Journal of Nematology 27(2):213-221. 1995. © The Society of Nematologists 1995. Two New Species of (Nemata) from Declining Red Pines in Maryland 1

TAFADZWA R. KAISA 2, AMY L. HARMAN 3, AND DAN M. HARMAN 4

Abstract: resinosi n. sp. and Ektaphelenchusjoyceae n. sp. are described and illustrated from red pines of the Allegheny plateau of Maryland, USA. The new species were found in trees infested with Bursaphelenchus xylophilus. Primary diagnostic characters of A. resinosi females are con- striction of the head, basal stylet knobs, tong postnterine sac, two incisures in the lateral field, and conical tail four to five anal body widths long with a simple terminal mucro. Diagnostic characters of the males are two pairs of subventral caudal papillae and spicule shape: Primary diagnostic charac- ters of E. joyceae females are a slight constriction of the head, six similar lips, conical tail, and short postuterine sac. Diagnostic characters of the males are spicule size and shape, a single row of spermatocytes, and one pair of caudal papillae. Within-tree distributions of A. resinosi and E. joyceae are presented. A total of 70% of both red-needled and chlorotic-needled trees in the study were positive for A. resinosi and E. joyceae. Branch hierarchy was related to the percentage of samples positive for A. resinosi. Key words: Aphelenchoides resinosi, Bursaphelenchus xylophilus, Ektaphelenchus joyceae, Maryland, nema- tode, new species, Pinus resinosa, red pine.

