European Journal of Taxonomy 156: 1-

http://dx.doi.org/10.5852/eit.2015.156

ISSN 2118-9773

www. europeani ournaloftaxonomy. eu

2015 • Espinasa L. et al.

This work is licensed under a Creative Commons Attribution 3.0 License.

Research article

um:lsid:zoobank.org:pub:F86D90F5-6C89-4521-9268-499D4160B475

A new epigean species of the genus Anelpistina (Insecta: Zygentoma:

Nicoletiidae) from Sierra de El Abra, Taninul, Mexico

Luis ESPINASA Nicole D. BARTOLO 2 & Solomon SLOAT 3

12 3 School of Science, Marist College, 3399 North Road, Poughkeepsie, NY 12601, U.S.A.

* Corresponding author: [email protected]

1 urn:lsid:zoobank.org:author:527A5737-8E85-4FCC-B5E5-C913EABA5012

2 urn:lsid:zoobank.org:author:FC9D487F-9C 14-4612-86AD-83AB517DDF52

3 um:lsid:zoobank.org:author:23364DDB-D92B-4246-8615-BEA6BC17ADB8

Abstract. Anelpistina taninuli sp. nov. (Insecta: Zygentoma: Nicoletiidae), a species from Taninul,

Sierra de El Abra, San Luis Potosi, Mexico, is described. We also report the 16S rRNA sequence of this

new species.

Keywords. Cubacubaninae, Thysanura, Anelpistina quinterensis, Cubacubana, Neonicoletia.

Espinasa L., Bartolo N.D. & Sloat S. 2015. Anew epigean species of the genus Anelpistina (Insecta: Zygentoma:

Nicoletiidae) from Sierra de El Abra, Taninul, Mexico. European Journal of Taxonomy 156: 1-7. http://dx.doi.

org/10.5852/eit.2015,156

Introduction

The caves in Sierra de El Abra and Sierra de Guatemala, in northern Mexico, are inhabited by the

cave adapted nicoletiid Anelpistina quinterensis (= Neonicoletia quinterensis Paclt, 1979). A recent

study (Espinasa et al. 2014) has shown that the species most likely migrated underground to reach both

mountain ranges within the last 12,000 years. An alternative hypothesis is that the species can survive

on the surface and have independently colonized both mountain ranges. The search for nicoletiids on

the surface has successfully resulted in collecting another species of nicoletiid, but has never turned

up a surface specimen of A. quinterensis (Espinasa et al. 2014). Further analysis showed that the

surface specimens collected in that study belonged to a previously undescribed species. We describe the

morphology of this new species and report on its 16S rRNA DNA sequence.

Material and methods

Dissections of the holotype and the largest female paratype were made with the aid of a Motic K series

stereo microscope and were mounted as fixed preparations with Cytoseal™ 60 solution (Richard-Allan

Scientific). The remaining samples were stored in a vial with ethanol. Type material will be deposited in

the Am erican Museum of Natural History (AMNH).

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European Journal of Taxonomy 156 : 1-7 ( 2015 )

Genomic DNA samples were extracted using Qiagen’s DNEasy® Tissue Kit by digesting a leg of the

male holotype and of the largest female paratype in lysis buffer. Amplification and sequencing of the

16S rRNA fragment was done as in Espinasa et al. (2007), following standard protocols and using

primers 16Sar and 16Sb for the 16S rRNA fragment (Edgecombe et al. 2002). Amplification was carried

out in a 50 pL volume reaction, with 1.25 units of AmpliTaq® DNA Polymerase (Perkin Elmer, Foster

City, California, USA), 200 pm of dNTPs and 1 pm of each primer. The PCR program consisted of an

initial denaturing step at 94 °C for 60 s, 35 amplification cycles (94 °C for 15 s, 49 °C for 15 s, 72 °C for

15 s), and a final step at 72 °C for 6 min in a Gene Amp® PCR System 9700 (Perkin Elmer). Success of

amplifications was checked with agarose gel electrophoresis and PCR amplified samples were purified

with the QIAGEN QIAquick PCR Purification Kit. Samples were then sent to SeqWright for direct

sequencing. The sequence editing software Sequencher™ 3.0 was used to read the chromatograms

obtained from the automated sequencer, make contigs and perform alignments of the sequences. External

primers were excluded from the analyses. The Basic Local Alig nm ent Search Tool (BLAST) from NCBI

was used as described in Young (2008) to find and identify the most similar sequences in GenBank.

