European Journal of Taxonomy 52: 1-17

http://dx.doi.org/10.5852/ejt.2013.52

ISSN 2118-9773

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2013 • Pena Santiago R. et al.

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Research article

um:lsid:zoobank.org:pub:4FFA63DB-C2AC-4732-9D28-B5B007F28F5F

Characterization of Iberian species of the genus Pungentus

Thorne & Swanger, 1936 (Nematoda, Dorylaimida, Nordiidae)

Reyes PENA-SANTIAGO'"^ *, Marcel CIOBANU'-^'* and Joaquin ABOLAFIA'-^

^ Departamento de Biologia Animal, Vegetal y Eeologia, Universidad de Jaen, Campus “Fas

Fagunillas” s/n, Edifieio B3, 23071- Jaen, Spain

^Institute of Biologieal Researeh, Department of Plant and Animal Taxonomy and Eeology, 48

Republieii Str., RO-400015 Cluj-Napoea, Romania

* Corresponding author: [email protected]

^ um:lsid:zoobank.org:author:15EDFBB8-F905-4379-B247-2C2ADBC596AE

" um:lsid:zoobank.org:author:237A3FlF-AB01-4B92-979B-77718B6D0ClE

^ urn:lsid:zoobank.org:author:D7E30782-F6F2-48CA-B26F-9CA512FEC078

Abstract. Several populations of four known speeies of the genus Pungentus (R clavatus, P. engadinensis,

P marietani and P. silvestris), eolleeted in the wild and in eultivated soils from the Iberian Peninsula,

are studied. Detailed redeseriptions and morphometries are presented for eaeh speeies. Illustrations are

provided, ineluding line drawings, light mieroseopy pietures of the four speeies as well as seanning

eleetron mieroseopy observations of P engadinensis. The Iberian populations are eompared to type and

other known populations, and new data are given that provide a better eharaeterization of these taxa.

Pungentus engadinensis is the most widely distributed speeies in the Iberian Peninsula.

Keywords. Free-living nematodes, Iberian Peninsula, morphology, Pungentus, taxonomy.

Pena-Santiago R., Ciobanu M. & Abolafia J. 2013. Characterization of Iberian species of the genus Pungentus

Thome & Swanger,1936 (Nematoda, Dorylaimida, Nordiidae). European Journal of Taxonomy 52: 1-17. http://

dx. doi.org/lQ.5852/eit.2013.52

Introduction

The genus Pungentus Thome & Swanger, 1936 is an interesting dorylaimid nematode genus. Its speeies

are eommon inhabitants of natural forest areas of the Holaretie range. So far, five speeies of Pungentus

have been reeorded for the Iberian fauna (see eompendium in Pena-Santiago et al. 2003). A sixth speeies,

namely P. intertextus (Thome & Swanger, 1936) was reeently transferred to Stenodorylaimus Alvarez-

Ortega & Pena-Santiago, 2011. Nevertheless, no morphologieal or taxonomieal study of these Iberian

taxa was ever earried out and their identity was never eonfirmed. General nematologieal surveys, mainly

eondueted in the southern Iberian Peninsula during the last three deeades to explore the dorylaimid

fauna of this region, yielded many populations of Pungentus.

European Journal of Taxonomy 52 : 1-17 ( 2013 )

Here, we present a detailed examination of this material, whieh allowed the eharaeterization of four

speeies and eonfirmation of their presenee in the Iberian Peninsula.

Material and Methods

Nematodes were eolleeted mainly in natural areas of the southern Iberian Peninsula during several

nematologieal surveys eondueted in the last three deeades. They were extraeted from soil samples

using the methods by Baermann (1917) and Flegg (1967), relaxed and killed by heat, fixed in 4%

formaldehyde, proeessed to anhydrous glyeerine following Siddiqi’s (1964) teehnique, and mounted on

permanent glass slides to allow handling and observation under light mieroseopy (LM). Morphometries

(Table 1) ineluded de Man’s indiees and most of the usual measurements; the loeation of pharyngeal

gland nuelei (DN, S^N and is provided after the seheme proposed by Loof & Coomans (1970).

Some of the best preserved speeimens were photographed with a Nikon Eelipse 80i mieroseope and a

Nikon DS digital eamera. Raw photographs were edited using Adobe® Photoshop® CS. Drawings were

made using a camera lucida attaehed to the light mieroseope. After their examination and identifieation,

a few speeimens of Pungentus engadinensis, preserved in glyeerine, were proeessed for observation

under SEM following the protoeol by Abolafia & Pena-Santiago (2005). The nematodes were hydrated

in distilled water, dehydrated in a graded ethanol and aeetone series, eritieal point dried, eoated with

gold, and observed with a JEOE JSM-5800 mieroseope.