Aphelenchoides species have been de- from Missouri (13) and the other in red scribed from conifers in the United States. pine from Maryland (9,10). To date, these Massey (19,20,21) described A. rhytium populations have remained unidentified. from eastern white pine (Pinus strobu~ L.), Another , Ektaphelenchus sp., loblolly pine (P. taeda L.), and red pine (P. was collected from the same red pines. resinosa Ait.) in New York; A. conophthori Massey (17"21) described eight Ektaphelen- from eastern white pine in Connecticut; chus species associated with conifers in the and A. hylurgi from eastern white pine in United States, three of which have been Maine. Aphelenchoides pityokteini (21) was synonymized with previous taxa (1). Ek- described from corkbark fir (Abies lasio- taphelenchus josephi and E. riograndensis carpa var. arizonica (Merriam) Lemm.) in were described from limber pine (P. flexilis New Mexico and A. polygraphi (21) from James) and red pine, respectively. Ek- Engelmann spruce (Picea engelmanni taphelenchus obtusus was described from En- Parry) in Arizona. Massey (19) found an gelmann spruce, E. prolobos from corkbark Aphelenchoides sp. of the A. parietinus group fir, and E. sandiaensis from white fir. associated with white fir (A. concolor Gord. The purpose of this paper is to describe & Glend. Lindl.) in New Mexico. In studies two previously undescribed taxa and to of pine wilt disease, two populations of an provide information on their distribution Aphelenchoides sp. were found occurring within dying red pines. with Bursaphelenchus xylophilus (Steiner & Buhrer, 1934) Nickle, 1970 in dying pines MATERIALS AND METHODS (9,10,13). One population was found in as- sociation with Scots pine (P. sylvestris L.) During the summer of 1987, dying red pines infested with Bursaphelenchus xylo- philus were collected from Savage River Received for publication 14 September 1993. State Forest in Allegany County, Mary- i Adapted from the first author's M.S. thesis, Frostburg land. Trees in that stand were 20--26 years State University. 2 P.O. Box 144, Syracuse, NY 13210. old. Although several stages of decadence s Professor of Biology, Frostburg State University, Frost- were noted among the afflicted trees in burg, MD 21532 and to whom correspondence should be red pine forests of Maryland (9,10), only addressed. 4 Associate Professor, University of Maryland, Center for two were chosen for collection: red- Environmental and Estuarine Studies, Appalachian Environ- mental Laboratory, Frostburg State University, Frostburg, needled trees and chlorotic-needled trees. MD 21532. Red-needled trees had uniformly red nee- 213 214 Journal of Nematology, Volume 27, No. 2,June 1995 dles and had died in the fall or winter prior er's method (11). Drawings were done with to the summer of collection. Chlorotic- a microscope fitted with a drawing attach- needled trees were in early stages of dying ment. All measurements are in microme- during midsummer of the year of collec- ters (~m) unless otherwise specified. tion. Ten red-needled trees were collected in May and June, and ten chlorotic- SYSTEMATICS needled trees were collected in July. Aphelenchoides resinosi n. sp. Samples from trunks and branches were (Fig. 1A-E) taken by means of the method of Harman (9,10). The lowest stem sample was re- Measurements moved approximately 20 cm from the Holotype (female in glycerine): L = 506; ground, and subsequent samples were stylet = 12;a-- 27;b = 9;c = 15;c' = taken at 1.5-m intervals. Terminal por- 2.4; V = 72. tions of stems were also collected. Branch Female (n = 40): L = 400-800 (mean = hierarchy (primary, secondary, tertiary) 530; SD = 110); stylet = 10-11 (mean = and branch decadence (three levels) were 10.6; SD = 0.6); ratios-a = 29-53 (mean utilized in sampling. One branch from = 35; SD = 5), b = 7-13 (mean = 9; SD each category on each 1.5-m trunk seg- = 2), c = 12-19 (mean = 15; SD = 2), c' ment was sampled. Samples were collected = 3-4 (mean = 3.4; SD = 0.4), V = 66- from the branches at 1.5-m intervals start- 79 (mean = 69; SD = 2.8). ing at the base of the branch and proceed- Allotype (male in glycerine): L = 484; stylet ing outward. Branch samples consisted of = 11; spicule = 15; a = 26;b = 7; c = 16; cylinders about 10 cm in length. c' = 2.3. were extracted by placing Male (n = 26): L = 360-480 (mean = small samples of wood in 1-oz plastic cups 450; SD = 60); stylet = 10 (mean = 10; and soaking in tap water for 24 hours. SD = 0); spicule = 13-15 (mean = 14; SD Specimens were heat-killed and preserved = 0.6); ratios-a = 31-48 (mean = 36; SD in glycerine-formaldehyde. Temporary = 4), b = 6-10 (mean = 8; SD = 1), c = mounts were made in lactophenol cotton 11-19 (mean = 14; SD = 2), c' = 2-3 blue after gently heating the nematodes in (mean = 2.4; SD = 0.1). a cavity slide containing a drop of tap wa- ter. Description Nematodes extracted from each sample Female: Body slender, slightly curved were counted. Final nematode counts were ventrally upon relaxation with heat. Cuti- expressed as number of nematodes per cle with fine, transverse striae. Lateral field gram of dry wood. Sample dry weight was with two incisures. Lip region and tail ter- determined following the method of minus not annulated. Body tapering pos- Brown and Bethel (4). Samples were oven- teriorly to conoid terminus. Terminus with dried at 100 C after nematode extraction small, fine mucro. Head set off by constric- and the weights recorded over a 48-hour tion (Fig. 1C); lips with guiding rods. Stylet period. Drying was stopped when weights delicate, basal knobs small. Stylet nar- remained constant over time, and the con- rowed anteriorly to fine tip. Tip of stylet stant weight was recorded as the dry very faint in preserved and fresh speci- weight of the sample. Eighty-three stem mens. Median bulb spherical with well- samples and 101 branch samples were an- developed, centrally situated valves. Duct alyzed from red-needled trees, and 77 pores into median bulb sometimes visible. stem samples and 97 branch samples from Nerve ring 3/4 of body width posterior chlorotic-needled trees. from base of median bulb, encircling Permanent mounts of nematodes were esophageal gland lobes and anterior end made in anhydrous glycerine after pro- of intestine. Esophageal gland lobes three cessing them slowly to glycerine by Hoop- body widths long. Excretory pore at level New Aphelenchoididae from Red Pine: Kaisa et al. 215

I ,

A 32 ,u,m B 28,u,m C 18,u,m D E 7/~m Fro. 1. Aphelenchoides resinosi n. sp. A) Full view of female. B) Full view of male. C) Head and neck of female. D) Stylet. E) Spicule. of nerve ring. Vulva slightly protuber- gland lobes. Oocytes in single row. Gonad ant. Vagina narrow, oblique. Ovary out- length 115 ~m. Postuterine sac 49 ~m stretched, not overlapping esophageal long, sometimes collapsed. 216 Journal of Nematology, Volume 27, No. 2,June 1995

