© 2024 The Au ho (s). 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), 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.
h ps://zoobank.o g/u n:lsid:zoobank.o g:pub:F1F9115F-7FB0-4B78-A5C1-83F26798CA5A
New eco ds o a u id wa e mi es om Po ugal, wi h he
in eg a i e desc ip ion o one new species
(Aca i, Hyd achnidia, A u idae)
VLADIMIR PEŠIĆ1,*, HARRY SMIT2,3, DINIS GIRÃO4,5,6, ALESSIO IANNUCCI7,
ELISABETH STUR8 & SÓNIA FERREIRA4,5,9
1 Depa men o Biology, Uni e si y o Mon eneg o, Ce injski pu b.b., 81000 Podgo ica, Mon eneg o;
h ps://o cid.o g/0000-0002-9724-345X
2 Na u alis Biodi e si y Cen e , P.O. Box 9517, 2300 RA Leiden, he Ne he lands;
h ps://o cid.o g/0000-0002-0376-6808
3 Museums Vic o ia Resea ch Ins i u e, Museums Vic o ia, GPO Box 666, Melbou ne, VIC 3001, Aus alia.
4 CIBIO, Cen o de In es igação em Biodi e sidade e Recu sos Gené icos, InBIO Labo a ó io Associado,
Campus de Vai ão, Uni e sidade do Po o, 4485-661 Vai ão, Vila do Conde, Po ugal;
h ps://o cid.o g/0009-0008-4842-0721; h ps://o cid.o g/0000-0002-6884-3966
5 BIOPOLIS P og am in Genomics, Biodi e si y and Land Planning, CIBIO, Campus de Vai ão, 4485-661
Vai ão, Vila do Conde, Po ugal.
6 Depa amen o de Biologia, Faculdade de Ciências, Uni e sidade do Po o, 4169-007 Po o, Po ugal.
7 Depa men o Biology, Uni e si y o Flo ence, Ses o Fio en ino, I aly;
h ps://o cid.o g/0000-0001-7729-4412
8 Depa men o Na u al His o y, NTNU Uni e si y Museum, No wegian Uni e si y o Science and Technology,
T ondheim, No way; h ps://o cid.o g/0000-0001-7796-8026
9 EBM, Es ação Biológica de Mé ola, P aça Luís de Camões, Mé ola, Po ugal.
* Co esponding au ho . E-mail: [email protected]
Recei ed 15 Decembe 2024│Accep ed by M. Miliša: 22 Decembe 2024│Published online 24 Decembe 2024
Abs ac
In his pape , h ee species o he wa e mi e amily A u idae collec ed in Po ugal we e iden i ied using an
in eg a i e app oach ha combines mo phology and DNA ba codes. Pa ab achypoda lusi anica sp. no . is
desc ibed as new o science. Two species, Hexaxonopsis omijni (K. Vie s, 1923) and Woolas ookia o undi ons
(K. Vie s, 1922) a e epo ed o he i s ime o he wa e mi e auna o Po ugal.
Key wo ds DNA-ba coding, cy och ome c oxidase subuni I (COI), axonomy, sys ema ics.
In oduc ion
The wa e mi e amily A u idae is a polyphyle ic axon including a high numbe o gene a widely
dis ibu ed on all con inen s excep An a c ica (Ge ecke e al. 2016; Smi 2020). In he Wes e n
Palaea c ic, he amily comp ises o h ee sub amilies, Albiinae K. Vie s, 1925, A u inae Tho , 1900
Ecologica Mon eneg ina 80: 209-219 (2024)
This jou nal is a ailable online a : www.bio axa.o g/em
h ps://dx.doi.o g/10.37828/em.2024.80.19
ATURID WATER MITES OF PORTUGAL
210
and Axonopsinae K. Vie s, 1929. A p esen , six species o he amily a e known om Po ugal (Can allo
e al. 2022, Pešić e al. 2024), one species o A u inae (A u us scabe K ame , 1875) and i e species
o Axonopsinae (Axonopsalbia medi e anea Lundblad, 1954, Ba baxonella o alis Lundblad, 1954,
B achypodopsis g acilis Pie sig, 1903, Hexaxonopsis se a a (Wal e , 1928) and Ljania bipapilla a
Tho , 1898). Two o hese, Axonopsalbia medi e anea and Ba baxonella o alis, o da a a e only known
om Po ugal.