Abbreviations

a = body length / greatest body diameter

b = body length / distanee from anterior end to pharyngo-intestinal junetion

e = body length / tail length

e’ = tail length / tail diameter at anus or eloaea

DN = distanee from body anterior end to the nueleus of pharyngeal dorsal gland expressed as

pereentage (%) of total neek length

DO = distanee from body anterior end to the outlet of pharyngeal dorsal gland expressed as

pereentage (%) of total neek length orifiee of dorsal gland

E = overall body length

n = number of speeimens on whieh measurements are based

SjNj = distanee from body anterior end to the anterior nueleus of first pair of ventro-sublateral

pharyngeal glands expressed as pereentage (%) of total neek length.

SjN 2 = distanee from body anterior end to the posterior nueleus of the first pair of ventro-sublateral

pharyngeal glands expressed as pereentage (%) of total neek length

S 2 N = distanee from body anterior end to the nuelei of the seeond pair of ventro-sublateral

pharyngeal glands expressed as pereentage (%) of total neek length

V = distanee from body anterior end to vulva expressed as pereentage (%) of the body length

PENA-SANTIAGO R., CIOBANU M. & ABOLAFIA J., Characterization of Iberian Pungentus

Table 1. Morphometries of Iberian populations of the genus Pungentus Thome & Swanger, 1936.

Measurements in pm (exeept L, in mm).

Species

Habitat

Province

Character n

P. clavatus

P. engadinensis

Several

13 $$ S

P marietani

Brnshwood

Granada

5??

P. silvestris

Pine forest

Granada

7??

Brnshwood

Hnesca

2??

Pine forest

Jaen

10??

Pine forest

Granada

10??

1.37-1.83

1.54, 1.45

0.77-1.21

0.90

1.15-1.56

1.47-1.90

1.65-2.10

35-42

33-39

25-38

34-40

35-42

3.5-4.4

3.5, 4.4

3.4-4.4

3.8

3.5-4.9

3.5-4.6

3.7-4.8

52-70

70, 56

46-62

53-71

51-71

55-70

c’

0.7-1.0

0.8-1.0

0.8

0.7-0.9

0.7-0.8

41-45

42, 48

42-54

44-49

41-45

42-47

Lip region diameter

13-14

8-10

11-13

14-15

Odontostyle length

30-34

33, 32

14-18

22-25

31-34

32-34

Odontophore length

30-34

13-17

22-28

26-32

28-35

Neck length

352-423

437, 408

194-280

240

320-337

338-451

366-507

Pharyngeal expansion length

190-206

221, 190

71-115

130-137

174-228

202-247

Body diameter at neck base

36-47

22-27

36-45

41-50

42-52

midbody

36-51

22-28

37-48

44-54

42-57

anns/cloaca

30-37

16-20

25-26

33-38

33-39

Prerectnm length

55-98

140, 131

42-64

72-108

108-200

130-187

Rectnm length

24-31

28, 24

21-25

24-28

23-40

26-33

Tail length

22-32

22, 26

14-21

19-22

26-34

25-31

Spicnle length

Ventromedian snpplements

Results

Phylum Nematoda Potts, 1932

Class Enoplea Inglis, 1983

Order Dorylaimida Pearse, 1942

Family Nordiidae Jairajpuri & Siddiqi, 1964

Genus Pungentus Thome & Swanger, 1936

Pungentus clavatus Ahmad & Jairajpuri, 1979

Figs lA-C, 2

Diagnosis

This speeies is distinguished by the length of its body (1.37-1.91 mm), the lip region being offset by

a eonstrietion and 13-15 pm wide, an odontostyle being 30-35 pm long and 2.3-2.5 times lip region

width or 1.7-2.1% of total body length, neek length of 352-437 pm, a pharyngeal expansion 190-221

pm long or 50% of total neek length, a monodelphie-opisthodelphie female genital system, with the

anterior genital braneh redueed to a very short prevulval sae up to 16 pm long or up to 0.4 times the body

diameter, a utems being a simple tube 101-129 pm long or 2.3-2.8 times the body diameter, V= 39-47,

and the female tail being rounded and somewhat elavate (22-31 pm, c = 41-75, e’ = 0.7-1.0).

Material examined

Nine females from two loeations, in various states of preservation.

European Journal of Taxonomy 52: 1-17 (2013)

Fig. 1. Iberian Pungentus species. - A-C. Pungentus clavatus Ahmad & Jairajpuri, 1979. - D-H.

Pungentus engadinensis (Altherr, 1950). - I-K. Pungentus marietani Altherr, 1950. - L-N. Pungentus

silvestris Coomans & Geraert, 1962. - A, D, 1, L. anterior region in lateral median view. B, E, J, M.

genital system. C, G, K, N. posterior body region. F. posterior body region. H. caudal region.