Male: Male shorter than female, body ages 450 p~m in length with a conoid, mu- J-shaped. Cuticle finely annulated, lip re- cronate tail bearing two pairs of subventral gion and tail terminus smooth. Head set caudal papillae. Spicules are 14 I~m long, off, lips bearing guiding rods. Excretory arcuate with a prominent condylus. pore, median bulb, and nerve ring same as female. Testis long, outstretched. Tail ta- Relationships pering to conoid terminus with small, In the female body length of 400-800 sharp mucro. Bursa and gubernaculum p,m, the presence of two incisures in the absent. Two pairs of subventral caudal pa- lateral field, and the presence of a simple pillae present: first pair adanal, second mucro, A. resinosi is similar to A. arachidis post-anal, about one body width anterior Bos, 1977 (3), A. fragariae (Ritzema-Bos, to tail tip. Spicules paired, typically aph- 1891) Christie, 1932 (5), and A. helicosoma elenchoid, arcuate and rose-thorn shaped; Maslen, 1979 (16). It differs from A. condylus rounded and prominent, ros- arachidis in the shorter male body (360- trum pointed and less prominent. Point of 480 ~m vs. 560-1040 p~m) and in the num- intersection of dorsal and ventral limbs not ber of caudal papillae (two vs. three pairs visible. in A. arachidis). Aphelenchoides fragariae is distinguished from A. resinosi by the longer Type host and locality male body (480-650 p,m) and the presence From the xylem of stems and branches of three pairs of caudal papillae. Aphelen- of 20-26-year-old dying red pines (Pinus choides helicosoma differs in the stouter fe- resinosa Ait.) infested with Bursaphelenchus male body (a = 25-32 vs. 29-53), stouter xylophilus, Savage River State Forest, Com- male body (a = 26-37 vs. 31---48), presence partment 42, Allegany County, Maryland, of a stylet guiding ring, longer male body USA. (505-925 ~m), and in the presence of three pairs of caudal papillae. Type specimens Aphelenchoides resinosi is similar to A. con- Holotype (female): Collected 6 June 1987, imucronatus Bessarabova, 1966 (2) and A. by T.R. Kaisa, A.L. Harman, and D.M. vigor Thorne & Malek, 1968 (26) in having Harman in Allegany County, Maryland, two incisures and a simple mucro. Aphelen- USA. Slide no. T-513t, deposited in the choides conimucronatus differs in the shorter U.S. Department of Agriculture Nema- female body of 280-440 p,m, shorter spi- tode Collection (USDANC), Beltsville, cule of 10 I~m vs. 13-15 p~m, and spicule Maryland. AUotype (male): Same data as ho- shape. The spicules of A. conimucronatus lotype. Slide no. T-514t, deposited in US- are smoothly curved with a non-prominent DANC, Beltsville, Maryland. Paratypes (7 condylus, whereas those of A. resinosi have females and 5 males): Same data as holotype, a distinct condylus and irregular curva- deposited in USDANC, Beltsville, Mary- ture. Aphelenchoides vigor differs in the land; Nematology Department, Rotham- longer female body of 900 p~m, longer sted Experimental Station, Harpenden, male body of 720 I~m, smaller V of 63 vs. Herts., England; and Frostburg State Uni- 66-75 for A. resinosi, the presence of three versity, Department of Biology, Frostburg, pairs of caudal papillae, and a bluntly Maryland. rounded spicule terminus versus a pointed terminus in A. resinosi. Diagnosis In the female body length of 410-550 The female of A. resinosi averages 530 ~,m, male body length of 380 ~m, stylet I~m in length with a slender body, annu- length of 10-12 p~m, and in the presence of lated cuticle, and two incisures in the lat- two pairs of caudal papillae, A. bicaudatus eral field. The tail is conoid with a simple (Imamura, 1931) Filipjev & Schuurmans mucro. The stylet is short (11 ~m) with Stekhoven, 1941 (8) is similar to A. resinosi. basal knobs. The male of A. resinosi aver- It can be distinguished from A. resinosi