A new species o Boulenoph ys (Anu a, Megoph yidae) om
no he n Jiangxi, China
Deming Shen1, Haiying Zhou1, Chaoyang Wang1, Jun Xu2, Shi Xu2, Yi eng Zhong1,
Zhenyu Wang1, Hao Cai1, Yahui Wu1, Meicai Wei1, Yankuo Li1
1 College o Li e Sciences, Jiangxi No mal Uni e si y, Nanchang 330022, China
2 Adminis a ion o Lushan Na ional Na u e Rese e, Jiujiang 332900, China
h ps://zoobank.o g/22135BBD-1048-4C9C-817F-2A2A2A96CC85
Co esponding au ho s: Meicai Wei ([email p o ec ed]); Yankuo Li ([email p o ec ed])
Academic edi o : Umilaela A i in ♦
Recei ed
25 July 2025 ♦
Accep ed
5 No embe 2025 ♦
Published
18 No embe 2025
Abs ac
A new species o he genus Boulenoph ys (Anu a, Megoph yidae), Boulenoph ys nebulosa sp. no ., is desc ibed om M . Lushan,
Jiangxi P o ince, China. Phylogene ic analyses based on mi ochond ial COI and 16S RNA genes show ha he new species o ms a
sis e lineage o B. mu umon ana. Mo phologically, B. nebulosa sp. no . di e s om all known congene s by a unique combina ion
o cha ac e s and om i s closes ela i e, B. mu umon ana, by he ollowing combina ion o cha ac e s: (1) ela i e inge leng hs
I < II < IV < III; (2) indis inc suba icula ube cle a he base o each inge ; (3) la ge male body size (SVL 31.8–32.3 mm);
(4) heels jus mee ing when hindlimbs a e olded; (5) ibio a sal a icula ion eaching he cen e o he eye when he leg is s e ched
o wa d; and (6) ome ine ee h p esen . This s udy expands he known dis ibu ion o Boulenoph ys in Jiangxi and p o ides base-
line da a o biodi e si y conse a ion e o s in M . Lushan.
Key Wo ds
Asian ho ned oads, biodi e si y, Boulenoph ys nebulosa sp. no ., axonomy
In oduc ion
The genus Boulenoph ys Fei, Ye & Jiang, 2016 (Me-
goph yinae) cu en ly comp ises 78 ecognized species
(Table 1). I s membe s a e dis ibu ed ac oss he Hima-
layas, no heas e n India, sou he n China, and Sou heas
Asia (Lyu e al. 2023; AmphibiaWeb 2025; F os 2025).
Despi e i s b oad ange, ecen in eg a i e s udies ha e
e ealed ex ensi e c yp ic di e si y wi hin Boulenoph ys,
much o which is no disce nible by mo phology alone
(Liu e al. 2018). These indings ha e spu ed axonom-
ic e isions wi hin he sub amily Megoph yinae, wi h
Lyu e al. (2023) p oposing a en-genus sys em ha a -
i ms Boulenoph ys as he mos species- ich lineage. In
Jiangxi, known Boulenoph ys species a e p ima ily de-
sc ibed om he Luoxiao Moun ains along he Hunan
bo de and he Wuyi Moun ains along he Fujian bo de .
Howe e , he no he n pa o Jiangxi, including he opo-
g aphically isola ed g ani ic massi o M . Lushan in he
Yang ze Ri e Basin, has ecei ed limi ed he pe ological
a en ion. This sugges s ha he species di e si y o Bou-
lenoph ys in his egion may s ill be unde es ima ed.
His o ically, wo Boulenoph ys popula ions ha e
been eco ded in M . Lushan: B. lushanensis Shen,
Zhou & Li (Shen e al. 2025), and ano he i s epo ed
by Yang e al. (2007), ini ially iden i ied as Megoph ys
mino b achykolos (Inge & Rome , 1961) (cu en ly B.
b achykolos). Howe e , once hough o be widely dis-
ibu ed in sou he n China and no he n Vie nam (Fei
e al. 2009), B. b achykolos has been es ic ed o Hong
Kong and Shenzhen based on ouche specimens and
molecula da a (Lyu e al. 2023). Fo ins ance, he e-
Zoosys . E ol. 101 (4) 2025, 2251–2266|DOI 10.3897/zse.101.166556
Copy igh Shen, D. e al. This is an open access a icle dis ibu ed unde he e ms o he C ea i e Commons A ibu ion License (CC BY 4.0), which pe mi s un es ic ed use,
dis ibu ion, and ep oduc ion in any medium, p o ided he o iginal au ho and sou ce a e c edi ed.
zse.penso .ne
Shen, D. e al.: A new species o Boulenoph ys (Anu a, Megoph yidae) om no he n Jiangxi, China
2252
Table 1. Re e ences o mo phological cha ac e s o 78 ecognized species o Boulenoph ys.
