Table Of ContentVol.34 No.1 Japanese Journal Nematologyof June, 2004
Cylindrocorpus anoplophorae n. sp. (Nematoda:
Cylindrocorporidae) isolated from the white-spotted longicorn
beetle, Anoplophora malasiana (Coleoptera: Cerambycidae)
Natsumi Kanzakil" and 2 Kazuyoshi Futail, 3
Cylindrocorpus anoplophorae n. sp. is described and figured. The dauer juveniles of C.
anoplophorae n. sp. were isolated from the genital capsule of the white-spotted longicorn
beetle, Anoplophora malasiana, and were reared on an Asparagine-Mannitol (A-M) agar.
Specimens of adult nematodes for description and measurements were collected from a 2
week-old culture on the A-M agar. The males of the new species have body length of ca.
830-1070ƒÊm, the value a of ca. 19-26, the value b of ca. 5-6, the value c of ca. 8-12, ca. 29-35
m length of slightly arcuate spicules, ca. 17-20 pm length of thorn shaped gubernaculum, ƒÊ
43-62 pm of tail conoid, nine pares of caudal papillae, and very narrow and tape like lep-
toderan bursa. The females of new species have body length of ca. 980-1260 pm, the value
a of ca. 17-23, the value b of ca. 6-7, the value c of ca. 7-9, ca. 114-161 ƒÊm length of elongat-
ed tail, and the values V of ca. 50-55. The new species is distinguished from the other
species within the genus by the morphometric values of male and female, number and
arrangement of male caudal papillae, shape and length of male bursa, and tail shape of
male and female. Jpn. J. Nematol. 34 (1), 11-19 (2004).
Key words - entomophilic nematode, Japan, morphology, taxonomy.
Cylindrocorporidae are defined as follows
INTRODUCTION
Head with (Andrassy, Goodey, 1963): six1984;
Dauer juveniles of a species of "nematode" lips each bearing a papilla of the internal circle,
were isolated from the genital capsule white-of whichpapillae six of or 10 and an external circle
malasianaAnoplophora beetle, spotted longicorn have not described in all genera. Amphids on
Thomson. The juveniles were reared on an lateral lips. Stoma with very long protostom.
agar, Asparagine-Mannitol then the adult nema- and Procorpus metacorpus fused form to a long
the culture, and identi-were obtained from todes and muscular cylindrical bulb. Isthmus slender,
fied as an undescribed species belonging to the terminal bulb glandular, but without valve
genus Cylondrocorpus Goodey, of 1939 family apparatus. Male with single testis, eight to 10
Cylindrocorporidae. pairs of caudal papillae, spicules paired, guber-
The morphological features of the family naculum present. Bursa present or absent.
or paired.single gonad(s) Female
Laboratory 1 of Environmental Mycoscience , Graduate Five genera are included in the andfamily,
of School Agriculture, Kyoto University, Sakyo-ku, Kyoto
each genus is distinguishable from the others
Japan. 606-8502,
Present: JSPS Post 2 Doctoral Fellow , Laboratory of Forest based on the male bursa and female (s)gonad
Protection, Faculty Agriculture, of Kagoshima University,
(Andrassy, 1984; Poinar et al., 2003):
Japan. 890-0065, Kagoshima Korimoto,
authorCorresponding 3 [email protected]: , has leptoderan bursa Cylindrocorpus and paired
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第34巻 第1号 日本線虫学会誌 2004年6月
gonads. Myctolaimus has no bursa and paired 100m1).
has peloderan bursa andMyctolaimellus gonads. Morphological observation:
paired gonads. Protocyrindrocorpus has lep- Adult nematodes were collected from a 2-
toderan bursa and single gonad. hasGoodeyus
week-old culture on Asparagine-Mannitol agar
peloderan bursa gonad.and single using Baermann funnel technique. The nema-
The had genus, Cylindrocorpus established
todes collected were killed by heat (60(cid:129)Ž for
by Goodey However, (1939). Andrassy (1984)
5 min.), fixed in TAF (formalin, 7 ml; tri-
syno-as a junior the genus Cylindrocorpus deleted
ethanolamine, 2ml; distilled water, 91ml),
nym andof the 1920, Cobb, genus Myctolaimus
processed through a glycerol-ethanol series
located following seven species to the genus
using Seinhorst's method (described in Hooper,
M M 1935; erectusMyctolaimus: Goodey, curzii
1986) and mounted in glycerin according to
Massey, M 1960; longistoma (Stefanski, 1922)
Maeseneer and d' Herde's method (described in
Andrassy, M 1984; macrolaimus (Schneider,
Hooper, 1986).
