Journal of Nematology 27(3):387-394. 1995. © The Society of Nematologists 1995. Noctuidonema dibolia n. sp. (Aphelenchida: Acugutturidae), an Ectoparasite of the latipes (: Noctuidae) 1

O. G. MARTI, JR, AND C, E. ROGERS 2

Abstract: Noctuidonema dibolia n. sp., an ectoparasite of adults of the noctuid moth (Guen6e) is described. The differentiating characters are a club-shaped body with a subterminal vulva in the female, spicules with a reduced matrix and sheath and closely apposed dorsal and ventral arms in the male, very long styler and conus, moderately prominent stylet knobs, a bluntly rounded head, and a large renette cell in both sexes. Lateral fields, rectum, anus, bursa, and gubernaculum are absent. Noctuidonema dibolia differs from the other species of the subfamily Noc- tuidonematinae in the size and robustness of the body, the length of the stylet and conus, the length of the tail, and the shape of the spicules. Key words: ectoparasite, parasite, Lepidoptera, Mocis latipes, moth, nematode, Noctuidae, Noctuidonema dibolia n. sp., .

Surveys of Lepidoptera in French Gui- MATERIALS AND METHODS ana (9,12), the tropical Americas (11,12), Mocis latipes adults were collected from and the southeastern United States (10) 1991-93 in a walk-in ultraviolet light trap documented the widespread occurrence of located on a research farm near Tifton, an ectoparasitic nematode, Noctuidonema Georgia. The external surfaces of the guyanense Remillet and Silvain, particularly moth abdomens were examined with a ste- among in the family Noctuidae. reoscopic microscope for the presence of Noctuidonema Remillet and Silvain is of in- nematodes. Infested moths were placed terest because it is one of only a few known into a 15-ml cup containing 2 ml of water metazoan parasites of adult moths (3) and plus a few drops of Tween 80 (polyoxyeth- is currently under study as a possible bio- ylenesorbitan monooleate) and shaken for logical control agent for the fall armyworm a few seconds to dislodge nematodes. moth, Spodoptera frugiperda (J. E. Smith), a Nematodes were concentrated by sedi- serious pest of corn, sorghum, and pasture mentation in distilled water, examined live grasses (12,13). or killed in hot water (60 C), and fixed Mocis latipes (Guen6e), whose larvae are and stored in TAF (Triethanolamine-For- known as striped grass loopers, is prima- malin-Water 7:2:91) (4) until processed rily a pest of pasture grasses (5,12). Speci- through an alcohol series to glycerin. The mens of the Noctuidonema, collected nematodes were then mounted on glass from M. latipes in Tift County, Georgia, slides and examined with differential in- were examined and found to differ from terference contrast or phase contrast op- N. guyanense, Vampyronema daptria (Ander- tics. Drawings are based on microscopic son and Laumond) (2,7), and the related examination and photographs of living as genus Acugutturus Hunt (6), and are here well as processed specimens. Values in the described as a new species, N. dibolia. The text are expressed as: range in p~m (mean specific name 'dibolia' is taken from Greek in txm -+ SD). (= pointed at both ends).

SYSTEMATICS Received for publication 19 September 1994. i Mention of a trade name, warranty, proprietary product, Noctuidonema dibolia n. sp. or vendor does not constitute an endorsement of a product and does not imply its approval to the exclusion of other (Figs. 1A-C;2A-E) products or vendors that may also be suitable. Microbiologist and Research Entomologist, USDA, ARS, Holotype ~emale): Length = 522 Ixm; Insect Biology and Population Management Research Labo- maximum width = 57 Ixm; width at vulva ratory, P.O. Box 748, Georgia Coastal Plain Experiment Sta- tion, Tifton, GA 31793. = 44 i~m; anterior end to excretory pore 387

...... [I ...... I ...... I 388 Journal of Nematology, Volume 27, No. 3, September 1995

= 22 ixm; anterior end to nerve ring = 16 ~m; anterior end to excretory pore = 137 Ixm; stylet length = 108 ~m; styler co- 25 >m; anterior end to nerve ring = 118 nus length = 91 p.m; metacorpus length ~m; stylet length = 89 p.m; stylet conus = 28 Ixm; metacorpus width = 23 ~xm; length = 73 Ixm; metacorpus length = 28 ovary length = 225 p.m; tail length = 66 Ixm; metacorpus width = 20 p~m; testis /~m. length = 225 p~m; tail length = 8 jxm; Allo~pe (male): Length = 464 p~m; max- spicule length = 44 Ixm; rostrum length = imum width = 41 ixm; width at cloaca = 12 ~xm.