ID Boulenoph ys Species Re e ences
1B. acu a Wang, Li & Jin, 2014 Li e al. 2014
2B. angka Wu, Suwannapoom, Poya ko , Pawangkhanan , Xu, Jin, Mu phy & Che, 2019 Wu e al. 2019
3B. anlongensis Li, Lu, Liu & Wang, 2020 Li e al. 2020
4B. baishanzuensis Wu, Li, Liu, Wang & Wu, 2020 Wu e al. 2020
5B. baolongensis Ye, Fei & Xie, 2007 Ye e al. 2007
6B. binchuanensis Ye & Fei, 1995 Ye and Fei 1995; Lyu e al. 2023
7B. binlingensis Jiang, Fei & Ye, 2009 Fei e al. 2009; Lyu e al. 2023
8B. boe ge i Boulenge , 1899 Boulenge 1899; Lyu e al. 2023
9B. b achykolos Inge & Rome , 1961 Inge and Rome 1961; Lyu e al. 2023
10 B. caobangensis Nguyen, Pham, Nguyen, Luong & Ziegle , 2020 Nguyen e al. 2020
11 B. caudop oc a Shen, 1994 Shen. 1994
12 B. cheni Wang & Liu, 2014 Wang e al. 2014
13 B. chishuiensis Xu, Li, Liu, Wei & Wang, 2020 Xu e al. 2020
14 B. congjiangensis Luo, Wang, Wang, Lu, Wang, Deng & Zhou, 2021 Luo e al. 2021
15 B. changyangensis Li, Shi, Liu, Zhao, Gao, & Wang, 2025 Li e al. 2025
16 B. daiyunensis Lyu, Wang & Wang, 2021 Lyu e al. 2021
17 B. dalaolingensis Wang, Chen & Liu, 2025 Wang e al. 2025a
18 B. daoji Lyu, Zeng, Wang & Wang, 2021 Lyu e al. 2021
19 B. daweimon is Rao & Yang, 1997 Rao and Yang 1997
20 B. daxuemon is Liu, Li, Cheng, Wei, Wang & Cheng, 2025 Liu e al. 2025b
21 B. dongguanensis Wang & Wang, 2019 Wang e al. 2019a
22 B. dupanglingensis Xiao & Mo, 2025 Xiao e al. 2025
23 B. elonga a Zeng, Wang, Chen, Xiao, Zhan, Li & Lin, 2024 Zeng e al. 2024
24 B. anjingmon is Zhang, Liang, Ran & Shen, 2012 Zhang e al. 2012
25 B. ansipanensis Tapley, Cu aja , Mahony, Nguyen, Dau, Luong, Le, Nguyen,Nguyen, Po way, Luong &
Rowley, 2018
Tapley e al. 2018
26 B. engshunensis Wang, Zeng, Lyu & Wang, 2022 Wang e al. 2022
27 B. igida Tapley, Cu aja, Nguyen, Po way, Mahony, Nguyen, Ha ding, Luong& Rowley, 2021 Tapley e al. 2021
28 B. gaolanensis Song, Wang,Qi, Wang, Wang,2024 Song e al.2024
29 B. gu u Kang, Wei & Shi, 2025" Kang e al. 2025
30 B. hengshanensis Qian, Hu, Mo, Gao, Zhang & Yang, 2023 Qian e al. 2023
31 B. hoanglienensis Tapley, Cu aja , Mahony, Nguyen, Dau, Luong, Le, Nguyen,Nguyen, Po way, Luong &
Rowley, 2018
Tapley e al. 2018
32 B. huangniushiensis Wang, Wu, Song, Huang, Zhuo, & Wang, 2025 Wang e al. 2025b
33 B. hung ai Wang, Zeng, Lyu, Xiao & Wang, 2022 Wang e al. 2022
34 B. insula is Wang, Liu, Lyu, Zeng & Wang, 2017 Wang e al. 2017a
35 B. jiangi Liu, Li, Wei, Xu, Cheng, Wang & Wu, 2020 Liu e al. 2020
36 B. jingdongensis Fei & Ye, 1983 Fei e al. 1983; Lyu e al. 2023
37 B. jinggangensis Wang, 2012 Wang e al. 2012
38 B. jiulianensis Wang, Zeng, Lyu & Wang, 2019 Wang e al. 2019a
39 B. kua unensis Pope, 1929 Pope 1929; Lyu e al. 2023
40 B. leishanensis Li, Xu, Liu, Jiang, Wei & Wang, 2018 Li e al. 2018
41 B. liboensis Zhang, Li, Xiao, Li, Pan, Wang, Zhang & Zhou, 2017 Zhang e al. 2017
42 B. lichun Lin, Chen, Li, Peng, Zeng, Wang, 2024 Lin e al.2024
43 B. lini Wang & Yang, 2014 Wang e al. 2014
44 B. lushanensis Shen, Zhou & Li, 2025 Shen e al. 2025
45 B. lishuiensis Wang, Liu & Jiang, 2017 Wang e al. 2017b
46 B. lushuiensis Shi, Li, Zhu, Jiang, Jiang & Wang, 2021 Shi e al. 2021
47 B. mino S ejnege , 1926 S ejnege 1926; Lyu e al. 2023
48 B. mi abilis Lyu, Wang & Zhao, 2020 Lyu e al. 2020
49 B. mu umon ana Wang, Lyu & Wang, 2019 Wang e al. 2019a
50 B. nankunensis Wang, Zeng & Wang, 2019 Wang e al. 2019a
51 B. nanlingensis Lyu, Wang, Liu & Wang, 2019 Wang e al. 2019a
52 B. obesa Wang, Li & Zhao, 2014 Li e al. 2014
53 B. omb ophila Messenge & Dahn, 2019 Messenge e al. 2019
54 B. omeimon is Liu, 1950 Liu 1950; Lyu e al. 2023
55 B. palpeb alespinosa Bou e , 1937 Bou e 1937; Lyu e al. 2023
56 B. pepe Wang & Zeng, 2024 Wang e al. 2024
57 B. puningensis Wang, Zeng, Lyu, Xiao & Wang, 2022 Wang e al. 2022
58 B. qianbeiensis Su, Shi, Wu, Li, Yao, Wang & Li, 2020 Su e al. 2020
59 B. ub ime a Tapley, Cu aja , Mahony, Chung, Dau, Nguyen, Luong & Rowley,2017 Tapley e al. 2017