La ely, as he esul s o ecen DNA ba coding ini ia i es conduc ed wi hin he amewo k o
he p ojec Biodi e si y Genomics Eu ope (BGE, h ps://biodi e si ygenomics.eu/), h ee species o he
amily A u idae we e collec ed, one o which is new o science and wo species a e epo ed o he i s
ime o he wa e mi e auna o Po ugal.
Ma e ial and Me hods
Wa e mi es we e collec ed by hand ne ing and p ese ed in 96% e hanol o he pu pose o he
molecula analyses. The specimens used o he molecula s udy a e lis ed in Table 1. A e DNA
ex ac ion, he specimen ouche s we e s o ed in 96% E OH and mo phologically examined. Some o
hese ouche s we e dissec ed and slide moun ed in Fau e’s medium, while he es was ans e ed o
Koenike’s luid.
Mo phological nomencla u e ollows Ge ecke e al. (2016). The holo ype and pa a ypes o he
new species a e deposi ed in he Na u alis Biodi e si y Cen e in Leiden (RMNH).
All measu emen s a e gi en in μm. The pho og aph o ejacula o y complex was made using a
came a on Samsung Galaxy sma phone. The ollowing abb e ia ions a e used: Ac-1 = i s ace abulum;
Cx-I = i s coxae; I-L-4-6 = ou h-six h segmen s o i s leg; P-1-P-5 = palp segmen s 1-5; RMNH =
Na u alis Biodi e si y Cen e , Leiden.
Table 1. De ails on DNA ba coded specimens, including coo dina es o sampling si es, he ba code index numbe
(N indica es a new BIN ha con ains only cu en sequences) and associa ed da a ob ained om BOLD. BOLD
da a p esen ed he e was las accessed on 10 h Decembe 2024.
Locali y
Coo dina es
Sample ID
P ocess ID
BIN
Family A u idae
Pa ab achypoda lusi anica Pešić & Smi sp. no .
Beja, Mé ola, São João dos
Caldei ei os
37.626°N,
7.810°W
BGE_00110_B01
BSNTN488-23
NBOLD:AFN8479
BGE_00228_H08
BSNTN947-23
Hexaxonopsis (Hexaxonopsis) omijni (K. Vie s, 1923)
B agança, Mi andela, To e de
Dona Chama
41.665°N,
7.146°W
BGE_00109_B07
BBIOP019-24
NBOLD:AFW0017
Woolas ookia o undi ons (K. Vie s, 1922)
Gua da, Seia, Rio Al a, P aia
Flu ial de Sabuguei o
40.401°N,
7.640°W
BGE_00227_A01
BSNTN951-23
NBOLD:AFV8119
BGE_00108_H06
BBIOP185-24
BGE_00108_H08
BBIOP187-24
Gua da, Man eigas, Poio do
Leão
40.399°N,
7.541°W
BGE_00108_D05
BBIOP136-24
Gua da, Man eigas, Pon e dos
F ades
40.403°N,
7.526°W
BGE_00108_D07
BBIOP138-24
Gua da, Man eigas, Co ão da
Pon e
40.443°N,
7.514°W
BGE_00108_G02
BBIOP169-24
Molecula and DNA ba code analyses
The molecula analysis was conduc ed a he Uni e si y o Flo ence (Flo ence, I aly). DNA was
ex ac ed using a non-des uc i e p o ocol. Fo he me hods used o cy och ome c oxidase subuni I
(COI) gene ampli ica ion and sequencing see Pešić e al. (2024). Raw eads we e demul iplexed using
PEŠIĆ ET AL.
Ecologica Mon eneg ina, 80, 2024, 209-219 211
he Paci ic Biosciences SMRT Link so wa e. Consensus sequences we e gene a ed wi h he PacBio
Amplicon Analysis (pbaa) ool. P ime imming, ansla ion and s op codon checking we e pe o med
using Geneious P ime 2024.0.1.
Consensus sequences we e made a ailable in he Ba code o Li e Da a Sys ems (BOLD).
Rele an ouche in o ma ion, pho os, and ecen ly gene a ed DNA ba codes a e publicly accessible
h ough he Da ase - DS-BGEPL03 BGE (a ailable a h ps:// 4.boldsys ems.o g/index.php-
/MAS_Managemen _Da aConsole?codes=DS-BGEPL03) in BOLD.
In his s udy DNA was ex ac ed om 9 specimens o he amily A u idae om Po ugal lis ed
in Table 1. Fo he o he species used in he analysis, COI sequence da a we e downloaded om he
espec i e sequence da a a chi es. In o al we used an COI da ase wi h 62 sequences o a u id wa e
mi es o phylogene ic analyses. Rep esen a i e o Hunga ohyd aca us sub e aneus Szalay, 1943 o
he amily Hunga ohyd aca idae was used as ou g oup.
