Proceedings of the Helminthological Society of Washington 35(1) 1968

Volume 35 January 1968 ^unriber 1

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ALLEN' , M.W:, AN' D E. M. NOFFSINGER. Reyision of the Genus Anaplectus 77 ; -•'.'

,, ,'.N, A',, -A-ND'-A^Ri'^AGfiENTi./'1 A. ' iPe*iph,eral^ ^ 'Nervous, 'S;ystem in Nenaa- todk'withv a Discussion, of its Furiptiqrialiand Phylpgeinetic tSignificancie . - '108 , iJXcoB H., ANb J. ^D. THOMAS. IMgenetic Tjeindtodes of -Am-';.":r:"c phibians ?md Reptiles frorh Ghana —_„.,—.._„,-.-_-„ .lV.._tr..J.i..T...___i..il. ' 1

J. ;JMivANDv°SV. R. NiGKLE; On the Classification and Life History /of Fergiisobia curfiei (Sphaerulafiidae: Nematoda) ._^_li-.-i-~~.^r'.-.^__i.J|.'i 40

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-lf --,;•> JESSE RvCHRISTrE,1968 A r: " s :g

JA^OB H. FISCHTHAL, 1972 - - ^-.::^ r ;1 WILLIAM R.NIGKLE,'1972 ^WILLIAM J. ^HARGIS^ JR., 1971- ' 1^4 7; GILBERT F. OTTO, 1969^ f

JOHN Tl-LUCKER, 1972. ; DEWEYJ. RASKL1970

SJC^fe-it A -; ^ PAULp3vElNSTEIN;i969 ^ •' y.C--" jfjj

VOLUME 35 JANUARY 1968 NUMBER 1

Digenetic Trematodes of Amphibians and Reptiles from Ghana1

JACOB H. FISCHTHAL AND J. D. THOMAS-

Most of the trematocles of this report were by loosely arranged parenchymal muscles; no collected by the junior author while on the apparent esophagus; cecal bifurcation 700 pre- faculty of the Department of Zoology, Uni- acetabular; cecal shoulders at bifurcation, pre- versity of Ghana, Legon; additional collec- acetabularly ceca narrow, postacetabularly ceca tions were made by the senior author during with more or less alternating narrow and lat- 1965-66 while a Fulbright lecturer in Zoology erally inflated areas, extending almost to pos- at University College of Cape Coast, Cape terior extremity. Coast. Specimens have been deposited in the Testes two, tandem, surfaces smooth to U. S. National Museum Helminthological Col- slightly wavy, in posterior three-fifths of hind- lection as indicated. All measurements are in body and in posterior two-fifths of body length; microns. The nomenclature adopted for the anterior testis U shaped, lying 1,865 postace- amphibian hosts from Ghana is based on the tabular, total length 635, total width 975, width work of Schi0tz (1964). right arm 235, width bend of U 140, width left arm 205; posterior testis Y shaped with Family Clinostomatidae widespread arms, lying 2,675 postacetabular, Clinostomum hylaranae n. sp., total length 510, total width 780, width right metacercaria (Fig. 1) arm 202, width left arm 183, length at stem 295. Vas deferens entering cirrus sac poster o- HOST: Hylarana albolabris (Hallowell) dorsally. Cirrus sac 410 by 298, thick walled, (Ranidae). muscular, within arms of anterior testis but HABITAT: Intestine. overlapping bend of U ventrally, lying 2,000 LOCALITY: Kade, Ghana. postacetabular, containing seminal vesicle, cir- DATE: 25 November 1961. rus, and prostate cells; proximal part of seminal SPECIMEN: USNM Helm. Coll. No. 62890. vesicle tubular, winding anteriorly, becoming DIAGNOSIS (based on single specimen): saccular and looping ventrally and posteriorly Body 6,875 by 2,175, elongate, widest at tes- in anterior part of cirrus sac, then becoming ticular level, tegument unspined. Forebody tubular again for short distance; cirrus thick 1,440, hinclbody 4,470; posttesticular space walled, muscular, winding; genital atrium shal- 1,285, postcecal space 272. Oral sucker 305 low, small; genital pore median, ventral to most by 470, subterminal ventral, anterior to ce- posterior part of cirrus sac. phalic collar; acetabulum 965 in diameter, Ovary 145 by 255, dextral, intercecal, inter- large, strong, anterior margin at about one- testicular, surface smooth to slightly wavy, fifth body length from anterior extremity; lying 2,480 postacetabular. Ootype complex sucker length ratio 1:3.16. Prepharynx 290 250 by 430, distinctly differentiated from sur- long; pharynx slightly muscular, surrounded rounding parenchyma, sinistral to and larger than ovary; oviduct much coiled within com- 1 Contribution from the Department of Biology, State University of New York at Binghamton, Binghamton, New plex and surrounded by profuse Mehlis' gland. York 13901 (J. H. Fischthal). -Address of J. D. Thomas: School of Biological Sci- Laurer's canal opening on dorsal surface dorsal ences, The University of Sussex, Falmer, Brighton, Sussex, to left part of ovary. Uteroduct thick walled, England. This study was supported in part by a State University muscular, surrounded by gland cells through- of New York Faculty Research Fellowship (FRF67-40-004) awarded to the senior author. out length, emerging from anteroventral part

Copyright © 2011, The Helminthological Society of Washington PROCEEDINGS OF THE HELMINTHOLOGICAL SOCIETY of ootype complex, arcs anterosinistrally over- long as adults), in the ootype complex being lapping left cecum ventrally, opening into uter- poorly differentiated, and in the vitelline folli- ine sac just anteromedial to left arm of anterior cles not being confluent in the midline ventral testis; uterine sac 1,690 long, thick walled, to the length of the uterine sac. Vitelline folli- extending from 380 postacetabular to just an- cles were not observed in C. pseudohetero- terior to cirrus sac; metraterm thin walled, stomum. emerging from posteroventral part of uterine sac, ventral to cirrus sac, opening into genital Family Haematoloechidae atrium. Vitelline follicles small, numerous, ex- Haematoloechus exoterorchis Rees, 1964 tending from level of posterior half of acetab- HOST: Dicroglossus occipitalis (Giinther) ulum to just around cecal ends, confluent in (syn. Rana o. G.) (Ranidae). midline from just postacetabular to short dis- HABITAT: Lungs. tance pretesticular as well as posttesticularly, LOCALITIES: Achimota, Cape Coast, Legon, lateral and ventral to ceca, ventral to uterine Nungua, Pokoasi; Ghana. sac. DATES: 1956-58, 1960-61, 1965. Excretory pore subterminal dorsal, 56 from SPECIMENS: USNM Helm. Coll. No. 62891. posterior extremity; bladder U shaped, with long extracecal arms sending branches through- Haematoloechus micrurus Rees, 1964 out body. DISCUSSION: Our form appears closest to HOST: Dicroglossus occipitalis. HABITAT: Lungs. C. pseudoheterostomum described by Tuban- gui (1933) from metacercariae from Rana LOCALITIES: Achimota, Cape Coast, Legon, Nungua; Ghana. magna Stejneger (Ranidae) from the Philip- DATES: 1956, 1960-61, 1965. pines and C. pyriforme described by Prudhoe SPECIMENS: USNM Helm. Coll. No. 62892. (1957) from adults from a mammal, Aomjx DISCUSSION: Rees (1964) described in detail capensis (Schinz) (Mustelidae), from the both species from the same host species from Congo. All have the anterior testis more or Achimota and Nungua. Our observations are less U shaped and the cirrus sac lies between essentially similar to hers. H. micnmis was the arms of the U (cirrus sac not described more frequently found at Cape Coast, while for C. pseudoheterostomum but probably is H. exoterorchis was more abundant in the other situated as indicated herein). The posterior localities about 100 miles to the east. testis is V shaped in the latter and sausage shaped in C. pyriforme. Fischthal and Kuntz Family Heterorchiidae (1963) noted that the shape of the testes varied in metacercariae of Euclinostomum Heterorchis ghanensis n. sp. (Figs. 2, 3) heterostomum (Rudolphi, 1809) Travassos, HOST: Hyperolius nitidulus Peters (Rha- 1928: anterior testis broadly U to crescent cophoridae). shaped, posterior Y to triangular shaped. Al- HABITAT: Small intestine. though the size of the body and acetabulum LOCALITY: Agbogba, Ghana. of C. pseudoheterostomum are similar to our DATE: 22 November 1961. species the former differs in having a much HOLOTYPE: USNM Helm. Coll. No. 62893. larger oral sucker (sucker length ratio 1:1.66); DIAGNOSIS (based on single specimen): Body also, the prepharynx in the former is very short, 1,695 by 790, elongate oval, widest just post- practically absent. C. pyriforme differs from acetabular; tegument spined, spines sparse pos- our species in being very small (2.5 to 3.5 mm teriorly, extending on ventral surface to short

Figures 1-11. 1. Clinostornuni hylaranae, mctaccrcaria, ventral view. 2. Heterorchis ghanensis, holotype, dorsal view. 3. Same. Terminal genitalia, dorsal view. 4. Metaplagiorchis biloborchis, holotype, ventral view. 5. Same. Terminal genitalia, ventral view. 6. Ostioloides rappiae, ventral view. 7. Same. Terminal genitalia, ventral view. 8. Ganeo africana, dorsal view. 9. Prosotocus exovitellosus, holotype, ventral view. 10. Same. Terminal genitalia, ventral view. 11. Halipegus ghanensis, holotype, dextrolateral view.

7/10

Abbreviations: C, cirrus; CS, cirrus sac; E, egg; GC, gland cells; CiP, genital pore; M, metraterm; PC, prostate cells; PP, pars prostatica; SV, seminal vesicle; U, uterus.

Copyright © 2011, The Helminthological Society of Washington PROCEEDINGS OF THE HELMINTHOLOGICAL SOCIETY distance postacetabular including entire oral posterior margin of acetabulum, overlapping sucker, on dorsal surface over oral sucker, also cecum ventrally. Oviduct thick walled, mus- on lateral and dorso- and ventrolateral body cular, from center of mesial margin of ovary. surfaces to posterior extremity. Forebody 310, Seminal receptacle 186 by 88, median to ovary, hindbody 1,030; posttesticular space 445, post- dorsal to Mehlis' gland. Laurer's canal not ob- vitellarian space 380, postcecal space 111. Oral served. Uterus ventral to excretory bladder, sucker 201 by 255, subterminal ventral, weakly posterior ends of testes, vitellaria and ceca, muscular; mouth opening very small transverse descending and ascending between testes, fill- slit closer to posterior part of sucker, suring posttesticular space. Metraterm 484 (longi- rounded by muscular sphincter. Acetabulum tudinal extent) by 56, as long as cirrus sac, 355 by 350, opening round, large, center at thick walled; proximal half with inner longi- level of anterior three-tenths of body length. tudinal and outer circular muscle layers, lumen Sucker length ratio 1:1.77. Prepharynx very large, gland cells surrounding metatermal walls short, conical; pharynx 112 by 110, anterior on outside, crossing cirrus sac and left cecum end trilobecl with two larger lateral lobes ventrally; distal half with strongly developed and one smaller, shorter midventral lobe, sur- muscle layers surrounded by gland cells lying rounded by large gland cells; esophagus 15 within metratermal wall, lumen narrow, as- long; cecal bifurcation just preacetabular; ceca cending left of cirrus sac. Vitelline follicles extending beyond posterior margin of excretory round to elongate, some lobed, relatively large, bladder to near posterior extremity before nar- few, in lateral fields, primarily ventral and lat- rowing abruptly and looping anteromeclianly eral to ceca but with few follicles dorsal, to open separately into excretory bladder. extending from level of posterior margin of Testes two, narrow, elongate, surfaces wavy, acetabulum to short distance posttesticular. intercecal to overlapping ceca ventrally, ven- Eggs yellow, operculate, eight measuring 19- tral to excretory bladder where they overlap 25 by 13-14. latter, diagonal with left testis more anterior, Excretory bladder Y shaped, stem 712 by right testis slightly overlapping ovary ventrally. 380, voluminous, saccular, may overlap parts Right testis 410 by 115, 210 postacetabular; of ceca dorsally, extending to 208 from pos- left testis 460 by 100, 80 postacetabular. Cir- terior extremity; arms short, rabbit earlike, rus sac 484 (longitudinal extent) by 100, thick postacetabular, right arm 140 by 17, left 160 walled, muscular, commencing 115 postacetab- by 65; excretory pore dorsal, very large, 205 ular at anterior margin of stem of excretory in diameter, lying 272 from posterior extremity. bladder, overlapping left excretory arm dor- DISCUSSION: The two known species in the sally, ascending sinistrolateral to acetabulum genus are H. crumenifer Baylis, 1915, from and ventral to left cecum, terminating at gen- Protoptems aethiopicus Heckel and P. annec- ital atrium and pore located sinistrolaterally at tens Owen (Lepidosirenidae) from Uganda, level between cecal bifurcation and anterior Cameroons and the Congo, and H. protopteri margin of acetabulum, containing seminal ves- described by Thomas (1958) from P. annectens icle, pars prostatica, prostate cells, and cirrus. from Ghana. Our species differs from them Seminal vesicle thick walled, muscular, bipar- in having the ceca opening into the excretory tite; posterior chamber 242 by 85, saccular; bladder and in occurring in an amphibian host anterior chamber 97 by 22, tubular. Pars pros- rather than lungfishes. It differs further from tatica 81 by 35, short, thick walled, cell lined. H. crumenifer in having a larger acetabulum, Cirrus long, muscular, winding to right, then testes nearly equal in size and pointed pos- anterosinistrally, looping posteriorly in ventral teriorly, the seminal receptacle at midovarian position to overlap distal part of pars prostatica level, and smaller eggs. Examination of new and its own. proximal end, then making broad material of II. protopteri indicates that the U loop to right to ascend without further wind- genital pore is not always marginal as originally ing as wider, more muscular terminal part. noted. The pharynx and ceca of our speci- Prostate cells surrounding anterior chamber of men contain erythrocytes; Dollfus (1950) re- seminal vesicle, pars prostatica and cirrus. ported blood in the ceca of H. crumenifer. Ovary 275 by 135, narrow, elongate, lobecl, The distribution of body spines in our speci- dextral, anterior margin 16 anterior to level of men may not be entirely correct as it appears

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 that some spines were lost; both Baylis (1915) 150-305 by 174-335; sucker length ratio and Dollfus (1950) noted that spines were 1:1.35-1.85; testis 160-710 by 174-835, trans- readily lost by H. cnimenifer. Odening (1964) versely elongate but somelimes almost round, indicated that the taxonomic position of the surface smooth to wavy or with very slight subfamily Heterorchiinae Dollfus, 1950, was indication of lobing; cirrus sac 60-150 by 36— uncertain. Dollfus (1963) erected the family 133, usually longitudinally elongate but some- Heterorchiidae within the superfamily Plagior- times almost round, slightly thick walled, mus- chioidea Dollfus, 1930. cular, containing sinuous, tubular internal seminal vesicle, ejaculatory duct and prostate cells; Family Paramphistomatidae genital pore median to submedian (right or Diplodiscus magnus (Srivastava, 1934) left), usually at level of cecal bifurcation, may be at level of esophageal bulb or just post- SYNONYMS: Diplodiscus amphichnis Tuban- bifurcal; ovary 87-355 by 97-440, usually gui, 1933 (in part), D. amphichnis magnus transversely elongate but may be round or Srivastava, 1934, D. amphichnis japonicus longitudinally elongate, surface smooth to Yamaguti, 1936, D. japonicus (Yamaguti, slightly wavy, usually submedian (right or 1936) Li, 1937, D. mehrai Pande, 1937. left) in position but may be median, usually HOSTS: Dicroglossus occipitalis (Giinther), separated from testis but may be in contact Rana galamensis Dum. and Bibr., Hylarana with or slightly overlap it dorsally; vitelline albolabris (Hallowell) (Ranidac); Kassina follicles large, surface smooth to wavy or some- senegalensis (Dum. and Bibr.) (Rhaeophor- what lobed, round to irregular in shape, 29-39 idae); Xenopus tropicalis (Gray) (Pipidae). in number, lateral and ventral to ceca with a HABITAT: Rectum. few intruding into intercecal space, usually LOCALITIES: Pokoasi (D. occipitalis), Legon extending to level of esophageal bulb or slightly (D. occipitalis, R. galamensis), Achimota (K. more anteriorly but in a few almost to oral senegalensis, X. tropicalis), Nungua (D. oc- diverticula, confluent dorsally posterior to cipitalis, R. galamensis), Kacle (H. albolabris), ovary and dorsal to anterior part of acetab- Cape Coast (D. occipitalis); Ghana. ulum, not confluent anteriorly; eggs one to DATES: 14 December 1957, 21 January more than 200 in number, usually fewer than 1958, 1 November 1960, 4 June 1961, 20 50, 40 measuring 106-136 by 68-90; excre- October 1965 (D. occipitalis); 4 June, 12 tory tubules in body proper as illustrated by November 1961 (R. galamensis); 12 Novem- Yamaguti (1936) for D. amphichnis japonicus, ber 1961 (X. tropicalis); 14 November 1961 acetabular tubules not observed. (K. senegalensis); 25 November 1961 (H. DISCUSSION: Only two species of Diplodiscus albolabris). Diesing, 1836, are known from Africa: D. SPECIMENS: USNM Helm. Coll. No. 62894 subclavatus (Pallas, 1760) Diesing, 1836, re- (from D. occipitalis); No. 62895 (R. gala- ported by Skrjabin (1916) from Bufo sp. mensis); No. 62896 (PI. albolabris); No. 62897 (Bufoniclae) from Central Africa; D. pal- (K. senegalensis); No. 62898 (X. tropicalis). lascatus described by Manter and Pritchard DESCRIPTION (based on 32 specimens from (1964) from Bufo regularis Reuss from the D. occipitalis, 12 measured; six from R. gala- Congo. Vercammen-Grandjean (1960) and mensis, four measured; one from H. albolabris, Pritchard (1964) noted that a record of D. measured; one from K. senegalensis, unmea- subclavatus from South Africa is without doubt sured; one from X. tropicalis, measured): Body Progonimodiscus doi/eri (Ortlepp, 1926) Ver- 1,140-4,325 by 590-1,910; living worms pink- cammen-Grandjean, 1960. We wonder if Skr- ish; eyespot pigment present; oral sucker (in- jabin's record of D. subclavatus may in reality cluding diverticula) 285-790 by 201-575, be D. magnus as herein reported by us or margins with sensory papillae, diverticula 128— D. pallascatus. 365 by 123-380; esophagus (including mus- Our specimens readily fit the description of cular bulb) 315-850 long, surrounded by Diplodiscus amphichnis magnus which was gland cells, bulb 73-182 wide; acetabulum originally described as a new subspecies by 385-1,250 by 520-1,640, margins with sen- Srivastava (1934) from Rana ct/anophlyctis sory papillae, conical sucker in acetabular cup Schneider and R. tigrina Merrem (Ranidae)