60 B. sangzhiensis Jiang, Ye & Fei, 2008 Jiang e al. 2008
61 B. sanmingensis Lyu & Wang, 2021 Lyu e al. 2021
62 B. shimen aina Lyu, Liu & Wang, 2020 Lyu e al. 2020
63 B. shuichengensis Tian & Sun, 1995 Tian and Sun 1995
64 B. shunhuangensis Wang, Deng, Liu, Wu & Liu, 2019 Wang e al. 2019b
65 B. spina a Liu & Hu, 1973 Hu e al. 1973; Lyu e al. 2023
66 B. ongboensis Wang & Lyu, 2021 Lyu e al. 2021
Zoosys . E ol. 101 (4) 2025, 2251–2266
zse.penso .ne
2253
co d om M . Dayaoshan, Guangxi, was e ised and de-
sc ibed as he new species B. yaoshanensis Qi, Mo, Lyu,
Wang & Wang, 2021, and he eco d om M . Hengshan,
Hunan, was e ised as he new species B. hengshanensis
Qian, Hu, Mo, Gao, Zhang & Yang, 2023. Consequen ly,
all p e ious eco ds o B. b achykolos om o he egions
a e now conside ed ques ionable and need o be ca e ully
e-examined. No ably, he popula ion o B. b achykolos
om M . Lushan has no ye been e alua ed using gene ic
o de ailed mo phological analyses, lea ing i s axonomic
iden i y un esol ed.
He pe ological su eys on M . Lushan since 2021
ha e led o he collec ion o se e al ho ned oad speci-
mens. Mo phological examina ions e ealed ha hese
indi iduals could no be classi ied as any known species
o he genus. While hey sha e a supe icial esemblance
o B. mu umon ana, hey exhibi dis inc di e ences in
a combina ion o mo phological cha ac e s, mos no-
ably in he ela i e leng h o hei inge s. To u he
elucida e hei axonomic s a us, we sequenced 16S and
COI mi ochond ial gene agmen s and pe o med phy-
logene ic analyses. In eg a i e e idence om phyloge-
ny and mo phology suppo s he ecogni ion o he M .
Lushan popula ion as a dis inc e olu iona y lineage.
Based on his e idence, we o mally desc ibe i as a new
species he ein.
Ma e ials and me hods
Sampling
Fieldwo k was conduc ed a M . Lushan, China. Se en
adul specimens and one adpole specimen we e collec -
ed in Ap il, June, and No embe 2021. A e cap u e, he
specimens we e eu hanized wi h icaine me hanesul on-
a e (MS-222). Li e o high muscle issues we e aken
o molecula expe imen s, p ese ed in 95% e hanol,
and s o ed a –70 °C. Whole specimens we e ixed in 75%
e hanol. All specimens and issue samples we e deposi -
ed in he Animal Museum o he Li e Sciences College,
Jiangxi No mal Uni e si y (JXNU), Nanchang, Jiangxi
P o ince, China. All animal p o ocols we e e iewed and
app o ed by he Animal E hical and Wel a e Commi ee
o Jiangxi No mal Uni e si y.
Mo phological analyses
Mo phological e minology and cha ac e de ini ions
ollowed Lyu e al. (2023) and subsequen axonomic
e isions o Boulenoph ys. Ex e nal measu emen s we e
aken using a digi al calipe (Neiko 01407A, s ainless
s eel, 6-inch) o he nea es 0.1 mm. The ollowing mea-
su emen s we e eco ded: snou – en leng h (SVL), head
leng h (HDL), head wid h (HDW), snou leng h (SNT),
in e nasal dis ance (IND), in e o bi al dis ance (IOD),
eye diame e (ED), ympanum diame e (TD), ympa-
num–eye dis ance (TED), hand leng h (HND), o ea m
leng h (RAD), ibial leng h (TIB), and oo leng h (FTL).
Sex was de e mined by he p esence o seconda y sexual
cha ac e is ics, including in e nal ocal sac openings o
nup ial pads in males, and by di ec obse a ion o cou -
ship calling when possible (Fei e al. 2009). The p esence
o absence o nup ial pads was e i ied unde a dissec ing
mic oscope. Compa a i e mo phological da a o conge-
ne s we e compiled om o iginal species desc ip ions
and axonomic e isions (Table 1), ensu ing consis ency
in cha ac e de ini ions and measu emen s anda ds.
Fo he single adpole o he undesc ibed species, he
ollowing mo phome ic cha ac e s we e measu ed: max-
imum body heigh (BH), maximum body wid h (BW), in-
e ocula dis ance (IOS), mou h wid h (MW), snou leng h
(SL), snou - o-spi aculum dis ance (SS), snou – en
leng h (SVL), ail heigh (TAH), ail leng h (TAL), maxi-
mum wid h o he ail base (TBW), and o al leng h (TOL).
