1
Eu opean Jou nal o Taxonomy 1011: 1–79
h ps://doi.o g/10.5852/ej .2025.1011.3023
eu opeanjou nalo axonomy.eu
ISSN 2118-9773
2025 · Webe D. e al.
This wo k is licensed unde a C ea i e Commons A ibu ion License (CC BY 4.0)
Recei ed: 8 Janua y 2025 • Accep ed: 11 June 2025 • Published: 25 Augus 2025
Topic edi o : Magalie Cas elin • Sec ion edi o : Fabio S och • Desk edi o : K is iaan Hoedemake s
Monog aph
u n:lsid:zoobank.o g:pub:09569CAD-967D-482F-A675-B4BCB7723F6F
Wa e di ine s desc ibed: six new species o he Nipha gus aquilex
complex (C us acea, Amphipoda)
Die e WEBER 1,*,† , T aian BRAD 2,*,† & Alexande M. WEIGAND 3
1 Senckenbe g Deu sches En omologisches Ins i u ,
Ebe swalde S aße 90, 15374 Münchebe g, Ge many.
1,3 Fonda ion aune- lo e, 25 ue Muns e , 2160 Luxembou g, Luxembou g.
2 Ins i u ul de Speologie “Emil Raco iță”, Depa men o Cluj-Napoca,
S . Clinicilo N . 5–7, 400006 Cluj-Napoca, Romania.
3 Musée na ional d’his oi e na u elle du Luxembou g, 25 ue Muns e ,
2160 Luxembou g, Luxembou g.
* Co esponding au ho s: die e [email p o ec ed], [email p o ec ed]
3 Email: Alexande .W[email p o ec ed]
† These au ho s con ibu ed equally o his wo k.
Abs ac . Wi h he onse o molecula axonomy, i has been ecognized ha g oundwa e c us aceans in
he amily Nipha gidae Bous ield, 1977 (C us acea, Amphipoda) can demons a e ei he highly a iable
o s a ic pheno ypes. Thus, i is ecommended ha new species desc ip ions ollow an in eg a i e
axonomic app oach, conside ing mo pho axonomic and molecula cha ac e is ics o es ablish new
axa. The mo phospecies Nipha gus aquilex Schiød e, 1855 has i s main dis ibu ion a ea in Cen al
Eu ope, spanning sou he n England, he no he n hal o F ance, he Benelux coun ies, Ge many and
he Czech Republic. Gene ic analyses ha e shown ha i comp ises a highly di e se species complex,
including amongs o he s he well- ecognized mo phospecies Nipha gus schellenbe gi S. Ka aman,
1932. In his s udy, we desc ibe six new species based on a ailable adul males and emales –
Nipha gus luxembu gensis Webe , Weigand & B ad sp. no ., N. pala inensis Webe & B ad sp. no .,
N. no mandiensis Webe & B ad sp. no ., N. wasgauensis Webe & B ad sp. no ., N. sa a iensis Webe &
B ad sp. no ., N. lo ha ingiensis Webe & B ad sp. no . – aking a e-e alua ed published gene ic
backbone as a u he e e ence in o accoun . The six newly desc ibed species appea ed pseudoc yp ic,
as mino mo phological di e ences we e no iced. A sho discussion o he mo phological and molecula
delimi a ions is p o ided.
Keywo ds. C yp ic di e si y, Nipha gus, in eg a i e axonomy, g oundwa e auna, species complex.
Webe D., B ad T. & Weigand A.M. 2025. Wa e di ine s desc ibed: six new species o he Nipha gus aquilex
complex (C us acea, Amphipoda). Eu opean Jou nal o Taxonomy 1011: 1–79.
h ps://doi.o g/10.5852/ej .2025.1011.3023
Eu opean Jou nal o Taxonomy 1011: 1–79 (2025)
2
In oduc ion
The amily Nipha gidae Bous ield, 1977 (C us acea, Amphipoda) comp ises mo e han 440 o mally
desc ibed species (Ho on e al. 2023) ound nea ly all o e Eu ope and pa s o Asia, om Spain
(Ka aman 2017) o I an (Esmaeili-Rineh e al. 2017). They a e usually s ygobion s, ha is, adap ed
and mos o en es ic ed o sub e anean li e in g oundwa e and g oundwa e -dependen habi a s (e.g.,
sp ings, in e s i ials). Species show ypical oglomo phic cha ac e is ics by lacking eyes and body
pigmen a ion, bu bea ing elonga ed appendages in mos cases. While he majo i y o he species we e
desc ibed based on hei mo phology alone, i was e ealed ha Nipha gidae ha bo s a ple ho a o c yp ic
o pseudoc yp ic species (Fiše & Zagmajs e 2009; Delić e al. 2017; Eme e al. 2018), which can be
a ibu ed o widely s able o la gely o e lapping mo phological cha ac e is ics. Desc ibing new species
by in eg a ing DNA sequences has become a s anda d o nea ly en yea s in Nipha gus axonomy (B ad
e al. 2015; Delić e al. 2017; Fiše e al. 2017, 2018).
The Nipha gus aquilex species complex (“aquilex” “wa e di ine ” in La in) includes mul iple c yp ic
species, and can se e as a p ime example o wi ness pas and cu en axonomic challenges (McIne ney
e al. 2014; Webe e al. 2023). Wes wood (1853) was he i s o ecognize N. aquilex Schiød e, 1855
as a species new o science, al hough he assigned his ma e ial o he i s eco ding o Nipha gus s ygius
(Schiød e, 1847) in England. Nipha gus aquilex was edesc ibed based on specimens om he neo ype
locali y C owbo ough (Uni ed Kingdom), which we e deposi ed in he B i ish Museum (Ka aman 1980).
Ano he axon o he N. aquilex species complex, Nipha gus schellenbe gi S. Ka aman, 1932, was
i s desc ibed om Gännsb ünnle, in he sou h o Sendelbach (Ba a ia, Ge many). In he same yea ,
N. schellenbe gi was edesc ibed as a subspecies o N. aquilex (Schellenbe g 1933), whe eas om 1972
onwa ds, i was again conside ed a species dis inc om N. aquilex sensu s ic o (S ašk aba 1972).
Nipha gus schellenbe gi was always ega ded as a single axonomic en i y (ei he a species o
subspecies le el), bu e en p io he age o DNA sequencing i was suspec ed ha N. aquilex migh
ac ually ep esen a species complex (Ge ecke e al. 2005). Se e al subspecies o N. aquilex we e
desc ibed: N. a. ejdowskyi (W ześniowski 1890; Schellenbe g 1932), N. a. au i (Schellenbe g 1933),
N. a. molda icus (Dob eanu e al. 1953), N. a. doba i (Ske 1999). While he sys ema ic posi ion o he
i s is s ill unce ain, he o he h ee axa ha e been ele a ed o he species le el (Fiše e al. 2009a). Thei
axonomic posi ion was co obo a ed by molecula analyses pe o med by McIne ney e al. (2014), who
delinea ed se e al gene ic lineages wi hin he N. aquilex species complex. Recen ly, Webe e al. (2023)
p o ided a comp ehensi e upda e on he axonomic si ua ion wi hin he N. aquilex species complex.
By in es iga ing he mi ochond ial cy och ome c oxidase subuni I (COI) and nuclea 28S ibosomal
RNA (28S DNA) genes and by applying a se o di e en molecula delinea ion me hods, he au ho s
we e able o conse a i ely de ine eigh molecula en i ies unde he axonomic umb ella o N. aquilex
(anno a ed as lineages A, B, NDJ, F, G, H, KL, I). Two axa we e iden i ied in he mo phospecies
N. schellenbe gi (R, S) (Webe e al. 2023). G oup B he eby e e ed o N. aquilex sensu s ic o, and
g oup S o N. schellenbe gi sensu s ic o. The nomencla u e o he molecula lineages used he ein ollows
ha o McIne ney e al. (2014) and ex ended by Webe e al. (2023). We ecommend ha all subsequen
s udies epo he anno a ions o axonomically undesc ibed molecula lineages.
In his s udy, and by e e ing o he molecula delimi a ion esul s o Webe e al. (2023) as a axonomic
backbone, we p o ide he e an in eg a i e axonomic amewo k and he o mal desc ip ion o six o
he molecula ly iden i ied species wi hin he N. aquilex species complex, ha is, g oup A, NDJ, F, G, I,
and R sensu Webe e al. (2023). A he same ime, because a much la ge se o DNA sequences was
a ailable (Webe e al. 2022; Webe & Weigand 2023), which was pa ly en iched by sequences ob ained
du ing ieldwo k conduc ed wi hin he scope o his s udy, we ha e e iewed all species delimi a ion
esul s o Webe e al. (2023), bu limi ed ou wo k o a minimum in e p e a ion necessa y o he o mal
desc ip ion o hese six new species.
WEBER D. e al., Six new species o he Nipha gus aquilex complex (Amphipoda)
3
We p o ide a lis o all a ailable gene ic sequences s o ed in GenBank (Saye s e al. 2021) o e e ence
pu poses, e e ing o he N. aquilex and N. schellenbe gi mo phospecies (Supp. ile 1). Fu he mo e,
gene ic sequences o se e al clades o he N. aquilex species complex we e named a e and published
using di e en anno a ion sys ems. We p o ide a compila ion o his his o ic e e encing (Supp. ile 2),
hus allowing eade s o compa e ou da a wi h hose in olde publica ions.
Ma e ial and me hods
Sampling
In addi ion o he published da a in GenBank (Webe e al. 2023; Webe & Weigand 2023), ex ensi e
sampling o Nipha gus specimens was pe o med in many egions o Ge many, Belgium, Luxembou g,
and F ance. Sampling o sp ings was pe o med by sie ing he sedimen and deb is p esen a he poin
whe e wa e eme ges om he subsu ace h ough a ious meshes. He e, specimens o Nipha gus, which
a e washed ou wi h he g oundwa e , hide and li e among sedimen pa icles, dead lea es, o o he ypes
o deb is. When he cu en o he sp ing wa e is no e y s ong, specimens o Nipha gus can mig a e
owa ds he su ace and back, mos p obably o eeding (Ku eck 1967; Husmann 1976). Sampling o
Nipha gus in ca es and a i icial ca i ies was pe o med by simple manual picking o specimens using
weeze s o by placing bai s wi h chicken li e o one day. The specimens we e indi idually sampled in
2 mL ials con aining 96% e hanol. To simpli y e ie ing he ype locali ies in he u u e, pho og aphs
o each o he six ype locali ies o he newly desc ibed species, as well as a pho og aph o he si e close
o he ype locali y whe e we collec ed N. schellenbe gi, we e made (Supp. ile 3).
To es ima e he p e e ed habi a o he indi idual species, we ca ego ized he collec ion si es in o sp ings,
la ge unde g ound ca i ies, and in e s i ial a eas (Supp. ile 4) o all species whe e we had mo e han en
occu ences. Among he sp ings, we dis inguished ou g oups (c . Thienemann 1922): 1. A heoc ene
sp ing had a localized ou le wi h a di ec ly isible discha ge. 2. A heloc ene sp ing has se e al small
ou le s o g oundwa e ha a e o en no di ec ly ecognizable; hey a e collec ed in a sump o o m an
e luen . 3. A limnoc ene sp ings om he bo om o a pond, and he wa e lows o e he wa e im.
4. The e m “basin” is no commonly used in he li e a u e. G oundwa e is o en channeled a i icially
ia a pipe in o an a i icial ough ha has been in es iga ed. La ge unde g ound ca i ies we e di ided
in o ca es, which a e o na u al o igin, mines, in which ma e ial was mined, wells, e ical pipes d i en
in o he ock o wa e ca chmen , and sou e ains, which e e o all o he human-made objec s no
used o mining. In e s i ial is a ine-g ained sys em be ween g a els, whe e we only sampled in e s i ial
habi a s close o a i e (Webe & Weigand 2023).
DNA analysis
DNA was ex ac ed om one pe eopod using he DNeasy Blood & Tissue Ki (Qiagen) o he NucleoSpin
Tissue Ki (Mache ey-Nagel, Dü en, Ge many), acco ding o he manu ac u e ’s ins uc ions. DNA
isola es we e s o ed a -20°C in he collec ions o he E olu iona y Biology and Ecology esea ch uni
o he Uni e si é lib e de B uxelles (Belgium), he Musée na ional d’his oi e na u elle du Luxembou g
(Luxembou g), and he Senckenbe g Ge man En omological Ins i u e (Ge many).
