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Nematodes of the Order Rhabditida from Andalucía Oriental, Spain. the Genera Protorhabditis (Osche, 1952) Dougherty, 1953 and D

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Nematodes of the Order Rhabditida from Andalucía Oriental, Spain. the Genera Protorhabditis (Osche, 1952) Dougherty, 1953 and D Journal of Nematology 39(3):263–274. 2007. © The Society of Nematologists 2007. Nematodes of the Order Rhabditida from Andalucía Oriental, Spain. The Genera Protorhabditis (Osche, 1952) Dougherty, 1953 and Diploscapter Cobb, 1913, with Description of P. spiculocrestata sp. n. and a Species Protorhabditis Key J. Abolafia,R.Pen˜a-Santiago Abstract: A new species of the genus Protorhabditis is described from natural areas in the SE Iberian Peninsula. Protorhabditis spiculocrestata sp. n. is distinguished by its body length 387–707 µm in females and 375–546 µm in males, lip very low and flattened, stoma 14–22 µm long, female tail conical-elongate (48–100 µm, c = 6.4–8.3, cЈ = 4.8–7.5), phasmid near anus, male tail conical (20–27 µm, c = 18.3–22.3, cЈ = 1.4–1.5), bursa peloderan closed anteriorly and bears eight papillae (1+2+1+1+3), spicules 23–26 µm long, and gubernaculum 10–16 µm long. Diploscapter coronatus is also presented. Description, measurements and illustrations, including SEM photographs, are provided. A key to species of Protorhabditis is also given as well a compendium of their measure- ments. Key words: description, Diploscapter, key, morphology, new species, Protorhabditis, Rhabditids, SE Spain, SEM, taxonomy. Rhabditid nematodes are an interesting zoological mis (Bütschli, 1873) Sudhaus, 1976, P. tristis (Hirsch- taxon. They are very abundant in all types of soil and mann, 1952) Dougherty, 1955 and D. coronatus (Cobb, sediments of freshwater bodies and play important eco- 1893) Cobb, 1913. Nevertheless, no relevant morpho- logical roles mainly as primary consumers—their free- logical or taxonomical information on them was pro- living forms display saprophagous or bacteriophagous vided in the corresponding publications. In this contri- feeding habits—but also as animal parasites, in particu- bution, part of the series on rhabditid species from lar enthomopathogenic forms. From a systematic point Andalucía Oriental (SE Iberian Peninsula, Spain), D. of view, rhabdits are a difficult nematode group whose coronatus and an undescribed species of Protorhabditis classification has been matter of long and strong dis- are reported. cussions and whose diversity is far from being well known. Materials and Methods Several years ago, a research project was planned to Nematodes were extracted from soil samples by study the Iberian rhabditid fauna, focused on both its Flegg’s (1967) method and a somewhat modified Baer- morphological characterization and its taxonomy, since mann’s (1917) funnel technique. Nematodes obtained no monographic study on this matter was available. were later relaxed and killed by heat, fixed in 4% form- The genera Protorhabditis (Osche, 1952) Dougherty, aldehyde, and processed to anhydrous glycerine ac- 1953 and Diploscapter Cobb, 1913 are two rhabditid taxa cording to Siddiqi (1964). Measurements were taken characterized by having a long stoma whose stegostom using an ocular micrometer, and drawings were made lacks glottoid apparatus. They were classified by An- using a drawing tube attached to a Leica microscope; drássy (1983) in separate families (Rhabditidae Örley, LM pictures were made using a Nikon Eclipse 80i mi- 1880 and Diploscapteridae Micoletzky, 1922, respec- croscope provided with a digital video camera Nikon tively) of the superfamily Rhabditoidea Örley, 1880, Digital Sight DS-5M. For SEM studies, fixed specimens mainly due to morphology of lip region: six rounded were hydrated in distilled water, dehydrated in a graded lips with similar aspect in Protorhabditis; subdorsal and ethanol and acetone series, critical point dried and subventral lips hook-like and lateral lips membranous coated with gold (Abolafia and Peña-Santiago, 2005), and rounded with dentate margin in Diploscapter. How- and observed with a JEOL JSM-5800 microscope. The ever, more recent evidence based on both morphologi- terminology used for morphology of stoma and spicules cal and molecular studies demonstrated a close rela- follows the proposals by De Ley et al. (1995) and Abo- tionship between both genera (Fitch, 2000; Sudhaus lafia and Peña-Santiago (2006), respectively. and Fitch, 2001). In addition, the D. coronatus specimens examined Three species belonging to these two genera have were cultured in vitro in petri dishes (2% agar and been recorded from several localities in the Iberian 1.5% glucose) and maintained under laboratory condi- Peninsula (Abolafia and Peña-Santiago, 2001): P. filifor- tions for several months. This culture is no longer avail- able. Received for publication July 25, 2007. Description Departamento de Biología Animal, Biología Vegetal y Ecología, Universidad de Jaén. Campus “Las Lagunillas” s/n. 23071-Jaén, Spain. The authors thank Dr. W. Sudhaus for his examination of type material of Protorhabditis spiculocrestata sp. n. Protorhabditis spiculocrestata, Research Technical Services of University of Jaén (Figs. 1–3) (Spain) for the assistance in SEM study of the material, and the financial support received from the project entitled “Fauna Ibérica VIII” (CGL2004– The specific epithet refers to the presence of a sub- 04680-C10–09 / BOS). E-mail: abolafi[email protected] dorsal crest in the spicules, a remarkable morphologi- This paper was edited by Zafar Handoo. cal feature of this species. 