European Journal of Taxonomy 146: 1-18

http://dx.doi.org/10.5852/ejt.2015.146

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

ISSN 2118-9773

www. europeanj ournaloftaxonomy. eu

2015 • Reboleira A.S.P.S. et al.

Research article

um:lsid:zoobank.org:pub:589D6A44-0761-4490-8039-007D72C38535

Highly disjunct and highly infected millipedes - a new cave-dwelling

species of Chiraziulus (Diplopoda: Spirostreptida: Cambalidae)

from Iran and notes on Laboulbeniales ectoparasites

Ana Sofia P.S. REBOLEIRA 1 , Mohamad Javad MALEK HOSSEINI 2 , Saber SADEGHI 3 &

Henrik ENGHOFF 4

14 Zoological Museum, Natural History Museum of Denmark, University of Copenhagen,

Universitetsparken 15, DK-2100 Copenhagen 0, Denmark.

2 Department of Biology, Payame Noor University (PNU), Yasouj, Iran.

2 > 3 Department of Biology, College of Sciences, Shiraz University, Iran.

1 Email: [email protected] (corresponding author)

1 urn:lsid:zoobank.org:author:338DE845-4839-4EF5-B684-587C021F076C

2 urn:lsid:zoobank.org: author: 54DE1791-C6F7-4C81-99DD-736BF C2F7A9F

3 urn:lsid:zoobank.org:author:B21098A7-8D55-43A9-96D9-D3C088219D63

4 um:lsid:zoobank.org:author:FBQ9A817-000D-43C3-BCC4-2BClE5373635

Abstract. Chiraziulus is a highly disjunct, hitherto monotypic genus of cambalid millipedes,

geographically isolated in Iran by more than 7000 km from its presumed closest relatives in East Asia

and North America. Recent fieldwork in caves of Iran has provided several specimens of this genus,

allowing the description of Chiraziulus troglopersicus sp. nov. The intraspecific variability of the type

species, C. kaiseri Mauries, 1983, is illustrated with scanning electron micrographs. Chiraziulus is

characterized by exceedingly long microtrichose gonopod flagella which from their insertion points on

the posterior face of the anterior gonopod coxites first point distad instead of basad or basad-posteriad

as in most other flagelliferous Cambalidea (and Julida), then traverse a groove on the mesal surface of

the anterior gonopod coxites, making a full (360°) loop. The same feature is also illustrated for the first

time in the genus Cambala. The patterns and prevalence of the infection with a species of ectoparasitic

fungus of the genus Rickia (order Laboulbeniales) in the type material of C. kaiseri is described. An

updated review of the cave-adapted fauna of Iran is given.

Key words. Cave-adapted, troglobiont, karst, Zagros, Iran, ectoparasitic fungus.

Reboleira A.S.P.S., Malek Hosseini M.J., Sadeghi S. & Enghoff H. 2015. Highly disjunct and highly infected

millipedes - a new cave-dwelling species of Chiraziulus (Diplopoda: Spirostreptida: Cambalidae) from Iran and

notes on Laboulbeniales ectoparasites. European Journal of Taxonomy 146: 1-18. http://dx.doi.org/10.5852/

ejt.2015,146

Introduction

The millipede order Spirostreptida is mainly distributed in the tropical and temperate zones of the

Southern Hemisphere, and in warm-temperate North America. In the Palaearctic subregion there are

European Journal of Taxonomy 146: 1-18 (2015)

some marginal occurrences in East Asia, and very few, disjunct occurrences in the western Palaearctic:

Odontostreptus spp. in Morocco, Archispirostreptus spp. in the Middle East, and the so far monotypic

genus Chiraziulus Mauries, 1983, in Iran (Shelley & Golovatch 2011: fig. 29). Whereas Odontostreptus

Attems, 1914 and Archispirostreptus Silvestri, 1895 belong to the family Spirostreptidae, suborder

Spirostreptidea, Chiraziulus is the only W Palaearctic representative of the suborder Cambalidea

(Shelley & Golovatch 20IE fig. 30).