Polyme ase chain eac ion (PCR) was pe o med o ampli y he s anda d ba coding agmen o he
cy och ome c oxidase subuni 1 gene (COI) (Folme e al. 1994) using he p ime pai HCO2198 and
LCO1490 (Folme e al. 1994) o he p ime pai HCO2198-JJ and LCO1490-JJ (As in & S üben 2008)
(10 pmol/μL), p emixed a a a io o 1 : 1. Fo COI, he PCR mix con ained 1.5 μL DNA ex ac (wi h
a ying concen a ions, no measu ed), 1 μL o p ime mix, 5 µL o “D eamTaq DNA Polyme ase”
(The mo Scien i ic, Macqua ie Pa k, Aus alia) and 5 µL o ul apu e wa e . PCR cycling condi ions we e
an ini ial 5 min dena u a ion s ep a 95°C, 38 cycles o 30 s o dena u a ion a 95°C, 90 s o annealing a
49°C, 60 s o ex ension a 72°C and a inal elonga ion s ep o 30 min a 68°C.
Eu opean Jou nal o Taxonomy 1011: 1–79 (2025)
4
A agmen o he nuclea 28S ibosomal RNA gene (28S DNA) was analyzed using he PCR p ime s
Niph15 and Niph16 (Ve o nik e al. 2005). The PCR mix u e o he 28S DNA ma ke con ained 2 μL
o DNA ex ac (wi h a ying concen a ion, no measu ed), 1 μL o each p ime (10 pmol/μL), 0.2 μL o
REDTaq polyme ase (Sigma-Ald ich, S . Louis, MI, USA), 5 µL o REDTaq eac ion bu e and 15.8 µL
o ul apu e wa e . PCR cycling condi ions o 28S DNA we e an ini ial 3-minu e dena u a ion a 95°C,
56 cycles o 30 s o dena u a ion a 94°C, 60 s o annealing a 45°C, and 90 s o ex ension a 72°C.
Bidi ec ional Sange sequencing was pe o med a Genosc een (Lille, F ance), Mac ogen (Ams e dam
and Maas ich , Ams e dam, The Ne he lands), o a he Musée na ional d’his oi e na u elle du
Luxembou g. Fo COI, he same p ime s used o PCR ampli ica ion we e used. Mo eo e , 28S DNA
was sequenced wi h h ee p ime s: Niph15 (Ve o nik e al. 2005), al e na i ely Niph15i (Webe e al.
2023), Niph20, and Niph21 (Flo e al. 2010). The sequences we e deposi ed in he NCBI GenBank
da abase and he accession numbe s a e p esen successi ely in Supp. ile 6 (COI) and Supp. ile 9 (28S
RNA). A lis o p ime s, oge he wi h hei nucleo ide sequence, o bo h PCR and sequencing is
p o ided in Supp. ile 5.
Molecula species delimi a ion and diagnosis
We used en iched molecula sequence da a (COI and 28S DNA) o e-e alua e he es ablished molecula
lineages wi hin he N. aquilex species complex sensu Webe e al. (2023), which se ed as he axonomic
backbone o ou in eg a i e species desc ip ions using molecula and mo phological da a. Molecula
sequence da a we e also analyzed o in e DNA cha ac e is ics ( ha is, indi idual pu e diagnos ic
nucleo ides o species-speci ic DNA sequence a ian s, ha is, DNA ba codes o nuclea alleles),
enabling a suppo ing di e en ial molecula diagnosis o he delinea ed species in he N. aquilex species
complex.
On 26 Augus 2023, all a ailable COI sequences o N. aquilex and N. schellenbe gi we e downloaded
om Genbank, en iched by he da ase om Webe , Weigand & Weigand (unpublished) and ou newly
ob ained sequences he ein. Thus, he ini ial COI da ase hus con ained 1284 en ies. These en ies
co esponded o 1319 indi idual COI sequences collec ed om 894 sampling si es (Supp. ile 6). This
disc epancy in he numbe o COI sequences exis s because McIne ney e al. (2014) s o ed a single COI
sequence in GenBank o specimens om di e en loca ions when hei COI sequences we e iden ical.
All incomple e sequences (i.e., sequences missing nucleo ides a one o bo h ends) we e emo ed om
da ase 1, as well as hose ha con ained any ambiguous cha ac e s (e.g., a single ‘N’)), esul ing in
1115 inal COI sequences (da ase 2). Da ase 1 was used o gene a e dis ibu ion maps and es ablish
a phylogene ic amewo k o he molecula iden i ica ion o specimens missing in da ase 2. The
phylogene ic ee hypo hesis was cons uc ed using MEGA-X (Kuma e al. 2018). Se en sho sequences
we e emo ed om da ase 1 and a neighbo -joining ee wi h 1000 boo s ap eplica ions was gene a ed
(Supp. ile 7 wi h desc ip ion o he me hod in Supp. ile 8). Da ase 2 was used o he dis ance-based
molecula species delimi a ion app oach ASAP (assemble species by au oma ic pa i ioning) (Puilland e
e al. 2021), e e ing o i s online e sion (h ps://bioin o.mnhn. /abi/public/asap/asapweb.h ml) and
s anda d se ings unde he Jukes-Can o (JC69) subs i u ion model o compu ing gene ic dis ances. We
p ima ily used pa i ion 1 o inal species delimi a ion, bu also compa ed hese esul s wi h pa i ions
2 and 3.
The da a o he 28S DNA da ase o igina ed om mul iple sou ces and comp ised 404 sequences
( e e ing o 415 specimens, since McIne ney e al. (2014) deposi ed a single sequence o specimens
wi h iden ical 28S DNA sequences), all o which a e p o ided in Supp. ile 9, which comp ises ull 28S
DNA agmen a ailable, 2 sequences (A in column N); only pa o 28S DNA agmen a ailable,
78 sequences (B in column N); da a om Webe e al. (2023), ull 28S DNA agmen and ull COI
a ailable, 195 sequences (C in column N); new da a wi hin his s udy, ull 28S DNA agmen a ailable,
WEBER D. e al., Six new species o he Nipha gus aquilex complex (Amphipoda)
5
138 sequences (D in column N); and new da a wi hin his s udy, only pa o 28S DNA agmen
a ailable, 2 sequences (E in column N).
The COI da ase 2 was used o in e molecula diagnos ic DNA cha ac e s and sequence
a ian s o he delinea ed species. The’ emo edup’ unc ion o he s na oolbox webse e
(h ps://a n.ug .es/s na oolbox/helpe / emo edup/; Apa icio-Pue a e al. 2022) was used o emo e
iden ical sequences (n = 674) in he COI alignmen , esul ing in 442 dis inc COI sequence a ian s
(= DNA ba codes). The alignmen was manually inspec ed o pu e diagnos ic nucleo ide posi ions
(Jö ge & Sch ödl 2014), i.e., conse ed nucleo ides wi hin a species, bu wi h dis inc cha ac e
s a es in all sequence a ian s o all he o he species. Howe e , in addi ion o elying on (a ew)
indi idual diagnos ic nucleo ide posi ions, species-speci ic se s o diagnos ic DNA ba codes a e used
in he di e en ial diagnosis sec ions o he espec i e species. Addi ionally, he 28S DNA da ase was
in es iga ed o iden i y diagnos ic nuclea sequences (alleles) o he newly desc ibed species. Molecula
diagnos ic cha ac e is ics a e p o ided in he diagnosis sec ion o each species sepa a ely.
Mo phological analysis
In he labo a o y, one male and one emale pa a ype we e selec ed o u he dissec ion, inspec ion, and
desc ip ion o he mo phological cha ac e s. Fo inspec ion o he in aspeci ic mo phological a iabili y,
we inspec ed 2–3 mo e pa a ypes o each species. The holo ype and pa a ype specimens we e no
gene ically analyzed bu o igina ed om a pool o specimens om a single locali y, o which specimens
ha e been gene ically analyzed o hei DNA sequence a ian s (COI and 28S). Special ca e was aken o
ensu e ha he gene ically analyzed specimens, he specimens selec ed o dissec ion, and he specimens
deposi ed as in ac holo ypes and pa a ypes did no exhibi any mo phological disc epancies. The
specimens selec ed o mo phological analysis we e imme sed in glyce ol, dissec ed body appendages
we e moun ed on slides in glyce ol, and he slide co e glasses we e sealed wi h a nish. The slides
we e inspec ed wi h an Olympus SZX16 s e eo mic oscope equipped wi h an Olympus SC180 came a,
and wi h an Olympus BX51 mic oscope equipped wi h an Olympus SC50 came a. The d awings we e
pe o med on p in ed pho og aphs ia manual inking and con inuous inspec ion o he slides unde
he mic oscope. The e minology used o body pa s and he choice o appendages conside ed o
measu emen s a e hose p e iously desc ibed (Fiše e al. 2009b). The mandibula palp chae o axy
nomencla u e ollowed Ka aman (1993). A able con aining all measu ed appendages o all species
desc ibed he ein is p esen ed in Supp. ile 10.
Reposi o y ma e ial
The specimens ( ypes and pa a ypes), o mic oscope slides con aining specimens appendages, used
o species desc ip ions in his s udy we e deposi ed in he Musée na ional d’his oi e na u elle du
Luxembou g (MNHNL) and in he Ins i u ul de Speologie “Emil Raco iță” (ISER) in Cluj-Napoca,
Romania. In he case o each desc ibed species, all specimens ecei ed an in en o y numbe and a e
p esen ed and explained in de ail in he designa ed chap e s in he esul s sec ion below.
Resul s
Molecula species delimi a ion
ASAP analysis o COI, pa i ion 1 wi h ecu sion and an ASAP-Sco e o 4.0, iden i ied 16 species;
pa i ion 2 wi h ecu sion and an ASAP-Sco e o 5.0, 21 species; and pa i ion 3 wi h ecu sion and an
ASAP-Sco e o 5.5, 23 species (Supp. ile 6). ASAP analysis o 28S DNA om a limi ed da ase esul ed
in he iden i ica ion o 10 pu a i e species (Webe e al. 2023). Species A, B, and I a e undispu ed in all
me hods.
Eu opean Jou nal o Taxonomy 1011: 1–79 (2025)
6
The si ua ion a ound species D + N is mo e complex. Eigh specimens o he molecula en i y D+N had
iden ical 28S DNA alleles, and he e o e belonged o he same species. Acco ding o all models, i is
undispu ed ha i is a sepa a e species om all o he species, bu pa i ions 2 and 3 o he ASAP o COI
de ine h ee species wi hin he D+N complex, iden ical in pa i ions 2 and 3, and he 28S DNA o all
specimens unde conside a ion a e iden ical.
Pa i ion 1 o ASAP o COI o he species F + F’ assumes a single species. Only he less likely pa i ions
2 and 3 assume wo sepa a e species. 28S DNA con ains wo he e ozygous indi iduals ha include bo h
F and F’.
Acco ding o all models, i is undispu ed ha species G is a sepa a e species om all o he species, bu
pa i ions 2 and 3 o he ASAP o COI de ine a single specimen as an addi ional sepa a e species.
Species R and S we e iden i ied as a single species by pa i ion 1, as wo sepa a e species by pa i ion 2,
and as h ee di e en species by pa i ion 3. The la ge numbe o 28S DNA sequences, 29 o species
R and 681 o species S, did no show any he e ozygous indi iduals be ween he wo pu a i e species.
In wo cases, F+F’ o N. luxembu gensis sp. no . and D+N o N. no mandiensis sp. no ., molecula
en i ies comp ised wo well-sepa a ed molecula clades each. Fo hese wo species, hei espec i e
molecula clades a e anno a ed wi h di e en symbols bu in he same species-speci ic colo when
p esen ing hei dis ibu ion maps.
Species desc ip ions
In his chap e , six species o he N. aquilex species complex a e o mally desc ibed, and in o ma ion on
N. aquilex sensu s ic o and N. schellenbe gi sensu s ic o is p o ided. Dis ibu ion maps we e gene a ed
in QGIS e . 3.34.8-P iz en (QGIS De elopmen Team 2025) and Inkscape e . 0.92 (2017), wi h colo s
co esponding o Webe e al. (2023) and ollow h ps://www.w3schools.com/colo s/colo s_names.asp
(Supp. ile 2). We we e able o de ine he ype o habi a o 855 sampling si es (Supp. ile 4), allowing
o a ough inspec ion o he species’ p e e ed ecology.
Class Malacos aca La eille, 1802
O de Amphipoda La eille, 1816
Family Nipha gidae Bous ield, 1977
Genus Nipha gus Schiød e, 1849
Nipha gus aquilex Schiød e, 1855
Fig. 1
Rema ks
Webe e al. (2023) ea ed his species as N. aquilex B, which can be conside ed N. aquilex sensu s ic o.