by New Aphelenchoididae from Red Pine: Kaisa et ai. 217 the bifurcation of the female tail, and in Male (n = 7): L = 400-470 (mean = the presence of a stylet guiding ring. 440; SD = 30); stylet = 14-16 (mean = Aphelenchoides appendurus Singh, 1967 15; SD = 0.8); spicule = 13-14 (mean = (25) is similar to A. resinosi in having two 14; SD = 0.3); ratios- a = 26-35 (mean = incisures and V of 66-69.8, but differs 32; SD = 3), b = 5-7 (mean = 6; SD = from it in the longer female body of 720- 0.6), c = 15-18 (mean = 16; SD = 1), c' = 880 ~m, longer male body of 810 ~m, 1.8-2.2 (mean = 2; SD = 0.2). longer stylet of 16.5-17.5 p~m vs. 10-11 Description p~m, and in the presence of a leaf-like mu- cro. Female: Body ventrally arcuate, tapering Aphelenchoides asterocaudatus Das, 1960 to conoid tail. Cuticle with fine, transverse (7) is similar to A. resinosi in the female striae. Head slightly set off by slight con- body length of 620 p~m, in having two in- striction. Tail terminus with lateral spike in cisures, and V of 68. It differs from A. resi- 31% of females. Stylet delicate, without nosi in the presence of a star-shaped mu- knobs. Stylet tip opening slanted ventrally cro. Males are unknown for A. asterocauda- (Fig. 2C). Median bulb oblong, longer than tus. wide, one body width long; valves well de- Aphelenchoides rosei Dmitrenko, 1966 (6) veloped, post-median. Excretory pore one- is similar to A. resinosi in having two in- half body width below nerve ring; hemi- cisures, but differs in the longer male stylet zonid just anterior to excretory pore. of 14 p~m, longer male body of 670 p~m, Nerve ring one body width posterior to and a nonmucronate male tail. Females are median bulb. Esophageal gland lobes long, unknown for A. rosei. about five times body width, overlapping Aphelenchoides brevistylus Jain & Singh, intestine dorsally. Vulva not protuberant, 1984 (12) and A. unisexus Jain & Singh, flap absent. Vagina short, narrow, oblique. 1984 (12) are similar to A. resinosi in having Rectum and anus absent; intestine termi- two incisures; in the female body lengths nating in blind sac. Ovary outstretched. of 390-630 p~m and 480-760 ~m, respec- Oocytes in two rows. Spermatheca oblong, tively; and in V of 63-71 and 62.3-67.2, sperm visible in 47% of females. Postuter- respectively. Aphelenchoides brevistylus males ine sac short, length about one-half corre- are similar to A. resinosi males in spicule sponding body width. length of 14-16 p~m and in having two Male: Male shorter than female. Body pairs of caudal papillae. Males are un- J-shaped or curved ventrally when re- known for A. unisexus. Both A. brevistylus laxed. Tail curved sharply through 180 °, and A. unisexus can be distinguished from tail region narrowing to pointed tip. Testis A. resinosi by their star-shaped mucros. outstretched, spermatocytes in single row. Aphelenchoides brevistylus also differs in hav- Spicules arcuate, condylus rounded, ing a shorter stylet of 6-8 ~m and in the prominent; rostrum small, pointed. One absence of stylet knobs. pair of ventral caudal papillae near tail tip. Bursa and gubernaculum absent. Ektaphelenchus joyceae n. sp. Type host and locality (Fig. 2A-G) From the xylem of stems and branches Measurements of 20-26-year-old dying red pines (Pinus resinosa Ait.) infested with Bursaphelenchus Holotype (female in glycerine): L = 710; xylophilus, Savage River State Forest, Com- stylet = 17; a = 36; b = 9; V = 83. partment 42, Allegany County, Maryland, Female (n = 32): L = 550-740 (mean = USA. 640; SD = 60); stylet = 14-18 (mean = 16; SD = 1); ratios- a = 29-44 (mean = Type specimens 34; SD = 3), b = 7-13 (mean = 9; SD = Holotype (female): Collected 6 June 1987, 1), V = 77-86 (mean = 82; SD = 2). by T.R. Kaisa, A.L. Harman, and D.M. 218 Journal of Nematology, Volume 27, No. 2,June 1995