Chiraziulus and its type species, C. kaiseri , were described by Mauries (1983) who erected it as a

subgenus of Nannolene Bollman, 1887. Golovatch (1983) tentatively suggested that full generic status

might be warranted for Chiraziulus , and Mauries (1987) listed it as a full genus. Nannolene , in its

current conception (Hoffman 1999), includes numerous nominal species from the western USA and the

Hawaiian Islands. Although relationships within Cambalidae are not at all clear, and the delimitation

of the family vis-a-vis other cambalidean families is equally uncertain (Enghoff et al. in press), the fact

remains that Chiraziulus is a highly disjunct genus, separated from all other Cambalidea by distances of

over 2000 km; if its closest relative is really Nannolene , the distance will surpass 7000 km.

Iran is located in a biogeographically interesting transition area, but the cave-adapted fauna of the

country is still poorly known (Malek Hosseini et al. 2015). The Zagros Mountains are rich in caves, and

many remain unexplored. This mountain range extends from Turkey and Iraq to Iran, and is oriented

from northwest to southeast, comprising nearly one-fifth of the surface area of Iran. Only eight cave-

adapted species (i.e., troglobionts for terrestrials and stygobionts for aquatics) have been recorded from

Iran so far: four amphipods, Niphargus alisadri Esmaeili-Rineh & Sari, 2013; N. daniali Esmaeili-

Rineh & Sari, 2013; A. khayyami Hekmatara, Zaksek, Heidari Baladehi & Fiser, 2013 and N. valachicus

Dobreanu & Manolache, 1933; one terrestrial isopod, Protracheoniscus gakalicus Kashani, Malek

Hosseini & Sadeghi, 2013; one species of goblin spider, Oonopidae, Trilacuna qarzi Malek Hosseini &

Grismado, 2015; and two cave fishes, the barb Gar a typhlops Braun & Kaiser, 1944 and the loach

Paracobitis smithi Smith, 1976 (Esmaeili-Rineh & Sari 2013; Hekmatara et al. 2013; Kashani et al.

2013; Malek Hosseini et al. 2015).

We revise the genus Chiraziulus , describe a new species from a cave in Iran and document intraspecific

variation of C. kaiseri. New observations on the flagellum in a species of the North American genus

Cambala Gray, 1832, are also included. Re-examination of the type material of C. kaiseri led to the

discovery of the presence of ectoparasitic fungi of the order Laboulbeniales; we also include information

about this infection.

Material and methods

New specimens of Chiraziulus (Fig. 1) were collected in two caves in Iran: Ghar Sarab Cave ( Ghar is

Persian for “cave”), located at 87 km northwest of Hamadan city, near Sarab village, altitude: 2200 m

asl, coordinates 35°14 , 41.7” N, 48 o 15’40.6” E, length: 2959.9 m and Neyneh Cave, located at 45 km

southwest of Dehdasht city, Kohgiluyeh and Boyerahmad Province, coordinates 30°40’ 18” N, 50°21 ’71”

E, 770 m asl (Fig. 2).

Specimens were studied using a Leica Wild M10 microscope equipped with an ocular micrometer. Parts

of specimens were mounted in glycerine on temporary slides for microscope study. Microphotographs

were taken using a Leica digital camera M205A mounted on a stereo microscope Leica DFC 420.

Images were processed with a Leica Application Suite program, and final stacking was made using

ZereneStacker software. For scanning electron microscopy, parts of specimens were critical point

dried in a Tousimis Autosamdi 815, series A. Other specimens were transferred to 96% ethanol, then to

acetone, air-dried, mounted on aluminium stubs, coated with platinum/palladium and studied in a JEOL

JSM-6335F scanning electron microscope. Images were processed with Adobe Photoshop CS6.

REBOLEIRA A.S.P.S. et al.. New Chiraziulus millipede from Iran

Material is deposited in the Zoological Museum, Natural History Museum of Denmark, University

of Copenhagen (ZMUC), the Collection of the Biology Department, Zoological Museum of Shiraz

University, Shiraz, Iran (ZM CBSU) and the Natural History Museum of Vienna (NHMW).