The species was egula ly ound in nea ly all ypes o bio opes, bu su p isingly o en in heloc ene
sp ings (Supp. ile 4.1). I is widesp ead and common in Wes e n Ge many, wi h mo e sca e ed eco ds
in Luxembou g, Belgium, and along he Channel Coas in F ance and G ea B i ain (Fig. 1). A neo ype
locali y was es ablished by Ka aman (1980) om C owbo ough (Uni ed Kingdom) wi hou men ioning
p ecise coo dina es. We sampled he egion o C owbo ough, and ou specimens and sequences can
he e o e be conside ed o o igina e om he neo ype locali y.
The d awing o Schiød e (1855) shows wo o he cha ac e is ics o N. aquilex, ha is, he single se a
on he dac ylus o he gna hopods and he ounded 3
d
epime al pla e. Ka aman (1980) e-desc ibed
N. aquilex in de ail, bu he did no ake in o conside a ion ha N. aquilex migh comp ise a species
WEBER D. e al., Six new species o he Nipha gus aquilex complex (Amphipoda)
7
complex o mo phological simila (pseudoc yp ic) o iden ical (c yp ic) axa. Ka aman’s specimens a e
now > 40 yea s old, and we hus did no y o sequence hem. Ne e heless, we ob ained h ee eshly
collec ed specimens om C owbo ough and sequenced wo COI and h ee 28S DNA sequences. All he
sequences co esponded o he o me species B senus Webe e al. (2023), which was al eady delinea ed
by McIne ney e al. (2014).
Nipha gus schellenbe gi Ka aman, 1932
Fig. 2
Rema ks
Webe e al. (2023) ea ed his species as N. schellenbe gi, which can be conside ed N. schellenbe gi
sensu s ic o, gi en ha we also included specimens e y close o he ype locali y in ou analyses.
The species is widesp ead om Cen al o Wes e n Ge many, Luxembou g, Belgium (Wallonia), he
Ne he lands (Limbu g), and Eas e n F ance (Fig. 2). While in he i s desc ip ion i was assumed ha
N. schellenbe gi mainly inhabi s sp ings (Ka aman 1932), we ound i egula ly in ca es, a i icial
ca e ns, meadow d ainages, sp ings, wells, and in he in e s i ial (Supp. ile 4.2). Ka aman (1932) se
Fig. 1. Dis ibu ion o Nipha gus aquilex Schiød e, 1855 sensu s ic o, con i med by molecula da a.
The neo ype locali y in C owbo ough (Uni ed Kingdom) is ma ked by a yellow s a , o e laying one si e
eco d. Ve y la ge eshwa e bodies a e indica ed; coun y codes ollow he ISO 3166 s anda d alpha-2
o ma .
Eu opean Jou nal o Taxonomy 1011: 1–79 (2025)
8
he Gänsb ünnle in he Sou h o Sendelbach (Ba a ia, Ge many) as ype locali y, e e ing o a well along
he sou he n ma gin o i s in e ed dis ibu ion ange in Cen al Ge many, bu he did no de ine any ype
ma e ial. We collec ed specimens in a sp ing e y close o Gänsb ünnle (Supp. ile 3.1). Mo e sou he n
dis ibu ion eco ds, pa icula ly in Cen al-Eas o Sou h-Eas e n F ance, mos likely belong o he newly
desc ibed sis e species Nipha gus lo ha ingiensis sp. no .
The d awing in Ka aman (1932) shows well he cha ac e is ics o N. schellenbe gi, i.e., he gna hopod II
nea ly in squa e- o m wi h i s 3 se ae on i s dac ylus. The wo d awings o he elson exhibi signi ican
disc epancies and a e o limi ed alue o axonomic de e mina ion. The same is ue o he wo d awings
o u opod I, in which he ou e amus is longe in one ins ance and he inne amus is longe in he o he .
A d awing o he en i e specimen is missing.
Fig. 2. Dis ibu ion o Nipha gus schellenbe gi S. Ka aman, 1932 sensu s ic o, con i med by molecula
da a. The ype locali y a Gänsb ünnle (Ge many) is ma ked by a yellow s a , o e laying wo si e eco ds.
Ve y la ge eshwa e bodies a e indica ed; coun y codes ollow he ISO 3166 s anda d alpha-2 o ma .
WEBER D. e al., Six new species o he Nipha gus aquilex complex (Amphipoda)
9
Nipha gus luxembu gensis Webe , Weigand & B ad sp. no .
u n:lsid:zoobank.o g:ac :1522BA40-BBC2-44B7-9E92-D0AE85085217
Figs 3–11
Diagnosis
Medium-sized Nipha gus species, poo ly se ose. Righ pos e o en al angle o epime al pla es. Ou e
6 spines o maxilla I on ou e lobe wi h 1 oo h each; 1 spine wi h se e al smalle ee h. Mandibula
palp wi h small numbe (1–2) o B se ae. Gna hopods wi h 1 se a along ou e ma gin o dac ylopodi es.
Pe eiopod VII, he longes leg, almos hal o o al body leng h. Pleopods e inaculum wi h 2 hooks.
U opod I, longe exopodi e. U opod II, longe endopodi e. U opod III sexually dimo phic, exopodi e
elonga ed in males. Telson wi h 3 apical spines on each lobe. The COI ma ke shows wo pu e diagnos ic
si es a posi ions 55 (C) and 358 (C). Six 28S DNA alleles a e diagnos ic (wi h 13 he e ozygous
specimens) as well as 79 COI ba codes.
E ymology
The species name de i es om he G and Duchy o Luxembou g, whe e i was i s disco e ed. In Webe
e al. (2023), his species was ea ed as N. aquilex F, F’.
Ma e ial examined
Holo ype
LUXEMBOURG • ♂; Gi s , Gu land, basin o a sp ing in he cen e o he hamle o Gi s e klaus;
49.7847° N, 6.4989° E; 19 Ma . 2018; Die e Webe leg.; kep in ac in 96% e hanol; 180319-10;
MNHNL130574.
Pa a ypes
LUXEMBOURG • 1 ♂; same collec ion da a as o holo ype; 30 Jan. 2018; dissec ed and appendages
d awn; 180130-32; ISER mic oscope slide DW180130-32 • 1 ♀; same collec ion da a as o p eceding;
dissec ed and appendages d awn; 180130-45; ISER mic oscope slide DW180130-45 • 1 ♂; same
collec ion da a as o holo ype; 27 Ma . 2017; ISER DW170327-02 • 1 ♀; same collec ion da a as o
holo ype; 19 Ma . 2018; ISER DW180319-09 • 1 ♀; same collec ion da a as o holo ype; 19 Ma . 2018;
ISER DW180319-13 • 1 ♀; same collec ion da a as o holo ype; 27 Ma . 2017; ISER DW 170327-06;
MNHNL130575.
Molecula da a
COI and 28S DNA sequences o specimens belonging o Nipha gus luxembu gensis sp. no . we e
deposi ed in GenBank. COI and 28S RNA accession numbe s a e p esen in Supp. ile 6 and Supp. ile 9,
espec i ely.
Desc ip ion (male pa a ype ISER DW180130-32)
Measu emen s
To al body leng h 7.7 mm (Fig. 3).
Head
Head (Fig. 3) 8.60% o o al body leng h. Eyes and os um absen .
An ennae
An enna I (Fig. 4A): wi h he main lagellum o med o 18 a icles ep esen ing 41% o o al body leng h.
Peduncle leng h ⅓ o o al leng h o an enna I. The accesso y lagellum (Fig. 4B) bia icula ed; p oximal
a icle sligh ly longe han i s a icle o main lagellum; dis al a icle ¼ o o al leng h o accesso y
lagellum, wi h 2 apical se ae o di e en leng hs and 1 aes he asc. Aes he ascs ⅓ o espec i e lagellum
a icles (Fig. 4C).
Eu opean Jou nal o Taxonomy 1011: 1–79 (2025)
16
U opods
U opod I (Fig. 10A): wi h wo do so-la e al ows o 4 spines on peduncle. Exopodi e sligh ly longe han
endopodi e, and a io o exopodi e leng h : endopodi e leng h 1.0 : 0.85. 1 s ong spine a base o u opod I.
U opod II (Fig. 10B): wi h 1 do sal spine and 3 apical spines. Endopodi e longe han exopodi e,
endopodi e leng h : exopodi e leng h a io 1.0 : 0.81, bo h ami wi h low numbe o spines.
U opod III (Fig. 10C): sexually di e en ia ed, longe in males. Peduncle wi h 4 apical spini o m se ae.
Sho endopodi e abou hal he leng h o peduncle, wi h 2 apical se ae. P oximal segmen o exopodi e
longe han dis al segmen ( a io 1.0 : 0.77). An e io ma gin o p oximal segmen o exopodi e wi h 5
Fig. 8. Nipha gus luxembu gensis Webe , Weigand & B ad sp. no ., pa a ype, ♂ (ISER DW180130-32).
A. Pe eopod V. B. Pe eopod VI. C. Pe eopod VII.
WEBER D. e al., Six new species o he Nipha gus aquilex complex (Amphipoda)
17
g oups o 2–3 se ae; pos e io ma gin o exopodi e wi h 3 g oups o 1–2 se ae; one g oup o 3 pos e o-
apical se ae. Dis al segmen o exopodi e wi h 3 g oups o 2 se ae on an e io ma gin, 3 g oups o 1–2
se ae on pos e io ma gin and 4 apical se ae o di e en leng hs.
Telson
Telson (Fig. 10E) nea ly as wide as long (wid h : leng h a io 0.95), wi h 1 subapical and 3 apical spines
o di e en leng hs on each lobe. Longes spine almos hal o elson leng h. 2 hin se ae o di e en
leng hs and plumose ip, 1 se a medially on 1 lobe.
Sexual dimo phism
The examined male and emale a e highly simila . Besides he usual sexual dimo phism (e.g., sligh ly
smalle gna hopods, p esence o oos egi es, and sligh ly deepe coxal pla es I–VI in emales), in
N. luxembu gensis sp. no ., emale u opod III (Fig. 10D) is sho e compa ed o ha o male. Female
elson simila o ha o he male, bu smalle (Fig. 10F).
In aspeci ic and in e speci ic a iabili y
The in aspeci ic mo phological a iabili y o N. luxembu gensis sp. no . appea s o be e y low. All
mo phological cha ac e s we e nea ly iden ical in all inspec ed specimens (da a no shown).
Type locali y, ecology and dis ibu ion
The ype locali y is he basin o a sp ing in he cen e o he hamle o Gi s e klaus (communi y o Gi s ,
Gu land, Luxembou g) a 49.7847° N, 6.4989° E (Supp. ile 3.2 & 3.3). The a i icial basin is made
o sands one. The wa e con ains algae; he bo om o he basin is co e ed wi h mud. Specimens o
Nipha gus luxembu gensis sp. no . we e ound in he mud as well as among he algae. The wa e has a
ha dness o 15°GH (whe e one deg ee equals o 17.9 mg/l CaCO
3
) and a sligh ly acidic pH o 6.7. The e
is no human access, nei he o he ac ual sp ing, no o he g oundwa e . We con i m he pe manen
Fig. 9. Nipha gus luxembu gensis Webe , Weigand & B ad sp. no ., pa a ype, ♂ (ISER DW180130-32).
A. Pleopod I, de ail o he e inaculum. B. De ail o he epime al pla es.
Eu opean Jou nal o Taxonomy 1011: 1–79 (2025)
18
p esence o his species in he ype locali y as i was ound on 10 July 2016, 4 Sep embe 2016,
30 Janua y 2018, 17 Feb ua y 2018 and 19 May 2018. In he same basin o he sp ing, N. schellenbe gi
sensu s ic o was also ound. We success ully collec ed 82 specimens o N. schellenbe gi in 15 sp ings
and 1 mine, a a dis ance o maximum 10 km a ound he ype locali y, in Luxembou g and in Ge many,
bu could no ind N. luxembu gensis he e. The ype locali y was ee o gamma ids (Amphipoda,
Gamma idae) du ing he pe iods o collec ion. Nipha gus luxembu gensis seems o be a ypical species
o he in e s i ial en i onmen (Supp. ile 4.3). I is widely dis ibu ed om he UK and No h o Cen al
F ance, Belgium, Ge many o he Czech Republic in he Eas (Fig. 11). The equen p esence in
Luxembou g and bo de ing Ge many (Rhineland-Pala ina e, Saa land) mus be mos likely a ibu ed o
a mo e in ense sampling in hose egions.