Copyright © 2011, The Helminthological Society of Washington PROCEEDINGS OF THE HELMINTHOLOGICAL SOCIETY from India. It was differentiated in part from Family Plagiorchiidac D. amphichrus Tubangni, 1933, in having the Metaplagiorchis biloborchis n. sp. vitelline follicles in separate fields anteriorly (Figs. 4, 5) rather than being confluent. Singh (1954) and Agrawal (1966) considered D. amphichrus HOST: Dicroglossus occipitalis (Giinther) magnus and D. mehrai Pande, 1937, synonyms (Ranidae). of D. amphichrus, citing the variable positions HABITAT: Duodenum. of the genital and excretory pores. D. mehrai LOCALITY: Nungua, Ghana. also has the vitelline follicles in separate fields DATE: 4 June 1961. anteriorly. There is no account in the literature HOLOTYPE: USNM Helm. Coll. No. 62900. of a single population from the same host spe- DIAGNOSIS (based on single specimen): Body cies being dimorphic with respect to the dis- 1,573 by 510, elongate, widest postacetabular, tribution of the vitelline follicles. Therefore, extremities rounded; tegument spined to short D. amphichrus magnus and D. mehrai cannot distance posttesticular. Parenchymal gland be considered synonyms of D. amphichrus. D. cells numerous lateral to pharynx and postero- amphichrus japonicus was described as a new lateral to oral sucker. Forebody 422, hindbodv subspecies by Yamaguti (1936) but was raised 1,004; preoral body 23, posttesticular space to specific rank (D. japonicus) independently 450, postcecal space 276, postvitellarian space by Li (1937) and Pande (1937) because of 220. Oral sucker 270 in diameter, ventral, the constant occurrence of the vitelline follicles opening longitudinally elongate. Acetabulum in separate fields anteriorly. Manter and Prit- 147 in diameter, at level of anterior two-fifths chard (1964) considered D. sinicus Li, 1937, of body length, opening transversely oval. a valid species rather than a synonym of D. Sucker length ratio 1:0.54. Prepharynx and amphichrus; they placed D. mclanosticti Yama- esophagus absent; pharynx 105 by 138, slightly guti and Mitunaga, 1943, in synonymy with overlapping oral sucker dorsally; cecal bifurca- D. sinicus. As a result of our comparative tion short distance preacetabular; ceca with studies we are raising the subspecies D. am- conspicuous internal cell layer, terminating phichrus magnus to specific rank, its designa- short of posterior extremity. Excretory pore tion becoming D. magnus (Srivastava, 1934). subterminal ventral; bladder not visible. We declare D. japonicus and D. mehrai syn- Testes two, diagonal, well separated from onyms of D. magnus. Additionally, we declare one another, deeply bilobed, lobes smooth, as synonymous with D. magnus those speci- overlapping ceca ventrally. Anterior testis mens of D. amphichrus in which the vitelline sinistral, 194 long, anterior lobe 100 by 138, follicles are in separate fields anteriorly [e.g., posterior lobe 102 by 126, latter more dorsal than anterior lobe and overlapping it; posterior D. amphichrus of Singh (1954) and Agrawal testis dextral, 206 long, anterior lobe 128 by (1966)]. 80, posterior lobe 133 by 78, latter more ventral than anterior lobe and overlapping it; Schizamphistomoides spinulosum anterior testis lying 225 and posterior testis (Looss, 1901) Stunkard, 1925 375 postacetabular. Cirrus sac 440 by 70, SYNONYM: Amphistomum spinulosum Looss, thick walled, muscular, commencing 127 post- 1901. acetabular and mesial to ovary, curving around HOST: Chelonia mi/das (L.) (Cheloniidae). right side of acetabulum to genital pore opening at anterosinistral margin of latter, overlap- HABITAT: Small intestine. ping margins of acetabulum and right cecum LOCALITY: Ada, Ghana. dorsally. Seminal vesicle saccular, bipartite, DATE: 21 October 1954. posterior chamber 235 by 67, anterior 58 by SPECIMENS: USNM Helm. Coll. No. 62899. 39. Pars prostatica long, cell lined. Cirrus DISCUSSION: The testes in our three worms muscular, long, protrusible. Prostate cells sur- are much transversely elongated. This trem- rounding anterior chamber of seminal vesicle atocle has been reported from the same host and pars prostatica. species from the Mediterranean Sea and Pacific- Ovary 131 by 121, smooth, dextral, lying coast of Panama. postacetabular, overlapping right cecum dor-

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 sally. Ootype complex posteromesial to ovary. did not carry conviction. Byrd and Maples Seminal receptacle absent. Uterus descending (1963) accepted Reynoldstrema. Manter and and ascending between testes, intercecal, filling Pritchard (1964) considered G. africana a postcecal space. Metraterm long, thick walled, second species of Haplometra Looss, 1899. muscular, surrounded by gland cells, com- Yamaguti (1958) characterized Haplometra as mencing at ovarian level just posterior to having the testes tandem or nearly so and the proximal end of cirrus sac, ascending ventral vitellaria extending the length of the ceca to latter along dextrodorsal part of acetabulum except for the posterior part, and as lacking to open into genital atrium. Vitelline follicles a seminal receptacle; Cheng (1959) character- extending from level of cecal bifurcation on ized Reynoldstrema as having diagonal testes, right, anterior margin of acetabulum on left, the vitellaria extending around the ends of the terminating slightly beyond cecal ends; dorsal, ceca, and a seminal receptacle. Manter and lateral and ventral to ceca, confluent post- Pritchard thought that the receptacle reported testicular dorsal to uterus. Eggs yellow, oper- by Dollfus (1950) was probably a uterine one; culate, eight measuring 33-37 by 18-20. however, Beverley-Burton (1963) reaffirmed DISCUSSION: Timofeeva (1963) created the the presence of a true receptacle. Therefore, genus Metaplagiorchis for five species of it is our opinion that G. africana cannot be Plagiorchis Liihe, 1899 [P. molini Lent and assigned to Haplometra. Nasir (1966) con- Freitas, 1940; P. himalayai (Jorclon, 1930); sidered Reynoldstrema a synonym of Glypt- P. lenti Freitas, 1941; P. momplei (Dollfus, helmins. We accept the genus Reynoldstrema 1932); P. ramlianus (Looss, 1896)] from am- and its single species R. africana (Dollfus, phibians and reptiles in which the ceca do not 1950) Cheng, 1959 [syn. Glypthelmins afri- extend to the posterior extremity, the vitellaria cana Dollfus, 1950; Plagiorchis (Plagiorchis) extend to the ends of the ceca or nearly so, and africanus (Dollfus, 1950) Vercammen-Grand- the uterine coils fill the body postcecally. Our jean, 1960; Haplometra africana (Dollfus, species appears closest to M. ramlianus but 1950) Manter and Pritchard, 1964]. Addi- differs from it and all other species of the tionally, we are assigning two other species genus in having both its testes deeply bilobed. to it: R. laurenti (Vercammen-Grancljean, Odening (1959a), in emending the diag- 1960) n. comb. [syn. Plagiorchis (Plagiorchis) nosis of Plagiorchis, removed all species (the laurenti Vercammen-Grandjean, I960]; R. above five of Timofeeva plus three others) in herghei (Vercammen-Grancljean, 1960) n. which the uterus fills the posterior end of the comb. [syn. Plagiorchis (Plagiorchis) berg- body and the vitellaria do not extend to the hei Vercammen-Grandjean, I960]. Odening posterior extremity; he suggested that they be (1964) placed Reynoldstrema in the family assigned to a new genus but did not do so. Macroderoididae (Plagiorchioiclea). Capron, Deblock, and Brygoo (1961) declared P. himalayai a synonym of P. ramlianus. Bev- Ostioloides rappiae (Szidat, 1932) erley-Burton (1963) noted in her redescription Odening, 1960 (Figs. 6, 7) of Glypthelmins africana Dollfus, 1950, that some of the species assigned to the genus SYNONYM: Haplometroides rappiae Sziclat, Plagiorchis from amphibians and reptiles (P. 1932. himalayai, P. momplei, P. ramlianus) show more HOSTS: Hyperolius concolor (Hallowell) similarity to G. africana than do the accepted (syn. Rappia c. H.), H. nasutns igbettensis species of Glypthelmins Stafford, 1905; she sug- Schi0tz, H. mtiduliis Peters (Rhacophoridae); gested that if a comparative survey was made Mabuia perrotetii (Dum. and Bibr.) (Scincidae). of plagiorchiids from these hosts some of the HABITAT: Small intestine. species concerned might possibly be assigned LOCALITIES: Tinkong (H. concolor), Ag- to a separate new genus. Cheng (1959) had bogba (H. concolor, H. nitidulus), Legon (H. already created the genus Reynoldstrema for nasutus), Cape Coast (H. concolor, M. perro- G. africana. Vercammen-Grandjean (1960) tetii) ; Ghana. transferred G. africana to Plagiorchis. Deblock DATES: 3 June (Tinkong), 11 July (Legon), and Capron (1962) noted that Cheng's dis- 22 November (Agbogba) 1961; 2 November cussion supporting creation of the new genus 1965, 17 April 1966 (Cape Coast).

SPECIMENS: USNM Helm. Coll. No. 62901 terior 68-177 by 38-77, anterior 26-41 by (from H. concolor); No. 62902 (H. nasutits 26-43. Pars prostatica thick walled, muscular, igbettcnsis); No. 62903 (H. nitidulus); No. cell lined, small, slightly elongate. Cirrus mus- 62904 (M. perrotetii). cular, protrusible, longer than pars prostatica. DESCRIPTION (based on 30 specimens, 14 Prostate cells surrounding anterior chamber of mature adults measured): Body 2,038-4,205 seminal vesicle, pars prostatica, and cirrus. by 830-1,360, elongate, widest at gonadal Genital pore clextrolateral, prececal, between level; entire tegument spined except for ex- esophagus and body margin. treme anterior tip in front of oral sucker, small Ovary 170-310 by 195-320, smooth, pre- papillae in unspined area. Parenchyma! gland testicular, overlapping acetabulum 26 to lying cells between acetabulum and oral sucker, fill- 105 postacetabular, sinistral to median in posi- ing almost entire prececal space, primarily tion in intercecal space. Seminal receptacle extracecal between acetabulum and cecal 75-175 by 75-157, posterior or posterodorsal bifurcation. Forebody 560-1,035, hindbody to ovary, dorsal to Mehlis' gland. Laurer's 1,260-2,920; preoral body 5-24, posttesticular canal muscular, winding to dorsal surface. space 890-2,130, postvitellarian space 495- Uterus voluminous, occupying most of hind- 1,110, postcecal space 520-1,215. Oral sucker body to posterior extremity, may coil extra- 163-219 by 184-240, transversely elongate, cecally toward level of cecal ends, single de- subterminal ventral. Acetabulum 172-250 by scending and ascending limbs passing between 184—265, usually transversely elongate but may testes, ascending limb passing to right of ovary. be round, center at level of anterior one- to Metraterm usually straight, 230-424 long, thick two-fifths body length. Sucker length ratio walled, muscular, surrounded by gland cells, 1:0.96-1.31. Prepharynx extremely short; crossing right cecum ventrally, passing dorsal pharynx 87-131 by 90-133, round to longi- to cirrus sac but may be on either side of it tudinally or transversely elongate; esophagus at times, usually longer than cirrus sac but 108—275 long; prepharynx, pharynx and esoph- may be same length or shorter, proximal end agus surrounded by gland cells; cecal bifurca- may begin short distance posterior or anterior tion 124—450 preacetabular; ceca conspicuously to proximal end of cirrus sac but distal end cell lined, length usually subequal with left always extending anterior to latter. Vitelline cecum usually shorter, terminating closer to follicles small, numerous, extending from short testes than to posterior extremity. distance prebifurcal or postbifurcal to short Testes two, smooth to slightly wavy, sym- distance anterior or posterior to cecal ends, metrical with left testis usually slightly an- right and left fields in any one individual may terior to right but reverse may occur, in contact start as well as end at different levels, follicles with or partly overlapping ceca ventrally, left lateral and ventral to ceca, some ventral fol- testis posterior to ovary but in one worm it licles intruding into intercecal space, a very is posterolateral to latter and in another antero- few lateral follicles intruding into dorsal extra- lateral, left testis usually separated from ovary cecal space. Eggs numerous, yellow, opercu- but may be in contact or overlap, testes larger late, 35 measuring 25-30 by 15-18. to smaller than ovary. Right testis 148-345 by Excretory bladder Y shaped, stem relatively 145-310, 133-535 postacetabular; left testis long and narrow, arms short, entirely post- 175-290 by 145-275, 135-608 postacetabular. testicular; bladder narrows just before subter- Vasa efferentia joining to form relatively long minal dorsal pore, narrow portion surrounded vas deferens. Cirrus sac 169-315 by 58-100, by gland cells. thick walled, muscular, commencing 28-275 preacetabular, diagonally oriented, crossing Ostioloides Odening, 1960 char, emend. right cecum ventrally, terminating at genital atrium lying between esophagus and right body DIAGNOSIS: Plagiorchiiclae. Body elongate, margin, containing seminal vesicle, pars pros- spined. Suckers about equal in size. Extremely tatica, prostate cells and cirrus. Seminal vesicle short prepharynx, pharynx and long esophagus saccular, bipartite, deep constriction separates present. Cecal bifurcation preacetabular; ceca chambers so that narrow channel connects conspicuously cell lined, extending posttesticular them, chambers thick walled, muscular, pos- but terminating well in advance of posterior

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 extremity. Testes two, symmetrical, intercecal, trolateral rather than dextrolateral, and sem- in middle body third. Cirrus sac preacetabular, inal vesicle saccular rather than coiled. passing ventral to right cecum, containing sac- Laiogonimus was created by Vercammen- cular, bipartite seminal vesicle, short pars pros- Grandjean (1960) for L. mariavirginiae from tatica and long cirrus. Genital pore clextro- Pti/chadena sp. (Ranidae) from the Congo. lateral between esophagus and body margin. Deblock and Capron (1962) described Astio- Ovary pretesticular, postacetabular, intercecal. trema (Biguetrema) tananarivense as a new Seminal receptacle and Laurer's canal present. subgenus and new species from frogs, Rana Uterus descending and ascending between mascareniensis Dum. and Bibr. (Ranidae), testes, filling most of inter- and postcecal por- Rhacophorus sp., R. goudoti (Tschudi) (Rha- tions of hindbody. Metraterm present. Vitel- cophoridae), and a snake, Liopholidophis lat- line follicles small, numerous, in lateral fields, eralis Dum. and Bibr. (Colubridae), from extending from cecal bifurcation to cecal ends. Madagascar. They indicated that additional Eggs operculate, numerous. Excretory bladder studies may possibly raise their new subgenus Y shaped with relatively long stem and short to generic level. In our opinion A. (B.) tana- arms, entirely posttesticular. narivense is a species of Laiogonimus, becom- TYPE SPECIES: O. rappiae (Sziclat, 1932). ing L. tananarivense (Deblock and Capron, DISCUSSION: Our collection consists of 13 1962) n. comb. The latter differs significantly mature adult, one young adult, and two im- from L. mariavirginiae in the testes not being mature worms from H. concolor, nine mature completely surrounded by the uterus, in having adults from H. nasutus igbettensis, and one a loop in the seminal vesicle, and in the oral mature adult each from PI. nitidulus and M. sucker being smaller than the acetabulum. perrotetii. Szidat (1932) originally described this form as Haplometroides rappiae from a Family Mesocoeliidae single specimen from Hyperolius (= Rappia) Mesocoelium monodi Dollfus, 1929 concolor from Liberia. Odening (1960) cre- HOSTS: Bufo cameronensis cameronensis ated the genus Ostioloides for H. rappiae, pro- Parker, B. reguhris Reuss (Bufonidae); Pty- visionally placing it in the subfamily Haema- chadena aequiplicata (Werner), Rana gala- toloechinae (Plagiorchiidae) until the species mensis Dum. and Bibr., Dicroglossus occipitalis was adequately described. We agree that this (Giinther), Conrana crassipes alleni (Barbour species does not belong in the genus Haplo- and Loveridge), Hylarana albolabris (Hallo- metroides Odhner, 1911. Ostioloides is closest well), Phrynobatrachus sp. (Ranidae); Hy- to Haplometroides, Laiogonimus Vercammen- perolius concolor (Hallowell) (Rhacophoridae); Grandjean, 1960, and Parahaplometroides Phrynomerus microps (Peters) (Phrynomer- Thatcher, 1963, differing from them in having idae); Mabuia perrotetii (Dum. and Bibr.) the vitellaria more extensively distributed, (Scincidae); Chamaeleo gracilis Hallowell testes symmetrical, and excretory bladder en- (Chamaeleonidae); Varanus niloticus (L.) tirely posttesticular [the extent of the bladder (Varanidae). is not indicated for Parahaplometroides by HABITAT: Small intestine. Thatcher (1963) or Stunkard and Gandal LOCALITIES: Achimota, Amedzofe, Cape (1966) but it probably extends pretesticular]. Coast, Kade, Nungua, west of Takoradi; Ostioloides differs further from Haplometroides Ghana. in having the genital pore extracecal and dex- DATES: 1956-57, 1961, 1965. trolateral between the esophagus and body SPECIMENS: USNM Helm. Coll. No. 62905 margin rather than intercecal; from Laiogoni- (from B. c. cameronensis); No. 62906 (B. mus in having the genital pore dextrolateral regularis); No. 62907 (P. aequiplicata); No. rather than sinistrolateral, and the ceca much 62908 (R. galamensis); No. 62909 (D. occipi- shorter; and from Parahaplometroides in hav- talis); No. 62910 (C. crassipes alleni); No. ing the ceca much shorter, and seminal vesicle 62911 (H. albolabris); No. 62912 (Phnjno- saccular rather than extensively coiled. There batrachus sp.); No. 62913 (H. concolor); No. is considerable similarity between Laiogonimus 62914 (P. microps); No. 62915 (M. perrotetii); and Parahaplometroides, the former differing No. 62916 (C. gracilis); No. 62917 (V. from the latter in having the genital pore sinis- niloticus).