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A B 40,u, m C 18/.Lm D 7u, m E 20u, m FG .... 5/~m FIG. 2. Ektaphelenchusjoyceaen. sp. A) Full view of female. B) Full view of male. C) Head and neck of female. D) Spicule. E) Female gonad. F) Female tail terminus with lateral spike. G) Simple female tail terminus.

Harman in Allegany County, Maryland, Maryland. Paratypes (8females and 3 males): USA. Slide no. T-515t, deposited in the Same data as holotype, deposited in US- U.S. Department of Agriculture Nema- DANC, BeltsviUe, Maryland; Nematology tode Collection (USDANC), Beltsville, Department, Rothamsted Experimental New Aphelenchoididae from Red Pine: Kaisa et al. 219

Station, Harpenden, Herts., England; and E. joyceae males in the longer stylet of 21 Frostburg State University, Department of p,m and in the presence of two pairs of Biology, Frostburg, Maryland. caudal papillae. A longer male body of 645-690 wm and two pairs of caudal pa- Diagnosis pillae distinguish E. tuerkorum from E. The female of E. joyceae averages 640 joyceae. p~m long with a moderately set-off head. With a stylet length of 16 Ixm, E. amitini The tail terminus is conoid at times with a (Fuchs, 1937) Rfihm, 1956 (24) is similar to small lateral spike. The stylet is 16 wm long E.joyceae, but differs by having stylet knobs and without basal knobs. Oocytes are in and a shorter body of 523 ~m vs. 550-740 two rows, the vagina is oblique, and the wm in E. joyceae. Males are unknown for E. postuterine sac is less than one body width amitini. long. The males orE. joyceae are short (440 Ektaphelenchus goffarti Rfihm, 1956 (24) wm) with pointed, conoid tails bearing one is similar to E. joyceae in the female body pair of ventral caudal papillae. Spermato- length of 547-701 ~m and male body cytes are in a single row, and spicules are length of 442-485 p,m. It differs from E. small (14 v~m) and smoothly curved with a joyceae in having one row of oocytes vs. two, prominent condylus and a small, pointed in the presence of two rows of spermato- rostrum. cytes vs. one, and in having three pairs of caudal papillae vs. one in E. joyceae. Relationships In the conical female tail and stylet with- In the female body length of 550-740 out knobs, E. joyceae is similar to E. betulae wm, stylet length of 14-18 p,m, and in hav- Rfihm, 1956 (24), E. obtusus Massey, 1956 ing two rows of oocytes, E. joyceae is similar (17), E. riograndensis Massey, 1964 (18), E. to E. larici Lazarevskaya, 1963 (15), E. mac- scolyti Rfihm, 1956 (24), E. skrjabini Laza- robulbosus Rfihm, 1956 (24), E. prolobos revskaya, 1962 (14), and E. zw61feri Rfihm, Massey 1964 (18), E. sandiaensis Massey, 1956 (24). Ektaphelenchus betulae, E. obtusus, 1964 (19), and E. tuerkorum Rfihm, 1956 E. riograndensis, E. scoltyi, and E. zw61feri all (24). It differs from E. larici in the short differ from E. joyceae in having longer fe- postuterine sac (less than one body width male bodies of greater than 740 Win, vs. six body widths long) and in the absence longer male bodies of greater than 600 of guiding rods in the lips. Males ofE. larici Win, and in the presence of three pairs of are unknown. Ektaphelenchusjoyceae differs caudal papillae. Ektaphelenchus skrjabini dif- from E. macrobulbosus in the arrangement fers in the shorter female body of 466-508 of spermatocytes (one vs. two rows), num- wm and in having three pairs of caudal ber of caudal papillae (one vs. two pairs), papillae. and in the rounded versus hooked spicule Ektaphelenchus josephi Massey, 1974 (21) terminus of E. macrobulbosus. Ektaphelen- is similar to E. joyceae in the conical female chus joyceae differs from E. prolobos in hav- tail, in having two rows of oocytes, and a ing a stylet without knobs, in the shorter short postuterine sac (less than one body male body of 400-470 wm vs. 610-660 width long). It differs from E. joyceae in the Win, in the presence of one vs. two rows of presence of styler knobs, longer female spermatocytes, and in the number of cau- body of 850-950 p~m, longer male body of dal papillae (one vs. two pairs). Ektaphelen- 670 wm, and in having two pairs of caudal chus sandiaensis differs in having a longer papillae and two rows of spermatocytes. stylet of 19.5-22 p~m vs. 14-18 p~m, in the bluntly rounded female tail terminus vs. DISCUSSION conical, and in having a postuterine sac that is twice as long as that of E. joyceae. The bionomics of Aphelenchoides resinosi Ektaphelenchus sandiaensis males differ from and its mode of transmission to red pines 220 Journal of Nematology, Volume 27, No. 2,June 1995

TABLE 1. Occurrence of Aphelenchoides resinosi, n. sp. (+) and its co-occurrence with Bursaphelenchus xylo- philus (B) and Ektaphelenchusjoyceae n. sp. (E) by sample category in Maryland red pines.