Fig. 10. Nipha gus luxembu gensis Webe , Weigand & B ad sp. no ., pa a ype, ♂ (ISER DW180130-
32) and pa a ype, ♀ (ISER DW180130-45). A. Male u opod I. B. Male u opod II. C. Male u opod III.
D. Female u opod III. E. Male elson. F. Female elson.
WEBER D. e al., Six new species o he Nipha gus aquilex complex (Amphipoda)
19
Nipha gus luxembu gensis sp. no . is he only species desc ibed he ein wi h wo s ongly di ided
subclades (F and F’). Al hough he gene ic dis ance is ela i ely la ge, he connec ion in he haplo ype
ne wo k iden i ies bo h as belonging o he same species. The wo clades canno be dis inguished
mo phologically. Ne e heless, hey a e geog aphically sepa a ed wi h only a small o e lap: Clade F’,
including he ype locali y, has i s dis ibu ion in he No h-Eas e n hal o F ance, Belgium, Luxembou g
and he e y Wes o Ge many (Fig. 11). Clade F is dis ibu ed in sou he n England, he e y No h
o F ance, Belgium, Luxembu g, all o e Ge many wi hou he No h and he Sou h, up o he Czech
Republic (Fig. 11). The geog aphic o e lap o bo h clades includes Belgium, Luxembou g, and he e y
Wes o Ge many. The highe numbe o si es o clade F migh be a sampling a e ac , as ou da ase
comp ised many mo e si es in Ge many han in F ance. I can be assumed ha he wo subclades o
N. luxembu gensis a e in he p ocess o ecological specia ion.
Nipha gus luxembu gensis sp. no . has a mode a e geog aphical o e lap wi h Nipha gus ejdo skyi
W ześniowski, 1890 in he e y Eas o i s dis ibu ion ange, wi h he la e ha ing been desc ibed
as N. pu eanus a . ejdo skyi om wells in P ague (W ześniowski 1890), he illage o Bechlín
close o Roudnice and om he wells be ween K alupy and Vodolka in Bohemia (Czech Republic).
W ześniowski’s d awings a e di icul o unde s and bu show some simila i ies o he N. aquilex species
Fig. 11. Dis ibu ion o Nipha gus luxembu gensis Webe , Weigand & B ad sp. no . con i med by
molecula da a. The ype locali y a Gi s e klaus (Luxembou g) is ma ked by a yellow s a , o e laying
one si e eco d. The wo molecula clades F and F’ wi hin N. luxembu gensis a e e e ed o by ci cles
and iangles, espec i ely. Ve y la ge eshwa e bodies a e indica ed; coun y codes ollow he ISO
3166 s anda d alpha-2 o ma .
Eu opean Jou nal o Taxonomy 1011: 1–79 (2025)
20
g oup. This axon was e-desc ibed as N. aquilex ejdo skyi and synonymised wi h N. aquilex a .
bohemica O mann (Schellenbe g 1932), based on ma e ial om Schneebe g in he O e Moun ains
(Saxony, Ge many), om B une sdo (Saxony, Ge many). Schellenbe g assumed ha h ee emales om
Heligoland (Schleswig-Hols ein, Ge many) also belong o his species. The inne lobe o he maxilla I o
he specimens om Heligoland has 3 o 6 apical se ae (Schellenbe g 1932), while N. luxembu gensis (as
well as all o he newly desc ibed species he ein) has 1 apical se a only. Nipha gus ejdo skyi is he e o e
no iden ical o N. luxembu gensis, bu DNA sequences o he o me a e no ye a ailable.
Nipha gus pala inensis Webe & B ad sp. no .
u n:lsid:zoobank.o g:ac :5DA10A74-7763-41C9-B0ED-76AD156C03D4
Figs 12–20
Diagnosis
Medium-sized Nipha gus species, poo ly se ose. Righ pos e o- en al angle o epime al pla es. Six
spines o maxilla I ou e lobe wi h 1–3 ee h each; 1 spine wi h se e al smalle ee h. Mandibula palp
wi h small numbe (1–2) o B se ae. Gna hopods wi h 1 se a along ou e ma gin o dac ylopodi es.
Pe eiopod VII, he longes leg, almos hal o o al body leng h. Pleopods e inaculum wi h 3 hooks.
U opod I, longe exopodi e. U opod II, longe endopodi e. U opod III sexually dimo phic, exopodi e
elonga ed in males. Telson wi h 4 apical spines on each lobe. The COI ma ke shows six pu e diagnos ic
si es a posi ions 31 (C), 184 (C), 223 (G), 353 (G), 358 (G) and 391 (A). A single 28S DNA allele is
diagnos ic as well as 21 COI ba codes.
E ymology
The species name de i es om he Ge man Fede al S a e o Rhineland-Pala ina e, whe e he ype locali y
is loca ed. In Webe e al. (2023), his species was ea ed as N. aquilex A.
Ma e ial examined
Holo ype
GERMANY • ♂; Rhineland-Pala ina e, Waldleiningen, Felsenb unnen in he Pala ina e Fo es in he
Middle Bun sands ein; heoc ene sp ing; 49.2881° N, 7.8467° E; 14 Dec. 2016; Die e Webe leg.; kep
in ac in 96% e hanol; 161214-02; MNHNL130574.
Pa a ypes
GERMANY • 1 ♂; same collec ion da a as o holo ype; 14 Dec. 2016; dissec ed and appendages d awn;
161214-01; ISER mic oscope slide DW161214-01 • 1 ♀; same collec ion da a as o holo ype; 8 Aug.
2017; dissec ed and appendages d awn; 170808-13; ISER mic oscope slide DW170808-13.
Molecula da a
COI and 28S DNA sequences o specimens belonging o Nipha gus pala inensis sp. no . we e deposi ed
in GenBank. COI and 28S RNA accession numbe s a e p esen in Supp. ile 6 and Supp. ile 9,
espec i ely.
Desc ip ion (male pa a ype ISER DW161214-01)
Measu emen s
To al body leng h 6.92 mm (Fig. 12).
Head
Head (Fig. 12) 8% o o al body leng h. Eyes and os um absen .
WEBER D. e al., Six new species o he Nipha gus aquilex complex (Amphipoda)
21
An ennae
An enna I (Fig. 13A): wi h main lagellum o med o 20 a icles, ep esen ing 50% o o al body leng h.
Peduncle leng h 35% o o al leng h o an enna I. The accesso y lagellum (Fig. 13B). Bia icula ed;
p oximal a icle sho e han i s a icle o main lagellum; dis al a icle 27% o o al leng h o accesso y
lagellum, wi h 4 apical se ae o di e en leng hs and 1 aes he asc. Aes he ascs ¾ o espec i e main
lagellum a icles (Fig. 13C).
An enna II (Fig. 13D): lagellum o med o 10 a icles and ep esen ing 50% o o al leng h o an enna II.
Mos lagellum a icles bea 1 aes he asc, longe han hal o espec i e lagellum a icles.
Mou hpa s
Lab um (Fig. 14A): ypical, subo oid shape. Labium (Fig. 14B). La ge inne lobes wi h 1 ow o ine
se ae on inne sides. Ou e lobes wi h 2 ows o ine se ae subapically on inne sides.
Maxilla I (Fig. 14C): wi h 3 apical se ae on dis al a icle o palp. 6 spines o ou e lobe wi h 1–3 ee h
each and 1 spine wi h se e al small ee h. Inne lobe wi h 1 apical se a.
Maxilla II (Fig. 14D): wi h inne lobe sligh ly sho e han ou e lobe. 1 apical ow o se ae and 4 se ae
on inne ma gin o each lobe.
Le mandible (Fig. 14E): 6 ee h on inciso p ocess. 5 ee h on lacinia mobilis. 8 se a ed se ae
in e cala ed wi h 4 i id se ae, be ween lacinia mobilis and mola p ocess.
Righ mandible (Fig. 14F): 5 ee h on inciso p ocess. Lacinia mobilis wi h se e al smalle ee h.
7 se a ed and 2 i id se ae be ween lacinia mobilis and mola p ocess. A long se a on mola p ocess.
Fig. 12. Nipha gus pala inensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW161214-01); gene al
appea ance.
Eu opean Jou nal o Taxonomy 1011: 1–79 (2025)
22
Fig. 13. Nipha gus pala inensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW161214-01). A. An enna I.
B. An enna I, de ails o accesso y lagellum. C. An enna I, de ails o aes he ascs. D. An enna II.
WEBER D. e al., Six new species o he Nipha gus aquilex complex (Amphipoda)
23
Fig. 14. Nipha gus pala inensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW161214-01). A. Lab um.
B. Labium. C. Maxilla I. D. Maxilla II. E. Le mandible. F. Righ mandible.
Eu opean Jou nal o Taxonomy 1011: 1–79 (2025)
24
Mandibula palps (Fig. 14E–F): highly simila and o same leng h. 3 a icles accoun o 17% (a icle 1),
39% (a icle 2) and 44% (a icle 3) o o al leng h o palp. P oximal a icle wi hou se ae, median a icle
wi h 8 en al se ae. Dis al a icle o palp wi h one g oup o 3 A se ae, h ee g oups wi h 1 B se ae,
17 D se ae and 5–6 E se ae.
Maxilliped (Fig. 15A): wi h palp o med o 4 a icles. A icle 1 wi h 2 se ae on inne ma gin. A icle 2
wi h 23 se ae aligned along inne ma gin. A icle 3 wi h 4 apical se ae, one g oup o 6 do sal se ae and
one g oup o 7 se ae on inne ma gin. A icle 4 wi h 1 se a loca ed on ou e ma gin and 2 ine se ae a
nail inse ion. Ou e lobe wi h 5 apical se ae and 9 la ened se ae on inne ma gin. Inne lobe p o ided
apically wi h 3 se ae, 1 la ened se a and 2 do sal se ae.
Gna hopods
Gna hopod I (Fig. 15B): coxal pla e in shape o ec angula apezoid, wi h dep h la ge han wid h ( a io
dep h : wid h 1.0 : 0.64). Basis leng h : wid h a io 1.0 : 0.44. Ischiopodi e wi h one pos e o en al g oup
o 3 se ae. Basis : ca pus leng h a io 1.0 : 0.58. Ca pus wi h ow o 7 se ae along en al ma gin, g oup
o 5 se ae loca ed an e odo sally and one g oup o 8 se ae on ca pus su ace close o en al ma gin.
P opodi e as long as wide, h ee g oups o 3–4 se ae on en al ma gin, one g oup o 4 se ae on do sal
ma gin, one g oup o 10 an e oapical se ae, 3 mesial se ae on la e al su ace, and one g oup o 3 se ae
close o palma co ne . 1 s ong palma spine, 1 suppo ing spine and 2 den icula e spines p esen in
palma co ne . Dac ylopodi e wi h claw 26% o e o al dac ylopodi e leng h and 1 se a along ou e
ma gin.
Gna hopod II (Fig. 15C): sligh ly la ge han gna hopod I, wi h coxal pla e in shape o apezoid; coxal
pla e wid h la ge han dep h ( a io wid h : dep h 1.0 : 0.7). Basis leng h : wid h a io 1.0 : 0.31. Ischiopodi e
wi h one pos e o en al g oup o 4 se ae. Basis : ca pus leng h a io 1.0 : 0.54. Ca pus wi h ow o 6 se ae
along en al ma gin, g oup o 4 se ae loca ed an e odo sally and 1 ow o 4 se ae on ca pus su ace close
o en al ma gin. P opodi e as long as wide, wi h 5 g oups o 3 se ae on en al ma gin, 4 se ae on do sal
ma gin, 1 mesial se a on la e al su ace, 8 an e oapical se ae, one g oup o 5 se ae on la e al su ace
close o en al ma gin, and 3 se ae close o palma co ne . 1 s ong palma spine, 1 suppo ing spine
and 2 den icula e spines p esen in palma co ne . Dac ylopodi e wi h claw 28% o o al dac ylopodi e
leng h and 1 se a along ou e ma gin.
Pe eopods
Pe eopod III (Fig. 16A): coxal pla e in shape o ec angula , wi h wid h la ge han dep h, wid h : dep h
a io 0.7. P opodi e : dac ylus leng h a io 1.0 : 0.45. Dac ylus, wi h nail measu ing hal o o al leng h o
dac ylus, 1 do sal se a wi h plumose ip, and 1 se a a nail base. Pe eopod III nea ly equal o pe eopod
IV (pe eopod III : pe eopod IV leng h a io 1.0 : 0.98).