PENA-SANTIAGO R., CIOBANU M. & ABOLAFIA J., Characterization of Iberian Pungentus

Morphometries

See Table 1.

Description

Female

Slender to very slender nematodes of medium size, 1.37-1.83 mm long. Habitus after fixation ventrally

eurved, espeeially in posterior body region, G-shaped. Body eylindrieal, tapering towards the anterior

end. Cutiele two-layered, 2.0-3.0 pm thiek in anterior region, 2.0-3.0 pm at mid-body and 4.0-6.0 pm

on dorsal side of tail; its outer layer with very fine transverse striation and thinner than inner one.

Lateral ehord 9-11 pm wide or oeeupying 21-28% of mid-body diameter, very granular but laeking any

differentiation. Ventral body pores pereeptible along the entire body. Lip region offset by eonstrietion,

2.2-2.4 times as wide as high and ca. one-third (30-34%) of body diameter at neek base. Lips moderately

angular and separate; labial and eephalie papillae distinet, but not espeeially protruding above eephalie

eontour. Amphid fovea eup-shaped, opening at level of eephalie eonstrietion, oeeupying 6.5-8.0 pm

or one-half to three-fifths of lip region diameter. Cheilostome almost eylindrieal, its walls distinetly

thiek and selerotised at the anterior half; perioral refraetive dots very eonspieuous. Odontostyle 14-18

times as long as wide, 2.3-2.5 times longer than lip region diameter or 1.7-2.1% of total body length;

Fig. 2. Pungentus clavatus Ahmad & Jairajpuri, 1979 ($, LM). A-B. Anterior region in lateral median

view. C. Pharyngo-intestinal junetion. D. Dorsal eell mass at level of uterus. E. Anterior region in lateral

surfaee view. F. Vagina. G. Caudal region. (Seale bars: A, B, E = 20 pm; C, D, F, G = 10 pm)

European Journal of Taxonomy 52: 1-17 (2013)

aperture 4.0-5.5 pm long or 12-17% of its length. Odontophore rod-like, almost equal (0.9-1.1 times)

to odontostyle. Guiding ring double, loeated at 20-22 pm or 1.5-1.6 times the lip region diameter from

anterior end. Pharynx enlarging very gradually; basal expansion 9.5-12.1 times as long as wide and 5.3-

5.4 times the eorresponding body diameter, oeeupying one-half (50%) of total neek length. Pharyngeal

gland nuelei loeated as follows: DO = 52-53, DN = 54-55, SjNj = 73, SjN^ = 74, S 2 N = 83-88. Nerve

ring at 144-156 pm from anterior end or 36-38% of total neek length. Cardia rounded eonoid, 10-11 x

10-11 pm. Genital system monodelphie-opisthodelphie; anterior braneh redueed to a short to vestigial,

uterine, prevulval sae up to 16 pm long or up to 0.4 body diameters; posterior braneh well developed,

180-305 pm long. Ovary 44-218 pm long, often reaehing and surpassing the sphineter level; ooeytes

first in two or more rows, then in one row. Oviduet 77-124 pm long, or 1.9-2.4 times the body diameter,

eonsisting of a slender portion with prismatie eells and a moderately developed pars dilatata. Sphineter

distinet between oviduet and uterus. Uterus a simple tube-like strueture, 101-129 pm long or 2.3-2.8

times the body diameter. Sperm eells absent within the genital traet. Uterine eggs 96-108 x 36-40 pm. A

distinet dorsal eell mass or eoelomoeyte (see Fig. 2D) is present at level of oviduet or uterus in two of the

speeimens examined. Vagina extending inwards 20-25 pm or about one-half (43-54%) of body diameter;

pars proximalis almost as long as wide, 13-18 x 12-16 pm, with eonvergent walls and enveloped by

weak eireular museulature; pars refringens with two weakly selerotized pieees; pars distalis 5.5-7.0 pm

long. Vulva a pre-equatorial, short transverse slit, whieh is preeeded by a depression of body surfaee.

Prereetum 2.5-2.6 anal body diameters long. Reetum 30-31 pm long, almost equal (0.9-1.0 times) to

anal body diameter. Tail short and rounded, more or less elavate. Caudal pores two pairs, one subdorsal,

another sublateral, at the middle of tail.

Male

Unknown.