Results

Systematics

Class Diplopoda Blainville-Gervais, 1844

Order Spirostreptida Brandt, 1833

Suborder Cambalidea Cook, 1895

Family Cambalidae Bollman, 1893

Genus Chiraziulus Mauries, 1983

Nannolene ( Chiraziulus ) Mauries, 1983: 250

Chiraziulus (as full genus) - Mauries 1987: 197.

Type species

Nannolene ( Chiraziulus ) kaiseri Mauries, 1983.

Fig. 1. Habitus of Chiraziulus troglopersicus sp. nov., paratype.

European Journal of Taxonomy 146: 1-18 (2015)

Diagnosis

A genus of medium-sized blind cambalid millipedes resembling the Nearctic genus Nannolene , with

longitudinal crests on the body rings and ozopores placed on prominent tubercles (Fig. 1). Sharing with

Nannolene remarkably long gonopodal flagella, which make a complete loop (360°) from their insertion

points on the anterior gonopods towards the interior part of the posterior gonopods, differing from

Nannolene by having the coxites of the anterior gonopods larger than the telopodites. First pair of male

legs not modified.

Distribution

Iran (Fig. 2).

Azarbaijan

Caspian sea

Tehran

Afghanistan

Kuwait

Oman sea

1:6.500.000

0 30 6090 1 20 150km

Persian gulf

i iy>

Qatar

Saudi Arabia

■ - Jr

United Arab

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Turkmenistan

Pakistan

Fig. 2. Distribution of species of Chiraziulus Mauries, 1983 in Iran (Mauries 1987). C. troglopersicus

sp. nov.: red dot (1); C. kaiseri : yellow dots (2-6). 1. Neyneh Cave. 2. “Chiraz” “montagne greseuse

au nord de la ville”. 3. 19 km W of Shiraz. 4. 5 km N of Persepolis. 5. Oasis 95 km N of Bandarabass.

6. Sarab Cave.

REBOLEIRA A.S.P.S. et al.. New Chiraziulus millipede from Iran

Chiraziulus kaiseri (Mauries, 1983)

Figs 3-8, 11-12

Nannolene ( Chiraziulus ) kaiseri Mauries, 1983: 252, figs 4-10.

Nannolene ( Chiraziulus ) kaiseri - Go 1 ovateh 1983: 166.

Chiraziulus kaiseri - Enghoff & Morawej 2005: 63.

Fig. 3. Chiraziulus kaiseri (Mauries, 1983), paratypes. Scanning electron micrographs. A. Head and first

body rings in lateral view. B. Last body rings (“tail”) in lateral view. C. Tip of antenna. D. Detail of the

limbus; notice lines of beadlike structures between cuticular scutes. E. Detail of labrum in frontal view.

F. Gnathochilarium in ventral view; the arrow shows the distomesal setae on stipes. Scale bars: A-B,

E-F = 100 pm; C = 10 pm; D = 1 pm.

European Journal of Taxonomy 146: 1-18 (2015)

Diagnosis

A species of Chiraziulus , differing from C. troglopersicus sp. nov. by having triangular anterior gonopods

in anterior view, the posterior branch of the telepodite smaller than the anterior branch, and the posterior

gonopod with a simple, sharp, spinelike process.

Type material examined

IRAN: Holotype ( f) and numerous paratypes, Shiraz, sandstone mountains N of the town, 1600 m asl.

Very dry, scattered vegetation including the first spring flowers, under stones and clods, 16 Feb. 1937,

E.W. Kaiser leg. (ZMUC).

Additional material examined

IRAN: 1 (J, Sarab Cave, 22 Jul. 2002, Alexandri leg. (ZMUC); 2 2 $ $, 19 Str. km W of Shiraz, G.

Pretz mann leg. (NHMW).