Pe eopod IV (Fig. 16B): ela i ely ec angula coxal pla e, wi h conca i y on pos e io ma gin,
wid h : dep h a io 1.0 : 0.7. P opodi e : dac ylus leng h a io 1.0 : 0.54. Robus dac ylus, wi h nail hal o
o al leng h o dac ylus; 1 do sal se a wi h plumose ip and 1 se a a nail base.
Pe eopod V (Fig. 17A): coxal pla e o i egula shape, wi h deep conca i y on en al side, 4 an e io
se ae and 1 pos e io se a. Basis ec angula , leng h : wid h a io 1.0 : 0.57, 4 se ae on an e io ma gin,
9 se ae on pos e io ma gin, 3 an e oapical se ae o di e en leng hs. Dac ylus wi h 1 do sal se a wi h
plumose ip, 1 se a a nail base, which ep esen s 45% o o al dac ylus leng h. P opodi e leng h : dac ylus
leng h a io 1.0 : 0.36.
Pe eopod VI (Fig. 17B): coxal pla e smalle han ha o pe eopod V, wi h conca i y on en al side less
deep han ha o pe eopod V, and 1 pos e io se a. Basis ec angula , leng h : wid h a io 1.0 : 0.55, 4 se ae
on an e io ma gin, 8 se ae on pos e io ma gin, 2 an e oapical se ae. Dac ylus wi h 1 plumose se a on
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ou e ma gin and 2 se ae o di e en leng hs nea nail base. Nail 36% o o al dac ylus leng h. Ra io
p opodi e : dac ylus leng h 1.0 : 0.33.
Pe eopod VII (Fig. 17C): longes leg (3.22 mm) o inspec ed pa a ype male. Coxal pla e apezoidal, wi h
1 se a on pos e io ma gin. Basis ec angula , a io leng h : wid h 1.0 : 0.5, 3 se ae on an e io ma gin, 14
se ae on pos e io ma gin and 2 small an e oapical se ae. Dac ylus wi h 1 plumose se a on ou e ma gin
and 2 se ae nea nail base. Nail leng h 31% o o al dac ylus leng h. Ra io p opodi e : dac ylus leng h
1.0 : 0.3.
Pe eopods V : VI : VII a io 1.0 : 1.35 : 1.42.
Fig. 15. Nipha gus pala inensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW161214-01).
A. Maxilliped. B. Gna hopod I. C. Gna hopod II.
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An ennae
An enna I (Fig. 22A): wi h main lagellum o med o 17 a icles, ep esen ing 55% o o al body leng h.
Peduncle leng h 40% o o al leng h o an enna I. Accesso y lagellum (Fig. 22B) bia icula ed; p oximal
a icle sligh ly longe han i s a icle o main lagellum; dis al a icle 28% o o al leng h o accesso y
lagellum, wi h 3 apical se ae o di e en leng hs and 1 aes he asc. Aes he ascs ⅓ o espec i e main
lagellum a icles (Fig. 22C).
An enna II (Fig. 22D): lagellum o med o 8 a icles and ep esen ing 42% o o al leng h o an enna II.
Mos lagellum a icles bea 1 aes he asc, ¼ leng h o espec i e lagellum a icles.
Mou hpa s
Lab um (Fig. 23A): ypical, subo oid shape. Labium (Fig. 23B). La ge inne lobes wi h 1 ow o ine
se ae on inne sides. Ou e lobes wi h 1 ow o ine se ae subapically on inne sides.
Maxilla I (Fig. 23C): wi h 3 apical se ae on dis al a icle o palp. Six spines o ou e lobe wi h 1 oo h
each and 1 spine wi h se e al small ee h. Inne lobe wi h 1 apical se a.
Maxilla II (Fig. 23D): wi h inne lobe sligh ly sho e han ou e lobe. 1 apical ow o se ae on each lobe.
Le mandible (Fig. 23E): 5 ee h on inciso p ocess. 3 ee h on lacinia mobilis. 10 se a ed se ae be ween
lacinia mobilis and mola p ocess.
Righ mandible (Fig. 23F): 4 ee h on inciso p ocess. 2 ee h on lacinia mobilis. 5 se a ed se ae be ween
lacinia mobilis and mola p ocess. Long se a on mola p ocess.
Mandibula palps (Fig. 23E–F): highly simila and o same leng h. 3 a icles accoun o 18% (a icle 1),
36% (a icle 2) and 46% (a icle 3) o o al leng h o palp. P oximal a icle wi hou se ae, median a icle
wi h 6–7 en al se ae. Dis al a icle o palp wi h one g oup o 5 A se ae, ou g oups wi h 1 B se ae each,
22–23D se ae and 6 E se ae.
Maxilliped (Fig. 24A): wi h palp o med o 4 a icles. A icle 1 ase ose. A icle 2 wi h 23 se ae aligned
along inne ma gin. A icle 3 wi h 6 apical se ae, one g oup o 6 do sal se ae and one g oup o 4 se ae
on inne ma gin. A icle 4 wi h 1 se a loca ed on ou e ma gin and 2 se ae a nail inse ion. Ou e lobe
wi h 5 apical se ae and 8 la ened se ae on inne ma gin. Inne lobe p o ided apically wi h 5 se ae and
1 la ened se a.
Gna hopods
Gna hopod I (Fig. 24B): coxal pla e in shape o ec angula apezoid, wi h dep h la ge han wid h ( a io
dep h : wid h 1.0 : 0.53). Basis leng h : wid h a io 1.0 : 0.64. Ischiopodi e wi h one pos e o en al g oup o
8 se ae. Basis : ca pus leng h a io 1.0 : 0.75. Ca pus wi h ow o 9 se ae o a ious leng hs along en al
ma gin, g oup o 3 se ae loca ed an e odo sally and one g oup o 2 se ae on ca pus su ace close o
en al ma gin. P opodi e nea ly as long as wide, i e g oups o 3–4 se ae on en al ma gin, wo g oups
o 2 se ae each on do sal ma gin, one g oup o 5 se ae on p opodi e su ace close o do sal ma gin, one
g oup o 10 an e oapical se ae, 2 mesial se ae on la e al su ace, 2 se ae on la e al su ace close o en al
ma gin and one g oup o 3 se ae close o palma co ne . 1 s ong palma spine, 1 suppo ing spine and 2
den icula e spines p esen in palma co ne . Dac ylopodi e wi h claw 30% o o al dac ylopodi e leng h
and 1 se a along ou e ma gin.
Gna hopod II (Fig. 24C): sligh ly la ge han gna hopod I, wi h coxal pla e in shape o apezoid; coxal
pla e wide han deep ( a io wid h : dep h 1.0 : 0.7). O oid gill and o same leng h as coxal pla e wid h.
Basis leng h : wid h a io 1.0 : 0.38. Ischiopodi e wi h one pos e o en al g oup o 6 se ae. Basis : ca pus
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Fig. 22. Nipha gus no mandiensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW180526-36).
A. An enna I. B. An enna I, de ails o he accesso y lagellum. C. An enna I, de ails o he aes he ascs.
D. An enna II.
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Fig. 23. Nipha gus no mandiensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW180526-36).
A. Lab um. B. Labium. C. Maxilla I. D. Maxilla II. E. Le mandible. F. Righ mandible.
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Fig. 24. Nipha gus no mandiensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW180526-36).
A. Maxilliped. B. Gna hopod I. C. Gna hopod II.
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leng h a io 1.0 : 0.57. Ca pus wi h ow o 9 se ae o a ious leng hs along en al ma gin, g oup o
6 se ae loca ed an e odo sally and 1 ow o 5 se ae on ca pus su ace close o en al ma gin. P opodi e
wid h la ge han leng h ( a io wid h : leng h 1.0 : 0.94), wi h 7 g oups o 3–5 se ae on en al ma gin,
one g oup o 6 se ae on do sal ma gin, one g oup o 2 se ae on la e al su ace close o do sal ma gin,
1 mesial se a on la e al su ace, one an e odo sal g oup o 8 se ae, and one g oup o 3 long se ae in
icini y o palma co ne . 1 s ong palma spine, 1 suppo ing spine and 3 den icula e spines p esen in
palma co ne . Dac ylopodi e wi h claw 30% o o al dac ylopodi e leng h and 1 se a along ou e ma gin.
Pe eopods
Pe eopod III (Fig. 25A): coxal pla e in shape o ec angula apezoid, wide han deep ( a io wid h : dep h
1.0 : 0.75). P opodi e : dac ylus leng h a io 1.0 : 0.4. Dac ylus, wi h nail measu ing hal o o al leng h
o dac ylus, 1 do sal se a wi h plumose ip, and 2 se ae a nail base. Pe eopod III sligh ly longe han
pe eopod IV (pe eopod III : pe eopod IV leng h a io 1.0 : 0.95).
Pe eopod IV (Fig. 25B): coxal pla e in shape o ec angula apezoid, wid h : dep h a io 1.0 : 0.68.
P opodi e : dac ylus leng h a io 1.0 : 0.4. Robus dac ylus, wi h nail measu ing almos hal o o al leng h
o dac ylus; 1 do sal se a wi h plumose ip and 2 se ae o di e en leng hs a nail base.
Pe eopod V (Fig. 26A). Sho es leg (2.64 mm) o inspec ed male pa a ype. Coxal pla e o i egula
shape, wi h deep conca i y on en al side, and 2 an e io se ae. Basis ec angula , leng h : wid h a io
1.0 : 0.62, 3 se ae on an e io ma gin, 6 se ae on pos e io ma gin, 3 an e oapical se ae o di e en leng hs.
Dac ylus wi h 1 do sal se a wi h plumose ip, 2 se ae o di e en leng hs a nail base, which ep esen ing
42% o o al dac ylus leng h. P opodi e leng h : dac ylus leng h a io 1.0 : 0.49.
Pe eopod VI (Fig. 26B): coxal pla e ela i ely simila o ha o pe eopod V, wi h 1 pos e io se a. Basis
ec angula , leng h : wid h a io 1.0 : 0.58, 4 se ae on an e io ma gin, 10 se ae on pos e io ma gin, 3
an e oapical se ae o di e en leng hs. Dac ylus wi h 1 plumose se a on ou e ma gin and 2 se ae o
di e en leng hs nea nail base. Nail ⅓ o o al dac ylus leng h. P opodi e : dac ylus leng h a io 1.0 : 0.42.
Pe eopod VII (Fig. 26C): longes leg (3.90 mm) o inspec ed pa a ype male. Coxal pla e apezoidal, wi h
1 se a on pos e io ma gin. Basis ec angula , a io leng h : wid h 1.0 : 0.52, 5 se ae on an e io ma gin, 11
se ae on pos e io ma gin and 2 an e oapical se ae o di e en leng hs. Dac ylus wi h 1 plumose se a on
ou e ma gin and 1 se a nea nail base. Nail leng h 35% o o al dac ylus leng h. Ra io p opodi e : dac ylus
leng h 1.0 : 0.38.
Pe eopods V : VI : VII a io 1.0 : 1.31 : 1.47.
Pleopods
Pleopods simila (pleopod III depic ed in Fig. 27A), wi h unequal ami and 5 hooks on e inaculum. One
o pleopods III la e ally wi h 2 addi ional hooks on e inaculum.
Epime al pla es (Fig. 27B)
Epime al pla e I wi h igh pos e o- en al angle, s aigh en al ma gin, ela i ely s aigh pos e io
ma gin wi h 4 se ae. Epime al pla e II wi h igh pos e o- en al angle, con ex en al ma gin wi h 2
spines, con ex pos e io ma gin wi h 3 spines. Epime al pla e III wi h igh pos e o en al angle, s aigh
en al ma gin wi h 2 spines, con ex pos e io ma gin wi h 5 spines.
U opods
U opod I (Fig. 28A): wi h wo do so-la e al ows o 3–4 spines on peduncle. Exopodi e longe han
endopodi e, exopodi e : endopodi e leng h a io 1.0 : 0.66. 1 s ong spine a base o u opod I.
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U opod II (Fig. 28B): wi h 4 do sal and 2 apical spines on peduncle. Endopodi e longe han exopodi e,
endopodi e : exopodi e leng h a io 1.0 : 0.84, bo h ami p o ided wi h 1–2 do sal and 5 apical spines.
U opod III (Fig. 28C) sexually di e en ia ed, longe in males. Peduncle wi h 4 pos e oapical se ae.