7(

5(

2

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Flo. 1. Noctuidonemadibolia n. sp. A) Female. B) Male. C) Spicules. Noctuidonema dibolia n. sp.: Marti and Rogers 389

a ~ C /V ~ ii ',

60um A

10urn B 10um C

,25urn D

30um E

FIG. 2. Noctuidonemadibolia n. sp. A) Fourth-stagejuvenile. B) Head end of female. C) Head end of male. D) Newly hatched second-stagejuvenile. E) Embryonatedegg with fully formed juvenile.

Paratypes: Measurements of 39 females 0.2) at midbody, anastomoses common. and 16 males in Tables 1-4. Lateral fields absent. Lip region low, bluntly rounded, not strongly set off. Description Styler flexible, delicate, elongate, repre- Female: Measurements in Tables 1-3. senting 19-22% of total body length. Co- Body club-like, slightly arcuate ventrally. nus representing 80-82% of total stylet Annules continuous, 1.0-1.6 wide (1.3 -+ length. Conus midpoint diameter 0.8-1.0 390 Journal of Nematology, Volume 27, No. 3, September 1995

TABLE 1. Measurements of 14 small paratype ing on host diet, containing amorphous in- (<500 p.m body length), non-gravid female Noctui- gesta, crystalline inclusions, and bacteria. donema dibolia n. sp. from Tift County, Georgia. Intestine ending in a blind, rounded sac Character ~ Mean SD Range extending posterior to vulva. Esophageal glands 45.3-65.7 long (59.7 +- 6.1) in small Measurements (p,m) non-gravid females, 43.8-77.4 long (66.7 Body length 455 37.2 396-495 Maximum body width 54 7.3 44-67 -+ 8.8) in large non-gravid females, and Body width at nerve ring 36 3.1 32-41 62.8-81.8 (72.8 +- 6.8) in large gravid fe- Body width at vulva 41 5.9 34-54 males. Body width at anterior 36 3.1 32-41 Ovary outstretched, .rarely reflexed, fre- end of intestine Anterior end to base 120 9.8 105-140 quently extending anterior to metacorpus of bulb in large specimens. Rachis present in large Metacorpus length 27 2.0 23-31 specimens. Oocytes progressively larger in Metacorpus width 23 1.8 20-26 posterior ovary, usually not more than one Excretory pore to 18 2.9 15-23 anterior end Stylet total length 99 6.1 83-108 TABLE 2. Measurements of 13 large paratype Stylet conus length 81 5.3 69-89 (>500 ~m body length), non-gravid female Noctui- Stylet shaftqength 18 3.0 15-23 donema dibolia n. sp. from Tiff County, Georgia. Head width 9 0.8 7-10 Head length 4 0.5 3-4 Character a Mean SD Range Ovary length 200 37.4 149-262 Vulva to end of tail 53 7.4 42-70 Measurements (~m) Vulva to anterior end 402 33.4 346-433 Body length 526 25.7 501-568 Vagina depth 24 4.1 18-31 Maximum body width 57 6.6 44---67 Anterior end to 129 25.4 85-187 Body width at nerve ring 36 4.6 28--44 anterior ovary Body width at vulva 44 6.7 32-54 Body width at anterior 36 5.3 26~4 Ratios and Percentages end of intestine a 8.5 0.7 7-10 Anterior end to bulb base 125 13.6 96-143 b 3.8 0.4 3-5 Metacorpus length 28 2.8 20-31 c 8.6 1.0 7-11 Metacorpus width 24 1.9 20-26 c' 1.3 0.2 1-2 Excretory pore to 19 2.5 15-23 V 88.3 1.3 86-91 anterior end M 82.0 2.6 76-86 Stylet total length 100 6.8 92-114 G 59.7 6.8 48-70 Stylet conus length 81 4.6 73-88 Ovary, % of body length 43.7 5.9 36-53 Head width 9 0.9 7-10 Stylet, % of body length 21.8 2.3 17-26 Head length 4 1.3 3-7 a a = body length/greatest body width; b = body length/ Ovary length 214 54.3 117-306 distance from anterior end to junction of esophagus and in- Vulva to end of tail 62 8.7 47-79 testine; c = body length/tail length; c' = tail length/body Vulva to anterior end 467 23.8 435-505 width at vulva; V = distance of vulva from anterior end x Vagina depth 27 5.4 15-34 100/body length; M = conus length x 100/styletlength; G = Anterior end to 136 28.6 67-175 distance from vulva to anterior end of ovary x 100/body anterior ovary length. Ratios and Percentages a 9.3 1.0 8-11 (1.0 -+ 0.1). Shaft diameter 1.0-1.6 (1.4 + b 4.2 0.4 4-5 0.3). Styler knobs rounded with concave c 8.7 1.1 7-11 anterior surfaces, 4.0-5.0 (4.6 + 0.4) wide, c' 1.5 0.3 1-2 V 88.7 1.8 86-92 6.0-20.0 (12.9 ± 4.9) from guide ring. M 81.1 4.7 73-89 Procorpus strongly reflexed at level of G 62.6 5.6 54-75 metacorpus when stylet is retracted. Meta- Ovary, % of body length 40.9 10.9 21-61 corpus ovoid, with a sclerotized valve Styler, % of body length 18.9 0.9 17-20 slightly offset posteriorly. Nerve ring im- a = body length/greatest body width; b = body length/ distance from anterior end to junction of esophagus and in- mediately posterior to metacorpus. Hemi- testine; c = body length/tail length; c' = tail length/body zonid position variable, 0-23.4 (10.4 ± 8.4) width at vulva; V = distance of vulva from anterior end x 1001body length; M = conus length x 100/stylerlength; G = posterior to nerve ring in gravid females. distance from vulva to anterior end of ovary x lOO/hody Intestinal contents usually yellow, depend- length. Noctuidonema dibolia n. sp.: Marti and Rogers 391