Description

Eyes absent, body depigmented, with prominent tergal crests. Ozopores on protruding tubercles, starting

on 5 th body ring (Fig. 3A, B). Antenna with four long apical cones and bacilliform sensilla seated in

small depressions on external part of the 5 th and 6 th antennomeres (Fig. 3C). Labrum and supralabral

chaetotaxy as in Fig. 3E. Gnathochilarial stipes with three setae, two distolateral and one distomesal

seta, the latter close to promentum. Lingual lamella with two setae. Promentum moderately setose

(Fig. 3F). Limbus with numerous tiny denticles (Fig. 3D). Borders between cuticular scutes on part of

body with lines of beadlike structures (Fig. 3D), similar to those observed by Akkari & Enghoff (2011)

and Enghoff (2014) in other millipedes.

Male

Anterior gonopods (Figs 4, 5A-F) with tongue-shaped, parallel-sided mid-sternal lobe. Each coxite

approximately like an equilateral triangle folded at right angles along vertical axis; resulting surfaces

facing anteriad and mesad; coxite hence narrow triangular in anterior view, ending in rather sharp point

(Fig. 5A); coxite in mesal view with a plane surface traversed by oblique groove for accommodation of

flagellum (Fig. 4); posterior margin sinuous, in apical view c. 30% of it with rugose-scaly microsculpture

Fig. 4. Chiraziulus kaiseri { Mauries, 1983), c?, paratype, anterior gonopod. Scanning electron micrographs.

A. Tip of flagellum. B. Flagellum insertion and loop across anterior gonopod coxite. C. Anterior gonopod

showing the entire extension of the flagellum. Scale bars: A= 10 pm; B-C =100 pm.

REBOLEIRA A.S.P.S. et al.. New Chiraziulus millipede from Iran

Fig. 5. Chiraziulus kaiseri (Mauries, 1983), <$, paratypes, gonopods. Scanning electron micrographs.

A. Anterior gonopods in anterior view. B. Anterior gonopods with one posterior gonopod in lateral

view. C. As B, in apical view. D. As B, in posterior view. E-F. Anterior gonopod with detail of the

processes on the tip of anterior gonopods. G-F Posterior gonopod. J. Detail of the mesal sternal part.

Abbreviations: C = coxal process; T = telepodite; Ta = anterior lobe of telopodite; Tb = posterior lobe of

telepodite. Scale bars: A, D, I = 100 pm; B-C, E-H = 10 pm.

European Journal of Taxonomy 146: 1-18 (2015)

Fig. 6. Chiraziulus kaiseri (Mauries, 1983), female vulva. Scanning electron micrographs. A. Vulva

in situ behind second pair of legs, posterior view. B. Details of right vulva behind second pair of legs.

C. Detail of microtubular structure under the vulva. D. Vulva in ventral view. E. Vulva in anterior view.

Abbreviations: o = operculum; b = bursa. Scale bars: A= 100 pm; B-E = 10 pm.

REBOLEIRA A.S.P.S. et al.. New Chiraziulus millipede from Iran

on lateral surface. Telopodite large, two-lobed; anterior lobe (regarded as part of the coxite (e) by Mauries)

larger ( Ta ), but not reaching tip of coxite (Fig. 5D-E), apical margin concave (Fig. 5F); in anterior view

Ta broadly visible lateral to coxite, with transverse “step”, distal to ‘step’ with several setae; in posterior

view Ta longitudinally concave, apically with several setae and with mesal margin (facing rugosity of

coxite) rugose; posterior lobe of telopodite ( Tp ) (T = telopodite of Mauries) much shorter than Ta , rod-

Fig. 7. Chiraziulus kaiseri (Mauries, 1983). Comparison of the paratype (A, C, E, G, I and K) and

specimen from Ghar Sarab Cave (B, D, F, H, J and L). A-B. Left anterior gonopods, mesal view.

C-D. Anterior gonopod, posterior view. E-F. Anterior gonopod, anterior view. G-H. Anterior gonopod,

lateral view. I-J. Anterior gonopod, apical view. K-L. Posterior gonopods.