M 1920; Cobb, M pellucidus Massey, 1960; 1866)
rifflei Massey and Hinds, 1970; M. zamithi Cylindrocorpus anoplophorae n. sp.
(Lordello, 1953) Andrassy, 1984. Poinar et al. 2)1, (Figs.
(2003), however, reestablished the genus
inevectus,a C. described species, Cylindrocorpus, Measurements:
and moved following four species to the genus See Table 1.
erectusC. 1935; Goodey, curzii C. Cylindrocorpus: Description:
Massey, C.C. longistoma Stefanski, 1922; 1960; Male. Cuticle thin, with fine transverse and
1866Schneider, macrolaimus longitudinal striations. Body cylindrical, curved
newly The found is nematode distinguished ventrally when heat-killed. Lip region separated
from all known species in some morphological into six protuiting lips, each possessing one tiny
characters and morphometrical values, and is papilla. Stoma long, narrow, flexible, with thick
anoplophoraeherein as Cylindrocorpus described and distinct cheilorhabdions. Pharyngeal sleeve
sp.n. surrounding 85-90% of total length of stoma.
Tooth or denticles absent. Corpus, typical of the
MATERIALS AND METHODS
family, procorpus and metacorpus fused to form
Isolation of nematodes from white-spotted a long and cylindrical muscular bulb, looks
beetle:longicorn roundish rectangle shape. Stoma and corpus
malasiana A. was An obtainedadult male of longer than isthmus and basal bulb. Isthmus
at Kitashirakawa Forest Research FieldStation, slender, with one-fourth body-diam. width,
Science Education and Research Center, Kyoto widening gradually at basal bulb. Basal bulb not
The was beetle dis-June 2001. University, 20 on muscular, lacking a valve apparatus. Nerve ring
and infection.sected its nematode examined encircles the isthmus at the midst of isthmus.
The dauer juveniles of the nematode Excretory pore between the nerve ring and basal
obtained from the beetle were transferred to bulb. Cardia well developed, muscular, darkened
Asparagine-Mannitol agar 0.1g;(K2HPO4, in color. Testis single, reflexed, sperm in single
MgSO4• 0.02g; 7H2O, 0.001g; CaC12, NaC1, or double row. Spicules paired, arcuate, distal
asparagine,g; 0.05 KNO3, g; 0.0002 FeCl3, g; 0.001 end pointed, manubrium distinct, knoblike, ter-
water,distilled g; 2.0 agar, g; 0.1 mannitol, g; 0.05 minal cucullus absent. Gubernaculum thorn like.
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Vol. 34 No. 1 Japanese Journal Nematologyof June, 2004
C
B
A
F
G
D E
Fig. 1. Cylindrocorpus anoplophorae n. sp. Male Female Anterior (A), (B), region of Lateral male (C), view of male tail (D),
Ventral view of male Female tail Female (E) tail reproductive (F), organ (G).
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第34巻 第1号 日本線虫学会誌 2004年6月
B C E G
A
F
D
Bar=
A, B: 100ƒÊm
C-G: 20#ƒÊm
Fig 2 C, adrocorpus a; oplophorae n sp Male Female (A), (B), Anterior region of male Body (C), surface of male (D),
Lateral view of male Female tail (E), vulva Female tail (F), (G)
Tail ventrally arcuate, with long pointed conoid narrow, tape like, covering between just besides
Nine pairs of caudal papillae present, first pair the fourth pair of the caudal papillae and just
anterior to manubrium, second pair at posterior before conoid
level of manubrium, third pair just anterior to Female. Body slightly curved when heat-
cloacal slit, fourth and fifth pairs at the mid way killed Anterior region similar to that of male.
of anus and the commencement of conoid, the Vulval lip conspicurously protuberant Vagina
others aggregating at the distal part of tail, third strongly musculated, transverse, darkened in
pair at sub ventral, fifth and ninth pairs at later- color Reproductive system didelphic Uterin sac
al, the others at ventral. Bursa leptoderan, very serving as spermatheca, with well developed
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Vol.34 No.1 Japanese Journal Nematologyof June, 2004
Table 1. Measurements of Cylindrocorpus anoplophorae n. sp. measurements (all in pm).