Sequence compa isons we e pe o med using MUSCLE alignmen (Edga 2004). In a and
in e speci ic gene ic dis ances we e calcula ed based on he Kimu a 2-pa ame e (K2P) model (Kimu a
1980), using MEGA 11 so wa e (Tamu a e al. 2021). The la e so wa e was used o calcula e
Neighbo -Joining (NJ) ees based on K2P dis ances (s anda d o ba coding s udies) using pai wise
dele ion o missing da a. B anch suppo was calcula ed using nonpa ame ic boo s ap (Felsens ein
1985) wi h 1000 eplica es and shown nex o he b anches. All codon posi ions we e conside ed in he
analyses.
Resul s
Sys ema ics
Family A u idae Tho , 1900
Genus Pa ab achypoda K. Vie s, 1926
Pa ab achypoda lusi anica Pešić & Smi sp. no .
h ps://zoobank.o g/u n:lsid:zoobank.o g:ac :FD9BD85E-9370-4E12-8103-14D7C1FA2BEC
Figs 1-4
Ma e ial examined — Holo ype ♂, dissec ed and slide moun ed (RMNH), Po ugal, Beja, Mé ola,
São João dos Caldei ei os, 37.626°N, 7.810°W, 17 May 2023, leg. Fe ei a, Beni ez-Bosco, Ek em, S u
& Tu accio, sequenced (Vouche ID: BGE_00110_B01; BOLD ID: BSNTN488-23). Pa a ype: 1♂
(RMNH) same da a as he holo ype, p ese ed in Koenike- luid (sequenced; BOLD ID: BSNTN947-
23). Diagnosis (Female unknown) — Do sal shield nea ly as wide as long, pos e io ma gin
medially p ojec ing; a pai o pos e io ly di ec ed, poin ed shea hs loca ed la e ally nea he pos e io
end o en al shield; gonopo e an e io ly su ounded by well scle o ized lips, Ac-1 loca ed on he
su ace o hese scle o ized lips, nume ous se ae an e io ly in ans e se line, and pos e io ly om he
le el o Ac-3; IV-L-4 dis ally wi h one cu ed, spa ula-shaped se a, IV-L-4 digi i o m en al ex ension
apically wi h h ee di e en se ae (one longe lanked by wo s ou se ae, one hicke , peg-like, he o he
hinne ).
Desc ip ion — Male (holo ype) — Idiosoma do sally 470 long and 493 wide, en ally 500
long. Colou yellowish wi h one unpai ed an e io and a pai o pos e ola e al iole ma ks (see Fig.
1A). Do sal shield nea ly as wide as long, pos e io ma gin medially p ojec ing and wi h exc e o y po e
placed e minally do sal on a p ojec ion (Fig. 1A). An e io ma gins o Cx-I ex ending sligh ly beyond
on al ma gin, gna hosomal bay 128 deep; dis ance be ween IV-L inse ion 271. A pai o pos e io ly
di ec ed poin ed shea hs loca ed la e ally nea he pos e io end o en al shield (Figs. 1A, 2). Pos e io
ma gin o he en e o ming a ounded conca e idge. Gonopo e in dis al pa lanked by well-
de eloped scle o ized lips, Ac-1 loca ed on he su ace o hese scle o ized lips, ace abula a anged in
an ob use iangle, nume ous ine se ae, an e io ly om he le el o Ac-2 longe , and pos e io ly om
he le el o Ac-3 sho e (Fig. 2). Ejacula o y complex 116 long.
ATURID WATER MITES OF PORTUGAL
212
Figu e 1. Pho og aphs o Pa ab achypoda lusi anica Pešić & Smi sp. no ., male pa a ype (BGE_00110_B01):
A – idiosoma, on al iew; B – A – idiosoma, en al iew. Pho og aphs by F. Ses i.
Palp as illus a ed in Figs. 2B-C: leng h/heigh : P-1, 49/19; P-2, 73/52; P-3, 36/33; P-4, 113/36; P-5,
36/20/; palpal segmen ela i ely s ou (leng h/heigh a io P-2, 1.4; P-3, 1.1; P-4, 3.1); P-1 wi h one
do sal se a; P-2 en al ma gin nea ly s aigh , dis ally o ming a sho blun p ojec ion; P-3 en al
ma gin conca e; P-4 p oximally na ow, in he cen e, nea he inse ion o wo en al se ae sligh ly
ele a ed o o m an ob use p ojec ion nea he inse ion o he s onge en al se a, a sligh ly hickened
se a and a ine se a la e ally on a u he blun ele a ion, do sal su ace (medially and en ally) o P-4
wi h se e al ine se ae. Capi ulum en al leng h 102; chelice a 134 long.