Distribution

Pungentus clavatus was eolleeted from two loealities: the provinee of Jaen (southern Iberian Peninsula),

Cazorla, Segura y Las Villas Natural Park Road to Segura river souree (38° 17’ N - 2°39’ W), in

assoeiation with pine forest; and the provinee of Huesea (northern Iberian Peninsula), road from Somport

to Huesea (42°34’ N - 0°32’58” W), assoeiated to Mediterranean brushwood, with dominant speeies

Buxus sempervirens LhmsiQus, 1153, Juniperus oxycedrus Lmnaeus, 1753, Rosa canina LirmaQus, 1753,

and Rubus sp.

Remarks

Although new data are provided and the ranges of many morphometrie features are extended, the present

deseription agrees very well with the original one. In addition to the type loeality in India, P. clavatus

has been reported from Italy (Vineiguerra & Giannetto 1987), Poland (Winiszewska-Slipinska 1987a)

and Spain (Hernandez et al. 1988). Winiszewska-Slipinska {op. cit.) also mentioned its oeeurrenee in

Canada. No signifieant interpopulational variation has been reported in this speeies. The most relevant

and eharaeteristie feature of the speeies is the elavate shape of its eaudal region. Pungentus clavatus

might be a widely distributed speeies in the Holaretie.

Pungentus engadinensis (Altherr, 1950)

Figs ID-H, 3-4

Diagnosis

This speeies is distinguished by its body length of 0.77-1.21 mm, a three-layered eutiele (espeeially

distinet at eaudal region), a lip region offset by a eonstrietion and being 8-10 pm wide, an odontostyle

14-20 pm long and 1.7-1.9 times the lip region width or 1.5-2.2% of the total body length, a neek 194-

PENA-SANTIAGO R., CIOBANU M. & ABOLAFIA J., Characterization of Iberian Pungentus

280 |am long, a pharyngeal expansion 71-115 pm long or 35-42% of total neek length, a monodelphie-

opisthodelphie female genital system, with anterior genital braneh absent, uterus a simple tube 39-44 pm

long or 1.4-1.8 times the body diameter, V= 41-54, a rounded tail (14-28 pm, c = 40-63, c’ = 0.8-1.1),

spieules 25-28 pm long and three to five widely spaeed, ventromedian supplements that laek hiatus.

Fig. 3. Pungentus engadinensis (Altherr, 1950) (LM). A. $, entire. B-D. Anterior region in median view.

E. Pharyngeal expansion. F. $, part of posterior genital braneh. G. Vagina. H, J. $, eaudal region. I. S,

eaudal region and spieules. (Seale bars: A = 200 pm; B, J, K = 10 pm; C, D, G, I = 5 pm; E, F = 20 pm)

European Journal of Taxonomy 52: 1-17 (2013)

Material examined

More than one hundred females and one male from more than 20 loeations, in various states of

eonservation.

Morphometries

See Table 1.

Description

General features of adults

Slender nematodes of small to medium size, 0.77-1.21 mm long. Habitus after fixation eurved ventrad,

C-shaped in females, G-shaped in male. Body eylindrieal, tapering towards the anterior end. Cutiele

three-layered, espeeially distinguishable at eaudal region, 1.5-2.5 pm thiek in anterior region and at

mid-body and 2.5-4.0 pm on dorsal side of tail; outer layer with very fine transverse striation. Lateral

ehord 5.5-6.5 pm wide or oeeupying 19-24 % of mid-body diameter, very granular but laeking any

differentiation. Ventral body pores pereeptible along the entire body. Lip region offset by eonstrietion,

2.4-3.2 times as wide as high and ca. one-third (31-38%) of body diameter at neek base. Under SEM, lips

mostly amalgamated and somewhat angular, laeking any distinguishable striation; labial and eephalie

papillae very distinet and visibly protruding, the inner labial papillae situated at the top of low but

pereeptible lobes; perioral area with (apparently) four small elevations (liplets) surrounding the short

dorsoventral, slit-like oral opening. Amphid fovea eup-shaped, opening at level of eephalie eonstrietion,

oeeupying 5.0-5.5 pm or about three-fifths of lip region diameter. Cheilostome almost eylindrieal, but

a little wider at its posterior half and with its walls distinetly thieker at the anterior half; eireum-oral

Fig. 4. Pungentus engadinensis (Altherr, 1950) (SEM). A. Eip region in lateral view. B. Same in frontal

view. C. Same in ventral view. D. Vulva in ventral view. E-F. Caudal region in ventral view. (Seale bars:

5 pm)

PENA-SANTIAGO R., CIOBANU M. & ABOLAFIA J., Characterization of Iberian Pungentus

refraetive dots very eonspieuous. Odontostyle 11-14 times as long as wide, 1.8-1.9 times longer than

lip region diameter or 1.5-2.2% of total body length; aperture 2.5-3.0 pm long or less than one-sixth

its length. Odontophore rod-like, but with small thiekenings at its base, nearly as long (0.9-1.1 times)

as odontostyle. Guiding ring double, loeated at 10-11 pm or 1.1-1.3 times the lip region diameter from

anterior end. Pharynx enlarging very gradually; basal expansion 5.5-7.4 times as long as wide and 3.1-

3.9 times the eorresponding body diameter, oeeupying about two-fifths (36-41%) of total neek length.