DISCUSSION: Thomas (1965a, b) has re- 165, mostly dorsal to acetabulum. Pars pros- ported in detail the ecology, anatomy, life tatica 201 by 75, very thick walled with rela- history and size allometry of this species from tively narrow lumen. Cirrus 230 by 40, long, Ghana. It has been reported from other parts thin walled, sinuous. Prostate cells very con- of Africa, including Madagascar. spicuous, numerous, compact, surrounding pars prostatica and cirrus. Genital atrium tubular, Family Lecithodendriidae thick walled, surrounded by gland cells. Gen- Ganeo africana (Skrjabin, 1916) ital pore sublateral, ventral, opening into body- Raw, 1950 (Fig. 8) depression. Ovary 140 by 158, smooth to slightly wavy, SYNONYM: Ganeo gloitoides ajricana Skr- median, 63 postacetabular, overlapping and jabin, 1916. diagonal to posterior testis. Ootype complex HOST: Dicroglossus occipitalis (Giinther) posterior to ovary; seminal receptacle 61 by (Ranidae). 39, dextral to Mehlis' gland, dorsal. Uterus HABITAT: Small intestine. winding postovarian, intercecal at vitellarian LOCALITY: Legon, Ghana. level, filling postvitellarian space, ventral to DATE: 11 May 1961. ceca. Metraterm thick walled, surrounded by SPECIMEN: USNM Helm. Coll. No. 62918. gland cells, opening into genital atrium left of DESCRIPTION (based on single specimen): cirrus. Vitelline follicles in lateral fields, lateral Body 2,225 by 432, elongate, sides from level and ventral to ceca, intruding slightly into of cecal bifurcation to posterior extremity intercecal space; right field 245 postacetabular, nearly parallel, tapering to narrower rounded 100 postovarian, left 141 postacetabular, at anterior extremity, posterior extremity bluntly midovary level; right field 575 from posterior rounded, tegument spined. Forebody 805, extremity, left 550; right field 440 long, left hindbody 1,265; preoral body 5, postovarian 576. Eggs numerous, yellow-brown, opercu- space 1,060. Oral sucker 90 by 87, subterminal late, 10 measuring 21-23 by 12-14. ventral. Acetabulum 155 by 138, center at Excretory bladder V shaped, thick walled, level of anterior two-fifths of body length. dorsal to uterus, anterior tips of arms 285 from Sucker length ratio 1:1.72. Prepharynx very posterior extremity; pore subterminal ventral. short; pharynx 73 by 68, slightly overlapping DISCUSSION: This trematode was originally oral sucker dorsally; esophagus 250 long; gland described by Skrjabin (1916) as a new variety, cells surrounding posterolateral margins of oral ajricana, of Ganeo glottoides Klein, 1905, from sucker, prepharynx and anterolateral margins Bufo sp. (Bufoniclae) from Central Africa. of pharynx; cecal bifurcation 375 preacetab- Kaw (1950) raised the variety to species rank; ular; ceca conspicuously cell lined, terminating Fotedar (1959) accepted the latter change. short of posterior extremity, right postcecal In the keys to the species of Ganeo Klein, space 198, left 326. 1905, given by Kaw and by Fotedar our speci- Testes two, tandem, smooth, longitudinally men could not be keyed to G. africana because to transversely elongate, clextral, partly over- the ceca do not extend to near the posterior lapping cecum dorsally. Anterior testis 148 by extremity. Additionally, Skrjabin's (1916) 151, anterodextral to and in contact with ace- illustration shows that the uterus does not tabulum, in contact with pseudocirrus sac; descend beyond the cecal ends, whereas it posterior testis 158 by 180, posterodextral to goes to the posterior extremity in our specimen. acetabulum, overlapping anterodextral part of All other characteristics of the latter are essen- ovary ventrally. Pseudocirrus sac 537 (longi- tially similar to the description and illustration tudinally extent) by 165, commencing 20 post- of G. africana given by Skrjabin. acetabular, sinistral, intercecal most of length, distal portion crossing left cecum ventrally, Prosthodendrium (Paralecithodendrium) terminating extracecally at genital atrium lying glandidosum (Looss, 1896) Dollfus, 1931 near body margin at bifurcal level, containing seminal vesicle, pars prostatica, prostate cells SYNONYMS: Distomum glandidosum Looss, and cirrus. Seminal vesicle tubular, thick 1896, D. obtusum Looss, 1896, probably D. walled, muscular, much coiled in area 160 by chefrenianiim Looss, 1896.

HOST: Chamacleo gmcilis Hallowell (Cha- 2—10 preacetabular, anterior to ovary, may maeleonidae). slightly overlap latter ventrally or be slightly HABITAT: Small intestine. separated, may overlap right cecum ventrally. LOCALITY: Achimota, Ghana. Left testis 78-123 by 97-116, submedian to DATES: 12 January 1954, 9, 15 March 1956. left of esophagus and right of proximal part SPECIMENS:' USNM Helm. Coll. No. 62919. of cirrus sac, may slightly overlap esophagus DISCUSSION: Three hosts harbored one, one, dorsally, posterior part of testis at or just an- and two worms, respectively. Dubois (1962) terior to cecal bifurcation, 51—90 preacetabular. reviewed the subgenus Paralecithodendrium Cirrus sac 375-441 (longitudinal extent) by Odhner, 1911, noting the presence of P. (P.) 90-99, sigmoid shaped in ventral view with glandulosum in a variety of bats and the larger proximal swollen part, thick walled, chameleon, Chamaeleo basiliscus Cope, from muscular, commencing just posterior to an- Egypt. Gupta (1986) reported this parasite terior margin of acetabulum or just preacetab- (as Paralecithodendrium obtusum and P. gland- ularly, ascending sinistrolateral, crossing cecum ulosum) from a bat from India. Our collection ventrally, opening into genital atrium, contain- represents new geographic distribution and ing seminal vesicle, pars prostatica, prostate host species records. cells and cirrus. Gland cells in parenchyma surrounding distal part of cirrus sac, metra- Family Pleurogenidae term, genital atrium and pore. Seminal vesicle Prosotocus exovitellosus n. sp. primarily saccular but parts may be tubular, (Figs. 9, 10) appearing bipartite in some but is actually coiled, fi'lling area 130-157 by 65-80, thick HOST: Chamaeleo gracilis Hallowell (Cha- walled but thinner than cirrus sac, muscular. maeleoniclae). Pars prostatica 145-172 by 56-75, club shaped, HABITAT: Small intestine. walls as for seminal vesicle, filled with large, LOCALITY: Achimota, Ghana. elongate, vesicular cells in proximal more DATE: 9 March 1956. swollen part and smaller, rounder cells in dis- SPECIMENS: USNM Helm. Coll. No. 62920 tal part. Cirrus in distal curve of cirrus sac, (holotype); No. 62921 (paratypes). thick walled, muscular, protrusible. Prostate DIAGNOSIS (based on three adult and two cells surrounding anterior part of seminal ves- immature worms; adults measured): Body icle, pars prostatica and cirrus. Genital atrium 648-770 by 502-595, oval; tegument com- short, very thick walled, muscular. Genital pletely spinecl. Forebody 293-382, hindbocly pore at sinistrolateral body margin in two, 29 250-265; preoral body 9-14. Parenchymal sublateral dorsal in one, at level of oral sucker. gland cells in forebody. Oral sucker 103-123 Ovary 78—97 by 115-148, transversely elon- by 103-126, round to slightly transversely gate, dextral, smooth to having very slight elongate, subterminal ventral. Acetabulum indentations, mesial part bluntly pointed with 103-123 by 103-136, round to slightly trans- oviduct emerging therefrom, dorsal to right versely elongate, center at level of 53-58 per cecum, anterior to right excretory arm. Ootype cent of body length from anterior extremity. complex entirely dorsal to acetabulum in two, Sucker length ratio 1:1.00-1.14. Prepharynx overlapping by one-half posterodextral part of very short; pharynx 61-68 by 56-65, usually acetabulum in one. Seminal receptacle 75- slightly longer than wide but may be wider 111 by 39—53. Laurer's canal not observed. than long, may overlap oral sucker dorsally; Uterus in most of hindbocly, coils to right of gland cells at posterolateral margins of oral ovary, to left vitelline mass, two transverse sucker, prepharynx and anterolateral margins loops, in ascent crossing left cecum ventrally of pharynx; esophagus 72-111 long, sur- and cirrus sac dorsally. Metraterm clextral to rounded by gland cells; cecal bifurcation 72- distal part of cirrus sac, short, thick walled, 82 preacetabular; ceca short, terminating lat- muscular, opening into genital atrium. Vitel- eral to acetabulum, conspicuously cell lined. line follicles in two separate clusters; right Testes two, smooth, usually transversely group extending from oral sucker level to right elongate, prececal. Right testis 75-111 by testis, mesially to esophagus, sometimes partly 128-172, lateral, at level of cecal bifurcation, overlapping right testis dorsally; left group

Copyright © 2011, The Helminthological Society of Washington 12 PROCEEDINGS OF THE HELMINTHOLOGICAL SOCIETY entirely sinistral to cirrus sac and left cecum, to level of vitellaria. Excretory arms uniting partly overlapping them dorsally. Right and dorsal to pharynx. left vitelline ducts crossing ceca dorsally, Testes two, smooth, diagonal; right testis uniting dorsal to Mehlis' gland to form small 415 by 330, more anterior than left, overlap- reservoir. Eggs light brown, operculate, 15 ping acetabular level 175; left testis 445 by measuring 21-26 by 13-16. 385, 120 postacetabular, overlapping level of Excretory bladder V shaped, dorsal to uterus, right testis 85. Seminal vesicle 260 by 92, lined with conspicuous cell layer, arms extend- commencing 760 preacetabular, lying free in ing lateral to acetabulum, apex of V sur- parenchyma, thick walled, muscular, narrow- rounded by gland cells; pore slightly sub- ing at distal end to ejaculatory duct opening terminal ventral. into shallow genital atrium; prostate cells free DISCUSSION: Our collection represents the in parenchyma, surrounding distal end of first records of the genus Prosotocus Looss, seminal vesicle and ejaculatory duct. Genital 1899, from a reptile and from Africa. Previous pore ventral to cecal bifurcation. records are for 11 species from ranid and Ovary 435 by 380, smooth, median, lying bufonid amphibians from India and Europe, 202 posttesticular. Seminal receptacle 97 by one from a vespertilionid bat from Europe, 111, postovarian. Mehlis' gland well de- and one from a mastacembelid freshwater fish veloped, compact, 205 by 210, posteroventral from India. In the key given by Murhar to ovary, sinistroventral to seminal receptacle. (1960) our species keyed to P. indicus Mehra Uterus descending short distance to position and Negi, 1928, in the key by Bhardwaj ventral to vitellaria, then ascending with much (1963) to P. mirabilis Grabda, 1959, and in coiling to posteroventral part of posterior testis, the key by Agrawal (1964) to P. mastacem- passing toward dorsal surface between ovary beli Agrawal, 1964. Our species differs sig- and testes, continuing ascent dorsal to testes nificantly from the above named and all others and acetabulum, preacetabularly coiling be- in the genus in having the left vitelline mass tween dorsal and ventral body surfaces to short lying lateral rather than median to the cirrus distance postbifurcal, descending to level pos- sac, hence the specific name exovitellosus. terior to seminal vesicle, becoming thick walled, Odening (1959b) placed the genus Prosotocus muscular, sinuous, ascending metraterm, latter in a new family Pleurogeniclae [sensu Pleuro- 365 in longitudinal extent. Vitellaria between geninae (Looss) ]. ovary and posterior extremity, five follicles in each group ranging between 210-260 in length. Family Halipegidae Eggs light brown, operculate, numerous, 12 Halipegus ghanensis n. sp. (Fig. 11) measuring 42-52 by 17-27, filament about twice as long as egg. HOST: Chamaeleo gracilis Hallowell (Cha- DISCUSSION: Beverley-Burton (1963) re- maeleonidae). viewed the genus Halipegus Looss, 1899, HABITAT: Stomach. recognizing eight species for which compara- LOCALITY: Achimota, Ghana. tive measurements are given; to these a ninth DATE: 9 January 1955. species should be added, H. insularis Capron, HOLOTYPE: USNM Helm. Coll. No. 62922. Deblock and Brygoo, 1961. Species of Hali- DIAGNOSIS (based on single specimen in pegus reported from Africa in the adult form dextrolateral view; measurements are length by are: H. ovocaudatus (Vulpian, 1858) Looss, 1899, from Rana fuscigula Dum. and Bibr. depth): Body 3,790 by 940, elongate; tegu- (Ranidae) from South Africa; H. sp. Porter, ment unspined. Forebody 1,565, hindbody 1938, from Bufo regularis Reuss (Bufonidae) 1,625; posttesticular space 1,065, postovarian from South Africa; H. africanus DoHfus, 1950, space 425. Oral sucker 317 by 345, subter- from Rana mascareniensis Dum. and Bibr. minal ventral. Acetabulum 600 by 540, equa- from the Congo; H. insularis from Chamaeleo torial. Sucker length ratio 1:1.89. Prepharynx oustaleti Macquard, C. verrucosiis Cuvier, C. absent; pharynx 120 by 115, overlapping oral latemlis Gray, Rana mascareniensis, Rhaco- sucker dorsally; esophagus very short, bifurcat- phorus goudoti (Tschudi) (Rhacophoridae), ing almost immediately into ceca which extend and an unidentified frog from Madagascar;

H. rhodesiensis Beverley-Burton, 1963, from HOST: Crocodilus niloticus Laurent (Croco- Xenopus laevis (Daudin) (Pipidae) from dilidae). Southern Rhodesia. H. sp. Porter resembles HABITAT: Small intestine. H. ovocaudatiis and may be synonymous with LOCALITY: Nungua, Ghana. it. Beverley-Burton (1963) noted that H. bulla DATES: December 1956, January, February Fain, 1953, described from immature worms 1958. from experimentally infected Rana fuscigula SPECIMENS: USNM Helm. Coll. No. 62923. from the Congo, superficially resembles H. DISCUSSION: This trematode has been re- rhodesiensis. Our species differs significantly ported from Osteolaemus tetraspis Cope from H. ovocaudatiis, II. sp. and H. africanus (Crocodilidae) from the Congo, Crocodilus in being less than one-third their lengths (13, cataphractus Cuvier from Gabon (as P. thom- 13, and 12.2 mm, respectively); it differs fur- asi gabonicum), and C. niloticus from Mada- ther from the first two species in having the gascar. Its presence in Ghana represents a testes close to the acetabulum rather than new geographic distribution record. Deblock, far removed, and from the third species in Capron, and Brygoo (1965) declared the having the vitelline follicles smooth rather variety gabonicum Dubois, 1948, a synonym than lobed. Although all specimens of H. of P. thomasi as their specimens from Mada- insularis are longer than our specimen the gascar possessed intermediate characteristics. metraterm in the former is less than one-half as long as in the latter and is shorter than the Order Strigeidida seminal vesicle rather than longer; the egg fila- Strigeidida Metacei caria Dollfus, 1950 ment in the former is 3.2-3.4 times longer than the egg rather than twice as long. Although HOST: Psammophis sibilans (L.) (Colu- H. rhodesiensis and our specimen are the same bridae). length the gonads in the former are one-half HABITAT: Encysted in viscera and mesen- as long as in the latter; the eggs in the former teries. are significantly longer (70-74), and the egg LOCALITY: Brimsu, Ghana. filament is three times longer than the egg. DATE: 15 November 1965. SPECIMENS: USNM Helm. Coll. No. 62924. Family Angiodictyidae DISCUSSION: Several hundred metacercariae Microscaphidium reticularis were present in transparent, membranous cysts in one of four African beauty snakes from the (Van Beneden, 1859) Looss, 1902 and Cape Coast area. Our specimens are similar to Microscaphidium aberrans Looss, 1899 metacercariae described (p. 91-94) and illustrated (Figs. 72, 73) by Dollfus (1950) from HOST: Chelonia mijdas (L.) (Cheloniidae). two different snakes, Lycophidion capense HABITAT: Esophagus. (Schmidt) (Colubridae) and Python sebae LOCALITY: Ada, Ghana. (Gmelin) (Boidae), from the Congo. DATES: 2, 21 October 1954. DISCUSSION: Our collection consists of im- Literature Cited mature specimens with one exception with a single egg. On both dates mixed infections Agrawal, V. 1964. A trematode, Prosotocus mas- occurred, with M. aberrans being considerably tacembeli n. sp. (subfamily Prosotocinae Yama- guti, 1958) from the intestine of a fresh water more common. fish, Mastacembelus armatus (Lacep.) from Lucknow. Proc. Helm. Soc. Wash. 31: 219- Family Proterodiplostomatidae 222. Pseudoneodiplostomum thomasi . 1966. Studies on some trematode parasites of frogs from Lucknow. Indian J. Helm. (Dollfus, 1935) Dubois, 1936 18: 82-90. SYNONYMS: Neodiplostoma thomasi Dollfus, Baylis, H. A. 1915. A trematode from Protop- terus. Ann. and Mag. Nat. Hist., Ser. 8, 16: 1935, Neodiplostoma sp. Dollfus, 1935, Pseu- 85-96. doneodiplostomum thomasi gabonicum Dubois, Beverley-Burton, M. 1963. Some digenetic trem- 1948. atodes from amphibians and reptiles from