European Journal of Taxonomy 146: 1-18 (2015)

shaped, apically broadly rounded, with several setae (Fig. 5D). Rugose tips of coxite and telepodite (Ta)

of anterior gonopod forming a pair of “forceps” (Fig. 5F). Flagellum exceedingly long, inserted below

mid-height of posterior margin of coxite, initially directed distad, then turning anteriad and traversing

mesal surface of coxite in slightly curved groove, then turning basad-posteriad, probably accommodated

in basal oblique groove (Fig. 4B-C); apical part of flagellum densely covered with c. 5 pm long hair¬

like, retrorse processes (Fig. 4A). Posterior gonopods (Fig. 5G-I) apically covered with setae and with

a long slender, spine-like, apically twisted process. Mesal sternal part of gonopods forming trough

densely covered with sharktooth-lilce denticles (Fig. 5J).

Female

Vulva in situ as in Fig. 6A-B. Peculiar, microtubular tissue present below insertion level of bursa

(Fig. 6C). Operculum (o) smaller than bursa, with 2 large apical setae; bursa massive, enveloping

operculum, with simple slit-shaped crest and 8 + 8 setae along distal margins (Fig. 6D, E).

Distribution

Known from several localities in Iran (Fig. 2).

Intrapecific variability of Chiraziulus kaiseri

In addition to the type locality and a few sites in its vicinity, Chiraziulus kaiseri has been recorded from

the south of Iran (Golovatch 1983). Examination of an adult male from an isolated cave (Sarab Cave) in

northwestern Iran, far from the type locality, leads us to include it in C. kaiseri as well and to regard the

differences in the male gonopods as intraspecific variability (Fig. 7).

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Podous rings

Fig. 8. Number of podous rings and midbody vertical diameter of the studied specimens. Chiraziulus

kaiseri (Mauries, 1983) in blue; Chiraziulus troglopersicus sp. nov. in red. A = adult • = juveniles

and $ $.

REBOLEIRA A.S.P.S. et al.. New Chiraziulus millipede from Iran

The anterior gonopods of the specimen from Sarah Cave shows some differences compared to the type

material, especially in the shape of the posterior branch of the telepodite (Fig. 7E-J). No remarkable

difference was observed in the posterior gonopods (Fig. 7K-L). Some variability was also observed in

the chaetotaxy of the gnathochilarial stipes (Fig. 3F). In the specimens from the type locality, Sarab Cave

and 19 km W of Shiraz, there are two distolateral and one distomesal setae, whereas in the specimen

illustrated by Golovatch (1983: 168, fig. 12), all three setae are distolateral.

Chiraziulus troglopersicus sp. nov.

um:lsid:zoobank.org:act:B8EF3E4B-E658-4CC6-B4DE-8045734547EE

Figs 1, 9-10

Diagnosis

A species of Chiraziulus , differing from C. kaiseri by anterior gonopods not presenting a triangular shape

in anterior view, by having equally sized anterior and posterior lobes of anterior gonopod telepodites,

and by posterior gonopod having a spine-like process ending in a halberd-like structure.

Etymology

From Greek troglo: cave, common prefix for terrestrial cave-dwelling animals, and Latin persicus :

Iranian.

Type material

Holotype

IRAN: S, Kohgiluyeh and Boyerahmad Province, Khaeiz protected area, Mt. Khaeiz, 45 km southwest

of Dehdasht city, Neyneh Cave, 30 o 40’ 18” N, 50 o 2E71” E, altitude 770 m (gonopods coated for SEM),

17 May 2012 (ZMUC).

Paratypes

IRAN: same data as the holotype: 4 $ $ (ZMUC); 1 $ and 3 SS (ZM CBSU).

Description

Males: 41M4 podous + 2 apodous rings, diameter 1.39-1.64 mm. Females: 39-51 podous + 2 apodous

rings, and 53 + 0 podous rings, diameter 1.38-1.75 mm. Habitus typical of the genus, eyes absent.