Pharyngeal gland nuelei obseure in the speeimens examined. Nerve ring at 80-91 pm from anterior end

or 32-38% of total neek length. Cardia rounded eonoid, 5.0-6.5 x 7-10 pm.

Female

Genital system monodelphie-opisthodelphie: anterior braneh absent and posterior one moderately

developed, 76-125 pm long. Ovary 53-110 pm long, often reaehing and surpassing the sphineter level;

ooeytes first in two or more rows, then in one row. Oviduet 39-63 pm long, or 1.5-2.4 times the body

diameter, eonsisting of a slender portion with prismatie eells and a moderately developed pars dilatata.

Sphineter distinet between oviduet and uterus. Uterus a simple tube-like strueture, 39-44 pm long or 1.4-

1.8 times the body diameter. Uterine egg 82-92 x 21-26 pm, 2.5-3.5 times as long as the eorresponding

body diameter. Vagina extending inwards 12-17 pm or one-half to three-fifths (50-60%) of body

diameter; pars proximalis 7-9 x 6-12 pm, with parallel or distally eonvergent walls and enveloped by

weak eireular museulature; pars refringens with two small, reetangular or triangular pieees measuring 1

X 3 pm and with a eombined width of 5-6 pm; and pars distalis 3-4 pm long. Vulva a nearly equatorial,

about 5 pm long, transverse slit. Prereetum 2.3-4.1, reetum 0.8-1.2 anal body diameters long. Tail short

and rounded, visibly more eonvex dorsally; the eutiele at its level eonsisting of a thiek inner layer, a

distinet intermediate layer and thinner outer layer, oeeasionally bearing blister-like struetures (saeeate

bodies). Two pairs of eausal pores, one subdorsal, another sublateral, at the middle of the tail.

Male

Very rare, only one speeimen found among several dozens of females. Genital system diorehie, with

opposite testes. In addition to the ad-eloaeal pair, situated at 6.5 pm from eloaeal aperture, there is a

series of five widely spaeed, 10-20 pm apart, ventromedian supplements, the most posterior of whieh is

loeated 9.5 pm from the ad-eloaeal pair, within the range of spieules. Spieules dorylaimoid, about 5.0

times as long as wide and 1.2 times the body diameter. Lateral guiding pieees obseure. Caudal region

similar to that of the female.

Distribution

Pungentus engadinensis is the most widely spread speeies of its genus in the southern Iberian Peninsula,

having been eolleeted from more than 20 loeations with many kinds of natural and eultivated systems,

whose dominant plant speeies are holm-oak {Quercus spp.), Mediterranean brushwood {Cistus spp.),

pine {Pinus spp.), poplar {Populus nigra Linnaeus, 1753), almond (Prunus amygdalus Batseh, 1801),

Mahaleb eherry {Prunus mahaleb Linnaeus, 1753), hazel {Corylus avellana Linnaeus, 1753), ete.

Remarks

The above deseription of Iberian populations perfeetly fits the available information about this speeies,

espeeially the eontributions by Andrassy (1962) and Coomans & Geraert (1962), who provided detailed

data and illustrations. Nevertheless, some relevant features sueh as the three-layered nature of the eutiele,

the morphology of the lip region and the strueture of the female genital system are here deseribed for the

first time. Besides, the range of some measurements (body length, tail length, ete.) and morphometries

(vulva position) are in general extended.

Pungentus engadinensis is eertainly the most eommon speeies in this genus, having been reported from

eleven eountries on three eontinents, all of them in the Holaretie: Belgium (Coomans & Geraert 1962),

European Journal of Taxonomy 52: 1-17 (2013)

Canada (Winiszewska-Slipinska 1987a; Andrassy 1991), Hungary (Andrassy 1962, 1991, 2009), Iraq

(Vinciguerra 2006), Italy (Zullini 1971, 1975), Poland (Brzeski 1963; Winiszewska-Slipinska 1987a,

b), Spain (Castillo et al. 1985), Switzerland (Altherr 1950b, 1952, 1953), the Netherlands (Loof &

Coomans 1970; Bongers 1988), United Kingdom (Wasilewska 1967) and USA (Vineiguerra 2006).