1) Mean (cid:129)} SD (range)
2) Curved median line
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第34巻 第1号 日本線虫学会誌 2004年6月
sphincter muscle, contains zero to six mature and relationships:Diagnosis
eggs. Ovaries reflex their entire length, oocytes anoplophorae n. Cylindrocorpus sp. is char-
in double lows for approximately half ovary acterised by very narrow, short, and leptoderan
Anus length promi-low. then a single slightly in bursa between the fourth covering papillae level
nent. Rectum short, half body-diam. long. Tail and proximal part of the conoid, nine pairs of
elongated conical shape tapering to a pointed caudal papillae preanal (3 and 6 postanal), long
terminus. spicule, and long tail conoid of males, and long
habitat Type and locality: females.tail of
from 2-week-were obtained Type specimens Five species have been included in the
old culture on Asparagine-Mannitol agar, 26 genus Cylindrocorpus: C. curzii, C. erectus, C.
The original July culture 2001. was started from inevectus, C. longistoma, and C. macrolaimus
the dauer juveniles from isolated the male geni- sp. is dis-n. anoplophorae C. 2003). et al., (Poinar
tal capsule of Anoplophora malasiana collected inevectus,C. erectus, C. curzii, from C. tinguished
at Kitahirakawa Forest Research Station, Field and longistoma by C. the number and arrange-
Science Education and Research Center, Kyoto ment of male caudal papillae and tail ofconoid
Japan. 606-8502 Kyoto, Sakyo-ku, University, males (Fig. 1 D, E, Table 2). Males of C.
Type specimens: anoplophorae n. sp. have nine pairs of caudal
number slide Cylindrocorpus(male) Holotype three pairs at anal pre i.e., and papillae, pairssix
Nematode Collectionanoplophorae M-01(USDA at post anal, and long tail conoid, while the
anoplophoraenumber: Cylindrocorpus deposition have males pairs curzii 10 of C. of caudal papil-
T- 573t); allotype (female) slide number lae (3 pre anal and 7 post anal) and short tail
Cylindrocorpus anoplophorae F-01 (USDA conoid C. (Goodey, 1935), longistoma have 10
Nematode Collection deposition number: pairs of caudal papillae (3 pre anal and 7 post
Cylindrocorpus anoplophorae T- 574t), USDA anal) and long tail conoid (Goodey, 1927;
Nematode Collection, Beltsville, MD, USA. and erectus the andmales of C. 1922), Stefanski,
slide Paratypes num-females and males): 15 (15 have pairs eight pre of inevectus them anal(2 C.
bers Cylindrocorpus anoplophorae M-2-10 and 6 post anal) and short tail conoid (Massey,
number:deposition Collection Nematode (USDA 2003).et al., Poinar 1960;
T- 5163p)- T- 5155p anoplophorae Cylindrocorpus The values morphometric are also available
Nematode Collection(USDA F-2-10 males), (nine for diagnostics for Cylindrocorpus species (see
anoplophoraenumber: Cylindrocorpus deposition n.anoplophorae The body of C. lengths Table 2).
females) (nine T- T-5172p) deposited 5164p- in sp. are larger than those of C. erectus (Massey,
USDA Nematode Collection, Beltsville, MD, Theand (Poinar et inevectus al., 2003). 1960) C.
USA; slide numbers Cylindrocorpus values a of C. anoplophorae n. sp. are smaller
(fiveF-11-15 males), (five M-11-15 anoplophorae than those of C. curzii (Goodey, 1935) and C.
The Herbarium females), and Insect ofMuseum longistoma (Stefanski, and 1922), larger than
the National Institute of Agro-Environmental those of C. The erectus spicule(Massey, 1960).