Endopodi e 67% o leng h o peduncle, wi h 4 small apical se ae. Dis al segmen o exopodi e sligh ly
longe han p oximal segmen ( a io 1.0 : 0.95). An e io ma gin o p oximal segmen o exopodi e wi h
Fig. 25. Nipha gus no mandiensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW 180526-36).
A. Pe eopod III. B. Pe eopod IV.
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Fig. 26. Nipha gus no mandiensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW180526-36).
A. Pe eopod V. B. Pe eopod VI. C. Pe eopod VII.
3 g oups o 3–4 se ae; pos e io ma gin o exopodi e wi h 4 g oups o 2–4 se ae; 1 an e o- and 1 pos e o-
apical se a. Dis al segmen o exopodi e wi h 3 g oups o 2 se ae on an e io ma gin, 4 g oups o 2 se ae
on pos e io ma gin, and 8 apical se ae o di e en leng hs.
Telson
Telson (Fig. 28E) wide (wid h : leng h a io 1.0 : 0.76), wi h 3 long apical spines, 2 spines and 2 hin se ae
wi h plumose ip loca ed medially on each lobe. Longes spine 78% o elson leng h.
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Sexual dimo phism
The examined male and emale a e highly simila . Besides he usual sexual dimo phism (e.g., sligh ly
smalle gna hopods, p esence o oos egi es, and sligh ly deepe coxal pla es I–VI in emales), in
N. no mandiensis sp. no ., he emale u opod III is sho e compa ed o ha o he male (Fig. 28D). The
emale elson is highly simila o ha o he male, bu smalle (Fig. 28F).
Type locali y, ecology and dis ibu ion
The species comp ises h ee dis inc gene ic clades (clade N, D and non-anno a ed) which a e dis ibu ed
in he No mandy (F ance), wi h u he sca e ed eco ds on he Channel Islands (Gue nsey, Je sey), on
wo islands o he B i any Coas o F ance and in mo e Cen al F ance (Fig. 29). The eby, clade D is
es ic ed o No mandy (F ance), whe eas bo h o he clades occu on islands in he A lan ic Ocean and
in mo e Cen al F ance. The ype locali y is he Ca iè e sou e ain de Sain -Vaas -en-Auge, Cal ados,
F ance a 49.2910° N, 0.00090° W, a sub e anean qua y in sands one ou side he illage (Supp. ile 3.6).
The 50 m long ca i y is comple ely d y wi h a small lake wi h s agnan wa e a 40–45 m dis ance
om he en ance, behind a collapsed mine galle y. The wa e le el a ies seasonally om 20–70 cm.
Specimens o N. no mandiensis sp. no . we e ound swimming on he clayey bo om o his lake. We
con i m he pe manen p esence o his species in he ype locali y as i was ound on 26 May 2018
as well as on 20
May 2019. We ound his species in se e al sp ings and in one sub e anean qua y.
Ecological de ails a e no in e ed due o he small numbe o sampling si es.
Fig. 27. Nipha gus no mandiensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW180526-36).
A. Pleopod III, de ail o he e inaculum. B. De ail o he epime al pla es.
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Fig. 28. Nipha gus no mandiensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW180526-36) and
pa a ype, ♀ (ISER DW180526-31). A. Male u opod I. B. Male u opod II. C. Male u opod III. D. Female
u opod III. E. Male elson. F. Female elson.
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Nipha gus wasgauensis Webe & B ad sp. no .
u n:lsid:zoobank.o g:ac :E36BC687-B276-47E9-B685-24FC0718F3DA
Figs 30–38
Diagnosis
Medium-sized Nipha gus species, poo ly se ose. Righ pos e o- en al angle o epime al pla es. 6
spines o maxilla I ou e lobe wi h 1–3 ee h each; 1 spine wi h se e al smalle ee h. Mandibula palp
wi h small numbe (1–2) o B se ae. Gna hopods wi h 2 se ae along ou e ma gin o dac ylopodi e.
Pe eiopod VII, he longes leg, almos hal o o al body leng h. Pleopods e inaculum wi h 2–3 hooks.
U opod I, longe exopodi e. U opod II, longe endopodi e. U opod III sexually dimo phic, exopodi e
elonga ed in males. Telson wi h 5 apical spines on each lobe. The COI ma ke shows a single pu e
diagnos ic si e a posi ion 586 (C). Two 28S DNA alleles a e diagnos ic (wi h 1 he e ozygous specimen)
as well as 12 COI ba codes.
Fig. 29. Dis ibu ion o Nipha gus no mandiensis Webe & B ad sp. no . con i med by molecula da a.
Clade D is ma ked by g een ci cles; clade N is ma ked by ligh -g een iangles; a so a non-anno a ed
clade is ma ked by black ci cles. The ype locali y a he Ca iè e sou e ain de Sain -Vaas -en-Auge
(F ance) is ma ked by a yellow s a , o e laying one si e eco d (clade D). Ve y la ge eshwa e bodies
a e indica ed; coun y codes ollow he ISO 3166 s anda d alpha-2 o ma .
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Pleopods
Pleopods I, II and III simila in shape (pleopod III, wi h 3 hooks on e inaculum, depic ed in Fig. 36A),
wi h unequal ami. Pleopods II and III wi h 2 hooks on e inaculum.
Epime al pla es (Fig. 36B)
Epime al pla e I wi h igh pos e o- en al angle, s aigh en al ma gin, con ex pos e io ma gin wi h
4 se ae. Epime al pla e II wi h igh pos e o- en al angle, con ex en al ma gin wi h 1 spine, con ex
pos e io ma gin wi h 5 spines. Epime al pla e III wi h igh pos e o en al angle, con ex en al ma gin
wi h 2 spines, con ex pos e io ma gin wi h 4 spines.
Fig. 35. Nipha gus wasgauensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW170101-22).
A. Pe eopod V. B. Pe eopod VI. C. Pe eopod VII.
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U opods
U opod I (Fig. 37A): wi h wo do sola e al ows o 2–4 spines on peduncle. Exopodi e sligh ly longe
han endopodi e, exopodi e : endopodi e leng h a io 1.0 : 0.91. 1 s ong spine a base o u opod I.
U opod II (Fig. 37B): wi h 3 do sal and 3 apical spines on peduncle. Endopodi e longe han exopodi e,
endopodi e : exopodi e leng h a io 1.0 : 0.89, bo h ami p o ided wi h 2 do sal and 5 apical spines o
di e en leng hs.
U opod III (Fig. 37C): sexually di e en ia ed, longe in males. Peduncle wi h 3 small apical se ae. Sho
endopodi e, 47% o leng h o peduncle, wi h 2 apical se ae. P oximal segmen o exopodi e longe han
dis al segmen ( a io 1.0 : 0.85). An e io ma gin o p oximal segmen o exopodi e wi h 3 g oups o 2–3
se ae; pos e io ma gin o exopodi e wi h 4 g oups o 1–2 se ae; 2 an e o- and 2 pos e o-apical se ae.
Dis al segmen o exopodi e wi h 2 se ae on an e io ma gin, 1 se a on pos e io ma gin, 2 subapical and
5 apical se ae o di e en leng hs.
Telson
Telson (Fig. 37E): wide (wid h : leng h a io 1.0 : 0.9), wi h 5 long apical spines alongside small se a wi h
plumose ip. Two hin se ae wi h plumose ip on ou e ma gin o 1 lobe. Longes spine 43% o elson
leng h.
Sexual dimo phism
The examined male and emale a e highly simila . Besides he usual sexual dimo phism (e.g., sligh ly
smalle gna hopods, p esence o oos egi es, and sligh ly deepe coxal pla es I–VI in emales), in
N. wasgauensis sp. no ., he emale u opod III is sho e han ha o male (Fig. 37D). The elson o he
Fig. 36. Nipha gus wasgauensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW170101-22).
A. Pleopod III, de ail o he e inaculum. B. De ail o he epime al pla es.
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inspec ed emale pa a ype (Fig. 37F) is smalle , wi h 6 spines on one lobe, and 2 ine se ae wi h plumose
ip on he ou e ma gin o bo h lobes.
Type locali y, ecology and dis ibu ion
The species shows a e y na ow dis ibu ion limi ed o he Wasgau egion, i.e., F anco-Ge man uplands
co e ing pa s o he Fede al S a e o Rhineland-Pala ina e (Ge many) and he Depa men s o Bas-
Rhin and Moselle (bo h F ance) (Fig. 38). The ype locali y is he Sülzlochquelle 2 in he communi y o
God ams ein (Rhineland-Pala ina e, Ge many) a 49.2152° N, 8.0842° E (Supp. ile 3.7). I is a na u al
heoc ene sp ing wi h a pe manen mode a e low, 21°GH ha dness and 380 ppm TDS. No o he species
Fig. 37. Nipha gus wasgauensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW170101-22) and
pa a ype, ♀ (ISER DW160716-25). A. Male u opod I. B. Male u opod II. C. Male u opod III. D. Female
u opod III. E. Male elson. F. Female elson.
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o ei he Nipha gus o gamma ids (Amphipoda; Gamma idae) we e ound in his sp ing. Ac oss i s
known dis ibu ion ange, i was ound in i e sp ings, wo abandoned mines and in one galle y o he
Sieg ied Line. The species seems o be absen in he in e s i ial en i onmen . P e ious published eco ds
om he S ollen am Sachsens ein (Ge many), which e e ed o N. aquilex aquilex (Webe 1988, 1989),
also belong o his species.
Fig. 38. Dis ibu ion o Nipha gus wasgauensis Webe & B ad sp. no . con i med by molecula da a.
The ype locali y a Sülzlochquelle 2 (Ge many) is ma ked by a yellow s a , o e laying one si e eco d.
Ve y la ge eshwa e bodies a e indica ed; coun y codes ollow he ISO 3166 s anda d alpha-2 o ma .
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Nipha gus sa a iensis Webe & B ad sp. no .
u n:lsid:zoobank.o g:ac :C7E58FC7-564C-410B-ADBE-EE2AC9BA06AD
Figs 39–47
Diagnosis
Medium-sized Nipha gus species, poo ly se ose. Righ pos e o- en al angle o epime al pla es. Six
spines o maxilla I ou e lobe wi h 1–2 ee h each; one spine wi h se e al smalle ee h. Mandibula
palp wi h small numbe (1–2) o B se ae. Gna hopods wi h 1 se a along ou e ma gin o dac ylopodi e.
Pe eiopod VII, he longes leg, almos hal o o al body leng h. Pleopods e inaculum wi h 2 hooks.
U opod I, longe exopodi e. U opod II, longe endopodi e. U opod III sexually dimo phic, exopodi e
elonga ed in males. Telson wi h 4 apical spines on each lobe. The COI ma ke shows wo single
pu e diagnos ic si es a posi ions 94 (A) and 357 (A). Fou 28S DNA alleles a e diagnos ic (wi h 7
he e ozygous specimens) as well as 23 COI ba codes.
E ymology
The species name de i es om he Ge man Fede al S a e o Saa land, in which he ype locali y is
loca ed. In Webe e al. (2023), his species was ea ed as N. aquilex G.
Ma e ial examined
Holo ype
GERMANY • ♂; Saa land, Zilla’s Kelle in Nunki chen; 49.4826° N, 6.8317° E; 30 Dec. 2017; Die e
Webe leg.; kep in ac in 96% e hanol; 171230-02; MNHNL130580.
Pa a ypes
GERMANY • 1 ♂; same collec ion da a as o holo ype; 30 Dec. 2017; dissec ed and appendages d awn;
171230-01; ISER mic oscope slide DW171230-01 • 1 ♀; same collec ion da a as o p eceding; dissec ed
and appendages d awn; 171230-05; ISER mic oscope slide DW171230-05 • 1 ♂; same collec ion da a
as o holo ype; 12 Ma . 2018; ISER DW180312-01 • 1 ♂; same collec ion da a as o holo ype; 12 Ma .
2018; ISER DW180312-06 • 1 ♀; same collec ion da a as o holo ype; 12 Ma . 2018; ISER DW180312-
03 • 1 ♀; same collec ion da a as o holo ype; 30 Dec. 2017; ISER DW171230-07 • 1 ♂; same collec ion
da a as o holo ype; 12 Ma . 2018; 180312-02; MNHNL130581 • 1 ♂; same collec ion da a as o
holo ype; 12 Ma . 2018; 180312-04; MNHNL130582 • 1 ♀; same collec ion da a as o holo ype; 30
Dec. 2017; 171230-04; MNHNL130583 • 1 ♀; same collec ion da a as o holo ype; 30 Dec. 2017;
171230-06; MNHNL130584.