Six supralabral setae. Ozopore starting on 5 th body ring (except for one female paratype that presents

teratological traits and in which ozopores start on 6 th body ring).

Male (Fig. 9)

Anterior gonopods (Fig. 9A-D) with remarkably long flagellum (Fig. 9A, D-E), making a full loop from

its insertion point on posterior face of anterior gonopod coxites pointing distad. Anterior gonopods with

blade-shaped and apically rounded coxite (not of a triangular shape in anterior view as in C. kaiseri )

(Fig. 9A-D); each coxite with 2 setae in mesal anterior view (s) (Fig. 9B) and a lateral ridge with 4 setae

at mid-height (dp); external lateral margin slightly concave, with rugose-scaly microsculpture (Fig. 9B).

Telepodite shorter than coxite, bi-lobed (Figs. 9A-D). Anterior and posterior lobes of telepodite (Ta

and Tp) about the same size; anterior lobe with 8 setae, slightly concave on side facing posterior lobe;

posterior lobe with 8 parallel setae. Flagellum (f) extremely long, covered with c. 5 pm long hair-like,

retrorse processes and a pointed tip (Fig. 9E). Posterior gonopods (Fig. 9F-J) apically covered with

setae and with a long slender spine-like process ending in a halberd-like expansion. Mesal sternal part

of gonopods forming a trough covered with sharktooth-like denticles (Fig. 9I-J).

European Journal of Taxonomy 146: 1-18 (2015)

Fig. 9. Chiraziulus troglopersicus sp. nov., $, paratype, gonopods. Scanning electron micrographs.

A. Anterior gonopods in posterior view. B. Detail of the process on the tip of an anterior gonopod in

posterior view. C. Anterior gonopods in anterior view. D. As C, in apical view. E. Tip of the flagellum.

F. Posterior gonopods in lateral view. G. Tip of the long spine-like process of the posterior gonopod.

H. Posterior gonopods in posterior view. I. As H, in anterior view. J. Detail of the mesal sternal part

of the gonopods. Abbreviations: C = coxal process; T = telepodite; f = flagellum; s = setae; dp = distal

process. Scale bars: A-D, F, H-I = 10 pm; E, G, J = 1 pm.

REBOLEIRA A.S.P.S. et al., New Chiraziulus millipede from Iran

Habitat

The new species was discovered in the Neyneh Cave (Fig. 10), located in the mountainous Khaeiz

protected area, in Zagros Mountains, southwestern Iran. Neyneh Cave is situated in the warmer part of

the province where the temperature at the surface reaches to 50°C during the summer. The temperature

inside the cave is 25°C; the relative humidity is very high, 95-99.9%, and the CCf level is 613-700 ppm.

There was no water flow in the cave. This cave is located in cliffs and has more than 700 m of horizontal

maze development. It is located in an area with difficult access, thus protected from human pressure;

however, there is some destruction due to the activities of treasure hunters. Chiraziulus troglopersicus

sp. nov. specimens were found in complete darkness, crawling on guano and cave walls. Some colonies

of bats produce enough guano to cover the cave soil, providing an important source of organic matter

for this subterranean trophic chain. Species richness in Neyneh Cave is relatively high and includes

several arthropod taxa (Acari, Araneae, Oniscidea, Collembola, Orthoptera, Coleoptera), as well as

some vertebrate taxa, such as gekkos, bats and hystricid rodents.

Notes on the gonopods of Cambala annulata (Say, 1821)

SEM study of one male of Cambala annulata (USA, NC, Surry Co., Elkin, 12 October 1975, R.M.

Shelley leg.) revealed that the anterior gonopod in this species, like that of Chiraziulus (and Nannolene),

has a remarkably long flagellum (Fig. 11 A) with a similar insertion, and with a similar full-loop course

in part involving a groove on the mesal side of the anterior gonopod coxite. The sharktooth-like structure

of the mesal gonopodal sternal part is also present (Fig. 11B).