TAaLE 3. Measurements of 12 gravid paratype and posteriorly toward the tail. Renette female Noctuidonema dibolia n. sp. from Tift County, cell nucleus large, 7.3-10.2 (8.8 + 1.2) in Georgia. gravid females, irregular in outline in fixed specimens, smooth in live specimens. Character ~ Mean SD Range Tail shape conical, with mucro small or Measurements (p.m) absent. Anus not visible with light micros- Body length 541 54.4 451-640 copy. Vulva subterminal (V = 88%), trans- Maximum body width 67 11.3 53-88 Body width at nerve ring 40 4.8 31--47 verse, with overlapping anterior lip. One Body width at vulva 52 8.5 42-67 pair of prevulval and one pair of postvul- Body width at anterior 40 4.8 31-47 val papillae. end of intestine Anterior end to bulb base 132 9.6 117-143 Males: Measurements in Table 4. Body Metacorpus length 28 2.8 23-32 hook-shaped, tapering gradually at ante- Metacorpus width 23 2.0 19-26 Excretory pore to 21 3.0 15-25 anterior end TABLE 4. Measurements of 16 paratype male Stylet total length 102 7.0 91-114 Noctuidonema dibolia n. sp. from Tift County, Georgia. Stylet cone length 82 3.3 76-86 Stylet shaft length 20 6.0 10-31 Character ~ Mean SD Range Head width 9 1.5 6-10 Head length 4 1.0 3-6 Measurements (v.m) Ovary length 258 38.2 198-320 Body length 422 48.6 343-524 Vulva to end of tail 68 5.4 60-79 Maximum body width 42 3.1 38-51 Vulva to anterior end 474 52.1 388-562 Body width at nerve ring 31 2.5 28-35 Vagina depth 30 7.3 20--45 Body width at cloaca 14 2.3 10-18 Intrauterine egg length 86 8.5 66-98 Body width at anterior 31 2.5 28-35 Intrauterine egg width 30 2.5 26-35 end of intestine Anterior end to 95 38.1 60-175 Anterior end to base 106 14.0 77-126 anterior ovary of bulb Metacorpus length 26 1.5 23-28 Ratios and Percentages Metacorpus width 20 2.1 16-23 a 8.2 1.1 7-10 Excretory pore to 21 4.2 15-31 b 4.1 0.3 4-5 anterior end c 8.0 0.7 7-10 Stylet total length 86 4.4 76-93 c' 1.3 0.2 1-2 Styler cone length 68 4.8 61-82 V 87.6 1.3 86-90 Stylet shaft length 18 5.1 16-26 M 80.7 4.7 73-89 Head width 8 1.0 7-10 G 69.7 6.3 57-77 Head length 4 0.9 3-6 Ovary, % of body length 47.8 3.7 41-54 Testis length 144 47.5 85-233 Stylet, % of body length 18.9 2.1 14-21 Spicule length 48 3.5 41-53 Spicule rostrum length 12 1.2 10-13 a a = body length/greatest body width; b = body length/ distance from anterior end to junction of esophagus and in- Cloaca to end of tail 9 1.1 6-10 testine; c = body length/tail length; c' = tail length/body Cloaca to anterior end 413 49,0 333-515 width at vulva; V = distance of vulva from anterior end x Anterior end to 132 21.8 98-174 100/body length; M = conus length x 100/styletlength; G = anterior testis distance from vulva to anterior end of ovary x 100/body length. Ratios and Percentages a 10.0 1.1 8--12 b 4.0 0.3 3-5 mature egg in uterus. Bacteria frequently c 50.8 1 I. 1 34-80 present, particularly in anterior ovary, and c' 0.7 0.2 0-1 V 98.0 0.4 97-99 sometimes seen in intestinal epithelial cells, M 78.7 5.5 70-93 intestinal lumen, and eggs. Uterine glands T 66.6 4.4 60-74 not seen. Spermatheca ovoid, thick-walled, Testis, % of body length 34.6 12.1 21-56 usually containing sperm. Intrauterine Stylet, % of body length 20.7 2.1 17-24 eggs 30 x 86 p,m, usually in 1-8 cell stage. a = body length/greatest body width; b = body length/ distance from anterior end to junction of esophagus and in- Excretory pore 14-25 p.m from anterior testine; c = body length/tail length; c' = tail length/body end. Renette cell large, about 20 x 60 jxm, width at cloaca; V = distance of cloaca frmn anterior end x 100/body length; M = conus length x 100/styletlength; T = consisting of a loosely coiled tubule ex- distance from cloaca to anterior end of testis x 100/body tending anteriorly to the excretory pore length. 