Results provided identification of previously described species of nicoletiids whose 16S sequence may

be closely related to the new species. The software SequencherTM 3.0 was also used to align these

Genbank sequences with the new specimens and to obtain the number of base pair differences and its %

of similarity (p value), estimated as bp differences / length of fragment.

Results

Molecular data

Molecular data was obtained from two individuals (GenBank# KR067394, KR067395). The 16S

rRNA fragments with primers excluded were 499 bp long. The two specimens differed from each

other by 1 bp (0.2 %). When the 16S rRNA fragments of the Taninul specimens were compared

against the other Cubacubaninae whose 16S rRNA has been sequenced, BLAST analysis shows

the new specimens most similar to A. nandalumii Espinasa et al., 2012 (GenBank# JQ340910) and

A. multispinata Espinasa & Boyko, 2009 (GenBank# KR067393), from which they differ by 102-104

bp (20.4% and 20.8%).

There are 52 species in six genera described within the subfamily Cubacubaninae and most have been

sequenced for the 16S rRNA. Currently there are 120 sequences available in GenBanlc. Using the 16S

rRNA fragment sequences of nicoletiid species across the subfamily Cubacubaninae, Espinasa et al.

(2012) concluded that pairs of specimens within a population differ by an average of 1.7 nucleotides

(0.3%; range 0-7 bp; n=29), by 3.4 nucleotides (0.7%; range 0-13 bp; n=22) in different populations

of the same species, and by 31.2 nucleotides (6.2%; range 10-64 bp; n=14) between sister species.

These parameters have subsequently been used several times (e.g., Espinsa & Mathes 2014; Espinasa

& Socci 2014) and found to be reliable indicators to support species recognition. A difference of at

least 102 bp (20.4%) between the Taninul specimens and any other described species, which had

its 16S rRNA sequenced, supports the hypothesis that they belong to a different species, described

below.

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ESPINASAL et al., Anew species of Anelpistina (Zygentoma: Nicoletiidae) from Mexico

Taxonomic description

Class Hexapoda Blainville, 1816

Order Zygentoma Borner, 1904

Family Nicoletiidae Escherich, 1905

Genus Anelpistina Silvestri, 1905

Anelpistina taninuli sp. nov.

urn:lsid:zoobank.org:act:92B2522E-739D-43D6-922F-F24A6D314E22

Figs 1A-I, 2A-I

Etymology

The species epithet, taninuli , refers to the locality, Taninul, where the species is found.

Material examined

Holotype

MEXICO: □, 7.5 mm, Taninul Hotel, Highway Cd. Valles Tampico km 15, San Fuis Potosi, Mexico,

under rocks (21 0 56’15.20” N, 98°53 , 25.50’ W, 75 masl), 16. Mar. 2013, F. Espinasa, A. Cahill and M.

Yurgel colls (ESP2013Taninull to be deposited in AMNH).

Paratypes

MEXICO: 3 0 0,5,4 and 3.3 mm long; 4 0 0,7, 5.5,4.7 and 4 mm long; collecting data as for holotype.

(ESP2013Taninul2-8 to be deposited in AMNH).

Description

Maximum body length 7.5 mm Maximum conserved length of ante nn a and caudal appendages 4.5 mm

General color light yellow to white. Pedicellus about % as long as first article and with unicellular glands

on ventral surface, clustered approximately in 4 groups and with a row of microchaetae bordering them

in form of a U. On outside lateral border a cluster of about four groups forming a row of unicellular

glands (Fig. 1A).