PEŠIĆ ET AL.
Ecologica Mon eneg ina, 80, 2024, 209-219 213
Figu e 2. Pa ab achypoda lusi anica Pešić & Smi sp. no ., male holo ype (BGE_00110_B01): A – idiosoma,
do sal iew; B – palp, la e al iew; C – palp, medial iew. Scale ba s = 100 μm.
Legs: I-L and II-L as shown in Figs. 4A-B, su ace o I-L-6 and II-L-6 wi h nume ous ine se ae; III-L
as illus a ed in Fig. 4C, III-L-5 dis ally enla ged, he e wi h ou do sal club-shaped se ae, one o hem
longe han he o he h ee (Fig. 4C inse ); IV-L-4-6 as shown in Figs. 4D-E; IV-L-4 dis ally enla ged,
he e bea ing six o se en se ae, wo o hem hea y (one cu ed, spa ula-shaped se a, shi ed mo e o
en odis al ma gin, he o he s aigh and longe ), he emaining slende and poin ed, and o ming a
p ominen digi i o m en al ex ension apically wi h h ee di e en se ae (one longe lanked by wo
s ou se ae, one hicke , peg-like, he o he hinne ); IV-L-5 hickened, leng h/heigh a io 3.3, do sal
ma gin cu ed, en al ma gin, wi h sca e ed se ae, in p oximal pa a peg-like se ae; IV-L-6 sligh ly
ATURID WATER MITES OF PORTUGAL
214
cu ed, leng h/heigh a io 5.3, en ally wi h a ow o longe se ae; claws modi ied as gi en in Figs.
3A(inse ), an e io claw longe han pos e io claw.
Numbe o swimming se ae: II-L-4. 2, II-L-5: 3; III-L-4: 5; IV-L-5. 6. Do sal leng h: I-L-2-6:
55, 81, 122, 139, 131; II-L-1-6: 72, 63, 91, 134, 144, 128; III-L-1-6: 66, 69, 94, 103. 98, 134; IV-L-1-6:
116, 88, 80, 69, 188, 166.
Female: unknown.
Figu e 3. Pa ab achypoda lusi anica Pešić & Smi sp. no ., male holo ype (BGE_00110_B01): en al shield.
Scale ba = 100 μm.
E ymology — The name e e s o he geog aphical p o enance ( om La in lusi anicus,
Lusi anian, Po uguese) om whe e he new species is desc ibed.
Species delimi a ion using DNA ba codes — We used a COI da ase wi h 79 sequences o
wa e mi e specimens o he amily A u idae lis ed in Table 1 and Supplemen ma e ial 1 o
phylogene ic analyses. Hunga ohyd aca us sub e aneus om Bosnia and He zego ina was used o
oo he ee. The NJ ee is p esen ed in Fig. 5. The COI ee sequences e ie ed om wo specimens
om Po ugal, which o m a unique clus e (BOLD:AFN8479), appea ed as a sis e g oup (albei wi h
a low suppo ) o he sequence belonging o Pa ab achypoda mon ii, a hi obion ic species widely
dis ibu ed in Eu ope. The mean K2P gene ic dis ance be ween COI sequences o Pa ab achypoda
lusi anica sp. no . and P. mon ii was es ima ed a 26.3%.
PEŠIĆ ET AL.
Ecologica Mon eneg ina, 80, 2024, 209-219 215
Figu e 4. Pa ab achypoda lusi anica Pešić & Smi sp. no ., male holo ype (BGE_00110_B01): A – I-L-2-6; B –
II-L (in inse : claw, enla ged 2x); C – III-L (in inse : claw and a club-shaped se a, enla ged 2x); D – IV-L an e io
(in inse : IV-L-4 digi i o m en al ex ension, and claws, enla ged 2x); E – IV-L pos e io . Scale ba = 100 μm.
ATURID WATER MITES OF PORTUGAL
216
Figu e 5. Neighbou -Joining ee o he amily A u idae, ob ained om 79 nucleo ide COI sequences lis ed in
Table 1 and Suppl. ma e ial 1. Hunga ohyd aca us sub e aneus Szalay, 1943 was used o oo he ee. BINs a e
based on he ba code analysis om 15 Decembe 2024.