The mo phome ic measu emen s we e s a is ical-
ly analyzed using Py hon 3.11 wi h he pandas, numpy,
sciki -lea n, and ma plo lib packages. Fo analyses, all
measu emen s we e ln- ans o med o no malize and
educe a iance, and hen scaled o emo e allome-
ic e ec s o body size using he ollowing equa ion:
Xa = Xln − β × (SVLln − SVLm), whe e Xa = adjus -
ed alue, Xln = ln- ans o med measu emen , β = un-
s anda dized eg ession coe icien o each species,
SVLln = ln- ans o med SVL, and SVLm = o e all a -
e age SVLln o all samples. The size-adjus ed measu e-
men s we e hen s anda dized using z-sco e no maliza-
ion o ensu e compa abili y ac oss a iables. P incipal
componen analysis (PCA) was pe o med o examine
mo phological a ia ion among species using he PCA
unc ion om sciki -lea n. Since sexual dimo phism is
ID Boulenoph ys Species Re e ences
67 B. ube og anula us Shen, Mo & Li, 2010 Mo e al. 2010
68 B. wugongensis Wang, Lyu & Wang, 2019 Wang e al. 2019a
69 B. wuliangshanensis Ye & Fei, 1995 Ye and Fei 1995
70 B. wushanensis Ye & Fei, 1995 Ye and Fei 1995
71 B. xiangnanensis Lyu, Zeng & Wang, 2020 Lyu e al. 2020
72 B. xianjuensis Wang, Wu, Peng, Shi, Lu & Wu, 2020 Wang e al. 2020
73 B. xue engmon is Lyu & Wang, 2023 Lyu e al. 2023
74 B. yangmingensis Lyu, Zeng & Wang, 2020 Lyu e al. 2020
75 B. yaoshanensis Qi, Mo, Lyu, Wang & Wang, 2021 Qi e al. 2021
76 B. yezhongensis Liu, Feng, Shen, Li, Cheng, Wei, Wang & Su, 2025 Liu e al. 2025a
77 B. yingdeensis Qi, Lyu, Wang & Wang, 2021 Qi e al. 2021
78 B. yunkaiensis Qi, Wang, Lyu & Wang, 2021 Qi e al. 2021
zse.penso .ne
Shen, D. e al.: A new species o Boulenoph ys (Anu a, Megoph yidae) om no he n Jiangxi, China
2254
common in ogs, we compiled a da ase using male spec-
imens om he ype se ies o B. mu umon ana (Wang e
al. 2019a) and he undesc ibed species o PCA.
DNA sequencing and molecula analyses
Genomic DNA was ex ac ed om issue samples using
he Uni e sal Genomic DNA Ki (CWBIO, Beijing, Chi-
na) ollowing he manu ac u e ’s p o ocol o animal is-
sue. Two mi ochond ial gene agmen s, he pa ial 16S
ibosomal RNA coding sequence (16S RNA) and he
pa ial cy och ome c oxidase subuni I gene (COI), we e
ampli ied in 50 µL eac ions. P ime pai s and PCR p o-
ocols ollowed Liu e al. (2018).
Fo he phylogene ic analyses, sequences o 78 Boule-
noph ys species we e ob ained om GenBank, excep o
B. changyangensis, B. dalaolingensis, and B. gu u due o
una ailable da a. Two ou g oup species, Xenoph ys glan-
dulosa and X. mangshanensis, we e also included (Suppl.
ma e ial 1: able S1). All sequences we e aligned using
he MUSCLE algo i hm wi h de aul pa ame e s. Phylo-
gene ic analyses we e conduc ed based on a conca ena -
ed mi ochond ial da ase comp ising COI and 16S RNA
genes ( o al leng h: 1,236 bp).
Bo h Maximum likelihood (ML) and Bayesian in e -
ence (BI) me hods we e employed o cons uc phyloge-
ne ic ees. The ML analysis was pe o med in IQ-TREE
3.0.1, wi h he bes - i subs i u ion model (TVM+F+R5)
selec ed using ModelFinde . B anch suppo was assessed
using 1,000 ul a as boo s ap eplica es. The BI analysis
was conduc ed in M Bayes 3.2 (Ronquis e al. 2012), us-
ing gene pa i ions de ined by Pa i ionFinde 2 (Lan ea
e al. 2017) (16S: 1–554; COI: 555–1236). Ma ko Chain
Mon e Ca lo (MCMC) sampling was un o 2,000,000
gene a ions, wi h ees sampled e e y 1,000 gene a ions
and he i s 25% disca ded as bu n-in. Con e gence was
conside ed achie ed when he a e age s anda d de ia ion
o spli equencies d opped below 0.01.
Acous ic analyses
Ad e isemen calls o he undesc ibed species we e e-
co ded om he holo ype (JXNU 21042833) on 15 June
2021 a a dis ance o 0.4 m on M . Lushan, Jiujiang, Ji-
angxi P o ince, China (ambien empe a u e: 22.3 °C, el-
a i e humidi y: 85%). Digi al eco dings we e made us-
ing a TASCAM DR05X eco de (96 kHz sampling a e,
24-bi dep h) wi h a buil -in mic ophone. The acous ic
analysis implemen ed a Py hon 3.9 pipeline using lib osa
o audio p ocessing, noise educe o spec al denoising,
and scipy.signal. ind_peaks o pulse de ec ion. Co e p o-
cedu es included ampli ude- h eshold segmen a ion a a
0.013 a io o isola e calls, spec al peak iden i ica ion o
pulse quan i ica ion, and dominan equency ex ac ion
ia a 512-poin Hann-windowed FFT. All pa ame e s
we e measu ed ollowing Tapley e al. (2021).