Southern Rhodesia including two new species Fotedar, D. N. 1959. On a new species of the and a new genus: Sarumitrema hystatorchis genus Ganeo Klein, 1905 and some notes on n. gen., n. sp. (Plagiorchiidae) and Halipegus the genus. J. Helm. 33: 151-160. rhodesiensis n. sp. (Halipegidae). Proc. Helm. Gupta, N. K. 1966. On some trematode para- Soc. Wash. 30: 49-59. sites of the family Lecithodendriidae Odhner, Bhardwaj, O. N. 1963. Morphology and tax- 1911. Res. Bull. (N.S.) Panjab Univ., Sci. 17: onomy of three new species belonging to the 121-129. subfamily Prosotocinae Yamaguti—1958 (Fam. Kaw, B. L. 1950. Studies in helminthology: Lecithodendriidae: Trematoda). Proc. Natl. Helminth parasites of Kashmir. Part I. Trem- Acad. Sci. India, Sec. B, 33: 345-354. atoda. Indian J. Helm. 2: 67-126. Byrd, E. E., and W. P. Maples. 1963. The Li, L. Y. 1937. Some trematode parasites of glypthelminths (Trematoda: Digenea), with frogs with a description of Diplodiscus sinicus a redescription of one species and the erection sp. nov. Lingnan Sci. J. 16: 61-70. of a new genus. Ztschr. Parasitenk. 22: 521- Manter, H. W., and M. H. Pritchard. 1964. 536. Mission de zoologie medicale au Maniema Capron, A., S. Deblock, and E. R. Brygoo. 1961. (Congo, Leopoldville) (P. L. G. Benoit, 1959). Miscellanea helminthologica madagascariensis. 5. Vermes-Trematoda. Ann. Mus. Roy. Afri- Trematodes de cameleons de Madagascar. que Centr., in-8", Sci. Zool. 132: 75-101. Arch. Inst. Pasteur Madagascar 29: 1-71. Murhar, B. M. 1960. On a new trematode, Cheng, T. C. 1959. Studies on the trematode Prosotocus dorsoponis (subfamily Prosotocinae family Brachycoeliidae, II. Revision of the Yamaguti 1958) from common Indian frog, genera Glypthelmins (Stafford, 1900) Staf- Rana tigrina Daud. Bull. Zool. Soc. Coll. Sci. ford, 1905, and Margeana Cort, 1919; and the Nagpur 3: 67-72. description of Reynoldstrema n. gen. (Gly- Nasir, P. 1966. Two new species of digenetic pthelminae n. subfam.). Am. Midi. Nat. 61: trematodes from Venezuelan amphibians. Proc. 68-88. Helm. Soc. Wash. 33: 166-170. Deblock, S., and A. Capron. 1962. Description Odening, K. 1959a. liber Plagiorchis, Ompha- d'Astiotrema (Riguetrema) tananarivense n. lometra und Allocreadium (Trematoda, Di- subgen., n. sp., Plagiorchiidae intestinal d'am- genea). Ztschr. Parasitenk. 19: 14-34. phibiens malagaches. Ann. Parasit. 37: 97- . 1959b. Das Exkretionssystem von Oin- 107. phalometra und Brachycoelium (Trematoda, , , and E. R. Brygoo. 1965. Trem- Digenea) und die Taxonomie der Unterord- atodes de reptiles (crocodiliens et sauriens) nung Plagiorchiata. Ztschr. Parasitenk. 19: de Madagascar et de Nossi-Be. An sujet de 442-457. sept especes, dont trois nouvelles des genres . 1960. Trematoda, Digenea. Plagior- Plagiorchis (Miiltiglandularis), Ommatobre- chiidae III (Haematoloechinae) und Om- phus et Cryptotropa. Bull. Mus. Natl. Hist. phalometridae. In Das Tierreich. Leiferung Nat, Paris, 2e Ser. 37: 503-522. 75, 77 p. Walter de Gruyter & Co., Berlin. . 1964. Zur Taxionomie der Trematode- Dollfus, R. P. 1950. Trematodes recoltes au nunterordnung Plagiorchiata. Monatsb. Deut- Congo Beige par le Professeur Paul Brien sch. Akad. Wissensch. Berlin 6: 191-198. (mai-aout 1937). Ann. Mus. Congo Beige, Pande, B. P. 1937. On some digenetic trem- C.-Zool., Ser. V, 1: 1-136. atodes from Rana cyanophlyctis of Kumaon . 1963. Hotes et lieux de recolte de quel- Hills. Proc. Indian Acad. Sci., Sec. B, 6: ques trematodes digenetique de vertebres cle 109-120. la collection du Musee royal de 1'Afrique cen- Pritchard, M. H. 1964. Notes on four hel- trale. Rev. Zool. et Bot. Africaines 68: 323- minths from the clawed toad, Xenopus laevw 357. (Daudin), in South Africa. Proc. Helm. Soc. Dubois, G. 1962. Contribution a 1'etude des Wash. 31: 121-128. trematodes de chiropteres. Revision du sous- Prudhoe, S. 1957. Exploration du Pare Na- genre Paralecithodendrium Odhner 1911. Rev. tional de 1'Upemba. Mission G. F. de Witte Svrisse Zool. 69: 385-407. (1946-1949). Trematoda. Inst. Pares Natx. Fischthal, J. H., and R. E. Kuntz. 1963. Trem- Congo Beige, Bruxelles, Fasc. 48, 28 p. atode parasites of fishes from Egypt. Part VI. Rees, G. 1964. Two new species of the genus The metacercaria of Euclinostomum lietero- Plaematoloechus ( Digenea: Plagiorchiidae) stomum (Rudolphi, 1809) Travassos, 1928 from Rana occipitalis- (Gunther) in Ghana. (Clinostomidae), with a review of the genus. Parasitology 54: 345-368. Tr. Am. Micr. Soc. 82: 335-342. Schiot/, A. 1964. A preliminary list of amphib-

ians collected in Ghana. Viclensk. Medcl. distomidae) and Acanthostomum bagri, n. sp. Dansk Naturh. Foren. 127: 1-17. (Acanthostomidae: Acanthostominae) from Singh, K. S. 1954. Some trematodes collected West Africa. Proc. Helm. Soc. Wash. 25: 8- in India. Tr. Am. Micr. Soc. 73: 202-210. 14. Skrjabin, K. I. 1916. Parasitic trematodes and . 1965a. Studies on some aspects of the nematodes collected by the expedition of Prof. ecology of Mesocoelium monodi, a trematode V. Dogiel and I. Sokolov in British East Africa. parasite of reptiles and amphibians. Proc. Sci. Results Zool. Exped. British E. Africa and Zool. Soc. London 145: 471-494. Uganda by Prof. V. Dogiel and I. Sokolov . 1965b. The anatomy, life history and Year 1914, 1: 1-157. (Russian and English size allometry of Mesocoelium monodi Dollfus, texts.) 1929. J. Zool. 146: 413-446. Srivastava, H. D. 1934. On new trematodes of Timoteeva, T. N. 1963. (Revision of the genus frogs and fishes of the United Provinces, India. Plagiorchis Liihe, 1899.) Trudy Gel'mintol. Part IV. The occurrence and seasonal inci- Lab. Akad. Nauk SSSR 12: 225-227. 1962. dence of infection of certain trematodes in (Russian text.) the above hosts. Proc. Acad. Sci. U.P. Agra Tubangui, M. A. 1933. Trematode parasites and Oudh, India 4: 113-119. of Philippine vertebrates, VI. Descriptions of Stunkard, H. W., and C. P. Gandal. 1966. A new species and classification. Philippine J. digenetic trematode, Parahaplometroides basil- Sci. 52: 167-197. iscae Thatcher, 1963, from the mouth of the Vercammen-Grandjean, P. H. 1960. Les trem- crested lizard, Basiliscus basiliscus. Zoologica atodes du lac Kivu sud (Vermes). Ann. Mus. (N. Y.) 51: 91-95. Roy. Afrique Centr., n.s. in 4", Sci. Zool. 5: Szidat, L. 1932. Parasiten aus Liberia und Franzosisch-Guinea. II. Teil: Trematoden. 1-171. Ztschr. Parasitenk. 4: 506-521. Yamaguti, S. 1936. Studies on the helminth Thatcher, V. E. 1963. The trematodes of the fauna of Japan. Part 14. Amphibian trem- basilisk lizard from Tabasco, Mexico. An. Inst. atodes. Jap. J. Zool. 6: 551-576. Biol. Mex. 34: 205-216. . 1958. Systema helminthum. Vol. I. Thomas, J. D. 1958. Two new digenetic trem- The digenetic trematodes of vertebrates. Parts atodes, Heterorchis protopteri, n. sp. (Fello- I and II. 1,575 p. Interscience Publ., N. Y.

Proteocephalus chologasteri sp. n. (Cestoda: Proteocephalidae) from the Spring Cavefish Chologaster agassizi Putman, 1782 (Pisces: Amblyopsidae) of Kentucky

FRED H. WHITTAKER AND LOREN G. HiLL1 Department of Biology, University of Louisville, Louisville, Kentucky

During recent studies by the junior author- the worms were identical and belonged to the on the ecology of the cavefish Chologaster genus Proteocephalus Weinland, 1858. agassizi Putman collected from Warren County, As far as it is known, there are no reports Kentucky, a small tapeworm was observed in the literature of tapeworms from C. agassizi. protruding from the vent of one of the pre- This cavefish is a member of the Amblyopsidae served fish. The worm was eventually re- which includes the small group of North Amer- moved intact, and morphological studies of ican freshwater cavefishes characterized by re- this and other cestode specimens from both duced or degenerate eyes, a thoracic vent in preserved and live C. agassizi indicated that older members and troglobitic or nocturnal habits. According to Hill (1966), C. agassizi 1 Present address: Department of Zoology, University of Oklahoma, Norman, Oklahoma. possesses pigment and hypertrophied sensory

Table 1. Occurrence of Proteocephahls chologasteri sp. n. in Chologaster agassizi collected at Rich Pond, Bowling Green, Kentucky.

A«e group No. fish No. fish Per cent frequency Intensity of of fish examined parasiti/.ed of infection infection

<)' 5 2 40 2 1 10 4 40 5 2 20 18 90 27 4 3^ 1 33.3 5 Totals 38 35 - 39

1 Less than 1 year old. - Two years old when collected and kept without feeding in laboratory for two years. receptors and is found in Kentucky, Tennessee, in length by 0.245-0.397 (0.344) in width. and southern Illinois. Scolex small and narrowest part of worm, from This paper presents the first description of 0.128-0.156 (0.137) in width; suckers well a cestode (Proteocephahls chologasteri sp. n.) developed, 0.053-0.088 (0.069) in diameter; from the spring cavefish Chologaster agassizi functional muscular apical sucker present, Putman in North America. 0.025-0.047 (0.033) in diameter. Relatively long unsegmented neck region from 0.510- Materials and Methods 0.782 (0.651) in length. Genital pores mid- Since tapeworms obtained from preserved marginal, slightly ventral, irregularly alternate. cavefish were unsuitable for critical taxonomic Cirrus pouch very small, ovoid or pyriforrn, studies, 38 live cavefish were collected from 0.062-0.074 (0.069) in length by 0.035-0.046 which 39 tapeworms in various stages of de- (0.040) in width in mature segments, only velopment were recovered (Table 1). The infrequently extending transversely beyond worms were placed in 0.8% saline to observe row of vitellaria. Everted cirrus unarmed. scolex and strobilar movements and then fixed Testes ovoicl, from 0.024-0.0387 (0.0388) in in Benin's fluid. length, numbering 33-47 (42) per mature The description of the adult tapeworm is segment. Testes medullary, distributed in based on hematoxylin-stained whole mounts layers throughout intervitelline field between and cross sections of 12 gravid specimens. anterior border of segment and ovary. Vas Drawings were made with the aid of a camera deferens coiled from near midline until its lucida, and all measurements are based upon entrance into cirrus pouch. Vagina, opening relaxed, unflattened specimens. into genital atrium anterior and slightly dorsal to cirrus pouch, passing medially from genital Proteocephahls chologasteri sp. n. atrium, paralleling cirrus pouch and passing (Figs. 1-3) posteriorly in dorsal medulla to ovary. Vagina, DESCRIPTION: (All measurements are in in region of ovary, curving ventrally and en- millimeters. Average measurements are given larging to form seminal receptacle just anterior in parentheses after the range.) Proteocephal- to ovarian isthmus. Ovary bilobed and situated idae, Proteocephalinae. Length of strobila 4.2- posteriorly in segment; lobes connected some- 9.7 (5.9), with a maximum number of 20 seg- what anteriorly by narrow isthmus. In gravid ments and a minimum of six for gravid speci- segments, isthmus arching anterior to ovarian mens; segmentation acraspedote and at times lobes. Follicular vitellaria medullary, in two indistinct. Strobila widening immediately pos- lateral fields extending from anterior border terior to scolex, gradually increasing in width of segment to ovary. Uterus in early mature throughout its length. Immature segments segments appearing as small, extremely thin- wider than long. Mature segments longer than walled tube extending only a short distance wide ranging in length from 0.368-0.546 anteriorly beyond anterior border of ovary. In (0.419) and in width from 0.234-0.379 mature and postmature segments, uterus in- (0.299). Gravid segments longer than wide, creasing in width with lateral branches and sometimes barrel-shaped, 0.546-0.978 (0.807) extending in dorsal medulla to near anterior

Figures 1-3. Proteocephaltis chologasteri sp. n. 1. Scolex. 2. Mature proglottid. 3. Gravid proglottid. Abbreviations: CP, cirrus pouch; O, ovary; OD, oviduct; T, testes; U, uterus; V, vitellaria; VA, vagina; VD, vas deferens. segmental border; numerous eggs now visible P. exiguus differs from P. chologasteri in in uterus. In gravid segments, uterus with 11 that its strobila is longer and proportionally to 15 branches on a side and occupying almost wider, and its cirrus pouch is considerably entire intervitelline field. Eggs approximately longer (0.280-0.340 mm), reaching to the spherical, 0.027-0.031 (0.029) in diameter. miclregion of the segment. Also the minimum HOST: Chologaster agassizi Putman. number of uterine branches recorded for P. HABITAT: Small intestine and pyloric caeca. exiguus is nine, while that observed in P. TYPE LOCALITY: Warren County, Kentucky. chologasteri is 11. TYPE SPECIMEN: USNM Helm. Coll. Nos.: holotype 61497, paratypes 61498. The longer strobila (30-50 mm), the greater number of testes (44-70) and the somewhat Discussion longer cirrus pouch (0.095-0.106 mm) of P. pusillus serve to differentiate this species from The size of the strobila and cirrus pouch in combination with the number of testes and P. chologasteri. uterine branches serve to distinguish P. cho- These differences are considered adequate logasteri sp. n. from the other members of the to justify designating the specimens from Ken- genus described from fishes. The only other tucky as a new species. members of the genus from fishes which are It is noteworthy that subsequent to the sufficiently similar to the present species to present investigation, several specimens of warrant differentiation are P. exiguus La Rue, Chologaster agassizi collected from cave re- 1914 and P. pusillus Ward, 1910. gions in Indiana and Illinois were examined

Copyright © 2011, The Helminthological Society of Washington 18 PROCEEDINGS OF THE HELMINTHOLOGICAL SOCIETY and also found to be infected with P. cholo- appreciation to Dr. J. Teague Self of the Uni- gasteri. versity of Oklahoma for his critical review of Summary the manuscript and helpful suggestions. Proteocephalus chologasleri, a new proteo- cephalan tapeworm from the small intestine Literature Cited and pyloric caeca of the spring cavefish Cholo- Hill, L. G. 1966. Studies on the biology of the gaster agassizi is described. The species is spring cavefish, Chologaster agassizi Putman. compared with two other members of the Doctoral thesis, University of Louisville. genus from fish. This represents the first re- LaRue, G. R. 1914. A revision of the cestode port of a cestode and probably the first for family Proteocephalidae. 111. Biol. Monog. 1: any helminth from a cavefish in North America. 1-349. Ward, II. B. 1910. Internal parasites of the Se- Acknowledgment bago salmon. Bull. U. S. Bur. Fisheries. (1908) The author wishes to express his sincere 28: 1151-1194.