Tsukuba, Science, Japan; Ibaraki, slide number n.and gubernaculum anoplophorae C. lengths of
F-male), (one M-16 anoplophorae Cylindrocorpus sp. are larger than those of C. curzii (Goodey,
EnvironmentalUniversity, Kyoto female), (one 16 C. inevectus (Poinar 1935), et and al., 2003), C.
Mycoscience Laboratory Collection, Kyoto, The longistoma (Stefanski, value 1922). V of C.
Japan. anoplophorae n. sp. is larger than those of C.
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Vol. 34 No. 1 Japanese Journal Nematologyof June,2004
Table 2. Morphometric comparisons of Cylindrocorj5us species (all measurements in „ˆm).
Morphometrics are not given for macrolaimus C. (Schneider, 1866).
After 1) Goodey (1935). After 2) Massey (1960). After 3) Poinar et al. 4) (2003). After Stefanski and (1922) Goodey (1927).
Sample ?: numbers 5) are given. not Values 6)*: are not given. Calculated 7) from the original figure.
erectus (Massey, 1960) and C. longistoma was found in a rotting yam from Nigeria and
1922).(Stefanski, rotting tissue of bitter cassava in Italian
Cylindrocorpus anoplophorae n. sp. could Somaliland (Goodey, Goodey, 1935; 1963), C.
by themacrolaimus be distinguished from not C. longistoma was reported from rat faeces in UK
number and arrangement of male caudal papil- and roots, ginger from imported (Goodey,China
and lae the shape of male tail Thus conoid. the 1927; Goodey, Stefanski, 1963; 1922), and C.
closest new to seems morphologically species C. was found in macrolaimus and moist soil decay-
macrolaimus. The morphometric values of C. ing matter (Schneider, 1866; Goodey, 1963).
macrolaimus are not given, and detailed mor- While for the entomophilic species, C. erectus
phological traits are not described, however, C. reported was as multi-"associated with Scolytus
be distinguished n. sp. can from C.anoplophorae striatus in American elm, americana" Ulmus in
based macrolaimus on the shape of male bursa. the United States and 1974), (Massey, 1960; C.
Males of anoplophorae n. C. sp. have very nar- was mature and described from decay-inevectus
tape row, like, and short bursa, which elongates guineensis and palm ing flowers, Elaeis fromoil
between just besides the fourth pair of the cau- the under surface of the elytra of the palmoil
dal papillae and just before conoid (Fig. 1E), Elaeidobius weevil, kamerunicus (Poinar et al.,
while the bursa of C. macrolaimus is flap like 2003).
and elongating between just besides the second The dauer juveniles of C. anoplophorae n.
and just conoidthe caudal before papillae of pair sp. were isolated from the male genital capsule
1866).(Schneider, Thus of white-spotted longicorn beetle. the new
species is assumed to inhabit in the xylem of
REMARKS
the beetle.plants, the trees host woody of
The Cylindrocorpus species have been that concluded We C n. sp. isanoplophorae
reported as the saplophagous or entomophilic to close macrolaimus C. based on the morpho-
free-living nematodes and have variable habi- logical traits, the i.e., number and the arrange-
tats. As for the saplophagous species, C. curzii caudal and ment papillae males.of conoid tail of
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第34巻 第1号 日本線虫学会誌 2004年6月
However, C. macrolaimus is described as the methods work for plant with and nema-soil
saplophagus soil nematode (Schneider, 1866), todes.(6th. Ed.) (Southey, J. F., ed.) Her
whereas, C. anoplophorae n. sp. was isolated pp. Majesty's 59-London, Stationary Office,
from a species of wood inhabiting beetle, A. 80.