263 264 Journal of Nematology, Volume 39, No. 3, September 2007 mus. Excretory pore at 82–92% of neck length, at level of isthmus base or basal bulb, behind the hemi- zonid. Deirid at basal bulb level, at 90–97% of neck length. Reproductive system didelphic-amphidelphic, with anterior branch dextral and posterior branch sinistral. Ovaries having a flexure. Uterus two times the corresponding body diameter long, having thicker walls at proximal portion and distinct lumen at distal one. Eggs 15–23 × 37–39 µm. Vagina extend- ing inwards to one-third of body diameter, with thicker walls at proximal portion. Rectum 1.1–1.3 times as long as anal body diameter. Tail conical- elongate, and terminus with acute mucro. Phasmid near anus. Male (n = 3): Similar to female in general morphol- ogy. Habitus ventrad curved after fixation. Reproduc- tive system monorchic. Testis reflexed ventrally anteri- orly. Tail conical. Bursa pelodera, closed anteriorly, and with eight papillae (1+2+1+1+3). Spicules free: ma- nubrium rounded; lamina bent at its middle, with well developed dorsal hump and ventral wing at its middle; terminus ventrad curved, with acute tip and a subdorsal crest. Gubernaculum sigmoid. Other material examined (see Table 1) Very similar to type population, but showing varia- tions in body length, shorter in the population from Almijara Mountain (387 µm in females and 395–405 µm in males), and longer in the populations from Al- FIG.1. Protorhabditis spiculocrestata sp. n. A: Neck. B: Anterior end. margen and Sierra de las Nieves (634 µm and 644–707 C: Female reproductive system. D: Entire male. E: Female posterior µm in females, respectively). end. F: Entire female. G, H: Male posterior end (lateral side). J: Idem (ventral side). I: Spicule. Diagnosis Measurements: Listed in micrometers in Table 1. Protorhabditis spiculocrestata sp. n. is distinguished by Type population collected near road from Fuente de Piedra its body length 387–707 µm in females and 375–546 µm to Alameda, province of Málaga in males, lips very low and flattened, stoma 14–22 µm, Female (n = 7): Small nematodes, 441–561 µm long. female tail conical-elongate (48–100 µm, c = 6.4–8.3, Body cylindrical, tapering towards both ends, more cЈ = 4.8–7.5), phasmid near to anus, male tail conical so posteriad. Habitus rather sigmoid after fixation. Cu- (20–27 µm, c = 18.3–22.3, cЈ = 1.4–1.5), bursa ticle with distinct transverse striations or annuli, about peloderan closed anteriorly and bears eight papillae 1 µm wide at midbody. Lateral field with five inci- (1+2+1+1+3), spicules 23–26 µm long, and gubernacu- sures or four ridged wings at midbody, two at posterior lum 10–16 µm long. region which fade out near anus. Lip region with six small lips, more or less rounded; papillae visible. Stoma Relationships rhabditoid, lacking glottoid apparatus. Cheilostom The new species resembles Protorhabditis oxyuroides with weakly refractive walls. Buccal ring (membrane Sudhaus, 1974, although it can be distinguished from it wedge ring) lower in longitudinal section. Buccal prism by its having lips lower and reduced (vs. more promi- with straight walls, very long and narrow. Pharyngeal nent), stoma shorter (14–22 µm vs. 21–27 µm), and collar very short. Procorpus cylindrical. Metacorpus spicules with different morphology (vs. terminus lack- swollen, almost ovoid, with its inner walls more re- ing subdorsal crest). fractive. Isthmus almost as long as procorpus. Basal bulb small, ovoid or pyriform. Cardia conoid to hemi- Type locality and habitat spherical, surrounded by intestinal tissue. Intestine lacking distinct specializations, but a cardiac portion Los Carvajales, near the road from Alameda to Fu- is sometimes differentiated at its anterior part. Nerve ente de Piedra, province of Málaga, Spain; GPS coor- ring at 71–80% of neck length, surrounding the isth- dinates: N 37°8.0Ј, E 4°43.0Ј. Soil in a Mediterranean Spanish Rhabditida: Abolafia and Peña-Santiago 265 FIG.2. Protorhabditis spiculocrestata sp. n. (LM). A: Neck. B: Female reproductive system (anterior branch). C: Female tail. D. Male posterior end (bursa view). E. Male posterior end (spicule view). plant community whose dominant species are Olea eu- carpa (L.) Boiss., Phlomis purpurea L., Thymus sp., Quer- ropaea L. var. sylvestris Brot., Daphne gnidium L., Rosmari- cus coccifera L., and Ulex parviflorus Pourret. Collected nus officinalis L., Cistus monspeliensis L., Retama sphaero- on 19 October 1991. 266 Journal of Nematology, Volume 39, No. 3, September 2007 FIG.3. Protorhabditis spiculocrestata sp. n. (SEM). A: Lip region (arrow indicates amphid opening). B: Lateral field at middle body. C: idem at posterior end. D: idem near to anus. E: Phasmid. F, G: Male posterior end.
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