392 Journal of Nematology, Volume 27, No. 3, September 1995 rior end, with posterior half strongly arc- similar to female. Renette cell nucleus 4.4- uate ventrally. Annules continuous, 1.2- 10.2 (7.6 +- 1.7), similar to female. 1.8 wide (1.4 +- 0.2) at midbody, anasto- moses common. Lateral field absent. Lip Type Host and Locality region low, bluntly rounded, not strongly Ectoparasitic on adults of Mocis latipes set off, similar to female. (Guen6e) (Lepidoptera: Noctuidae), par- Stylet flexible, delicate, elongate, atten- ticularly on intersegmental membranes of uated at tip, 17-24% of total body length. the abdomen. Type locality is the Bel- Conus representing about 79% of total flower Farm, located 5 km NNW of stylet length. Stylet knobs rounded, with Tifton, Tift County, Georgia. concave anterior surfaces. Stylet conus di- ameter 0.7-1.0 (0.9 + 0.1), shaft diameter Type Designations 1.0-1.6 (1.2 +- 0.2), knob width 2.6-5.0 Holotype (female) (slide T-519t), allo- (3.8 -+ 0.8). Knobs to guide ring 6.8-17.0 type (male) (slide T-520t), 5 paratype (13.9 -+ 3.0). slides (slides T-4526p-T-4530p), and one Metacorpus ovoid, with a sclerotized val- vial of paratypes in glycerin (vial T-370p) vular apparatus offset slightly posteriorly. deposited in the U.S. Department of Agri- Procorpus strongly flexed at level of meta- culture Nematode Collection, Nematology corpus. Esophageal glands 36.5-71.5 (58.1 Laboratory, USDA ARS, BARC-West, + 11.38) long, overlying anterior intestine. Beltsville, Maryland 20705. Four paratype Nerve ring immediately posterior to meta- slides deposited at each of the following: corpus. Hemizonid position variable, 7.3- Museum National d'Histoire Naturelle, 33.6 (15.9 -+ 8.5) posterior to nerve ring. Laboratoire des Vers, Paris, France; Insti- Intestine a blind sac, terminating well an- tuut voor Dierkunde, Gent, Belgium; Bio- terior to cloaca. Intestinal contents usually systematics Research Centre, Canadian yellow, similar to female. Tail short, blunt, National Collection of Nematodes, Canada with one pair of precloacal and one pair of Agriculture, Ottawa, Ontario, Canada; postcloacal papillae. Tail mucro absent or Rothamsted Experimental Station, Nema- low. Spicules paired, sclerotized, nearly tode Collection, Entomology and Nem- equal in size and shape, ventrally arcuate, atology Department, Harpenden, Herts, with dorsal and ventral arms not well sep- United Kingdom. arated, ending in fine points closely ap- posed at tips. Matrix and sheath present, Diagnosis but greatly reduced. Rostrum slender, ar- All stages of N. dibolia n. sp. are ectopar- cuate, distal end slightly enlarged. Manu- asitic on adult Mocis latipes. Stylet length is bria well developed, wedge-shaped, with 83-114 Ixm in females, 76-93 ~m in males, right slightly more rounded