Pungentus marietani Altherr, 1950

Figs 11-K, 5

P. alpinus Vineiguerra & Giannetto, 1984: 49, figs 1-3.

Diagnosis

This speeies is distinguishable by the length of its body (1.15-1.66 mm), the lip region being offset by a

eonstrietion and 11-15 pm wide, an odontostyle being 22-28 pm long and 1.9-2.3 times lip region width

or 1.5-2.1% of total body length, neek length of 320-337 pm, a pharyngeal expansion 130-176 pm long

or 39-43% of total neek length, a didelphie-amphidelphie female genital system, a uterus being tripartite

and 56-68 pm long or 1.4-1.6 times the body diameter, V= 42-52, and the female tail being rounded

eonoid (19-26 pm, c = 52-74, c’ = 0.7-1.0).

Material examined

Five females from one loeation, in various states of preservation.

Morphometries

See Table 1.

Description

Female

Moderately slender to slender nematodes of medium size, 1.15-1.56 mm long. Habitus after fixation

ventrally eurved, espeeially in the posterior body region, C- or G-shaped. Body eylindrieal, tapering

towards the anterior end. Cutiele three-layered, espeeially pereeptible at the tail, 1.5-2.5 pm thiek in

anterior region, 2.0-2.5 pm at mid-body and 4.0-5.0 pm on dorsal side of tail; outer layer with very fine

transverse striation, intermediate layer thieker than the outer and inner layers. Lateral ehord 5.5-7.0 pm

wide or oeeupying 12-19% of mid-body diameter, very granular, but laeking any differentiation. Lateral

pores eomparatively eoarse, in two rows at the margins of lateral ehord; ventral pores well pereeptible

along entire body (Fig. IK), apparently eonneeting with sub-epidermal gland bodies. Lip region offset

by eonstrietion, 2.4-2.5 times as wide as high and ca. one-third (28-35%) of body diameter at neek

base. Lips moderately angular and separate; labial and eephalie papillae distinet, but not espeeially

protruding above eephalie eontour. Amphid fovea eup-shaped, opening at level of eephalie eonstrietion,

oeeupying 6.5 pm or about one-half (52%) of lip region diameter. Cheilostome almost eylindrieal,

its walls distinetly thiek and selerotized at the anterior half; perioral refraetive dots or platelets very

prominent. Odontostyle 10.0-12.5 times as long as wide, 1.8-2.3 times longer than lip region diameter

or 1.5-2.1% of total body length; aperture 3.0-5.0 pm long or 14-19% of its length. Odontophore rod¬

like, almost equal (0.9-1.1 times) to odontostyle. Guiding ring double, loeated at 15-16 pm or 1.2-1.5

times the lip region diameter from anterior end. Pharynx enlarging gradually; basal expansion 5.7-7.1

times as long as wide and 2.9-3.7 times the eorresponding body diameter long, oeeupying about two-

fifths (39-43%) of total neek length. Pharyngeal gland nuelei loeated as follows: DO = 52, DN = 56-

61, SjNj = 77, SjN 2 = 79, S 2 N = 87-90. Nerve ring at 120-128 pm from anterior end or 38% of total

neek length. Cardia rounded eonoid, 7-9 x 10-14 pm. Genital system didelphie-amphidelphie, with both

branehes well and equally developed, the anterior 169-182, the posterior 144-210 pm long. Ovaries 43-

162 pm long; ooeytes first in two or more rows, then in one row. Oviduet 82-118 pm long, or 2.2-2.9

PENA-SANTIAGO R., CIOBANU M. & ABOLAFIA J., Characterization of Iberian Pungentus

times the body diameter, eonsisting of a slender portion with prismatie eells and a moderately developed

pars dilatata. Oviduet-utems junetion marked by a distinet sphineter. Uterus 56-68 pm long or 1.4-1.6

times the body diameter, apparently tripartite, i.e., eonsisting of a proximal seetion with thiek walls and

distinet lumen, an intermediate narrowing, and a spheroid pars dilatata uteri. Vagina extending inwards

18-22 pm or about one-half (44-59%) of body diameter; pars proximalis almost as long as wide, 13-18 x

12-16 pm, with eonvergent walls and enveloped by weak eireular museulature; pars refringens not very

distinet in the females examined, but two pieees might be present; pars distalis 5.5-7.0 pm long. Vulva

a pre-equatorial, short transverse slit, whieh is preeeded by a depression of body surfaee. Prereetum 2.9-

4.2, reetum 0.9-1.1 anal body diameters long. Tail short and rounded eonoid, somewhat more straight

ventrally. Caudal pores two pairs, subdorsal, at the anterior half of tail.

Fig. 5. Pungentus marietani Altherr, 1950 ($, LM). A-C. Anterior region in median view. D. Posterior

genital braneh. E. Posterior body region. F. Phayngeal expansion. G. Phayngo-intestinal junetion. H.

