological
data. Y. He performed the CT-scan. F. Wang performed the molecular experiments, conducted morphological
measurements, analyzed the data, and prepared the manuscript. All authors edited and approved the final version of
the manuscript.

378 · Zootaxa 5477 (3) © 2024 Magnolia Press Wang et al.

 Acknowledgements

We thank Mr. Jiantao Hu and Mr. Lei Jiang for their constructive suggestions during preparation of this work. C. Li
was supported by “Science and Technology Commission of Shanghai Municipality (19410740500)”.

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A new species of the fine-toothed sleeper, Microdous hanlini Zootaxa 5477 (3) © 2024 Magnolia Press · 379