Resul s
Phylogene ic ees econs uc ed using BI and ML me hods
showed highly consis en opologies. The ML ee is shown
in Fig. 1, wi h node suppo conside ed s ong when boo -
s ap alues (BS) ≥ 70 o Bayesian pos e io p obabili ies
(BPP) ≥ 0.90. Samples o he undesc ibed species o med
a s ongly suppo ed monophyle ic clade in bo h analyses
(BS = 100; BPP = 1.0). This clade was consis en ly eco e ed
as sis e o B. mu umon ana (BS = 97; BPP = 1.0). Gene ic
dis ances based on he COI and 16S genes, calcula ed using
he unco ec ed p-dis ance model, showed ha di e gence
wi hin he undesc ibed species was below 0.6% o COI and
below 0.4% o 16S. The COI gene dis ance be ween he
undesc ibed species and i s closes ela ed species, B. mu-
umon ana, was 8.2%; 10.8% wi h B. caudop oc a; 11.4%
wi h B. boe ge i; and 12.1% wi h B. liboensis. Fo he 16S
gene, he closes gene ic dis ance was 1.5% wi h B. boe -
ge i, 1.7% wi h B. mu umon ana, 2.7% wi h B. caudop oc a,
and 3.2% wi h B. liboensis (Suppl. ma e ial 1: ables S2, S3).
Mo phologically, in he PCA analysis, he ex ac ed
componen s PC1 and PC2 explained 62.30% and 25.92%
o he a iance, espec i ely, accoun ing o 88.23% cu-
mula i ely. As shown in he sca e plo o PC1 and PC2
(Fig. 2), he undesc ibed species and B. mu umon ana
a e sepa a ed wi hou o e lapping. Mo eo e , he unde-
sc ibed species can be dis inguished om all ecognized
congene s by a unique combina ion o mo phological
cha ac e s (see Taxonomic accoun below). Suppo ed by
bo h phylogene ic and mo phological e idence, we o -
mally desc ibe i below as a new species.
Taxonomic accoun
Boulenoph ys nebulosa Shen & Li, sp. no .
h ps://zoobank.o g/6BC99CDF-720B-41EA-AE1E-C0216AD3EF6B
Fig. 3, Table 2
Type ma e ials. Holo ype. • JXNU 21042833 (Fig. 3A–F),
adul male, collec ed by Deming Shen, Haiying Zhou, Yi eng
Zhong, Chaoyang Wang, Hao Cai, Jun Xu and Shi Xu on 28
Ap il 2021 om M . Lushan (29°32.56'N, 115°57.58'E; ca.
970 m a.s.l.), Lushan Ci y, Jiujiang, Jiangxi, China.
Pa a ypes. (n = 6) • JXNU 21042832 (Fig. 4A–C),
JXNU 21042834, adul males, and JXNU 21042810, adul
emale, collec ed by Deming Shen, Haiying Zhou, Yi eng
Zhong, Chaoyang Wang, Hao Cai, Jun Xu and Shi Xu on
28 Ap il 2021 om M . Lushan (29°31.87'N, 115°55.87'E;
ca. 660 m a.s.l.); • JXNU 21061501(Fig. 4D–F), JXNU
21061502, JXNU 21061504, adul emales, collec ed by
same collec o s on 15 June 2021 om he ype locali y
(29°32.56'N, 115°57.58'E; ca. 970 m a.s.l.).
O he ma e ial examined. • One adpole (JXNU
21110702; Fig. 5) collec ed by Haiying Zhou on 2 No-
embe 2021.
E ymology. The speci ic epi he nebulosa ( eminine o
La in nebulosus) e e s o he equen ly mis -sh ouded
Zoosys . E ol. 101 (4) 2025, 2251–2266
zse.penso .ne
2255
mon ane o es s abo e 600 m on M . Lushan, Jiangxi
P o ince, China. This name no only e lec s he species’
habi a p e e ence bu also alludes o he egion’s amous
“Cloud & Fog Tea” (云雾茶, yún wù chá) speciali y.
The English e nacula name “Yunwu Ho ned Toad” is
he e p oposed, wi h he co esponding Chinese name
“云雾角蟾” (Yúnwù Jiǎochán).