Vagina. I-J. Caudal region. (Seale bars: A, C, D, G, J = 20 pm; B, H, I = 10 pm; E, F = 50 pm)

European Journal of Taxonomy 52: 1-17 (2013)

Male

Unknown.

Distribution

This species was collected from one location in the southern Iberian Peninsula: the province of Granada,

Sierra Nevada National Park, near road to Veleta summit (37°7’ N - 3°32’ W), in association with

Mediterranean brushwood Retama sphaerocarpa (Linnaeus, 1753) Boissier 1840 as dominant species.

Remarks

The specimens from the Iberian population of this species are nearly identical to those of the types

as described by Altherr (1950a) as well as to the type material of P. alpinus Vinciguerra & Giannetto,

1984, the latter regarded as a junior synonym by Ahmad et al. (2000). Some morphological features,

for instance the nature of the cuticle and the female genital system, are more accurately described here.

Pungentus marietani is certainly another widely distributed species in Europe, where it has been

reported from Germany (Ahmad et al. 2000), Italy (Vinciguerra & Giannetto 1984, as P. alpinus),

Poland (Brzeski 1963; Wasilewska 1967; Winiszewska-Slipinska 1987a, b, but see comments below),

Switzerland (Altherr 1950a) and the Netherlands (Bongers 1988, as P. alpinus). Besides, Vinciguerra

(2006) mentioned its occurrence in Serbia, Slovakia, Macedonia and Canada. The true identity of the

specimens studied by Winiszewska-Slipinska (1987a) remains uncertain, because of their larger general

size: body 1.5-1.9 mm long (n=27) and odontostyle 28-31 pm; this material probably belongs to another

species.

Pungentus silvestris (de Man, 1912)

Figs IL-N, 6

P. thornei Goodey, 1943: 6, figs 1-6.

Diagnosis

This species is characterized by the length of its body (1.50-2.20 mm), the lip region being offset by a

constriction and 12-18 pm wide, an odontostyle being 30-38 pm long and 2.3-2.7 times lip region width

or 1.7-1.9% of total body length, neck length of 366-507 pm, a pharyngeal expansion 160-247 pm long

or 46-56% of total neck length, a mono-opisthodelphic female genital system, a uterus being a simple

tube 98-124 pm long or 2.1-2.7 times the body diameter, V= 39-46, and the female tail rounded (26-34

pm, c = 43-90, c’ = 0.7-1.0).

Material examined

Twenty females from two locations, in various states of preservation.

Morphometries

See Table 1.

Description

Female

Slender nematodes of medium size, 1.65-2.10 mm long. Habitus after fixation ventrally curved,

especially in posterior body region, C- or G-shaped. Body cylindrical, tapering towards the anterior

end. Cuticle three-layered, 2.0-3.0 pm in anterior region, 2.5-3.0 pm at mid-body and 6.0-8.0 pm on

dorsal side of tail; its outer layer with very fine transverse striations and intermediate layer distinctly

thicker than inner and outer ones. Lateral chord 10-13 pm wide or occupying one-fifth to one-fourth

PENA-SANTIAGO R., CIOBANU M. & ABOLAFIA J., Characterization of Iberian Pungentus

Fig. 6. Pungentus silvestris (de Man, 1912) ($, LM). A-C. Anterior region in median lateral view. D.

Lip region in surfaee lateral view. E, I. Vagina and pre-vulval uterine sae. F-G: Posterior genital braneh.

H. Pharyngeal expansion. J. Caudal region. (Seale bars.- A-E, I, J = 10 pm; F-H = 50 pm)

European Journal of Taxonomy 52: 1-17 (2013)

(19-24%) of mid-body diameter, very granular but laeking any differentiation. Lateral pores obseure

in the speeimens examined; ventral pores well pereeptible (Fig. IN) along the entire body. Lip region

offset by eonstrietion, 2.1-2.6 times as wide as high and ca. one-third (29-35%) of body diameter at neek

base. Lips mostly amalgamated and rounded; labial and eephalie papillae distinet, but not espeeially

protruding above eephalie eontour. Amphid fovea eup-shaped, opening at level of eephalie eonstrietion,

oeeupying 8-9 pm or about three-fifths (55-62%) of lip region diameter. Cheilostome almost eylindrieal,

its walls distinetly thieker and selerotized at the anterior half; perioral refraetive dots or platelets very

prominent. Odontostyle 12.2-14.8 times as long as wide, 2.1-2.4 times longer than lip region diameter

or 1.7-1.9% of total body length; aperture 4.0-5.5 pm long or 13-18% of its length. Odontophore rod¬

like, hardly shorter (0.9 times) than odontostyle. Guiding ring double, loeated at 21-23 pm or 1.6 times

the lip region diameter from anterior end. Pharynx enlarging gradually; basal expansion 7.3-9.6 times

as long as wide and 4.4-5.6 times the eorresponding body diameter long, oeeupying about one-half

(48-56%) of total neek length. Pharyngeal gland nuelei loeated as follows: DO = 48-52, DN = 50-54,

SjNj = 71, SjN 2 = ?, S 2 N = 88-89. Nerve ring at 124-160 pm from anterior end or 31-37% of total

neek length. Cardia rounded eonoid, 13-18 x 14-17 pm. Genital system monodelphie-opisthodelphie.