than left. Gu- with a conus 73-89% of total styler length bernaculum and bursa absent. Cloacal cyl- in females and 70-93% in males. Cephalic inder thin-walled, projecting 4-6 Ixm end similar in males and females. Tail nearly perpendicular to tail axis during sp- length is 42-79 ixm in females and 6-10 icule protraction. txm in males. Spicules have dorsal and ven- Testis single, outstretched anteriorly, tral arms closely apposed, not separated; without rachis. Anterior end of testis usu- the matrix and sheath are present but ally not extending anterior to metacorpus. much reduced. A large renette cell is Seminal vesicle containing spermatids and present in both sexes. set off from remainder of testis by a mod- erate constriction. Bacteria commonly Relationships present in testis. Males of N. dibolia n. sp. are smaller and Excretory pore 15-30 ~m from anterior more slender than those of N. guyanense, end. Renette cell large, about 20 × 50 txm, have shorter stylets and shorter styler coni, Noctuidonema dibolia n. sp.: Marti and Rogers 393 and have spicules that differ in size and also reported the presence of stylet knobs structure. The spicules of N. dibolia n. sp. in N. daptria. Stylet knobs are present in N. are smaller and simpler than those of dibolia. We examined Noctuidonema speci- N. guyanense and have a greatly reduced mens from several species of Spodoptera (S. matrix and sheath. In N. dibolia n. sp., frugiperda, S. ornithogalli, S. latifascia, and S. the spicules have well-developed, wedge- androgea) from the United States and shaped manubria, not reduced as in V. French Guiana and found that styler knobs daptria, and have dorsal and ventral arms were present in all Noctuidonema specimens closely apposed, not widely separated as in examined. During processing of N. dibolia V. daptria. Males ofN. dibolia n. sp. are sim- specimens for this report, we observed that ilar in size to V. daptria males, but have the basal part of the styler frequently be- longer stylets and longer styler coni. comes transparent, rendering the knobs Female N. dibolia n. sp. are smaller and difficult to see. However, the knobs are more slender than those of N. guyanense easily visible in living specimens. Our ob- and have a shorter stylet, shorter stylet co- servations indicate that stylet knobs are nus, and longer tail. Female N. dibolia n. sp. present in all species of Noctuidonema and differ from those of V. daptria in having a that this feature unites the known species longer stylet, longer styler cone, and of the subfamily Noctuidonematinae and shorter tail. The excretory pore is farther aids in differentiating them from Acugut- from the anterior end in N. dibolia than in turus. Spicule structure in N. guyanense and V. daptria. N. dibolia are similar enough to justify grouping them within a single genus, and DISCUSSION different enough to identify them as sepa- rate species and separate them from the Hunt (7) erected the family Acuguttu- related genus Vampyronema. ridae to include the genera Acugutturus, Noctuidonema, and the new genus Vampy- LITERATURE CITED ronema. Remillet and Silvain (8) placed 1. Anderson, R. V., and C. Laumond. 