Occurrence of ectoparasitic fungi on Chiraziulus kaiseri

Studies of the obligate ectoparasitic fungi of the order Laboulbeniales associated with millipedes are

scarce (e.g., De Kesel et al. 2013; Santamaria et al. 2014; Enghoff & Santamaria 2015; Reboleira &

Fig. 10. Chiraziulus troglopersicus sp. nov., habitat; Neyneh Cave in Zagros Mountains, Iran.

European Journal of Taxonomy 146: 1-18 (2015)

Fig. 11. Cambala annulata (Say, 1821), anterior gonopod. A. Mesal view. B. Detail of the mesal sternal

part. C. Apical part of coxal processes and telepodite. Scale bars: A= 100 pm; B-C = 10 pm.

Fig. 12. Chiraziulus kaiseri (Mauries, 1983), paratypes infected with ectoparasitic fungi of the genus

Richia Cavara, order Laboulbeniales. A. Arrows indicate black dots on the body rings corresponding to

the insertion of the fungi. B. Scanning electron micrograph of the collum with one fungus (arrow). Scale

bars: A = 1 mm; B = 100 pm.

REBOLEIRA A.S.P.S. et al., New Chiraziulus millipede from Iran

Table 1 . Numbers of Chiraziulus kaiseri (Mauries, 1983) specimens infected with the ectoparasitic

fungus Rickia sp.

Infected

Non-infected

Male

Female

Male

Female

Type locality

19 km W of Shiraz

—

Sarab Cave

—

Enghoff 2015). These fungi live only on the surface of living animals, and during the revision of the type

material of Chiraziulus kaiseri , deposited in ZMUC (Fig. 12), almost all type specimens were found

to be infected with an undescribed species of the genus Rickia , which in spite of its small size can be

detected due to the dark coloration of the foot and appendages (Fig. 12A).

Some Laboulbeniales have a preference for growing on specific parts of the millipede body, and the

transmission mechanisms play an important role for the position of the fungus on the host (Enghoff &

Santamaria 2015; Reboleira & Enghoff 2015). In the case of Rickia sp. on C. kaiseri , the fungus is

randomly distributed along the millipede (Fig. 12): it was found on body rings, legs, head, antennae and

surprisingly also on the paraprocts (anal valves), both in adults and juveniles, suggesting that transmission

does not exclusively take place during copulation as occurs in other species (Santamaria et al. 2014).

Of the 70 specimens studied, 66 where infected, which means that the infection rate may be around

94% (Table 1). This is significantly higher than the 64% infection rate recorded for populations in

caves of the callipodidan Lusitanipus alternans (Verhoeff, 1893), infected with Diplopodomyces

lusitanipodos Santam., Enghoff & Reboleira in central Portugal, and than the 63% recorded for the

blaniulid Archib ore oiidus pallidus (Brade-Birks, 1920) infected with Troglomyces triandrus Santam. &

Enghoff, but still lower than the 100% infections reported for Troglomycespusillus Santam. & Enghoff

on the blaniulid Iberoiulus cavernicola Ceuca, 1967 (Enghoff & Santamaria 2015; Reboleira & Enghoff

2015). The four specimens from 19 km W of Shiraz were also all infected.

More remarkably, the specimen of Chiraziulus kaiseri from Sarab Cave, despite being part of a disjunct

population in the north of Iran, was also infected with the same species of Rickia.

Discussion

Chiraziulus kaiseri presents a considerable degree of intraspecific variation even within the type material.

The new population found isolated in a cave in the northwest of Iran is also parasitized by the same

Rickia (morpho)species as the type material. These fungi are normally highly host specific (De Kesel

1996), and the presence of the same ectoparasitic species in disjunct populations is herewith recorded for

the second time for millipedes, as it was recently reported for Diplopodomyces lusitanipodos Santam.,

Enghoff & Reboleira and Troglomyces manfrediae Colla (Santamaria et al. 2014).