Red-needled trees Chlorotic-needled trees

Tree Primary Secondary Tree Primary Secondary number Trunk branches branches number Trunk branches branches

1 - - - 11 - + - 2 - - - 12 - - + ,B 3 - - + 13 - + + 4 - - - 14 - - - 5 - + + ,B 15 - + + 6 +,E + + 16 + + - 7 - +,E + 17 + + + 8 - +,B + 18 - - - 9 +,B + + 19 - - + 10 - + - 20 + + ,B +

are unknown. Possibly, it is a mycophagous than primary branches (Table 2). In both species that is carried by insects associated types of trees, average nematodes per sam- with pines, as Massey (21), Nickle (22), and ple weight from primary branch samples Nickle and Hooper (23) have reported as- were about one-third the number from sociations of aphelenchs with pine bark secondary branch samples. Branch hierar- beetles. chy was related to percentages of samples Distributions ofA. resinosi, E.joyceae, and containing A. resinosi. The number of sam- B. xylophilus were determined and com- ples containing the nematode was greater pared (Table 1). Aphelenchoides resinosi co- in secondary branches than in primary occurred with B. xylophilus in both trunk branches (P = 0.002). Ektaphelenchus and branch samples from red-needled joyceae was sparsely distributed in the sam- trees, but only in branch samples from pled trees, occurring in two of 10 red- chlorotic-needled trees. Aphelenchoides resi- needled trees and in none of the chlorotic- nosi co-occurred with E. joyceae only in red- needled trees. needled trees. The respective roles of A. resinosi and E. Seventy percent of red-needled trees joyceae in the bioecology of dying red pines contained A. resinosi. Occurrence was more was not determined in this study. Their frequent in branch than in trunk samples frequencies were relatively low in all sam- (Table I). Eighty percent of chlorotic- ple categories compared to that of B. xylo- needled trees contained A. resinosi and had philus. Massey (21) stated that aphelen- similar frequency of occurrence in pri- choid nematodes were often associated mary and secondary branch samples. Av- with bark beetles phoretically, and it seems erage A. resinosi per-gram sample weight likely that A. resinosi and E. joyceae are also was higher among secondary branches insect associates, not tree pathogens.

TABLE 2. Occurrence ofAphelenchoides resinosin. sp. by numbers in red-needled and chlorotic-needled red pine branches.

Total dry weight Total number Average number of samples (g) of nematodes of nematodes/g

Primary Secondary Primary Secondary Primary Secondary branches branches branches branches branches branches

Red-needled trees 189 5 821 63 4 13 Chlorotic-needled trees 63 3 44 68 7 22 New Aphelenchoididae from Red Pine: Kaisa et al. 221