Diagnosis. This species can be dis inguished om i s
congene s by he combina ion o ollowing mo phological
cha ac e is ics: (1) adul males, SVL 31.8–32.3 mm (n = 3);
mu umon ana
sp. no .nebulosa
glandulosa
mangshanensis
acu a
gaolanensis
lushuiensis
ansipanensis
chishuiensis
mino
daxuemon is
jiangi
caudop oc a
jinggangensis
lushanensis
leishanensis
palpeb alespinosa
igida
hoanglienensis
JXNU 21042810
boe ge i
liboensis
ongboensis
xue engmon is
ube og anula a
congjiangensis
wuliangshanensis
JXNU 21061504
SYS a006390
SYS a006392
SYS a006419
SYS a006391
lishuiensis
b achykolos
elonga a
mi abilis
shunhuangensis
baolongensis
wushanensis
ub ime a
JXNU 21061501
daoji
xianjuensis
lichun
yaoshanensis
hengshanensis
JXNU 21110702
wugongensis
dupanglingensis
yunkaiensis
jiulianensis
caobangensis
jingdongensis
JXNU 21042832
dongguanensis
nankunensis
baishanzuensis
sanmingensis
engshunensis
shimen aina
yangmingensis
angka
anlongensis
JXNU 21042833
hung ai
insula is
cheni
daiyunensis
lini
nanlingensis
yingdeensis
binchuanensis
daweimon is
omeimon is
JXNU 21061502
JXNU 21042834
omb ophila
kua unensis
puningensis
huangniushiensis
xiangnanensis
binlingensis
obesa
pepe
qianbeiensis
yezhongensis
spina a
sangzhiensis
anjingmon is
100/1
93/1
94/-
-/-
90/-
78/-
100/1
-/-
100/1
99/1
95/-
100/1
94/-
83/1
99/-
97/1
80/-
100/1
85/-
-/-
-/-
94/0.98
91/-
100/1
100/1
93/-
94/1
99/1
99/1
95/0.93
-/-
-/-
96/-
-/-
-/-
99/-
89/-
-/-
-/-
98/1
-/-
72/-
97/1
74/-
-/-
78/-
-/-
-/-
100/1
98/-
-/-
-/-
100/1
99/1
-/-
86/-
100/1
99/-
-/-
-/-
100/1
85/-
70/-
99/-
100/1
90/0.97
-/-
100/1
94/-
100/-
100/1
100/1
100/1
99/-
100/1
100/0.98
Xenoph ys
Boulenoph ys
0.02
T ee scale
Figu e 1. The maximum likelihood opology in e ed om pa ial DNA sequences o he mi ochond ial 16S RNA and COI genes.
Boo s ap suppo alues and Bayesian pos e io p obabili ies a e shown a he ee nodes. A dash deno es boo s ap suppo below
70 o Bayesian pos e io p obabili y below 0.9.
zse.penso .ne
Shen, D. e al.: A new species o Boulenoph ys (Anu a, Megoph yidae) om no he n Jiangxi, China
2256
Figu e 2. P incipal componen analysis based on he mo phome ic measu emen s o Boulenoph ys nebulosa sp. no . and B. mu u-
mon ana. A. Sca e plo o PC1 and PC2; B. Loadings plo o PC1 and PC2.
Figu e 3. Holo ype o Boulenoph ys nebulosa sp. no . (JXNU 21042833) in li e: A. La e al iew; B. Do sal iew; C. Ven al iew;
D. Vola and do sal iews o le hand; E. Plan a iew o le oo ; F. Edge o ongue. Pho os by Deming Shen.
Table 2. Measu emen s (mm) o adul specimens in he ype se ies o Boulenoph ys nebulosa sp. no .
JXNU 21042832 JXNU 21042833 JXNU 21042834 JXNU 21042810 JXNU 21061504 JXNU 21061502 JXNU 21061501
Sex Male Male Male Female Female Female Female
SVL 31.8 32.2 32.3 37.1 36.5 35 37.3
HDL 10.3 10.6 10.4 12.1 11.5 11.6 11.7
HDW 9.9 9.6 10.3 11.1 11.5 11 11.6
SNT 3.1 3.8 3.4 4.1 4.2 3.9 3.9
IND 3.2 3.3 3.3 3.5 3.6 3.2 3.6
IOD 3.2 2.8 2.8 3 3.6 3.2 3.7
ED 3.8 3.7 4.1 4.1 4.3 4 4.4
TD 1.8 1.5 1.5 2.2 2.1 1.7 1.8
TED 1.1 1.2 1.1 1.4 1 1.4 1.9
HND 6.7 7 7 7.9 7.5 7.5 7.4
RAD 7.3 7.7 7.7 7.9 8.1 8.5 8.6
TIB 14.3 14.5 14.6 16 15.2 16.3 16.7
FTL 13.7 12.7 14.4 14.3 14.4 14.8 14.6
Zoosys . E ol. 101 (4) 2025, 2251–2266
zse.penso .ne
2257
adul emales, SVL 35.0–37.3 mm (n = 4); (2) can hus
os alis well de eloped, ongue no no ched pos e io ly;
(3) ympanum dis inc ; (4) ome ine idges weak; om-
e ine ee h p esen ; (5) ou e ma gin o he uppe eyelid
bea s a small, p ominen ho n-like ube cle; sup a ym-
panic old dis inc , na ow, cu ing pos e o en ally o
abo e he a m; (6) ela i e inge leng hs I < II < IV < III;
indis inc suba icula ube cle on base o each inge ;
Figu e 4. Pa a ypes o Boulenoph ys nebulosa sp. no . in li e. Male pa a ype (JXNU 21042832): A. La e al iew; B. Do sal iew;