Molecula da a
COI and 28S DNA sequences o specimens belonging o Nipha gus sa a iensis sp. no . we e deposi ed
in GenBank. COI and 28S RNA accession numbe s a e p esen in Supp. ile 6 and Supp. ile 9,
espec i ely.
Desc ip ion (male pa a ype ISER DW171230-01)
The o al body leng h is 7.91 mm (Fig. 39).
Head
Head (Fig. 39) 6.95% o o al body leng h. Eyes and os um absen .
An ennae
An enna I (Fig. 40A): wi h main lagellum o med o 18 a icles, ep esen ing 50% o o al body leng h.
Peduncle leng h ⅓ o o al leng h o an enna I. The accesso y lagellum (Fig. 40B) bia icula ed; p oximal
a icle almos wice as long as i s a icle o main lagellum; dis al a icle ¼ o o al leng h o accesso y
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lagellum, wi h 2 apical se ae o di e en leng hs and one aes he asc. Aes he ascs sligh ly mo e han hal
o espec i e lagellum a icles (Fig. 40C).
An enna II (Fig. 40D): lagellum o med o 6 a icles and ep esen ing 40% o o al leng h o an enna II.
Mos lagellum a icles bea one sho aes he asc.
Mou hpa s
Lab um (Fig. 41A): ypical, subo oid shape. Labium (Fig. 41B). Rela i ely small inne lobes wi h 1 ow
o ine se ae loca ed subapically on inne sides. Ou e lobes each wi h 1 ow o ine se ae on ou e sides,
1 ow o ine se ae subapically on inne sides and 1 ow o ine apical se ae.
Maxilla I (Fig. 41C): wi h 4 apical se ae on he dis al a icle o palp. Six spines o ou e lobe wi h 1–2
ee h each and one spine wi h se e al smalle ee h. Inne lobe wi h 1 apical se a.
Maxilla II (Fig. 41D): wi h inne lobe sligh ly sho e han ou e lobe. 1 apical ow o se ae each lobe.
Inne lobe wi h 2 subapical se ae on inne ma gin. Ou e lobe wi h 3 smalle se ae on ou e ma gin.
Le mandible (Fig. 41E): 5 ee h on inciso p ocess. 3 ee h on lacinia mobilis. 6 se a ed se ae be ween
lacinia mobilis and mola p ocess.
Righ mandible (Fig. 41F): 5 ee h on inciso p ocess. 5 small ee h on lacinia mobilis. 4 se a ed se ae
al e na ed wi h 4 i id se ae be ween lacinia mobilis and mola p ocess. Long se a on mola p ocess.
Mandibula palps (Fig. 41E–F): highly simila and o same leng h. 3 a icles accoun o 21% (a icle 1),
37% (a icle 2) and 42% (a icle 3) o o al leng h o palp. P oximal a icle wi h 1 apical se a. Median
Fig. 39. Nipha gus sa a iensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW171230-01); gene al
appea ance.
Eu opean Jou nal o Taxonomy 1011: 1–79 (2025)
54
a icle wi h 7 en al se ae. Dis al a icle o palp wi h one g oup o 3 A se ae, wo g oups wi h 1 B se a
each, 23–24 D se ae and 6–7 E se ae.
Maxilliped (Fig. 42A): wi h palp o med o 4 a icles. A icle 1 ase ose. A icle 2 wi h 14 se ae aligned
along inne ma gin. A icle 3 wi h 5 apical se ae, one g oup o 4 do sal se ae and one g oup o 5 se ae
on inne ma gin. A icle 4 wi h 1 se a loca ed on ou e ma gin and 2 se ae a nail inse ion. Ou e lobe
wi h 3 apical se ae and 9 se ae on inne ma gin, o which 7 la ened. Inne lobe p o ided apically wi h
1 la ened se a and 5 se ae.
Fig. 40. Nipha gus sa a iensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW171230-01). A. An enna I.
B. An enna I, de ails o accesso y lagellum. C. An enna I, de ails o aes he ascs. D. An enna II.
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Gna hopods
Gna hopod I (Fig. 42B): coxal pla e in shape o ec angula apezoid, wi h dep h la ge han wid h ( a io
dep h : wid h 1.0 : 0.62). Basis leng h : wid h a io 1.0 : 0.40. Ischiopodi e wi h one pos e o en al g oup
o 3 se ae. Basis : ca pus leng h a io 1.0 : 0.56. Ca pus wi h ow o 6 se ae along en al ma gin, g oup
o 5 se ae loca ed an e odo sally and wo g oups o 2–6 se ae on ca pus su ace close o en al ma gin.
P opodi e nea ly as long as wide, 4 g oups o 1–4 se ae on en al ma gin, one g oup o 3 se ae on do sal
ma gin, one g oup o 5 an e odo sal se ae, one g oup o 6 an e oapical se ae, 1 mesial se a on la e al
su ace, 1 se a on la e al su ace close o en al ma gin and one g oup o 2 se ae close o palma co ne .
Fig. 41. Nipha gus sa a iensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW171230-01). A. Lab um.
B. Labium. C. Maxilla I. D. Maxilla II. E. Le mandible. F. Righ mandible.
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One s ong palma spine, one suppo ing spine and wo den icula e spines p esen in palma co ne .
Dac ylopodi e wi h claw 33% o o al dac ylopodi e leng h and 1 se a along ou e ma gin.
Gna hopod II (Fig. 42C): sligh ly la ge han gna hopod I, wi h coxal pla e in shape o apezoid; coxal
pla e sligh ly wide han long ( a io wid h : dep h 1.0 : 0.80). Basis leng h : wid h a io is 1.0 : 0.29.
Ischiopodi e wi h one pos e o en al g oup o 4 se ae. Basis : ca pus leng h a io 1.0 : 0.56. Ca pus wi h
Fig. 42. Nipha gus sa a iensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW171230-01). A. Maxilliped.
B. Gna hopod I. C. Gna hopod II.
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ow o 8 se ae o a ious leng hs along en al ma gin, g oup o 5 se ae loca ed an e odo sally and one
g oup o 5 se ae on ca pus su ace close o en al ma gin. P opodi e almos as long as wide, 5 g oups
o 2–3 se ae on en al ma gin, 1 se a on do sal ma gin, 1 se a on la e al su ace close o en al ma gin
and 5 an e oapical se ae o di e en leng hs. 1 mesial se a on he la e al su ace, 3 se ae close o palma
co ne . 1 s ong palma spine, 1 suppo ing spine and 2 den icula e spines p esen in palma co ne .
Dac ylopodi e wi h claw 40% o o al dac ylopodi e leng h and 1 se a along ou e ma gin.
Pe eopods
Pe eopod III (Fig. 43A): coxal pla e in shape o apezoid, a io wid h-dep h 1.0 : 0.73. P opodi e : dac ylus
leng h a io 1.0 : 0.4. Dac ylus wi h nail measu ing hal o o al leng h o dac ylus, 1 do sal se a wi h
plumose ip. Pe eopod III equal sligh ly longe han pe eopod IV (pe eopod III : pe eopod IV leng h a io
1.0 : 0.95).
Pe eopod IV (Fig. 43B): ela i ely ec angula coxal pla e, wid h : dep h a io 1.0 : 0.75.
P opodi e : dac ylus leng h a io 1.0 : 0.41. Robus dac ylus, wi h nail measu ing hal o o al leng h o
dac ylus; 1 do sal se a wi h plumose ip.
Pe eopod V (Fig. 44A): coxal pla e o i egula shape, wi h deep conca i y on en al side, 2 an e io
se ae and 1 pos e io se a. Basis ec angula , leng h : wid h a io 1.0 : 0.64, 6 se ae on pos e io ma gin, 3
se ae on an e io ma gin, 2 an e oapical se ae o di e en leng hs. Dac ylus wi h do sal se a wi h plumose
ip, 2 se ae a nail base, which ep esen s 44% o o al dac ylus leng h. P opodi e leng h : dac ylus leng h
a io 1.0 : 0.31.
Pe eopod VI (Fig. 44B): coxal pla e smalle han ha o pe eopod V, bu simila in shape and 1 pos e io
se a. Basis ec angula , leng h : wid h a io 1.0 : 0.55, 6 se ae on pos e io ma gin, 3 se ae on an e io
Fig. 43. Nipha gus sa a iensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW171230-01).
A. Pe eopod III. B. Pe eopod IV.
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Fig. 49. Nipha gus lo ha ingiensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW160703-02).
A. An enna I. B. An enna I, de ails o accesso y lagellum. C. An enna I, de ails o aes he ascs.
D. An enna II.
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Fig. 50. Nipha gus lo ha ingiensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW160703-02).
A. Lab um. B. Labium. C. Maxilla I. D. Maxilla II. E. Le mandible. F. Righ mandible.
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wi h 9–11 en al se ae. Dis al a icle o palp wi h one g oup o 4A se ae, 4 g oups wi h 1–2 B se ae,
17–22 D se ae and 7 E se ae.
Maxilliped (Fig. 51A): wi h palp o med o 4 a icles. A icle 1 ase ose. A icle 2 wi h 31 se ae aligned
along inne ma gin. A icle 3 wi h 6 apical se ae, one g oup o 6 do sal se ae and one g oup o 9 se ae
on inne ma gin. A icle 4 wi h 1 se a loca ed on ou e ma gin and 2 se ae a nail inse ion. Ou e lobe
wi h 4 apical se ae and 15 la ened se ae on inne ma gin. Inne lobe p o ided apically wi h 3 se ae, 1
la ened se a and 2 plumose se ae.
Gna hopods
Gna hopod I (Fig. 51B): coxal pla e in shape o ec angula , wi h dep h la ge han wid h ( a io
dep h : wid h 1.0 : 0.8). Basis leng h : wid h a io 1.0 : 0.41. Ischiopodi e wi h one pos e o en al g oup
o 5 se ae. Basis : ca pus leng h a io 1.0 : 0.56. Ca pus wi h ow o 14 se ae o a ious leng hs along
en al ma gin, g oup o 7 se ae loca ed an e odo sally and one g oup o 5 se ae on ca pus su ace close
o en al ma gin. P opodi e nea ly as long as wide, se en g oups o 2–5 se ae on en al ma gin, one
g oup o 6 se ae on do sal ma gin, one g oup o 3 se ae on p opodi e su ace close o do sal ma gin, one
g oup o 5 an e oapical se ae, 3 mesial se ae on la e al su ace, 2 se ae on la e al su ace close o en al
ma gin and one g oup o 4 se ae close o palma co ne . One s ong palma spine, one suppo ing spine
and wo den icula e spines p esen in palma co ne . Dac ylopodi e wi h claw 26% o o al dac ylopodi e
leng h and 3 se ae along ou e ma gin.
Gna hopod II (Fig. 51C): sligh ly la ge han gna hopod I, wi h coxal pla e in h shape o ec angula
apezoid; coxal pla e wid h equal o i s dep h ( a io wid h : dep h 1.0 : 1.0). O oid gill and o same
leng h as basis. Basis leng h : wid h a io 1.0 : 0.27. Ischiopodi e wi h one pos e o en al g oup o 3 se ae.
Basis : ca pus leng h a io 1.0 : 0.54. Ca pus wi h ow o 12 se ae o a ious leng hs along en al ma gin,
g oup o 4 se ae loca ed an e odo sally and 1 ow o 10 se ae on ca pus su ace close o en al ma gin.
P opodi e as long as wide, wi h 8 g oups o 1–5 se ae on en al ma gin, 5 se ae on do sal ma gin, 3
mesial se ae on la e al su ace, 1 se a on la e al su ace close o do sal ma gin, 6 an e oapical se ae and 4
se ae close o palma co ne . 1 s ong palma spine, 1 suppo ing spine and 2 den icula e spines p esen in
palma co ne . Dac ylopodi e wi h claw 33% o o al dac ylopodi e leng h and 3 se ae along ou e ma gin.
Pe eopods
Pe eopod III (Fig. 52A): coxal pla e in shape o ec angula apezoid, as deep as wide. P opodi e : dac ylus
leng h a io 1.0 : 0.44. Dac ylus, wi h nail measu ing hal o o al leng h o dac ylus, 1 do sal se a
wi h plumose ip, and 1 se a a nail base. Pe eopod III sligh ly longe han pe eopod IV (pe eopod
III : pe eopod IV leng h a io 1.0 : 0.92).