LITERATURE CITED 14. Lazarevskaya, S. L. 1962. New species of nema- todes from the banded pine weevil (Pissodes pini L.). 1. Baujard, P. 1984. Remarques sur la sous-famille Trudy Germintologicheskoi Laboratorii Akademii des Ektaphelenchoides Paramonov 1964 et proposition Nauk SSSR 12:144--152. [In Russian]. d' Ektaphelenchoides n. gen. (Nematoda: Aphelen- 15. Lazarevskaya, S.L. 1963. Nematode fauna of choididae). Revue de N~matologie 7:147-171. Orthotomicus laricis and O. proximus (Coleoptera: Ip- 2. Bessarabova, L. M. 1966. A new nematode spe- idae). Helminthologia 4:254-265. [In Russian]. cies, Aphelenchoides conimucronatus, found on legumes 16. Maslen, N. R. 1979. Six new nematode species in the Moscow region. Zoologicheskii Zhurnal 45: from the maritime Antarctic. Nematologica 25:288- 1569-1570. [In Russian]. 308. 3. Bos, W.S. 1977. n. sp. (Nematoda: Aphelenchoidea), an endoparasite of the 17. Massey, C.L. 1956. Nematode parasites and testa of groundnuts in Nigeria. Journal of Plant Dis- associates of the Engelmann spruce beetle (Dendrocto- eases and Protection 84:95-99. nus engelmanni Hopk.). Proceedings of the Hel- 4. Brown, N. C., and J. S. Bethel. 1958. Lumber, minthological Society of Washington 23:14-24. 2nd ed. New York: John Wiley. 18. Massey, C.L. 1964. Two new species of the 5. Christie, J. R. 1932. Recent observations on the nematode genus Ektaphelenchus (Nematoda: Aphelen- strawberry dwarf nematode in Massachusetts. Plant choididae) parasites of bark beetles in the southwest- Disease Reporter 6:113-114. ern United States. Proceedings of the Helmintholog- 6. Dmitrenko, M.A. 1966. Two new phytohel- ical Society of Washington 32:37-40. minth species. Zoologicheskii Zhurnal 45:764-766. 19. Massey, C.L. 1964. The nematode parasites [In Russian]. and associates of the fir engraver beetle, Scolytus ven- 7. Das, V. M. 1960. Studies on the nematode par- tralis LeConte, in New Mexico. Journal of Insect Pa- asites of plants in Hyderabad (Andra Pradesh, India). thology 6:133-155. Zeitschrift fiir Parasitenkunde 19:553-605. 20. Massey, C.L. 1971. Nematode associates of 8. Filipjev, I. N., andJ. H. Schuurmans Stekhoven. several species ofPissodes (Coleoptera: Curculionidae) 1941. A manual of agricultural helminthology. Lei- in the United States. Annals of the Entomological So- den: E.J. Brill. ciety of America 64:162-169. 9. Harman, A. L. 1985. Insect-nematode-red pine 21. Massey, C. L. 1974. Biology and of associations in western Maryland with major empha- nematode parasites and associates of bark beetles in sis on Bursaphelenchus xylophilus and Monochamus spp. the United States. Agricultural Handbook no. 446. Ph.D. dissertation, University of Maryland, College USDA Forest Service, Washington, DC: U.S. Govern- Park. ment Printing Office. 10. Harman, A. L., L. R. Krusberg, and W. R. 22. Nickle, W. R. 1970. A taxonomic review of the Niclde. 1986. Pinewood nematode, Bursaphelenchus xy- genera of the Aphelenchoidea (Fuchs 1937) Thorne lophilus, associated with red pine, Pinus resinosa, in 1949 (Nematoda: ). Journal of Nematolo- western Maryland. Journal of Nematology 18:575- gy 2:375-392. 580. 11. Hooper, D.J. 1970. Handling, fixing, staining, 23. Nickle, W. R., and D.J. Hooper. 1991. The Aphelenchina: bud, leaf, and insect nematodes. Pp. and mounting nematodes. Pp. 39-54 inJ. F. Southey, 465-507 W. R. Nickle, ed. Manual of Agricultural ed. Laboratory methods for work with plant and soil in Nematology. New York: Marcel Dekker. nematodes, 5th Ed. London: Her Majesty's Stationery Office. 24. Rtihm, W. 1956. Die nematoden der Ipiden. 12. Jain, V. K., and S. P. Singh. 1984. Two new Parasitologische Schriftenreihe no. 6. Jena. species of Aphelenchoides (Nematoda: Aphelenchoid- 25. Singh, S. D. 1967. On two new species of the idae) from India. Bulletin of Entomology 25:135- genus Aphelenchoides Fischer 1894 (Nematoda: Aph- 142. elenchoididae) from North India. Journal of Hel- 13. Kondo, E., A. Foudin, M. Linit, M. Smith, R. minthology 61:63-70. Bolla, R. Winter, and V. Dropkin. 1982. Pine wilt 26. Thorne, G., and R. B. Malek. 1968. Nematodes disease: Nematological, entomological, and biochem- of the Northern Great Plains. Part I. Tylenchida ical investigations. University of Missouri Agricul- (Nematoda: ). Technical Bulletin 31. tural Experiment Station Bulletin SR 282. Columbia. South Dakota Agricultural Experiment Station.