C. Ven al iew. Female pa a ype (JXNU 21061501): D. La e al iew; E. Do sal iew; F. Ven al iew. Pho os by Deming Shen.
zse.penso .ne
Shen, D. e al.: A new species o Boulenoph ys (Anu a, Megoph yidae) om no he n Jiangxi, China
2258
(7) heels mee ing when hindlimbs olded; ibio a sal a -
icula ion eaching cen e o eye when leg s e ched o -
wa d; (8) oes wi h na ow inges and only udimen a y
webbing; (9) ough do sal skin, a V-shaped, X-shaped o
Y-shaped idge is p esen on cen al back o unk, do sal
su ace bea s sho , i egula and weak idges, wi h sca -
e ed ube cles and g anules, do sola e al idges absen ;
se e al ube cles on lanks and do sal hindlimbs; en al
skin smoo h wi h g anules; (10) do sal su ace yellowish
b own; hollow b own iangula ma king be ween eyes;
b own V-shaped, X-shaped, o Y-shaped ma king on
cen al back o unk; do sal limbs and digi s wi h da k
b own ans e se bands; and (11) dense nup ial spines on
do sal bases o inge s I and II in b eeding adul males,
subgula ocal sac p esen in males.
Desc ip ion o holo ype. JXNU 21042833, adul
male, body size small, SVL 32.2 mm; head wid h sligh -
ly smalle han head leng h, HDW/HDL 0.91; snou
ounded in do sal iew, p ojec ing, sloping backwa d o
mou h in p o ile, p o uding well beyond ma gin o lowe
jaw; op o head la ; eyes la ge, ED 0.35 o HDL, pupil
e ical, o oid in na u al s a e; nos il obliquely o oid;
can hus os alis well de eloped; lo eal egion sligh ly
oblique; in e nasal dis ance la ge han in e o bi al dis-
ance; ympanic egion oblique, ympanum mode a e in
size, ma gin clea , uppe ma gin in con ac wi h sup a-
ympanic old, TD/ED 0.41; la ge o oid choanae a base
o maxilla; ome ine idge weak, ome ine ee h p esen ,
maxilla y ee h p esen ; ma gin o ongue ounded, no
no ched dis ally; p esence o subgula ocal sac.
Figu e 5. Tadpole specimen o Boulenoph ys nebulosa sp. no . (JXNU 21110702) in p ese a i e: A. La e al iew; B. Ven al iew;
C. Do sal iew. Pho os by Deming Shen.
Zoosys . E ol. 101 (4) 2025, 2251–2266
zse.penso .ne
2259
Fo ea m leng h 0.24 o SVL, hand 0.22 o SVL; web-
bing absen be ween inge s, la e al inges absen , ela-
i e inge leng h I < II < IV < III; ips o inge s sligh ly
dila ed, ound; indis inc suba icula ube cles p esen ; in-
ne me aca pal ube cle obse ably enla ged, ou e one in-
dis inc ; single nup ial pad bea ing nup ial spines p esen
on do sal su ace o i s and second inge s, espec i ely.
Hindlimbs sho , heels mee ing when lexed hindlimbs
a e held a igh angles o he body axis; ibio a sal a ic-
ula ion eaching he cen e o he eye when he hindlimb
is s e ched o wa d along he body; ibio ibula leng h
0.45 o SVL, oo leng h 0.39 o SVL; ela i e oe leng h
I < II < V < III < IV; ips o oes ounded and sligh ly
dila ed; oes wi h na ow la e al inges and udimen a y
webbing; suba icula ube cles indis inc a he base o
each oe; inne me a a sal ube cle long and o oid; ou e
me a a sal ube cle absen .
Do sal skin smoo h wi h g anules; a discon inuous
Y-shaped idge p esen on he cen al do sum o unk,
discon inuous longi udinal idges p esen la e ally; se e al
ounded ube cles p esen on he lanks and do sal su ac-
es o he hindlimbs; a p ominen , small ho n-like ube cle
loca ed on he uppe eyelid; sup a ympanic old dis inc ,
ex ending om he pos e io co ne o he eye o abo e
he inse ion o he o elimb; ympanum dis inc ; en al
skin smoo h o weakly g anula , wi h spinose ube cles
su ounding he cloacal egion; pec o al gland small and
dis inc , loca ed close o he axilla; a single la ge emo al
gland p esen on he pos e io su ace o he high, si ua ed
app oxima ely midway be ween he knee and he cloaca.
Colo a ion o holo ype. In li e, do sum yellowish
b own, wi h a hollow b own iangula in e o bi al
ma king be ween eyes and a dis inc Y-shaped b own
ma king a he cen e o he do sum, su ounded by in-
dis inc b own ne like pa e n; do sal su aces o limbs
yellowish b own, each bea ing se e al b own ans-
e se ba s; axilla y egion and adjacen la e al lank
ligh yellowish b own; ube cles on he uppe eyelids
o ange; a e ical b own s ipe p esen below he eye,
ex ending om he lowe ma gin o he eye o he
uppe lip; en al su ace o h oa and ches g eyish
b own, bea ing i egula da k b own and whi e blo ch-
es; en al su aces o limbs, manus and pes pu ple,
wi h whi e blo ches; abdominal su ace g eyish whi e,
sca e ed wi h c eamy whi e and o ange spo s; inne
and ou e me a a sal ube cles and inne me aca pal u-
be cle pale pu ple; pec o al whi e, digi s and emo al
glands g eyish whi e; i is ligh yellowish b own.
In p ese a i e, he do sum ades om yellowish
b own o g eyish, while b own ma kings, including a
Y-shaped in e scapula ma king, a iangula in e o bi -
al ma king and ans e se bands on he do sal su aces
o he o elimbs and hindlimbs, da ken o g eyish b own
and become mo e conspicuous. The en al su aces ade
o g eyish whi e, making he o iginal whi e spo s and
blo ches mo e p ominen .