Two New Species of Harpyrhynchus from Herons in North America (Acarina, Trombidiformes, Harpyrhynchidae)

ELIZABETH M. Bo YD1 Department of Biological Sciences, Mount Holyoke College, South Hadley, Massachusetts

During a survey of parasites of herons be- The mites were mounted either in Hover's tween 1959 and 1964, two new species of medium or stained in eosin and mounted in Harpijrhijnchitfi were obtained by the detergent Permount. Larvae and nymphs as well as adult washings of the skin of the great blue heron, males and females were collected; most of the Ardea h. herodius and the Eastern green heron, larvae were in the process of molting into Butorides v. virescens. Of the 34 great blue nymphs and one female from the green heron herons examined by the author, two of the contained an egg, which measured 80 by 70 ^.. five shot in Ontario, Canada, and six of the 29 killed in Massachusetts, harbored one spe- Harpyrhynchus herodius n. sp. cies of Harpyrhynchus, while four of the 23 (Figs. A-J) green herons, all from Massachusetts, were FEMALE (Figs. A-E): Body a translucent parasitized by the other species. Both species pale green; cuticle with fine striations on the are very similar but differ markedly from H. peripheral region of dorsum and over most of longipilus Banks (1905) and H. brevis Ewing venter. Length 232 p (range 208-256 /,,), 1911, the only two species previously described width approximately equal to length, 238 p. from North American birds, all passerines. (range 224-248 /j,}. Rostrum, pointed in mid- By separating the powder down areas of the dorsal line, covers posterior third of palps, plumage from the rest of the skin and sub- sclerotized basally. Gnathosoma and pharynx jecting them to the detergent technique, it was visible ventrally. found that these mites inhabit the former Palp (Figs. C, D): The most proximal of region (powder down). Only in heavy infes- the three segments bears two setae ventrally; tations were they collected from the remainder the middle segment, one anterolateral seta of the plumage. clorsally. Terminal segment has three regular setae and three elaborately developed ones, 1 This work was supported in part by grant No. AI 05483-01 from the National Institutes of Health, U. S. of which one is a terminal claw, flat with five Public Health Service and was presented in part at the First International Congress of Parasitologists, Rome, 1964. teeth, and extends forward and inward. The

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 19 other two are etenoform, and arise dorsally. tarsi I and II less elaborate, shorter, 41 ^. The The more mesial has nine serrations and curves two longest setae of leg III measure 423 //, posterolaterally; the second one with four to (inner) and 327 ^ (outer). Leg IV (Fig. G) five serrations stretches laterally and slightly two segments, only distal one with setae, two posteriorly. One seta arises at the base of the of the three are longer than the body, inner latter; the other two are ventral, the more 345 /t,, outer 230 /,.. medial borders the bifid palpal thumb and HOST: Great blue heron, Ardea h. herodius. stems from a swollen base. LOCATION: Skin of powder down plumage Leg I: Six segments; the second bears one areas. and the third three setae ventrally. The fourth LOCALITY: Massachusetts, USA. segment has three ventral and one dorsal; the TYPE SPECIMENS: Holotype female No. 3258 fifth four ventral and one dorsal setae. Tarsus and paratype male USNM. (Fig. E) dorsally, a short sensory seta (10.5 REMARKS: The male, as in other species of by 2.2 p.) flanked by a simple seta each side; Harpyrhynchus, differs from the female in its ventrally, a featherlike pulvillus, 112 ^ long, smaller size and dorsal position of the genital between two claws, two short clawlike setae opening with its three pairs of setae. In addi- and four regular setae of which three are tion tarsi I, II have better developed sensory short and delicate. Leg II: Similar to leg I setae but less elaborate pulvilli and leg IV except tarsus has a narrower sensory seta and consists of two segments, the distal segment lacks the second clawlike seta. Leg III: Three with three setae. segments, devoid of setae dorsally. Segment NYMPH: Similar to adults except in the two with one seta; terminal segment with five, following respects: Palpi with poorly de- of which three are as long or longer than the veloped mesial and lateral etenoform setae; body; the two terminal are thick at their base the former with five serrations bends slightly and extend posteriorly for up to 400 and 460 p., posteriorly. Legs I and II: Pulvilli of tarsi less the medial is the longer. Leg IV: Three seg- well developed; segments three and four with ments; setae on ventral surface, one on middle fewer setae, one on segment three; two (one segment, two of three on the terminal segment ventral, one dorsal) on segment four of leg I, extend posteriorly, the lateral for 290 p., the a single (ventral) on segment four of leg II. medial for 480 ju. Leg IV with two setae. Anus ventral, incon- Body setae: Dorsum (Fig. A): Five pairs spicuous. of long simple setae. The first pair is close to LARVA (Figs. I, J): In all three larvae col- outer edge of stigma, where palp and leg I lected, the structures of the nymph were ob- join the body; it extends anteriorly for 150 p. served within the larval body. The larva is (Fig. C). The next two pairs form a straight similar to the adult except in the following line across the propodosomal region; the re- features. Body considerably smaller, 130 by maining two pairs are lateral, the outer, the 150 ,0,; legs, palpi and setae proportionately longest, extends posteriorly for 145 p.. Venter smaller. Palps lack the lateral etenoform setae (Fig. B): Five pairs short setae; lateral and and its associated regular seta; mesial eteno- median propodosomal, median metapodosomal, form seta directed first anteriorly then slightly and two very small genito-anal setae on the posterolaterally. Palpal thumb's two blades lightly sclerotized genito-anal plate. face forwards then backwards rather than lat- MALE (Figs. F—H): Similar to female ex- erally. Omitting those on the tarsi, legs I and cept in following respects: Body much smaller, II possess only four setae (one on segment 190 by 204 p. so that the outer lateral seta of three, three on segment five, of which one is clorsum, 110 ^ long, extends beyond the pos- dorsal). Leg III of one segment with three terior end of body. Legs, setae and palpi pro- setae; leg IV absent. Anus ventral. portionately shorter. Genito-anal region on dorsal surface; genital area provided with three Harpyrhynchus butorides n. sp. pairs short setae (Fig. F). Sensory setae of (Figs. K-N) tarsi I and II (Fig. H) better developed (for- FEMALE (Figs. K, L): Body translucent mer measures 12.5 by 4.5 p.; latter same dimen- pale green; cuticle finely striated along the sions as that of tarsus I female). Pulvilli of edge of dorsum, and between the legs and

Figures A-E. Harpyrhynchus herodius female from Ardea h. herodius. For Figures A-D, all the ap- pendicular setae are inserted in one-half of the figure; only those seen in the respective view are shown in the other half of the figure. A. Dorsum. B. Venter. C. Capitulum dorsum. D. Capitulum venter. E. Tarsus I venter. Figures F-H. Harpyrhynchus herodius male. F. Dorsum. All the setae are inserted in right half of figure; only those seen in dorsal view are shown in left half of figure. G. Posterior part of body in ventral view. H. Tarsus I venter. Figures I, J. Harpyrhynchus herodius larval stage. I. Venter. Indications of nymph forming within shown on right half. J. Dorsum. Indications of nymph shown in left half; setae of leg III cut off. Figures K-N. Harpyrhynchus butorides from Butorides v. virescens. K. Tarsus I of female: 1, venter; 2, dorsum, processes on ventral surface shown as dotted lines. L. Capitulum of female; 1, right hall venter; 2, left half dorsum. M. Portion of capitulum of nymph to show terminal segment palp: 1, right half venter; 2, left half dorsum. N. Portion of capitulum of larva to show last two segments of palp: 1, right half venter; 2, left half dorsum.

Copyright © 2011, The Helminthological Society of Washington 22 • PROCEEDINGS OF THE HELMINTHOLOGICAL SOCIETY posterior region of venter. Length average HOST: Eastern green heron, Butorides v. 230 ^, width 235 p.. Rostrum pointed in mid- virescens. line covering posterior region of palps, sclero- LOCATION: Skin of powder down plumage tized at base. Pharynx and gnathosoma visible areas. ventrally. LOCALITY : Massachusetts. Palp (Fig. L) three-segmented; basal with TYPE SPECIMENS: Holotype female No. 3259 two setae ventrally; middle with one antero- and paratype male USNM. lateral seta dorsally. Distal segment with two NYMPH: Similar to adults except for the regular and three ctenoform setae. Each cteno- following: terminal segment of palp (Fig. M) form seta possesses eight to nine paired proc- with distal modified seta as a flat claw of five esses and extends laterally and posteriorly for teeth and extending forward and inward; 16 p.. Palpal thumb relatively prominent, 7 p. mesial ctenoform seta small with few processes wide, bifid, outer blade more heavily sclero- projecting anterolaterally and only at its tip tized. Lateral to the thumb and arising from slightly backward; lateral ctenoform seta rep- a swollen base is one regular seta. The second resented as a bristle, 14 /j, long. Legs I and II regular seta lies anterior to the most lateral lack setae on segments one to four. Fifth seg- ctenoform seta on the dorsal surface. ment and tarsi as in adult except pulvilli with Leg I: Six segmented; one ventral seta on four-plumed processes. Leg III, two segments, segment two, three ventral on three, three proximal lacks setae, distal five setae, as in ventral and one dorsal on four, and four ven- adult, but shorter; the inner longest, 100 p.. tral and one dorsal on five. Tarsus I (Fig. K): Leg IV, single segment with two setae, the Dorsally one sensory seta, 10 //,, tapers distally inner longest, 95 p.. Anus ventral, inconspic- with one simple seta on either side; ventrally uous; genital opening and setae absent. tierlike arrangement of processes ending in an LARVA: Like adult except for the following: eight-plumed pulvillus 90 p, long between a Body including legs, palpi and setae much pair of claws, and more proximal one pair of smaller. Palp (Fig. N) with modified terminal clawlike setae and two pairs of simple setae. seta as in nymph; mesial ctenoform replaced Leg II like leg I, but tarsus has a smaller sen- by four-toothed flat claw directed posteriorly sory seta, and lacks one of the clawlike seta lateral ctenoform setae absent; only one seta, and its associated simple one. Leg III: three dorsal, borders the minute palpal thumb. Legs segments, one ventral seta on middle segment, I and II possess only one seta on segment three, five on distal one, two of which short; the two and two ventral and one dorsal on segment longest measure 385 and 365 ^ (outer). Leg five; none on segments two and four. Leg III IV: three segmented, one ventral seta on with three setae on its single segment; leg IV middle and three on terminal segment, the absent. Anus ventral. two long setae 290 ^ (outer) and 480 p, in REMARKS: These two new species from the length. great blue and green herons are closely related Body setae: Dorsum: Five pairs long simple judging from their marked similarity for they setae; the most anterior between base of palp appear to be identical at first glance. The and leg I approximately 95 p.. The second and adult of H. butorides is distinctive from H. third pairs form a straight line on the propo- herodius in the terminal segment of the palp; dosomal; the remaining two pairs lateral (the tarsi I and II, and the shorter lengths of two outer, longer, 120 p.). Venter: Three pairs setae on the dorsum (outer lateral and one body setae (lateral and median propoclosomal adjacent to stigma), and the shorter setae on and median metapodosomal) and two pairs legs III and IV. The palp has a terminal claw, minute genito-anal setae. two ctenoform and three regular setae on its MALE: Like female except: body 190 by terminal segment in PI. herodius, but three 200 p. with proportionately shorter legs, setae, ctenoform and two regular setae in H. bu- and palps. Sensory seta on tarsus I, 10.5 by torides. In the latter, the terminal segment of 3 p.; pulvilli of tarsi I and II six-plumed. Leg tarsi I and II possesses a sclerotized rodlike IV two-segmented; distal one with three setae ridge, with parts arranged in tiers, less elab- of which the two long ones measure 345 and orate with smaller pulvillus and sensory seta; 230 p. (outer). tarsi II has four instead of five ventral setae.

Their immature stages may also be distin- is clearly identified by its body which is wider guished from each other. The nymph of H. than long, its relatively long posterior genital butorides differs in the terminal palpal seg- seta and by leg III which terminates in one ment, particularly the lateral ctenoform seta short and four long setae and leg IV in four which is replaced by a bristle; the number and setae (in both, the inner and the outer are length of setae on the legs; and leg III com- long). The heron species resembles most posed of two and leg IV of one segment. Its closely H. tracheatus but this has a different larval stage is similar to the larva of H. herodius arrangement of the five pairs of dorsal setae, except for its terminal palpal segment. This the last segment of leg III has two short and resembles that of the nymph, but its mesial two long setae of equal length, and that of ctenoform seta is represented as a posteriorly leg IV has three setae, of which the innermost directed flat claw with four teeth; the lateral is extremely long. ctenoform seta is absent and only one dorsal Males have been described from only five seta borders the minute palpal thumb. species, not from H. capelhe nor H. tracheatus. The heron species are readily distinguished It is interesting that the Harpyrhynchus col- from the species previously described from lected from the herons included males, nymphs, America in three major ways: (1) the nature and larvae as well as females. of the modified setae of the palp; (2) the num- Harpyrhynchus mites occur on or in the skin ber and arrangement of the setae on the dor- (Lawrence, 1959c) of birds; some in the sum; and (3) the number of the long setae feather follicles and in some cases have caused on legs III and IV. In H. longipilus, the palp tumors (Fritsch, 1954). In North American has four feathered setae, the dorsum nine pairs hosts the mite has been responsible for causing of setae (five on the shield; four outside); the a pathological condition of the skin. Banks long setae of leg III number four, on leg IV (1905) obtained H. longipilus from a tumor five. The palp has four recurved hooks and under the wing of a crossbill, Loxia sp. Ewing one long seta in H. brevis, the dorsum two (1911) has referred to H. brevis, which he pairs of setae; leg III with seven and leg IV described from the evening grosbeak, as the with five long setae. tumor-forming mite and it has since been The 13 species of Harpyrhynchus reported reported from Wilson's thrush, Eastern song from the Old World including the one from sparrow, redwinged blackbird (Morley and Australia (Fritsch, 1954; Lawrence, 1959a, b, Shillinger, 1937) and rusty blackbird (Chad- c) represent the following ten orders of birds: dock, 1941). In the redwing, it produced ex- Anseriformes (H. plumaris Fritsch), Falconi- tensive orangy nodules on the breast, thigh, formes (H. tracheatus Fritsch), Galliformes and under the neck and wing. No such tumor (H. numidae Lawrence), Charadriiformes (H. formations were visible in the herons, and the capellae Fritsch), Coliiformes (H. cristagalli two new species, H. herodius and H. butorides, Berlese et Trouessart), Cuculiformes (H. ver- inhabit primarily the powder region of the cammeni Lawrence), Apodiformes (H. reduc- plumage. tus Fritsch), Psittaciformes (H. rosellacinus Summary Lawrence), Piciformes (H. pectinifer Law- Harpyrhynchus herodius and H. butorides rence), and Passeriformes (H. cylindripalpus are described from Ardea h. herodius and Fritsch, H. monstrosus Fritsch, H. nidulans Butorides v. virescem. They differ markedly (Nitzsch), H. pilirostris Berlese et Trouessart, from the two species reported from North H. plumaris Fritsch). The new species from American birds—all passerines. The new the herons are the first records for the genus species constitute the first records from the in the order Ciconiiformes. Ciconiiformes. Larvae, nymphs, and adults The dorsal setae are smooth in eight of the were present. They were associated with the 13 species; three of them have leg IV consist- powder down portions of the skin and ap- ing of a single segment. Only two of the peared to cause no pathological condition. remaining five species, H. capellae and H. tracheatus, possess a three-segmented leg IV, Literature Cited and like the heron species they also have five Banks, N. 1905. Descriptions of some new pairs of dorsal setae. The former, H. capellae, mites. Proc. Ent. Soc. Wash. 7: 133-142.

Chaddock, T. T. 1941. Rare mites found on nus, parasites d'oiseanx et de mammiferes. Wisconsin blackbird. Wise. Conservat. 6: 33— Acarologia 1: 106-118. 34. . 1959b. A new mite parasite (Harpy- Ewing, H. E. 1911. New predaceous and para- rhynchus} from the roselle Parakeet (Trom- sitic Acarina. Psyche 18: 37-43. bidiformes, Acari). Proc. Linn. Soc. N.S. Fritsch, W. 1954. Die Milbengattung Harpij- Wales 84: 238-241. 1959c. New mite parasites of African rhynchus Megnin 1878 (Subordo Trombidii- birds ( Myobiidae, Cheyletidae). Parasitology formes, Fam. Myobiida e Megn. 1877). Zool. 49: 416-438. Anz. 152: 177-198. Morley, L. C., and J. E. Shilliiiger. 1937. Para- Lawrence, R. F. 1959a. Acariens (Harpyrhyn- sitic tumors in wild birds. |. Amer. Vet. Med. chidae, Listropboridae) nouveaux on pen con- Ass. 91; n.s. 44: 94-97.