Head with macrochaetae and microchaetae as shown in Fig. 1B-C, with approximately 8 + 8 macrochaetae

on border of insertion of antenna. Mouthparts short, labial palp as in Fig. ID. Fength of apical article

equal to width and distinctly larger than penultimate article. Penultimate article with bulge containing

two macrochaetae. Fabium and first article of labial palp with macrochaetae. Maxilla as shown in Fig.

IE. Ultimate article of maxillary palp approximately equal in length to penultimate article. Apex of galea

with two conules of different widths (Fig. IF). Facinia as in Fig. IF. Mandible chaetotaxy as in Fig. 1G,

with approximately four macrochaetae heavily sclerotized plus several macrochaetae less sclerotized.

Nota with about 6 macrochaetae on lateral borders apart from several setae of varied sizes (Fig. 1H-I).

Fegs as shown in Fig. 2A. Hind tibia approximately 3 times longer than wide and slightly shorter than

tarsus. Claws with a hairy appearance (Fig. 2B), similar to other Anelpistina (Espinasa et al. 2007).

Abdominal sterna II-VII subdivided into coxites and sternites and sterna VIII and IX of male entire

(Fig. 2E-H), as in other members of subfamily. No apparent modifications in urosternum III of male.

Urostemum IV of adult male with 1 + 1 articulated submedian appendages, their length being about 2x

their width (Fig. 2C). Appendages acute, pointy and narrow, except at their base where they are broad,

with a bulge on edge towards stylets. Appendages surpass stylets of this segment by half length of stylets

(Fig. 2C). Apex of appendages slightly curved, but not quite hooked. Urosternum VIII of adult male

deeply emarginated, its projections rounded (Fig. 2E). Urosternum IX of adult male as in Fig. 2D-E.

Point of insertion of parameres in urosternum IX very deep. Coxal processes with a few slightly more

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European Journal of Taxonomy 156 : 1-7 ( 2015 )

sclerotized setae (Fig. 2D-E). Stylets IX slightly larger than others, with two macrochaetae and extra

subapical pair. Other stylets with one macrochaeta plus subapical pair (Fig. 2E, H). Terminal spine with

small teeth. Stylets IX otherwise without modifications in males. Penis and parameres of adult males as

in Fig. 2D-E. Parameres attaining about half length of stylets IX, bend distinctly outwards, tapering on

0.02 mm

0.1 mm

G

0.2 mm

0.1 mm

H

0.2 mm

0.1 mm

0.2 mm

0.5 mm

Fig. 1 . Anelpistina taninuli sp. nov. Holotype, adult □. A. Pedicellus (outside lateral view). B. Head.

C. Border of insertion of antenna (arrows point at alveoli where macrochaetae have fallen). D. Labium.

E. Maxilla. F. Apex of lacinia and galea. Galea with two conules of different widths. G. Mandible.

H. Nota. I. Border of nota.

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ESPINASAL et al., Anew species of Anelpistina (Zygentoma: Nicoletiidae) from Mexico

B C

0.5 mm

Fig. 2. Anelpistina taninuli sp. nov. Holotype, adult □ and paratype, □. A. Hind leg. B. Claws with

enlarged section showing hairy appearance. C. Urosternum IV. D. Male genital area. E. Urostemum

VIII-IX. F. Urotergite X. G. Cercus. H. Ovipositor and subgenital plate. I. Apex of ovipositor.

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European Journal of Taxonomy 156 : 1-7 ( 2015 )

the inner side to create an acute and slightly hooked apex. Distal portion somewhat sclerotized and with

small, specialized hairs (Fig. 2D).

Urotergite X shallowly emarginated, posterior angles with two macrochaetae, one distinctly longer,

plus a few relatively strong setae (Fig. 2F). Length of inner macrochaetae slightly longer than distance

between them. Cercus of adult male with longer than wide basal annulus, followed by three annuli of

about equal length and width, and then a very long annulus with spines. Composition of spines includes

4 spines; a long, acute and slightly curved spine, a small one, a strong, subacute spine, and a small one.