Pe eopod IV (Fig. 52B): ela i ely ec angula coxal pla e, wi h conca i y on pos e io ma gin,
wid h : dep h a io 1.0 : 0.94. P opodi e leng h : dac ylus leng h a io 1.0 : 0.4. Robus dac ylus, wi h nail
measu ing hal o o al leng h o dac ylus; 1 do sal se a wi h plumose ip and 2 se ae o di e en leng hs
a nail base.
Pe eopod V (Fig. 53A): coxal pla e o i egula shape, wi h deep conca i y on en al side, 6 an e io
se ae and 2 pos e io se ae. Basis o oid- ec angula shaped, leng h : wid h a io 1.0 : 0.62, 5 g oups o 1–2
se ae on an e io ma gin, 12 se ae on pos e io ma gin, 4 an e oapical se ae o di e en leng hs. Dac ylus
wi h 1 do sal se a wi h plumose ip, 2 se ae o di e en leng hs a nail base, which ep esen s 40% o
o al dac ylus leng h. P opodi e leng h : dac ylus leng h a io 1.0 : 0.28.
Pe eopod VI (Fig. 53B): coxal pla e smalle han ha o pe eopod V, bu highly simila in shape and 2
pos e io se ae. Basis o oid- ec angula shaped, leng h : wid h a io 1.0 : 0.51, 6 g oups o 1–2 se ae on
an e io ma gin, 12 se ae on pos e io ma gin, 5 an e oapical se ae o di e en leng hs. Dac ylus wi h 1
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plumose se a on ou e ma gin and 2 se ae o di e en leng hs nea nail base. Nail 35% o o al dac ylus
leng h. Ra io p opodi e : dac ylus leng h 1.0 : 0.34.
Pe eopod VII (Fig. 53C): longes leg (5.65 mm) o inspec ed pa a ype male. Coxal pla e apezoidal, wi h
1 se a on pos e io ma gin. Basis o oid- ec angula , a io leng h : wid h 1.0 : 0.5, 5 g oups o 1–2 se ae
Fig. 51. Nipha gus lo ha ingiensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW160703-02).
A. Maxilliped. B. Gna hopod I. C. Gna hopod II.
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on an e io ma gin, 14 se ae on pos e io ma gin and 4 an e oapical se ae o di e en leng hs. Dac ylus
wi h 1 plumose se a on ou e ma gin and 2 se ae o di e en leng hs nea nail base. Nail leng h 40% o
o al dac ylus leng h. Ra io p opodi e : dac ylus leng h 1.0 : 0.3.
Pe eopods V : VI : VII a io 1.0 : 1.22 : 1.52.
Pleopods
Pleopods: simila (pleopod I depic ed in Fig. 54A), wi h unequal ami, 3 (pleopods I and II) and 4
(pleopods III) hooks on e inaculum.
Fig. 52. Nipha gus lo ha ingiensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW160703-02).
A. Pe eopod III. B. Pe eopod IV.
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Fig. 53. Nipha gus lo ha ingiensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW160703-02).
A. Pe eopod V. B. Pe eopod VI. C. Pe eopod VII.
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Epime al pla es (Fig. 54B)
Epime al pla e I wi h igh pos e o- en al angle, ela i ely s aigh en al ma gin, con ex pos e io
ma gin wi h 2 se ae. Epime al pla e II wi h igh pos e o- en al angle, con ex en al ma gin wi h 1
an e o- en al spine, con ex pos e io ma gin wi h 8 se ae. Epime al pla e III wi h igh pos e o en al
angle, ela i e s aigh en al ma gin wi h 2 an e o- en al spines, s aigh pos e io ma gin wi h 9 se ae.
U opods
U opod I (Fig. 55A): wi h wo do so-la e al ows o 4–5 spines on peduncle. Exopodi e sligh ly longe
han endopodi e, a io exopodi e : endopodi e leng hs 1.0 : 0.95. 1 s ong spine a base o u opod I.
U opod II (Fig. 55B): wi h 2 do sal spine and 3 apical spines on peduncle. Endopodi e sligh ly longe
han exopodi e, endopodi e : exopodi e leng h a io 1.0 : 0.96, bo h ami p o ided wi h 3–4 do sal and
5–6 apical spines.
U opod III (Fig. 55C): sexually di e en ia ed, longe in males. Peduncle wi h 3 apical and 3 pos e io
se ae. Sho endopodi e, 40% o leng h o peduncle, wi h 1 apical se a. P oximal segmen o exopodi e
longe han dis al segmen ( a io 1.0 : 0.7). An e io ma gin o p oximal segmen o exopodi e wi h 6
g oups o 2–4 se ae (including plumose se ae); pos e io ma gin o exopodi e wi h 3 g oups o 3–4 se ae;
4 an e o- and 4 pos e o-apical se ae. Dis al segmen o exopodi e wi h 3 g oups o 1–2 se ae on an e io
ma gin, 3 g oups o 1–2 se ae on pos e io ma gin, 4 apical and 4 subapical se ae o di e en leng hs.
Fig. 54. Nipha gus lo ha ingiensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW160703-02).
A. Pleopod I, de ail o he e inaculum. B. De ail o he epime al pla es.
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Telson
Telson (Fig. 55E): nea ly as wide as long (wid h : leng h a io 0.95), wi h 2 subapical and 5 apical spines
o di e en leng hs on each lobe. Longes spine sligh ly longe han hal o elson leng h. 2 hin se ae o
di e en leng hs and plumose ip, 1 se a medially on 1 lobe.
Sexual dimo phism
The examined male and emale a e highly simila . Besides he usual sexual dimo phism (e.g., sligh ly
smalle gna hopods, p esence o oos egi es, and sligh ly deepe coxal pla es I–VI in emales), in
Fig. 55. Nipha gus lo ha ingiensis Webe & B ad sp. no ., pa a ype, ♂ (ISER DW160703-02) and
pa a ype, ♀ (ISER DW160703-01). A. Male u opod I. B. Male u opod II. C. Male u opod III. D. Female
u opod III. E. Male elson. F. Female elson.
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N. lo ha ingiensis sp. no ., he emale u opod III (Fig. 55D) is sho e han ha o he male, and he
elson o he inspec ed emale pa a ype is p o ided apically wi h 4 se ae only on each lobe (Fig. 55F).
Type locali y, ecology and dis ibu ion
The species is widely dis ibu ed in eas e n F ance (Fig. 56), wi h a sligh geog aphical o e lap wi h i s
sis e species N. schellenbe gi sensu s ic o in he no heas e n ma gin o i s dis ibu ion ange (Fig. 2).
I comp ises wo dis inc gene ic clades, o which one is e y a e and cu en ly only known om wo
si es in he eas e n pa o he species’ ange. The ype locali y is he Tunnel du Col des C oix (connec ing
Vosges and Hau e-Saône, F ance) a 47.8632° N, 6.7464° E, in he e y eas o he species known
dis ibu ion (Supp. ile 3.10). I is a 1087 m long abandoned ailway unnel wi h a 1 m ail gage once
connec ing Lu e (Hau e-Saône, F ance) wi h Le Thillo (Vosges, F ance). A e i s closu e in 1962, i was
used o he supply o po able wa e and was he e o e locked.
Nipha gus lo ha ingiensis sp. no . can be ound in a a ie y o sub e anean bio opes (Supp. ile 4.6).
Howe e , he in e ed absence in he in e s i ial en i onmen is mos likely due o he ac ha he species
does no occu in he a ea whe e he in e s i ial en i onmen was in ensi ely sampled.
Fig. 56. Dis ibu ion o Nipha gus lo ha ingiensis Webe & B ad sp. no . con i med by molecula da a.
The ype locali y a he Tunnel du Col des C oix (F ance) is ma ked by a yellow s a , o e laying one
si e eco d. Ci cles and diamonds e e o wo dis inc gene ic clades. Ve y la ge eshwa e bodies a e
indica ed; coun y codes ollow he ISO 3166 s anda d alpha-2 o ma .
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Discussion
No es on c ypsis and pseudoc ypsis in he N. aquilex species complex
Wi h he ecen in eg a ion o DNA cha ac e s, Webe e al. (2023) p o ided he i s solid molecula
amewo k o shed u he ligh on he complex axonomic si ua ion wi hin he N. aquilex species
complex. In he p esen wo k, by adding new molecula da a as well as an in-dep h mo phological
( e)in es iga ion, we newly desc ibed six o he molecula en i ies delinea ed by Webe e al. (2023).
Mo phologically, he six species desc ibed he ein can be conside ed pseudoc yp ic because se e al mino
mo phological di e ences we e obse ed. In gene al, species a e coun ed as c yp ic i hey canno be
dis inguished by hei mo phology, ha is, i he mo phological a ia ion wi hin a species is ha high
(i.e., a iable) o low (i.e., s a ic) ha i widely o e laps wi h i s congene s. C yp ic species o Nipha gus
can only be dis inguished by means o me hods o he han mo phology, which a e cu en ly mainly
molecula cha ac e s in hei DNA. By inco po a ing o he lines o e idence (e.g., DNA sequences) and
mo phologically e-examining he specimens a hand acco ding o he p oposed gene ic clus e ing, i may
become appa en ha o he mo phological cha ac e s can se e as diagnos ic ai s o allow obus species
delinea ions (Knowl on 1993; Lu ikhuizen & Dekke 2010). The N. aquilex species complex is an
excellen example o a combina ion o c yp ic and pseudoc yp ic species pai s. The six species desc ibed
he ein can be bes dis inguished by he nucleo ide sequences o hei COI and 28S DNA gene agmen s,
along wi h a ew mo phological di e ences. Pu e diagnos ic nucleo ide posi ions in hei COI ba codes
– allowing he unambiguous iden i ica ion o a single species om one o a ew single nucleo ide
polymo phisms (SNPs) – we e sca ce o e en non-exis ing o some species (N. schellenbe gi sensu
s ic o, N. no mandiensis sp. no ., N. lo ha ingiensis sp. no .). A no able excep ion was N. pala inensis
sp. no ., which showed six pu e diagnos ic nucleo ide posi ions. The educed numbe o in o ma i e
(diagnos ic) SNPs o iden i y single species can be bes explained by he combina ion o he ollowing
aspec s: a) COI comp ises a a he as -e ol ing p o ein-coding gene, wi h he degene acy o he gene ic
code allowing codons o mu a e mo e easily while s ill coding o an iden ical amino acid, b) nucleo ide
da a has only ou cha ac e s a es (A, T, G, C), and homoplas ic cha ac e s can a ise o e ime due o
pa allel mu a ions a homologous si es, c) he numbe o unique DNA sequence a ian s was high in
se e al species (213 o N. schellenbe gi sensu s ic o, 79 o N. luxembu gensis sp. no ., and 51 o
N. aquilex sensu s ic o), and d) he analysis o sis e species pai s wi hin a hype di e se species complex
wi h mo e simila gene ic sequence a ian s when compa ed o mo e dis an ly ela ed congene s. All
hese aspec s educe he likelihood o obse ing pu e diagnos ic SNPs o a single species.
No es on molecula species delimi a ions
In a mo e comp ehensi e ASAP species delimi a ion app oach, he new gene ic da a yielded be ween
16 and 21 po en ial species, while ini ially only 10 o 15 po en ial species we e delinea ed (Webe e al.
2023). Howe e , no all po en ial species g oups we e a ec ed by ASAP analysis o ou comp ehensi e
molecula da ase . As such, he newly desc ibed species N. pala inensis sp. no . ( o me species A sensu
Webe e al. 2023) and N. wasgauensis sp. no ( o me species I sensu Webe e al. 2023) always o med
a single dis inc molecula clus e (Table 1). The same si ua ion holds ue o he o me species B
sensu Webe e al. (2023), which, based on h ee specimens ound in he s a ed a ea o he ype locali y
(C owbo ough, UK) (Ka aman 1980), can be assigned o N. aquilex sensu s ic o. This axon has no
been e-desc ibed he e because a comp ehensi e e-desc ip ion is a ailable (Ka aman 1980).
Fo he o he axa mo phologically examined in he p esen s udy, he gene ically en iched ASAP analysis
p o ided a mo e complex si ua ion. The wo molecula lineages, S and R+R* sensu Webe e al. (2023),
o he mo phospecies N. schellenbe gi we e clea ly sepa a ed by hei 28S DNA ma ke s, bu ASAP
analysis ailed o clea ly sepa a e hem. This esul , howe e , was easonable gi en ha ASAP is a gene ic
dis ance-based app oach, and only wo mu a ional s eps a e p esen be ween hei nuclea 28S DNA
alleles. Howe e , COI pa i ions also di e ed in hei esul s. Pa i ion 1 clus e ed bo h po en ial species