Postembryonic Development and Reproduction in Diploscapter coronata (Nematoda: Rhabditidae)

HELEN CAROL HECHLER1

Introduction coverslip, a drop of a gar medium added, and Diploscapter coronata (Cobb, 1893) Cobb, the coverslip inverted over a depression slide 1913 has been known for over seven decades, and sealed with petroleum jelly. The nema- and seven other species of this genus have todes were examined every 4 hours until de- since been described. However, comparatively velopment was complete. For staining to study little is known about the biology and develop- gonad development and chromosomes, petri ment of the species of Diploscapter. Yokogawa dishes in which an abundance of eggs or the (1936) reported the presence of D. coronata larval stages desired was present were flooded in human urinary sediment and Chandler with water, the resulting suspension centri- (1938) found the same species in the stomachs fuged, and the supernatant discarded. About of nine human patients with a deficiency of 3 ml of Carnoy fixative was poured into the gastric hydrochloric acid. Kampfe (1962) centrifuge tube with agitation to suspend the reported on the effect of desiccation and CCX nematodes. Thirty minutes later the tube was concentration on D. coronata, while Wahab centrifuged, the Carnoy fluid discarded, and (1962) found D. lycostoma in the pharyngeal the nematodes suspended in a few drops of glands of ants and cultured the nematocles on acetic orcein. After about 12 hours, drops of raw potato. Hechler (1967) studied molting the suspension of nematodes in the stain were in D. coronata, and the following is a report placed on microslides and coverslips were on postembryonic development and reproduc- added and sealed. Specimens to be measured were fixed in FAAGO, dehydrated according tion in the same species. to the method of Baker, and mounted in glyc- Materials and Methods erine. Measurements for the de Man formulae were made on camera lucida drawings, whereas The nematodes were established and main- the genital primordia, stoma, and lips, were tained in culture as described by Hechler measured with an ocular micrometer. In molt- (1967). To determine the number of molts, ing specimens the nematode within the exu- newly hatched nematodes were placed indi- vium was measured, not the loose cuticle. vidually in a drop of bacterial slime on a Postembryonic Development 1 Formerly Department of Plant Pathology, University of Illinois, Urbana; present address Nematology Investigations, Four molts occur during the life cycle of USDA, Agricultural Research Service, Beltsville, Maryland 20705. D. coromita, all after the first stage larva

emerges from the egg. All stages feed, and SECOND STAGE (Fig. 1C): Dimensions: L = development stops if the nematocle is removed 0.20-0.23 mm; a = 16-20; b = 2.9-3.8; c = from a source of food. The de Man values are 4-5. Genital primordium 7-9 ^ long at 52— similar for each stage, except for a somewhat 54% of body length, stoma 11-13 ^ long, ham- larger "c" value for the adult. Thus, although uli 4—5 p, wide. In the second stage the sub- the nematocles increase in size, there is little median lips, or hamuli, are hook-shaped and change in body shape except for the compara- sclerotizecl, and the lateral lips, or laciniae, are tively shorter tail in the adult. Development thin, oval, with fringed margins, extending an- of gonads is uniform throughout the popula- teriorly on either side of the oral aperture as tion; nematodes of the same age have gonads in the adult. The cheilorhabdions and lateral at the same stage of development. fields are as in the first stage. The genital FIRST STAGE (Fig. 1A): Dimensions: L = primordium is slightly larger than in the first 0.18-0.19 mm; a = 16-19; b = 2.7-3.0; c = stage, with the two terminal nuclei nearly as 4-5. Genital primordium 4-5 ^ long, with its large as the two central germinal nuclei and of center located at 53-57% of body length from almost the same staining character. No ventral anterior end. Stoma 11 ^ long. Lips all chord nucleus was seen to divide in second rounded, not set off from body or modified stage specimens. However, in a few specimens as in subsequent stages. Cheilorhabdions form one, or more rarely two, larger, more lightly a narrow ring at the anterior end of the stoma. stained nuclei were seen in the ventral chord Lateral fields consist of a single ridge which opposite the position of the genital primordium. extends from the base of the stoma to the anus. They usually did not appear until the second The genital primordium is oval in shape and molt. situated obliquely in the body in ventral view. SECOND MOLT: Dimensions: L = 0.25-0.26 It contains two large centrally located germinal mm; a = 15-18; b = 2.8-3.2; c = 4-5. Gen- nuclei which appear lightly stained except for ital primordium 7-9 /j, long at 50-52% of body numerous, small, discrete, densely stained par- length. The terminal somatic nuclei in the ticles distributed within them. Two smaller genital primordium divide once or twice during somatic nuclei which stain uniformly are lo- the molt, to make a total of two large germinal cated at the posterior and anterior ends of nuclei and four to six smaller somatic nuclei by the genital primordium. The somatic nuclei the time the nematode emerges from the exu- are appressed to the wall so that they appear vium (Fig. ID). One or two ventral chord crescent shaped in lateral view. Rarely both nuclei, larger and lighter in color than the somatic nuclei occur at the same end of the other ventral chord nuclei, are present opposite genital primordium. In the ventral chord, in the genital primordium. a single row between the clusters of nuclei at THIRD STAGE (Fig. IE): Dimensions: L = the base of the esophagus and at the anus, 0.24-0.32 mm; a = 16-20; b = 2.7-3.9; c = there are always 15 nuclei. They stain densely 5-6. Genital primordium 8-15 ^ long at 50- and uniformly with orcein and are always 54% of body length. Stoma 13-15 ^ long, spaced as shown in Figure 1A. hamuli 5-6 ^ wide. Cheilorhabdions and lat- FIRST MOLT: Dimensions: L = 0.19-0.21 eral fields as in the first stage. At the begin- mm; a = 13-18; b = 2.9-3.1; c = 4-5. Genital ning of the stage the two central, larger primordium 4-6 //, long at 50-55% of body germinal nuclei in the genital primordium are length from anterior end. The ventral chord easily distinguished from the smaller somatic nuclei divide during the molt until about 48 nuclei arranged around the periphery. Later both types of nuclei divide, until about 16 are present between the base of the esophagus nuclei are present. It is then difficult to dif- and the anus, usually in a single row, although ferentiate the two types of nuclei because they occasionally a few overlap each other. There is all become similar in size and staining char- a gap with no nuclei opposite the genital pri- acter. As the genital primordium begins to mordium. The genital primordium changes elongate an anterior and a posterior lobe are little in size or shape, but the two somatic formed. As the nematode body elongates the nuclei within it become round and less densely small, dark staining ventral chord nuclei be- stained (Fig. IB). come more widely spaced, but none was seen

Figure 1. Stages in the development of Diploscapter coronata. A. First stage, lateral view; B. First molt, genital primordium, ventral view; C. Second stage, lateral view; D. Second molt, genital primordium, ventral view; E. Third stage, lateral view; F. Third molt, genital primordium, ventral view; G. Fourth stage, lateral view; H. Fourth molt, genital primordium, ventral view; I. Adult, lateral view.

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 27 to divide. The larger, more lightly stained nuclei are arranged around the periphery of a nuclei opposite the genital primordium increase round to ovate structure which becomes con- to four by the time the third molt begins. siderably expanded in the adult. Proximal to THIRD MOLT: Dimensions: L = 0.29-0.32 this structure the tubular form of the uterus mm; a = 14-18; b = 2.7-3.7; c = 5. Genital becomes apparent. The vaginal nuclei origi- primordium 15-36 p. long at 49-53% of body nating within the gonad become smaller and length. The gonad lobes elongate, and by the more densely stained than they were in the end of the molt there are seven to nine nuclei fourth stage and most of them are grouped on in each lobe: one at the terminus, and six to either side of the flattened vagina, with a very eight nuclei arranged in two rows. The central few located dorsally. During the molt the part of the genital primordinm consists of two remaining specialized ventral chord nuclei layers of wedge-shaped cells arranged around move within the vaginal tube. They are similar the future vaginal opening. Early in the molt in size and staining character to the lateral there are four large, lightly stained, ventral nuclei, but located centrally and ventrally. chord nuclei in a single row opposite the mid- ADULT (Fig. II): Dimensions: L — 0.46- dle of the genital primordium; by the end of 0.54 (0,50)* mm; a = 16-20 (17.7); b = 3.8- the molt six to eight specialized nuclei are 5.1 (4.5); c = 6-8 (6.6); V = 45-54 (50)%. present, arranged in two rows (Fig. IF). Stoma 20 ^ long, hamuli 8-9 p, wide. Head FOURTH STAGE (Fig. 1G): Dimensions: L = framework consists of two bowed sclerotized 0.32-0.36 mm; a = 16-20; b = 3.4-3.9; c = pieces, one dorsal and one ventral, surround- 5—6. Genital primordium 25-98 ^ long, center ing the stoma at their centers and extending at 50—55% of body length. Hamuli 6 p. wide, anteriorly and laterally at each end. Gheilor- stoma 14-16 ^ long. Cheilorhabclions form a habdions forming a shallow inverted funnel narrow ring and lateral fields consist of a at the anterior end of the stoma. Lateral fields single ridge, as in the first stage. The genital composed of two ridges between the base of primordium elongates considerably until, just the stoma and the anus. Excretory pore oppo- before the final molt, the two lobes recurve. site median bulb, hemizonid anterior and con- At this time there are about 20 germinal cells tiguous to it. Nerve ring just behind middle in each lobe. Proximal to the germ cells in of isthmus. Amphids located just posterior to each lobe is a slightly constricted section with laciniae. Papillae in center of laciniae easily four rows of three or four small, densely stained seen. Papillae on submedian lips, seen on only nuclei. This is followed by a less constricted two favorable en face views, located lateral to section with four rows of two or three larger, the centers of the hamuli. Phasmids about 15 p, more lightly stained nuclei. The vaginal pri- posterior to anus, small and inconspicuous. mordium consists of two rows of five or six Gonads recurved at ends, each lobe 105-170 p, dorsal nuclei and two rows of six or seven ven- long, germ cells four to five to a cross section tral nuclei. The two ventral rows are farther at widest part of ovary. Wall of ovary thin and apart at their centers, surrounding the vaginal seems to be anucleate. An oval, hyaline struc- opening. The lightly stained ventral chord ture with about 14 small, densely stained nuclei nuclei increase to about 16, with eight just spaced around its periphery is present at the posterior and eight just anterior to the vaginal junction of ovary and uterus (Fig. 2C). This primordium. One or two of each group move would probably function as a spermatheca in dorsally within the vaginal opening by the time the presence of spermatozoa. Proximal part of of the beginning of the fourth molt. gonad with thick, convoluted walls containing FOURTH MOLT: Dimensions: L = 0.41-0.42 nuclei which are larger and stained lighter in mm; a = 14-17; b = 3.7-3.9; c = 4-6. Genital color than those in the oval structure. Vulva primordium 98-110 ^ long, center at 50-52% a crosswise slit. Both anterior and posterior of body length. During the final molt the to the vagina about 10 densely stained nuclei gonad lobes increase in length and width and of ventral chord origin are located centrally the germinal cells divide until there are about and ventrally, while on either side of the vagina 30 in each lobe, arranged in three or four rows are a posterior and anterior group of about in the widest part (Fig. 1H). At the junction nine similar nuclei originating from the gonad. of ovary and uterus the small densely stained * Value in parentheses is the mean.

There are about 56 ventral chord nuclei ar- so old that meiosis was no longer proceeding ranged in a single row between the base of in their testes, although spermatozoa were pres- the esophagus and the anus, excluding those ent. Therefore, nothing was learned about the involved in the structure of the vagina. process of spermatogenesis. Reproduction Discussion Although mitotic divisions occur throughout Hirschmann (1962) reported that in Dity- the distal half of the ovary, it is very difficult lenchus triformis the ventral chord nuclei, to identify and count individual chromosomes whose descendents will eventually form the in the dividing nuclei. In cells which are not lining of the vagina, can be identified in young dividing the nuclei contain many small discrete second stage females because they are larger particles which stain deeply with orcein. In and less deeply stained than the other ventral the proximal part of the ovary the deep stain- chord nuclei. Similar nuclei could also be ing particles disappear and the nuclei appear detected in a few second stage D. coronata lighter in color than the surrounding cytoplasm. females, and they were always present by the Prophase of meiosis becomes evident when the second molt. Most of the nuclear divisions in cell reaches the proximal end of the ovary. At the ventral chord occur during the first molt that time the nucleolus and two long, double, in D. coronata, at the time the nuclei increase twisted strands appear in the nucleus (Fig. from a constant number of 15 to about 48. 2A). Before the oocyte passes into the uterus Possibly the specialized nuclei arise at this the nucleolus disappears (Fig. 2B). The time and differentiate by the time of the sec- chromatin then begins to contract to deeply ond molt. staining rods which appear double (Fig. 2C), During the first stage the epithelial nuclei and meanwhile the oocyte moves into the in the genital primordium are smaller and more uterus. The rods continue to contract until uniformly stained than the germinal nuclei and they are of the shape shown in Figure 2D and the two types of nuclei are easily distinguished. they separate completely. There is no pairing As development of the nematodes progresses, of the chromosomes, either side by side or end the two types of nuclei become increasingly to end. One maturation division occurs, usually similar in size, shape, and staining characteris- near the middle of the long axis of the egg, tics. During the third stage they could not be and one polar body is formed which remains differentiated. By the third molt it is again visible until the end of the second or third possible to recognize some of the somatic cleavage division. Both the polar body and nuclei because of their location within the the egg nucleus contain two rods (Figs. 2D, developing genital primordium, but the nature 2E). No second meiotic division takes place. of each nucleus is not absolutely clear until The egg nucleus moves to the center of the late in the fourth stage. Therefore, it was not egg and the two rods elongate and become possible to determine the contribution of each double (Fig. 2F). The first cleavage division of the early somatic nuclei to the final struc- occurs either before or after the egg is laid. ture of the gonads or to determine whether the At the cleavage metaphase two double strands terminal cell of each lobe was of somatic or of chromatin are present and at telophase two germinal origin. Since in hundreds of stained short chromosomes can be seen at each pole females the terminal nucleus was never seen to (Fig. 2G). They elongate again before the divide, it is probably somatic. next cleavage (Fig. 2H). The phenomenon of All four molts occur after the nematode chromosome diminution was not detected as emerges from the egg. As noted by Thomas late as the eight cell stage. Subsequent nuclei (1965) for Acrobeles complexus, the lip region were so small that the chromosomes were diffi- of the first stage is of a different form in D. cult to see. coronata than in subsequent stages, with the No sperm cells are present within the body lips all rounded rather than being modified to of females and, among several hundred thou- hamuli and laciniae. This change was noted sand females seen during this investigation, previously by Hechler (1967), as well as the only three males were found. These males change in the stoma during the final molt, were treated with Carnoy-orcein. They were when the adult cheilostom becomes wider and

Figure 2. A. Prophase I showing nucleolus (arrow) and two dyads; B. Prophase I after disappearance of nucleolus; C. Late prophase I; D. Telophase I showing polar nucleus; E. Egg nucleus, same egg as shown in Figure D; F. Prophase of first cleavage, nucleus migrating to center of egg, polar body (arrow) at side; G. Telophase of first cleavage; H. Prophase of second cleavage. funnel-shaped. In addition there is an increase ther development of the gonad. Growth seems from one to two ridges in the lateral fields at to proceed at the same rate during the larval the final molt. stages as during the molts. Yuen (1965) stated that in Helicotylenchus During maturation of the egg in D. coronata vulgaris the principal growth of the gonacls doubling of the chromatin could be seen during occurs during the molts whereas there is little prophase, with two dyad groups in the nucleus change in them between molts. In D. coronata by metaphase. These groups could never be the gonad changed little from hatching through resolved as tetrads. Pairing of the two dyad the second molt. However, beginning early in groups, either side by side or end to end, was the third stage dividing nuclei could be seen not observed; although occasionally the groups in the genital prirnordiiim throughout the fur- seemed to lie side by side at metaphase it was

probably coincidental since this behavior was gen zur Autokologie von Heterordera rosto- not consistent. After the first maturation di- chiensis Wollenweber uncl Heterodera schach- vision the inner nucleus was found at the tii Schmidt sowie einiger anderer Nematodar- center of the egg, with two doubled chromo- ten Abweichender Lebenstiitten. Parasitol- somes, and only two chromosomes were found ogische Schriftenreihe. Jena. Number 14, 205 in each telophase nucleus of the first cleavage. pp. Thomas, P. R. 1965. Biology of Arcobeles It is therefore concluded that there is a somatic complexes Thorne, cultivated on agar. Nema- number of two unpaired chromosomes in this tologica 11: 395-408. isolate of D. coronata. No second maturation Triantaphyllou, A. C. 1962. Oo'genesis in the division was detected, the chromosomes did root-knot nematode Meloidogyne javanica. not pair before meiosis, and no sperm cells Nematologica 7: 105-113. were seen within the females. Hence this iso- Triantapthyllou, A. C. 1963. Polyploidy and late reproduces by mitotic parthenogenesis, as parthenogenesis in the root-knot nematode reported by Triantaphyllou (1962, 1963, 1966) Meloidogyne arenaria. ]. Morphol. 113: 489- for Meloidogyne javanica, M. arenaria, and 499. Triantaphyllou, A. C. 1966. Polyploidy and some populations of M. hapla. reproductive patterns in the root-knot nema- Literature Cited tode Meloidogyne hapla. J. Morphol. 118: Chandler, A. C. 1938. Diploscapter coronata as 403-413. a facultative parasite of man with a general re- Wahab, A. 1962. Untersuchungen liber Nema- view of vertebrate parasitism by rhabditoid toden in den Driisen des Kopfes der Ameisen worms. Parasitology 30: 44-45. (Formicidae). Zeitschrift fiir Morphologie und Hechler, Helen Carol. 1967. Morphological Okologie der Tiere. 52 (1): 33-92. changes during the molt of Diploscapter coro- Yokogawa, S. 1936. A human case of accidental nata (Nematoda: Rhabditidae). Proc. Helm. parasitism of Diploscapter coronata (Cobb, Soc. Wash. 34: 151-155. 1893) Cobb, 1913. Zoological Magazine 48 Hirschmann, Hedwig. 1962. The life cycle of (8/10): 507-511. (In Japanese: English sum- Ditylenchus triformis (Nematoda: Tylenchida) mary p. 507). with emphasis on post-embryonic develop- Yuen, Pick. 1965. Further observation of Hcli- ment. Proc. Helm. Soc. Wash. 29: 30-43. cott/lenchus vulgar is Yuen. Nematologica 11: Kiimpfe, L. 1962. Vergleichende Untersuchun- 623-637.