The two large spines inserted in tubercles (Fig. 2G).

Adult female genital area as in Fig. 2H. Subgenital plate rounded, half as long as wide. Ovipositor

surpassing stylets IX by about 2.75 times the length of stylus and gonapophyses with about 25

pseudoarticles. Distal portion with modified setae (Fig. 21).

Postembryonic development

The holotype male was 7.5 mm long and had glands in the pedicellus, an articulated appendage on

urosternum IV, and spines on the cerci. All three other smaller males (5, 4 and 3.3 mm) lacked all these

secondary sexual characters. In the largest female (7 mm), the ovipositor surpasses stylets IX by about

2.75 times their length and the gonapophyses have about 25 pseudoarticles. In the female which is 5.5

mm long, it surpasses them by once their length and has about 22 pseudo articles. At a length of 4.7 mm

they barely reach the tip of the stylets and no subdivisions are evident. At a length of 4 mm the ovipositor

is just starting to form.

Distribution

Specimens of this species have been collected only from the type locality.

Discussion

This species is a member of the subfamily Cubacubaninae. It has stylets on urosternite II, but lacks

scales, sensory pegs in the appendix dorsalis, and conspicuous lateral lobes bearing numerous glandular

pores in the labium. As such, its generic allocation is within Anelpistina Silvestri, 1905 (= Cubacubana

Wygodzinsky & Hollinger, 1977 = Neonicoletia Paclt, 1979) as defined by Espinasa et al. (2007).

Males of Anelpistina taninuli sp. nov. can be differentiated from all previously described A nelpistina by

the unique articulated appendages of their urosternum IV, that are broad at the base, with a bulge on the

edge towards the stylets, which then become acute, pointed and narrow. These appendages somewhat

resemble those of A. multispinata and A. boneti Wygodzinsky, 1946 by being acute and narrow, but they

differ in that they are less pointed and, most importantly, lack a bulge at their base. Another defining

character of the new species shared with only two other species, A. bolivari Wygodzinsky, 1946 and

A. musticensis Espinasa et al ., 2009, is that their parameres, which are not extremely long, bend distinctly

outwards, tapering on the inner side. The new species is easily differentiated from these two species

because A. bolivari has broad articulated appendages on urosternum IV and A. musticensis does not have

the articulated appendages, while the new species has pointed and narrow appendages. Furthermore,

both species have a very different spine disposition on the cerci from that of the new species.

Finally, the new species can be differentiated from any other described species within the genus

Anelpistina by the cercus of adult males. The typical cercus has a longer than wide basal annulus,

followed by a very long annulus with spines. Sometimes, between these two, there is an annulus which

is wider than long. In the new species, there are three annuli of about equal length and width between

the two aforementioned annuli. Caution should be used with this character because the number of basal

annuli can vary among individuals of the same population.

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ESPINASAL et al., Anew species of Anelpistina (Zygentoma: Nicoletiidae) from Mexico

The assignment of the specimens to a previously undescribed species is supported by the BLAST analyses

which show that their 16S rRNA sequence is very different from that of other Anelpistina species.

Acknowledgments

This project was partially supported by the Marist College through a VPAA grant and by the School of

Science. Melany Cruz is thanked for the drawings.

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Manuscript received. 18 May 2015

Manuscript accepted. 28 September 2015

Published on: 25 November 2015

Topic editor: Koen Martens

Desk editor: Kristiaan Hoe demakers

Printed versions of all papers are also deposited in the libraries of the institutes that are members of the

EJT consortium: Museum national d’Histoire naturelle, Paris, France; Botanic Garden Meise, Belgium;

Royal Museum for Central Africa, Tervuren, Belgium; Natural History Museum, London, United

Kingdom; Royal Belgian Institute of Natural Sciences, Brussels, Belgium; Natural History Museum of

Denmark, Copenhagen, Denmark.

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