1990. Noc- Noctuidonema in the subfamily Acugutturi- tuidonema guyanense Remillet & Silvain, 1988: Mor- nae and distinguished it from Acugutturus phologie du spicule, redescription du mille et diag- by differences in body shape, stylet length, nose amend6e du genre Noctuidonema. Revue de N6matologie 13:433---436. shape of head and tail, position of the ex- 2. Anderson, R. V., and C. Laumond. 1992. Noc- cretory pore, shape and size of the spi- tuidonema daptria, n. sp. (Nematoda:Aphelenchoi- cules, and presence of a bursa. Anderson didae), an ectoparasite of the moth Lesmone porcia and Laumond (2) described Noctuidonema (Stoll). Journal of Nematology 24:16-22. 3. Ashley, T. R. 1979. Classification and distribu- daptria, which Hunt (7) later redescribed as tion of fall armyworm parasites. Florida Entomologist Vampyronema and placed with Noctuidonema 62:114-123. in the newly created subfamily Noctui- 4. Courtney, W. D., D. Polley, and V. L. Miller. donematinae. t955. TAF, an improved fixative in nematode tech- nique. Plant Disease Reporter 39:570-571. Hunt (7) noted that basal swellings and 5. Dean, T.W. 1985. Behavioral biology of the knobs were absent on the stylet in A. para- striped grass looper, Mocis latipes (Guen6e), in north- siticus, and Remillet and Silvain (8) re- central Florida. Ph.D. dissertation, University of Flor- ported that stylet knobs were present in N. ida, GainesviUe. guyanense. Anderson and Laumond (1) 6. Hunt, D.J. 1980. Acugutturus parasiticus n. g., n. sp., a remarkable ectoparasitic aphelenchoid nema- provided more details on the morphology tode from Periplaneta americana (k.), with proposal of of the spicule and redescribed the male of Acugutturinae n.subf. Systematic Parasitology 1:167- N. guyanense. However, they also reported 170. that the basal part of the styler of N. gu- 7. Hunt, D.J. 1993. Aphelenchida, Longidoridae, and Trichodoridae: Their systematics and bionomics. yanense is uniformly tubular, without swell- CAB International, Wallingford, UK. ings or knobs. Anderson and Laumond (2) 8. Remillet, M., and J.-F. Silvain. 1988. Noc- 394 Journal of Nematology, Volume 27, No. 3, September 1995 tuidonema guyanense n. g., n. sp., (Nematoda: Aph- 11. Simmons, A. M., and C. E. Rogers. 1990. Dis- elenchoididae) ectoparasite de noctuelles du genre tribution and prevalence of an ectoparasitic nema- Spodoptera (Lepidoptera: Noctuidae). Revue de tode, Noctuidonema guyanense, on moths of the fall ar- Nfmatologie 11:21-24. myworm (Lepidoptera: Noctuidae) in the tropical 9. Rogers, C. E., O. G. Marti, A. M. Simmons, and Americas. Journal of Entomological Science 25:510- J.-F. Silvain. 1990. Host range of Noetuidonema guya- 518. nense (Nematoda: Aphelenchoididae): An ectopara- 12. Silvain, J.-F. 1984. Premihres observations sur site of moths in French Guiana. Environmental En- l'4cologie de Spodopterafrugiperda (J. E. Smith) et Mo- tomology 19:795-798. cis latipes (Guen4e), noctuelles d4pr4datrices des 10. Rogers, C. E., A. M. Simmons, and O.G. gramin6es fourraghres en Guyane frangaise. Prairies Marti. 1990. Parasitism of Lepidoptera adults by Noc- guyanaises et elevage bovin. Les Colloques de I'INRA tuidonema guyanense Remillet and Silvain (Nematoda: 24:243-272. Aphelenchoididae) in southeastern United States. 13. Sparks, A. N. 1979. A review of the biology of Journal of Agricultural Entomology 7:241-245. the fall armyworm. Florida Entomologist 62:82-87.