Helicometra antarcticae sp. nov. from Antarctic Coastal Fishes1

HARRY L. HOLLOW AY, JR., AND JEFFREY W. BIER Department of Biology, Roanoke College, Salem, Virginia

Introduction Cable, 1960; H. markwitschi Pogoreltzeva, The following species ascribed to the genus 1954; H. insolita Polyanski, 1955; H. pteroixi Helicometra Odhner, 1902 may be added to (Gupta, 1956) Fischthal and Kuntz, 1965; the list of Manter (1954), subsequent taxo- H. boseli Nagaty, 1956; H. dochmosorchis nomic changes are indicated: li. torta Lin ton, Manter and Pritchard, 1960; H. marmotatae 1910 (Siddiqi and Cable, 1960 synonymized Nagaty and Abdel Aal, 1962; //. nasae Nagaty H. pretiosa Bravo-Hollis and Manter, 1957 and Abdel Aal, 1962; H. indica Agrawal, 1964; with this concept); H. plovmornini Isaychikov, H. rectisacciis (Fischthal and Kuntz, 1964); 1928; H. equilata (Manter, 1933) Siddiqi and H. boreoensis Fischthal and Kuntz, 1965 bring- ing the total number of species to 22 including 1 Supported in part by Grants GA 145 and 495 from the species described in this paper and acceptance United States Antarctic Research Program, National Science Foundation. of synonymy suggested.

Manter (1954) employed two names for a anterior to acetabulum. Ceca extend laterally single taxon in the genus, Helicometra gran- to near posterior end of body, ending blindly. dora and Helicometra magnora. The former is Testes two, in posterior half of body, slightly more prominent, whereas the latter was em- to moderately lobed, oblique, overlapping in ployed in the explanation of the figures. After 49 specimens, maximum separation 0.078. consultation (H. W. Manter, personal com- Posttesticular space 0.446-1.491 (0.878). An- munication) and for reasons previously con- terior testis 0.212-0.725 (0.372) long by sidered, Helicometra magnora is synonymized 0.223-0.959 (0.408) wide, left of midline in with Helicometra grandora. 64, right of midline in five specimens. Pos- terior testis 0.223-0.860 (0.394) long by Materials and Methods 0.234-0.892 (0.439) wide, near midline in The present paper describes a new species 58, right of midline in seven, left of midline of digenea collected from two species of fishes in four specimens. Vasa efferentia arise from around McMurdo Station, 77°51' S latitude; anterior borders of testes. One from anterior 166°38' E longitude, during the austral sum- testis passes anteriad left of midline and be- mer of 1964-65. Byrd (1963) considered lit- tween left lateral portion of uterine coils. Vas erature on trematodes of marine fishes of the efferens from posterior testis passes anteriad southern seas. right of midline, dorsal to ovary and between The description is based on specimens fixed, right lateral portions of uterine coils. At pos- unflattened, with warm Gilson's mercuric- terior border of acetabulum vasa efferentia acetic-nitric mixture or Romeis' AFA. The converge toward midline and enter posterior mercurial salts were removed by the method end of cirrus pouch. Seminal vesicle in pos- of Ebbett and Holloway, 1967. Whole mounts terior portion of cirrus pouch. Dilated portion were stained with Semichon's carmine or borax of seminal vesicle extends anteriad one-fourth carmine, while sections were stained with com- to one-half length of cirrus sac, contracts and bination hematoxylin. flexes posteriad toward posterior border of HOSTS: Dissostichiis mawsoni Norman, 1937 cirrus pouch, then flexes anteriad and mediad and Rhigophila dearborni DeWitt, 1962. to exit through cirrus as ejaculatory duct. An- HABITAT IN HOST: Intestine and ceca. terior portion of cirrus sac filled with prostatic LOCALITY: McMurdo Sound, Antarctica. cells. Cirrus sac 0.413-1.070 (0.623) long by TYPES: USNM Helm. Coll. No. 63113 (holo- 0.089-0.357 (0.173) wide, about midacetab- type from D. mawsoni}; No. 63114 (paratype ular in posterior extent, opens ventral to cecal from R. dearborni). Opecoelidae Ozaki, 1925: bifurcation. Extrusible cirrus 0.118-0.263 Plagioporinae Manter, 1947. (0.195) long by 0.066-0.176 (0.105) wide. Ovary 0.134-0.903 (0.299) long by 0.212- Description 0.725 (0,371) wide, variously lobed: 3(1), Based on 80 specimens with the characters 4(38), 5(12), 6(13), 7(1), 8(1); slightly of the genus (all measurements in millimeters right of midline, confluent with or separated unless noted): Body (Fig. 1) 2.33-5.85 (mean from anterior testis. Muscular oviduct (Figs. 3.50) long by 0,56-1.90 (1.17), maximum 7 and 8) arises from anterior lobe of ovary, width just posterior to acetabulum, unarmed, passes dorsiad to flexure where wall of oviduct lanceolate. Oral sucker 0.156-0.412 (0.258) becomes thinner and ciliated. Seminal recep- long by 0.290-0.602 (0.430) wide, terminal. tacle confluent with flexure by a short stalk, Acetabulum 0,334-0.892 (0.535) long by has ciliated walls, dorsal to ovary, and of 0.290-0.870 (0.525) wide about one-third variable size. Thick-walled Laurer's canal body length from anterior end. Sucker ratio (Figs. 7 and 8) passes from anterio-dorsal end 1:1.22. Prepharynx 0.039-0.118 (0.075) long of seminal receptacle opposite stalk, dorsally, by 0.021-0.108 (0.047) wide. Pharynx 0.105- laterally, and, in all but one case, posteriorly 0.226 (0.146) long by 0.116-0.255 (0.175) to open on dorsal surface. A short distance wide, subglobular. Esophagus 0.105-0.379 from flexure, oviduct receives common vitelline (0.239) long by 0.024-0.074 (0.040) wide. duct (Figs. 6 and 8) and soon expands into Cecal bifurcation 0.250-0.647 (0.412) pos- ciliated ootype (Figs. 6, 7, and 8). Diffuse terior to oral sucker and 0.071-0,569 (0.255) Mehlis' glands surround ootype which com-

Figures 1-8. 1. Helicometra antarcticae, ventral view. Scale = 0.50 mm. 2. Reconstruction of terminal male genitalia. Scale = 0.10 mm. 3. Egg. Scale = 0.05 mm. 4-7. Serial sections, anterior to posterior, of female genital complex. Scale below Figure 7 equals 0.10 mm and applies to Figures 4-7. 8. Diagrammatic reconstruction of female genital complex. Scale = 0.10 mm.

Table 1. Differentiation of Helicometra antarcticae from closely related species.

Characteristic Helicometra fasciata Helicometra antarcticae Helicometra insolita

Overall length: 1.30-4.30 2.30-5.90 (3.50) 3.00-4.50 width: 0.40-0.83 0.60-1.90 (1.20) 0.87-1.00 Oral sucker length: 0.15-0.23 0.16-0.41 (0.26) 0.42-0.48 width : 0.15-0.23 0.29-0.60 (0.43) 0.42-0.52 position: sub terminal terminal terminal Acetabulum diameter: 0.22-0.36 0.29-0.89 (0.53) 0.40-0.46 Sucker width ratio: 1:1.50 1:1.22 1:1.00 Prepharynx: present, short 0.07 by 0.05 not developed (pubescent worms) Pharynx: 0.09-0.10 0.15 by 0.17 0.15-0.17 Esophagus length: 0.15-0.33 0.11-0.38 (0.24) 0.12-0.17 Cirrus sac length : variable 0.41-1.07 (0.62) 0.44-0.58 extent: just pre- to midacetabular midacetabular midacetabular Excretory pore: dorsal near posterior end terminal vesicle : tubular anterior to ovary tubular, near acetabulum Vitelline follicles extent: near genital pore lateral between genital pore and posterior edge acetabulum to ceca near posterior end acetabulum laterally sur- lateral near posterior end contiguous posttesticular rounding ceca near pos- contiguous posttesticular terior end, contiguous or not posttesticular Ovary length : 0.13-0.90 (0.30) 0.33-0.42 width : 0.25-0.35 0.21-0.72 (0.37) lobes: 3-8 deep 4, 3-8 deep 3 shallow Eggs length : 0.060-0.084 0.065-0.110 (0.079) 0.071-0.084 width: 0.023-0.037 0.026-0.055 (0.042) 0.032-0.038 filaments : 0.20-0.67 2.1 yes operculum : yes yes Testes position : tandem, midbody anterior left posterior mid- tandem, midbody line lobation : variable variable, shallow to mod- regular, deep erate Posttesticular space: 0.50 0.45-1.49 (0.88) Anterior testis length: 0.13-0.35 0.21-0.72 (0.37) 0.31-0.42 width: 0.20-0.35 0.22-0.96 (0.41) Posterior testis length: 0.23-0.35 0.22-0.86 (0.39) 0.37-0.44 width: 0.25-0.35 0.23-0.89 (0.44)

municates with nonciliated uterus. Vitelline 0.065-0.110 (0.079) long by 0.026-0.055 reservoir (Figs. 7 and 8) dorsal to ovary. (0.042) wide and possessing hollow filament Vitelline follicles (Fig. 1), lateral to and sur- 2.1 long which arises from anopercular end. rounding ceca, extend from near posterior end Excretory pore terminal. Excretory bladder of body to region between anterior border of tubular, extending to near posterior edge of acetabulum and cecal bifurcation; posttestic- acetabulum, possessing sphincter muscle just ularly contiguous or not. Uterus (Fig. 1) coils anterior to excretory pore. to posterior border of acetabulum, turns Helicometra antarcticae differs from all abruptly anteriad and becomes thicker and members of the genus except II. insolita in pos- more muscular. Metraterm closely adherent to sessing a terminal oral sucker. H. antarcticae left side of cirrus pouch; female genital pore differs significantly from H. insolita in sucker beside male pore. Egg capsules (Fig. 3) ratio, oral sucker length, anteriad extent of

Copyright © 2011, The Helminthological Society of Washington 34 PROCEEDINGS OF THE HELMINTHOLOGICAL SOCIETY vitellaria, lobation and arrangement of testes, of ovary. H. antarcticae keys, Skrjabin and and lobation of ovary (Table 1). H. antarcticae Koval, 1958 key, to H. fasciata (Rudolphi, keys to H. fasciata (Rudolphi, 1819) in the 1819) from which it differs in position and key of Skrjabin and Koval, 1958. It differs width of oral sucker, diameter of acetabulum from H. fasciata in position and width of oral and in morphology of cirrus and Laurer's canal. sucker (lengths not comparable) and diameter Literature Cited of acetabulum (Table 1). Cirrus, elongate and having terminal bulbular process (Palombi, Byrd, M. A. 1963. Helminth parasites of Ant- arctic vertebrates. Part 1. Digenetic trem- 1929) in H. fasciata, is stout and plain in H. atodes of marine fishes. Proc. Helm. Soc. antarcticae. Normal direction of Laurer's canal Wash. 30: 129-148. is anteriad to near acetabular level (Odhner, Ebbett, R., and H. L. Holloway, Jr. 1967. Re- 1901) in H. fasciata and posteriad to about moval of mercurial precipitates from digenetic midovarian level in H. antarcticae. trematodes stored in Gilson's fluid. Stain Tech- nol. 42: 261-262. Summary Manter, H. W. 1954. Some digenetic trematodes from fishes of New Zealand. Roy. Soc. Species ascribed to the genus Helicometra N. Z., Trans. 82: 475-568. Odhner, 1902 since 1954 are listed, subsequent Odhner, T. 1901. Revision einiger Arten der taxonomic changes are also indicated and //. Distomengattung Allocreadium Lss. Zool. magnora Manter, 1954 is synonymized with Jahrb. Abt. Syst. Geogr. Biol. 14: 483-520. H. grandora Manter, 1954. Helicometra ant- Palombi, A. 1929. Ricerche sul ciclo evolutivo arcticae sp. nov. from Dissostichtis maivsoni di Helicometra fasciata (Rud.). Revisione Norman, 1937 and Rhigophila dearborni De- delle specie del genere Helicometra Odhner. Witt, 1962 caught in McMurdo Sound, Ant- Publ. Staz. Zool. Napoli 9: 1-56. Siddiqi, A. H., and R. M. Cable. 1960. Di- arctica (77°51/ S latitude; 166°38' E longitude) is described from unflattened specimens. genetic trematodes of marine fishes of Puerto Rico. Sci. Surv. Porto Rico and Virg. Is. 17: H. antarcticae like H. insolita Polyanski, 1955 257-369. possesses a terminal oral sucker but differs Skrjabin, K. I., and V. P. Koval. 1958. Sub- significantly from H. insolita in sucker ratio, family Plagioporinae Manter, 1947. In Skr- oral sucker length, anteriad extent of vitellaria, jabin, K. I., Trematodes of Animals and Man. lobation and arrangement of testes and lobation Moskva. Vol. 15: 424-811 (Russian text).

Hatching Response of Hypsoperine otter soni to Canary grass Root Emanations1

A. J. WEBBER, JR., AND K. R. BARKERL>

Host root emanations have been shown to under favorable environmental conditions increase greatly larval emergence from eggs (Godfrey, 1931; Bergeson, 1959). However, of many Heterodera species (Triffitt, 1930; Viglierchio and Lownsbery (1960) demon- Shepherd, 1962). Root-knot larvae, Meloido- strated that larval emergence of Meloidogyne gijne spp-, have been found to emerge readily species was increased in the presence of germinating tomatoes. No hatching stimulant has 1 Research supported in part by Hatch funds. Published with approval of the Director of the Agricultural Experi- been demonstrated to occur in host plants of ment Station. Project 904. 2 Formerly Gradviate Assistant, Department of Plant Pa- Meloidodera or Hypsoperine species. thology, University of Wisconsin, Madison, now with the U. S. Army Biological Laboratories, Ft. Detrick, Maryland; Low larval emergence from eggs of Hypso- and Assistant Professor, Plant Pathology Department, Uni- perine ottersoni, a new species which is being versity of Wisconsin, Madison, now with North Carolina State University, Raleigh, North Carolina. described by Thorne (personal communica-

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 35 tion), under laboratory conditions suggested were added to each crock 1 week before the that in nature a hatching stimulant might be first leachate collection and immediately after operative. Experiments were designed to de- each succeeding collection. Leachates were termine the effect of root emanations from collected by refluxing distilled water into each the primary host, canarygrass, and a nonhost crock through the pinchcock and draining tomato, upon the rate of larval emergence of through the pinchcock. Sufficient water was this nematode. added to each crock to cover the surface of the sand. Leachates were bottled and stored at Methods 4 C for later use. Hatching experiments with Hypsoperine Leachate activity was tested by the same ottersoni were conducted using 2-week-old technique used with seedlings. Eggs contain- Reeds canarygrass and Bonny Best tomato ing fully formed second stage larvae and seedlings. Seedlings of both test plants were leachates from 6-week-old seedlings were used. germinated in steamed silica sand and watered Five ml of canarygrass, tomato, or control with distilled water as needed. Inoculum was leachate were added per Syracuse watch glass. obtained from infected canarygrass from the Two and one-half ml of leachate were added to field and the greenhouse. Galls were har- the respective treatments at 2-day intervals. vested, washed and teased apart in sterile dis- Rates of larval emergence were determined in tilled water. Eggs containing fully formed sec- the manner previously described. ond stage larvae were transferred with a fine Similar experiments were conducted with capillary pipette to sterile distilled water in a leachates concentrated 5 times. Concentration Syracuse watch glass and thoroughly mixed. was performed at 30 C with a Rinco vacuum Fifty of the washed eggs were then transferred distillation unit, equipped with a condenser. to each of several gridded Syracuse watch glasses. Less than 0.1 ml of water was in- Results and Discussion cluded with the transfer. Five ml of sterile Emanations from young canarygrass roots distilled water and 5 canarygrass or tomato significantly increased the larval emergence of seedlings were placed in each watch glass. H. ottersoni (Fig. 1A). Egg hatch was gen- Controls consisted of similar units without erally increased two- to four-fold over water seedlings. Four replicates were incubated sep- controls. An increase greater than four-fold arately in 1-pint baking dishes covered with was obtained in an experiment in which the paper bags at 22 ± 1 C. Larval emergence eggs had been collected from the field in the was determined under a stereomicroscope at fall of the year. However, another trial with daily intervals for 7 clays. a different egg source resulted in only a 30% Root leachates for hatching experiments increase in egg hatching with canarygrass seed- were obtained from canarygrass and tomato lings. Generally, larval emergence from eggs seedlings as follows. A one-hole rubber stopper collected in the fall was four to six times greater was placed in the hole in the base of each of than from eggs collected in the spring. several 8-inch glazed crocks. Glass wool was Similar hatching experiments showed to- placed over the inner end and an L-shaped mato, a nonhost plant, to have little effect on piece of glass tubing fitted into the outer end larval emergence. Most trials with tomato of each stopper. A short piece of rubber tubing with a pinchcock was attached to the seedlings gave no increase in egg hatch. How- protruding end of the glass tube. Crocks were ever, one trial gave an apparent increase of filled with washed and steamed silica sand and 60% over the control, but this difference was 20 2-week-old tomato or 100 2-week-old not statistically significant (Fig. 1A). canarygrass seedlings were transplanted to Unconcentratecl crude canarygrass leachate each crock. Controls consisted of similar units increased the rate of larval emergence by two- without seedlings. Crocks were placed in an to four-fold (Fig. IB). All trials with the air conditioned greenhouse at 22 ± 2 C. canarygrass leachate resulted in significant in- Leachates were collected at weekly intervals creases in larval emergence. Tomato leachates, beginning 2 weeks after transplanting. Two however, gave little response. Experiments hundred ml of Hoaglancl's nutrient solution with concentrated crude leachates failed to