Anterior braneh redueed to a very short pre-vulval, uterine sae 8-17 pm long or less than half (0.2-0.4

times) the body diameter. Posterior braneh well developed, 176-498 pm long: ovary 98-240 pm long,

with ooeytes first in two or more rows, then in one row; oviduet 75-122 pm long, or 1.7-2.3 times the

body diameter, eonsisting of a slender portion with prismatie eells and a moderately developed pars

dilatata. Sphineter distinet between oviduet and uterus. Uterus apparently a simple tube 98-124 pm long

or 2.1-2.7 times the body diameter. Vagina extending inwards 23-27 pm or about one-half (50-58%) of

body diameter; pars proximalis almost as long as wide, 14-17 x 12-14 pm, with eonvergent walls and

enveloped by weak eireular museulature; pars refringens with two weakly selerotized pieees, 3.5-4.0 x 5

pm, and with a eombined width of about 9-10 pm; pars distalis 3.0-3.5 pm long. Vulva a pre-equatorial,

short transverse slit, preeeded by a depression of the body surfaee. A prevulval, gland-like strueture

(‘paravulva’) present in one speeimen (Fig. 61). Prereetum 3.1-3.7, reetum 0.8-1.2 anal body diameters

long. Tail short and rounded. Caudal pores two pairs, one sublateral, another subdorsal, at the middle of

the tail.

Male

Not found.

Distribution

This speeies has been eolleeted from two loeations in the southern Iberian Peninsula: the provinee

of Jaen, Cazorla, Segura y Las Villas Natural Park, near the road to the Segura river souree (38° 17’

N - 2°39’ W), in assoeiation with pine forest; and the provinee of Granada, Sierra Nevada National Park,

near the road from Capileira to Veleta (36°57’ N - 3°21’ W), in a pine forest and among herbaeeous

plants where the dominant speeies v^?iS Asphodelus albus Miller, 1768.

Remarks

The present deseription fits those available from the literature. It agrees espeeially well with the re-

deseription of the speeies by Coomans & Geraert (1962). These authors regarded P. thornei Goodey,

1943 as a junior synonym of P. silvestris, an aetion that seems to be totally justified and herein aeeepted.

Pungentus silvestris might be a eosmopolitan speeies. It has been reported from many European eountries

sueh as Belgium (Coomans & Geraert 1962), Bulgaria (llieva et al. 1993), Denmark (Yeates 1972, as

P. thornei), Germany (Altherr 1968, as P. thornei), Italy (Marinari et al. 1980), Poland (Winiszewska-

Slipinska 1987), Russia (Skarbilovieh & Ovsjukova 1989), Spain (Momeal & Campoy 1982, as P.

thornei). The Netherlands (de Man 1912; Coomans & Geraert 1962; Bongers 1988) and United Kingdom

(Goodey 1943, as P. thornei).

PENA-SANTIAGO R., CIOBANU M. & ABOLAFIA J., Characterization of Iberian Pungentus

Acknowledgements

The authors espeeially thank the finaneial support reeeived from the projeets Fauna Iberica: Nematoda,

Dorylaimoidea (excepto Longidoridae) (Spanish Ministry of Seienee and Innovation, ref CGL2007-

66786-C08-08; eo-finaneed FEDER) and Revision de la familia Nordiidae (Consejeria de Innovaeion,

Cieneia y Empresa, Andalusian Regional Government, Spain; ref RNM-475). SEM pietures were

obtained with the assistanee ofteehnieal staff and the equipment of‘Servieios Teenieos de Investigaeion’,

University of Jaen.

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Manuscript received: 5 April 2013

Manuscript accepted: 1 July 2013

Published on: 5 August 2013

Topic editor: Rudy Jocque

Desk editor: Kristiaan Hoedemakers

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

the FJT eonsortium: Museum National d’Histoire Naturelle, Paris, Franee; National Botanie Garden

of Belgium, Meise, Belgium; Royal Museum for Central Afriea, Tervuren, Belgium; Natural History

Museum, Fondon, United Kingdom; Royal Belgian Institute of Natural Seienees, Bmssels, Belgium;

Natural History Museum of Denmark, Copenhagen, Denmark.