Va ia ion. Mo phome ic da a a e gi en in Ta-
ble 2. All pa a ypes con o m o he holo ype in key
diagnos ic cha ac e s bu di e in do sal pa e ning,
anging om specimens wi h an indis inc b own ne -
like pa e n as in he holo ype o hose bea ing only an
X-shaped in e scapula ma king and a iangula in e -
o bi al ma king (e.g., JXNU 21042832). Female pa a-
ypes lack nup ial pads and ke a inized spines, which
a e p esen in males.
Tadpole desc ip ion. The adpole was con i med as
Boulenoph ys nebulosa sp. no . by molecula phyloge-
ne ic analyses. The ollowing adpole desc ip ion is based
on he specimen JXNU 21110702 a Gosne S age 29
(Fig. 5). Body dep essed and longi udinally o al; TOL
28.0 mm; body wid h longe han body heigh , BW/BH
1.3; eyes do sola e al, pupils ound; nos il nea eyes;
spi acle on he le side o he body and dis inc ; ail ap-
p oxima ely 2.3 imes as long as snou - en leng h; ail
heigh g ea e han body heigh , TAH/BH 1.2; do sal in
a ising jus behind ail base, highes nea mid-leng h, a-
pe ing g adually o blun ly poin ed ip; o al disk e minal;
lips expanded, di ec ed upwa d, o ming umbelli o m
s uc u e; la e al co ne s o o al disc old medially, u n-
ing upwa d when no ully ex ended.
In p ese a i e, he backg ound colo o he head and
unk ades om b own o da k b own, in e spe sed wi h
da k-b own ch oma opho es, and becomes p og essi ely
pale owa d he ail ip. A eas lacking whi e pigmen a ion
con ain ewe melanocy es. The belly bea s dense whi e
spo s, especially concen a ed on he ches , while he la -
e al su aces o he ail a e co e ed wi h yellowish-b own
spo s. BH 2.7; BW 3.6; IOD 2.5; MW 1.6; SL 1.8; SS 3.2;
SVL 8.5; TAH 3.3; TAL 19.5; TBW 1.4; TOL 28.0.
Ad e isemen call. Call desc ip ions a e based on
he ocaliza ions o he male holo ype JXNU 21042833.
Spec og ams and wa e o ms a e shown in Fig. 6. A o al
o 13 call g oups we e analyzed. Each call had an a -
e age du a ion o 120.3 ± 7.4 ms (110.3–135.6 ms) and
an a e age in e -call in e al o 487.6 ± 48.8 ms (415.9–
573.4 ms). Calls we e epea ed a a mean a e o 2.05
calls/s (cons an epe i ion a e). Each call consis ed o
31.7 ± 2.1 pulses (28–35 pulses). The dominan equen-
cy a e aged 3.70 ± 0.08 kHz (3.59–3.84 kHz).
Compa isons. Mo phological compa isons be ween
he p oposed new species and o he ecognized species
o he genus Boulenoph ys a e p o ided (Table 3). Bou-
lenoph ys nebulosa sp. no . is phylogene ically mos
closely ela ed o B. mu umon ana, o ming a well-sup-
po ed sis e clade. Howe e , he new species can be
eadily dis inguished om B. mu umon ana by he
ollowing combina ion o cha ac e s: 1) ela i e in-
ge leng hs I < II < IV < III ( s. II = IV < I < III); 2)
indis inc suba icula ube cle on base o each inge
( s. dis inc suba icula ube cle); 3) la ge male body
size, SVL 31.8–32.3 mm ( s. 30.1–30.8 mm); 4) heels
jus mee ing when hindlimbs olded ( s. heels o e lap-
ping); 5) ibio a sal a icula ion eaching he cen e o
eye when leg s e ched o wa d ( s. eaching egion be-
ween pos e io ma gin o ympanum and cen e o eye);
6) ome ine ee h p esen ( s. ome ine ee h absen ).
zse.penso .ne
Shen, D. e al.: A new species o Boulenoph ys (Anu a, Megoph yidae) om no he n Jiangxi, China
2266
Supplemen a y ma e ial 1
Addi ional in o ma ion
Au ho s: Deming Shen, Haiying Zhou, Chaoyang Wang,
Jun Xu, Shi Xu, Yi eng Zhong, Zhenyu Wang, Hao
Cai, Yahui Wu, Meicai Wei, Yankuo Li
Da a ype: xlsx
Explana ion no e: able S1. Sample lis o genus Bouleno-
ph ys and ou g oups o phylogene ic analysis in his
s udy. able S2. Unco ec ed p-dis ance (%) among
Boulenoph ys species om he COI. able S3. Un-
co ec ed p-dis ance (%) among Boulenoph ys spe-
cies om he 16S.
Copy igh no ice: This da ase is made a ailable unde
he Open Da abase License (h p://openda acommons.
o g/licenses/odbl/1.0/). The Open Da abase License
(ODbL) is a license ag eemen in ended o allow us-
e s o eely sha e, modi y, and use his Da ase while
main aining his same eedom o o he s, p o ided
ha he o iginal sou ce and au ho (s) a e c edi ed.
Link: h ps://doi.o g/10.3897/zse.101.166556.suppl1