The highly relict character of the genus Chiraziulus had already been pointed out by Golovatch (1983),

due to its remarkable disjunct distribution relative to other Cambalidae, which are distributed mainly in

North and Central America, Hawaii and Australia. The closest relative of Chiraziulus may be Nannolene

(including Hawaicambala Mauries, 1983), as suggested by the original classification of C. kaiseri in

Nannolene (Mauries 1983). A synapomorphy could be the exceedingly long gonopodal flagellum which

from its insertion point on the posterior face of the anterior gonopod coxites points distad instead of

basad or basad-posteriad as in most other flagelliferous Cambalidea (and Julida), and then makes a full

European Journal of Taxonomy 146: 1-18 (2015)

(360°) loop. Illustrations of the flagellar loop in some Hawaiian and Californian species of Nannolene

are provided by Silvestri (1904: figs 28-29) and Chamberlin (1922: pi. 1, fig. 6), respectively. However,

not all Nannolene species have such extraordinary flagella (see, e.g., Silvestri 1904: fig. 19); also a

specimen of Nannolene keiferi Chamberlin, 1943, has flagella of normal length which do not make the

360° loop. On the other hand, we could show that this peculiar type of flagellum is also present in the

genus Cambala. Clearly, the genus-level taxonomy and phylogeny of Cambalidae needs a closer study.

Mauries (1983) mentioned the following characters in support of his new subgenus Chiraziulus :

1. Anterior gonopod coxites with much longer parasagittal lobes than in Nannolene s.s.

2. First pair of male legs unmodified (although there is some uncertainty about the first pair of legs in

Nannolene s.s.)

3. Lack of eyes

4. Presence of longitudinal crests and grooves on the metaterga

5. The geographical isolation

We can support the first and fourth of these characters, although we have re-interpreted the “parties

coxales parasagittales” as belonging to the telopodites. The second, third and fifth characters are less

convincing: the male of N. keiferi that we have examined has unmodified first legs, several (sub)genera

of, e.g., Julidae and Blaniulidae include eyed as well as blind species, and finally, geographical isolation

per se should not be used as an argument for creating a new taxon, as this could easily lead to circular

argumentation. For the time being, we maintain Chiraziulus as a full genus, following Mauries (1987).

The microtrichose flagellum in both species of Chiraziulus is probably used for sperm competition, cf.

Barnett & Telford (1996), Enghoff (1985, 1992, 1995) and Frederiksen & Enghoff (2015).

The anterior gonopod coxite and telopodite in Chiraziulus seem to form a “forceps” with the opposing

scaly-rugose edges. This is probably an analogue of the “forceps” formed by the promerite (= anterior

gonopod) and mesomerite (part of posterior gonopod) in several Julidae, which has been shown to

grip the operculum of the vulva during copulation (Haacker & Fuchs 1970; Tadler 1996; see also Read

1990). Mauries interpreted both parts of the forceps as belonging to the coxite, but the SEM images

clearly show that it is composed of apical parts of the coxite and the anterior branch of the telopodite.

We here add a cave-adapted millipede to the short list of Iranian cave fauna by describing a new species

of Chiraziulus , raising to ten the number of cave-adapted species known for the country.

Acknowledgements

This project was funded by the Danish Council for Independent Research, ref. DFF-FNU 4002-00269

and the Iran National Science Foundation (INSF-89002421). Dr. Carles Ribera donated the specimens of

C. kaiseri from Sarab Cave, Dr. R.M. Shelley, Radford, North Carolina, donated specimens of Cambala

annulata and Nannolene keiferi , and Dr. Nesrine Akkari kindly loaned specimens deposited in NHMW.

Dr. Sergi Santamaria, Barcelona, kindly confirmed the generic affiliation of the ectoparasitic fungus.

We are indebted to Mr. Gholam Reza Haghbin and the environmental monitoring station officers of the

Khaeiz protected area that guided MJMH to the cave with warm hospitality.

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Manuscript received: 22 June 2015

Manuscript accepted: 12 August 2015

Published on: 22 October 2015

Topic editor: Rudy Jocque

Desk editor: Charlotte Thionois

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, De nm ark