Rema ks — In ega d o he posi ion o he geni al ield (loca ed close o he pos e io ma gin
o he en al shield) and simila shape o palp (P-2 wi h a e y sho , ounded dis o en al p ojec ion)
and se a ion o IV-L, he new species om Po ugal is placed in genus Pa ab achypoda. Cu en ly, he
la e genus, includes i e species known om he Wes e n Palaea c ic, i.e. P. modes a (Koenike, 1911),
known om Cen al and eas e n Eu ope, P. bade i Di Saba ino & Cicolani, 1991, known om cen al
Medi e anean (I aly: Ab uzzo, Sicily, Sa dinia; F ance: Co sica; Saba ino & Cicolani, 1991, Ge ecke
e al. 2014) and Tu key (Esen e al. 2014), Pa ab achypoda mu ila (Wal e , 1928), known om he
Wes Medi e anean a ea (Alge ia, Ty henian islands - Ge ecke e al. 2014), P. mon ii (Maglio, 1924)
known om cen al and sou he n Eu ope and P. o ien alis Pešić & Esen, 2013. The la e species was
o iginally desc ibed om Tu key (Esen e al. 2014), and includes also popula ions unde he (e oneous)
name o P. mu ila om C imea (Tuzo skij 1978, including also he ea ed la al s age) and Tu key
(Pešić e al. 2006). F om all hese species, he new species om Po ugal can be sepa a ed by he unique
shape o do sal shield (nea ly as wide as long, pos e io ma gin medially p ojec ing) and geni al ield
(gonopo e an e io ly su ounded by well scle o ized lips, Ac-1 loca ed on su ace o hese scle o ized
lips, nume ous se ae a ound he gonopo e, an e io ly in ans e se line, and in a ield pos e o-medially
o Ac-3. Mo eo e , he p esence o a pai o pos e io ly di ec ed poin ed shea hs loca ed la e ally nea
he pos e io end o en al shield is an unique ea u e and no p esen in o he species o he genus.
In ega d o IV-L se a ion, males o Pa ab achypoda mu ila esemble he new species in he
p esence o h ee di e en apical se ae on he en al digi i o m ex ension o IV-L-4 (one ine, longe
PEŠIĆ ET AL.
Ecologica Mon eneg ina, 80, 2024, 209-219 217
se a lanked by wo s ou se ae, one o hem hicke han he o he ). Howe e , in he la e species IV-
L-5 is much slende e , wi h subpa allel do sal and en al ma gins, bea ing nume ous longe en al
se ae. In P. lusi anica IV-L-5 is hickened and wi h mo e cu ed do sal ma gin, mo e simila o B.
o ien alis, en al se ae in p oximal pa peg-like. Mo eo e , he dis al ma gin o IV-L-4 o P. mu ila
lacks he cha ac e is ic spa ula-like se a p esen in he new species.
Dis ibu ion — Po ugal; known only om he ype locali y (Fig. 6) in he Alen ejo egion
(Sou he n Po ugal).
Figu e 6. Pho og aph o he ype locali y (São João dos Caldei ei os) o Pa ab achypoda lusi anica sp. no .
Pho og aph by P. Gadawski.
Genus Hexaxonopsis K. Vie s, 1926
Hexaxonopsis (Hexaxonopsis) omijni (K. Vie s, 1923)
Ma e ial examined — Po ugal, B agança, Mi andela, To e de Dona Chama, 41.665°N, 7.146°W, 256
m asl., 13 Jul. 2023 leg. Fe ei a & Padilha, 1♂ (sequenced), dissec ed and slide moun ed (RMNH).
Rema ks — The Po uguese specimen molecula ly analyzed in his s udy keys ou as
Hexaxonopsis omijni wi h he key o Ge ecke e al. (2016). This specimen o ms a unique BIN
(BOLD:AFW0017). In he phylogene ic ee, he BIN is posi ioned as a sis e clade o H. se a a om
Mon eneg o.
Dis ibu ion — Wes e n and cen al Eu ope (Pešić e al. 2010; Ge ecke e al. 2016); Tu key
(Esen e al. 2011). New o Po ugal.
Genus Woolas ookia Habeeb, 1954
Woolas ookia o undi ons (K. Vie s, 1922)
Ma e ial examined — Po ugal, Gua da: Seia, Rio Al a, P aia Flu ial de Sabuguei o, 40.401°N,
7.64°W, 1021 m a.s.l., 24 Aug. 2023, leg. Fe ei a, Beni ez-Bosco & Padilha, 2♂, 1♀