malasiana. Therefore the biological features of conheci-ao Contrbbuioao E.(1953) G. L. Lordello,
to sp. might C.be rather close n. anoplophorae C. mento dos do solo de nematOdeos algumas
erectus and C. inevectus, the associate of bark regiOes do Estado de Sao Paulo. Doctoral
and (Massey, that beetle 1960), of the palmoil 75pp.Piracicaba. Thesis,
lifeThe respectively. 2003), et al., (Poinar weevil of nematoda,A species new L.(1960) C. Massey,
cannot species be gen-history of Cylindrocorpus Cylindrocorpus erectus associated with
the because habi-by descriptions, above eralized Scolytus multistriatus Marsh in American
tats and infor-among species significantly differ elm. Proceedings of the Helminthological
mation about vectors are not given for longi-C. 42-44.Washington 27, of Society
the exami-Therefore stoma macrolaimus. and C. Massey, L.(1974) C. Biology and taxonomy of
nations on the life history of each species are nematode parasites and associates of bark
needed to understand characteris-the biological beetle in the United States. USDA
and the systematic genus.tics relationship of the 233pp.446, No. Handbook Agriculture
Nematodesand E.(1970) T. Hinds, L. Massey, C.
LITERATURE CITED
from aspen canker in Colorado and New
Andrassy, I.(1984) Klasse Nematoda. Gustav 97-48, Journal of Zoology Canadian Mexico.
Stuttgart, Fischer Verlag, 509pp. 108.
Cobb, N. A.(1920) One hundred new nemas. Poinar, G. O., Jackson, T. A., Bell, N. L. and
Nematology 9,to of Contributions a Science Wahid, M. inevec-B.(2003) Cylindrocorpus
217-343. tus sp. n. associated with the palm oil wee-
Goodey, J. B.(1963) Soil and Freshwater vil, Elaeidobius kamerunicus (Faust)
Nematodes Methuen ed.). (Rev. & Ltd.Co. with a synopsisCurculionidae), (Coleoptera:
544pp.London, of and the family Cylindrocorporidae estab-
Goodey, T.(1927) Cylindrogaster coprophaga lishment of Longibuccidae n. fam.
gen. et sp. nov. a nematode found in a cul- Nematology 5,Nematoda). (Diplogastroidea:
Journalrat. brown from a wild of ture faeces -190.183
25-32.5, Helminthology of Schneider, A. F.(1866) Monographie der
n. sp.,curzii Cylindrogaster T.(1935) On Goodey, 357pp.Berlin, Reimer, G. Nematoden.
a saprophagous nematode. Journal of Stefanski, W.(1922) Excretion chez les
13,19-24.Helminthology nematodes libres. Archiwum Nauk
T.(1939) nom, Goodey, nov. forCylindrocorpus Biologicznych Towarzystwa Naukowego
Cylindrogaster Goodey, 1927 (Nematoda). Warszawaskiego 1,1-33.
17,149-150.Helminthology Journal of
Hooper, J.(1986) D. Handling, fixing, staining Received October 23, 2003.
and mounting nematodes. In: Laboratory
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Vol.34 No.1 Japanese Journal of Nematology June, 2004
和文摘要 ギ ンーマ ンニ トール培地上 で2週 間生育 したカル
ゴマダラカミキリ チ ャーか ら得 られた雌雄成 虫 を用 いた。本種 雄
Anoplophora malasiana から 成 虫は、体長約830-1070μm、a値 約19-26、b値 約
検出された線虫 5-6、c値 約8-10で あ り、長 さ約28-35μmの や
Cylindrocorpus anoplophorae n. sp. や湾 曲 した交接刺、長 さ約17-20μmの 爪状 の副刺 、
長 さ約43-62μmの 尾 端突起 、9対 の尾乳頭 、非
神崎 菜摘 ・二井 一禎 常 に幅 の狭 い尾翼 を有 してい た。雌 成虫 は、体
長約980-1260μm、 尾長約114-161μm、a値 約17-23、
Cylindrocorpus 属の未記載種をC. anoplophorae b値 約6-7、c値 約7-9、V値 約50-56で あった。
n.sp.と して記載 した。本種 の耐久型幼 虫は、 ゴ 本種 は、雌 雄成 虫 の形態 計測 値 、尾部 の形 状 、
マ ダラカ ミキ リ、Anoplophora malasiana の雄成 雄成虫 の尾 翼 の形 態、尾乳 頭 の配 列 によ り、本
虫の生殖器から検出された。記載にはアスパラ 属既知種 と識別可能 であった。
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