20- 20 CANARY GRASS SEEDLINGS B CANARY GRASS LEACHATE TOMATO SEEDLINGS TOMATO LEACHATE CONTROL 5- CONTROL LEACHATE

o o- o o CL

Figure 1. Effect of canarygrass and tomato emanations on larval emergence of H. ottersoni. A. Re- sponse of spring-collected eggs to seedlings (5% Isd on 7th day 6.15%). B. Larval emergence as affected by nonconcentrated leachates (5% Isd on 7th day 6.42%). accentuate larval emergence regardless of the are known to influence the emergence of material used. Heterodera species. The pH (8.4) of the Larval emergence of many Heterodera spe- canarygrass and tomato root leachates was cies is increased as much as 100—1,000 times higher than that of the control leachate (pH by root emanations from their primary hosts 7.0). However, no significant increases in (Shepherd, 1962). However, larval emergence emergence were obtained with tomato root of Heterodera rostochiensis Wollenweber and leachate. Apparently, the difference in pH Heterodera schachtii Schmidt is not increased was not responsible for the observed increases by emanations from nonhost grasses (Hesling, with canarygrass root leachate. Since all crocks Pawelska, and Shepherd, 1961). Eggs of received equal quantities of Hoagland's nutri- Heterodera avenae Filipjev, a nematode that ent solution, it is unlikely that differences in attacks only grass species, apparently are not ionic concentration could explain the increase stimulated to hatch by host-root emanations in larval emergence. Variation in the concen- (Hesling, 1957). Hatching of H. ottersoni, tration of elemental constituents due to uptake which also attacks only grasses, was stimulated by test plants and their possible effect on larval by emanations from roots of its native host. emergence need further investigation. Increases of approximately 100 per cent have Variation in the magnitude of larval emer- been obtained with Meloidogyne species with gence between trials with canarygrass seedlings germinating tomato seedlings (Viglierchio and or their root leachate may be due to differences Lownsbery, 1960). Although the hatching re- in egg maturity and permeability. Hetero- sponse of H. ottersoni to host-root emanations geneity in cyst permeability is known to influ- is less than that of most Heterodera species, it ence larval emergence of Heterodera species appears to be greater than that obtained with (Ellenby, 1955a, b). The low rate of emer- Meloidogyne species. gence of larvae from eggs collected in the Variations in the pH of the hatching medium spring may also be related to egg maturity and (Robinson and Neal, 1956) and the ionic con- permeability. centration of certain mineral elements (Ellenby Hypsoperine species are described as pos- and Gilbert, 1958; Robinson and Neal, 1959)', sessing morphological characters intermediate

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 37 between Heterodera and Meloidogyne species potato root eelworm, Heterodera rostochiensis (Sledge and Golden, 1964). Present data on Wollenweber. Nature 182: 925-926. the hatching response of H. ottersoni indicate Godfrey, G. H. 1931. Some techniques used in that stimulation of larval emergence of this the study of the root-knot nematode, Heterod- species by host root emanations is likewise era radicicola. Phytopathology 21: 323-329. Hesling, J. J. 1957. The hatching response of intermediate between Heterodera and Meloido- Heterodera major (O. Schmidt) to certain root gyne species. diffusates. Nematologiea 2: 123-125. , K. Pawelska, and A. M. Shepherd. 1961. Summary The response of the potato-root eelworm, Het- Larval emergence of Hypsoperine ottersoni erodera rostochiensis Wollenweber and the beet eelworm, H. schachtii Schmidt to root Thorne was increased two- to four-fold by ema- diffusates of some grasses, cereals, and Tage- nations from the roots of canary grass seedlings. tes minuta. Nematologiea 6: 207-213. Similar increases were obtained with canary- Robinson, T., and A. L. Neal. 1956. The in- grass root leachate. Nonhost tomato seedlings fluence of hydrogen ion concentration on the or their root leachate did not significantly in- emergence of golden nematode larvae. Phy- crease hatch. The observed increases are not topathology 46: 665-667. believed to be due to variation in pH or ionic- , and A. L. Neal. 1959. The influence concentration of the hatching medium, but to of certain mineral elements on emergence of the chemical properties of the leachate itself. golden nematode larvae. Proc. Helm. Soc. Wash. 26: 60-64. Literature Cited Shepherd, A. M. 1962. The emergence of larvae from cysts in the genus Heterodera. Tech. Bergeson, G. B. 1959. The influence of tem- Comm. No. 32 of the commonwealth Bureau perature on the survival of some species of of Helminthology, St. Albans, Herts., C.A.B. the genus Meloidogyne, in the absence of a Farnham Royal, Bucks. 90 p. host. Nematologica 4: 344-354. Sledge, E. B., and A. M. Golden. 1964. Hyp- Ellenby, C. 1955a. The seasonal response of the soperine graminis (Nematoda: Heteroderidae), potato-root eelworm, Heterodera rostochiensis a new genus and species of plant-parasitic Wollenweber: emergence of larvae through- nematode. Proc. Helm. Soc. Wash. 31: 83-88. out the year from cysts exposed to different Triffitt, M. J. 1930. On the bionomics of Het- temperature cycles. Ann. Appl. Biol. 43: 1-11. erodera schachtii on potatoes with special ref- . 1955b. The permeability of the hatch- erence of mustard on the escape of larvae from ing factor of the cyst wall of the potato-root cysts. J. Helm. 8: 19-48. eelworm, Heterodera rostochiensis Wollen- Viglierchio, D., and B. F. Lownsbery. 1960. The weber. Ann. Appl. Biol. 43: 12-18. hatching response of Meloidogyne species to , and A. B. Gilbert. 1958. Influence the emanations from the roots of germinating of certain inorganic ions on hatching of the tomatoes. Nematologiea 5: 153-157.

IN M E M O R I A M

Gordon M. Clark F. Ben Struble 1929-1967 1913-1966 Member since 1957 Member since 1959

George R. LaRue Arthur C. Walton 1882-1967 1892-1967 Member 1913-1959 Member since 1942 Elected Honorary Member 1959 Recipient of Anniversary Award 1966

Rotylenchus pini n. sp. (Nematoda: Hoplolaimidae) from Forest Nurseries in Japan

YASUHAHU MAMIYA Division of Forest Pathology, Government Forest Experiment Station, Meguro, Tokyo, Japan

During a nematocle survey of forest nurseries areolated only in region of esophagus. Tail in Japan conducted in 1964 and 1965, an terminus hemispherical, slightly more curved undescribed species of Rotylenchus was found dorsally, with ten annules (paratypes range associated with coniferous seedlings in two 6-13). Ratio of tail length to body width at nurseries. This nematocle was also recovered anus 0.8 (paratypes range 0.7-1.0). from soil samples collected in 1965 by Mr. Y. MALE (allotype): L = 0.96 mm; a = 36; Suto, of Shimane Forest Experiment Station, b = 6.1; c = 36; spear = 28 p.; O = 17; guber- from the root zone of pine seedlings in a nur- naculum = 10 p; spicnles = 27 p.. sery in Shimane Prefecture, Honshu. Body curved ventrally when relaxed. Lip This new species is described herein as region broadly hemispherical, slightly higher Rotylenchus pini n. sp. than female, not set off, six annules without Measurements were made on specimens longitudinal striations. Anterior portion of killed by gentle heat, fixed in TAF + F.A. spear slightly longer than posterior portion. 4:10 and mounted in glycerine (modified Spear knobs round, slightly flattened ante- Baker's method by J. B. Goodey, 1957). riorly. Basal portion of esophageal glands compact, slightly overlap intestine. Phasmids Rotylenchus pini n. sp. 22 annules anterior to anus (paratypes range 12-23). MEASUREMENTS: FEMALES (25) (paratypes): HOLOTYPE: Female, collected by Y. Suto, L = 0.93 mm (0.80-1.05); a = 32 (26-35); b = 6.6 (5.6-7.6); c = 53 (42-62); V = 56 10 December 1965. Slide number Rotylenchus 1-1, Government Forest Experiment Station, (52-60); spear = 29 p. (28-31); O = 13-20. Laboratory of Forest Pathology Collection, To- MALES (9) (paratypes): L = 0.91 mm kyo, Japan. (0.83-1.01); a = 35 (31-38); b= (5,5-6,3); ALLOTYPE: Male, same data as holotype. c = 30 (28-36); spear = 29 p. (27-31); O = Slide number Rotylenchus 1-2, Government 17-19; gubernaculum = 11 p. (10-11); spic- Forest Experiment Station, Laboratory of For- ules = 26 p (25-28); capitulum = 9 ^ (8-10). est Pathology Collection, Tokyo, Japan. FEMALE (holotype): L = 0.96 mm; a = 30; PARATYPES: Thirty-three females, nine males, b = 6.5; c = 56; V = 56; spear = 30 ^ O same data as holotype and allotype, deposited = 17. with Government Forest Experiment Station, Body in spiral shape when relaxed by gentle Laboratory of Forest Pathology, Tokyo, Japan. heat. Lip region broadly hemispherical, not TYPE HABITAT: Soil around roots of Japa- set off, six annules without longitudinal stria- nese red pine (Pinus densiflora Sieb. et Zucc.). tions. Anterior portion of spear slightly longer TYPE LOCALITY: Forest nursery, Sakurae- than posterior portion. Spear knobs round, cho, Ochi-gnn, Shimane, Japan. slightly flattened anteriorly. Excretory pore at DIAGNOSIS: Rotylenchus pini can be distin- level of posterior part of isthmus. Hemizonid guished from the other species of this genus just above excretory pore. Nerve ring at pos- except R. breviglans Slier, 1965 by the compact terior part of isthmus. Esophageal glands basal portion of esophageal glands and its short, occupying nine body annules, compact overlapping the intestine very slightly. This at basal portion, slightly overlap intestine two species is most closely related to R. breviglans body annules. Epiptygma present. Sperma- but differs in the presence of males, having theca \vell developed with sperm. Phasmids distinct spermatheca in females, more anterior 25 annules anterior to anus (paratypes range position of the phasmids, and the shape of tail 11—30). Lateral field marked by four incisures, without ventral projection.

i 1 A 50/* i HB,E,F,G,H 20;* CD

Figure 1. Rotylenchus pini n. sp. A. Female; B. Female, anterior portion; C. Female, face view; D. Female, cross-section through basal annule of lip region; E-F. Female, posterior portion; G. Male, anterior portion; H. Male, posterior portion.

Discussion Acknowledgments With some specimens of R. pini it is very The author wishes to thank Dr. S. A. Sher hard to determine whether the greatest esoph- for confirming the identification of the species, ageal gland overlap is dorsal or ventral. Sher and for the loan of paratypes of R. breviglans, (1965) discussed this gland overlap feature for and Dr. J. L. Ruehle and Dr. A. Morgan R. breviglans and additional closely related but Golden, for their reading and comments on the undescribed species available to him. Despite manuscript. Thanks are also due to Mr. Y. the nature of esophageal gland overlap, R. Suto for his sending soil samples from the type pini is still considered a Rotylenchus species locality. because it closely resembles R. breviglans and has so many characters typical of the genus. Literature Cited Additional specimens of R. pini have been Goodey, J. B. 1957. Laboratory methods for found in the following habitats and localities: work with plant and soil nematodes. Technical hinoki (Chamaecyparis obtusa Endl.), Iwa- Bulletin No. 2, Ministry of Agriculture, and mura National Forest Nursery, Iwamura-cho, Fisheries and Food, London. Third Edition, Ena-gun, Gifu, Japan; sugi (Cryptomeria 47 p. japonica D. Don.), Muramatsu National Forest Sher, S. A. 1965. Revision of the Hoplolaiminae Nursery, Muramatsu-cho, Nakakanbara-gun, (Nematoda) V. Rotylendius Filipjev, 1936. Niigata, Japan. Nematologica 11: 173-198.

On the Classification and Life History of Fergusobia curriei (Sphaerulariidae: Nematoda)

J. M. FlSHER1 AND W. R. NlCKLE2

The original description of Fergusobia curriei occurs in some species of Allantonematidae. (Johnston, 1938) Christie, 1941, a parasite of Geraert (1966) accepted the family status of Eucalyptus gall flies in Australia, included sev- Fergusobiidae, but although recognizing the eral unusual features which have caused diffi- lack of information, did not accept the relation culty in understanding the classification and with Criconematidae. Nickle (1967), on the biology of this nematode. Goodey (1963) basis of insect parasitism and the present proposed a new subfamily, Fergusobiinae, in studies, placed Fergusobiinae in the Sphaeru- Allantonematidae to include this species. Later lariidae. Siddiqi and Goodey (1963) proposed a new In view of these transfers and the need for family, Fergusobiidae, with a suggested rela- more morphological and biological information, tionship with Criconematidae because of the the nematode has been studied further and in fused and swollen metacorpus and procorpus, more detail. Although nematodes associated crescentic valve-plates in the anterior portion with a number of different flies of the genus of the metacorpus, and the presence of a basal Fergusonina were investigated, only data on bulb. Nickle (1965) questioned the interpre- the nematode associated with Fergusonina tation of the valve in the median bulb and tillyardi Tonn. are recorded here. This species suggested that the crescentic shape may have of fly was found in flower bud galls on Eu- been due to accumulation of secretions as calyptus camaldulensis Dehn. near Mt. Bold, S. Australia, and its development from egg to 1 Department of Plant Pathology, Waite Agricultural Re- search Institute, Adelaide, South Australia. adult with the associated stages of the nema- - Nematology Investigations, Crops Research Division, Agricultural Research Service, USDA, Beltsville, Maryland. tode has been studied over the last 4 years.

All measurements were made from nema- framework lightly sclerotizecl. Stylet tylen- tocles killed by hot acetic acid (Seinhorst, choicl. Dorsal gland orifice approximately 3 ^ 1962) and mounted in fixative F.A. 4: 10. posterior to stylet knobs. Ampulla large, just posterior to gland orifice; gland duct broad, Redescription of Fergusobia curriei except for portion between ampulla and esoph- (Johnston, 1938) Christie, 1941 ageal lumen and for passage through the Currie (1937) described the nematode isthmus. Anterior portion of esophagus swol- Anguillulina (Fergusobia) tumifaciens, and a len, nonmuscular, with valvelike structure sit- year later Johnston found this taxon to be pre- uated anteriorly, not criconematidlike. Lumen occupied by Anguillulina tumefaciens (Cobb, of esophagus narrower anterior to valvelike 1932) T. Goodey, 1933, and proposed a structure, broader posterior to it. Anterior part new name, Anguillulina (Fergusobia) curriei. of esophagus narrows abruptly to short isthmus, Though Cobb's tumefaciens was transferred encircled by nerve ring. Esophagus expands temporarily to the genus Pratylenchus by Filip- abruptly posterior to nerve ring to contain large jev in 1936, it is still a valid case of homonymy. glands which overlap intestine; dorsal gland Christie (1941) raised the subgenus Fergusobia contains prominent nucleus, 10 by 13 p. with to generic rank, but gave Currie credit for the nucleolus approximately 5 p. in diameter. Ex- species instead of Johnston. The combination, cretory pore located at level of anterior end of Fergusobia tumifaciens (Currie, 1937) Chitw. intestine; hemizonid anterior to pore. Intestine and Chitw., 1950 used by many authors since packed with granules. Testis single, extending 1950 is not correct nor did it appear in Chit- anteriorly to region of nerve ring, may have wood and Chitwood (1950). flexure in older specimens. Reproductive tract packed with small, round sperm. Spicules Fergusobia curriei (Johnston, 1938) Christie, paired, with diagnostic right-angled bend ap- 1941. proximately halfway along their length. Gu- Anguillulina (Fergusobia) tumifaciens Currie, bernaculum absent. Caudal alae narrow, 1937. smooth, peloderan. Tail tip bluntly rounded. Anguillulina (Fergusobia) curriei Johnston, Phasmids not seen. 1938. DESCRIPTION OF INFECTIVE STAGE FEMALE Fergusobia curriei (Currie, 1937) Christie, (Fig. IB): Occurs in gall. Larger than par- 1941. thenogenetic female. Ventral side of body con- Fergusobia tumifaciens Currie, 1917. (Error vex when killed by heat. Cuticle finely in Wachek, 1955). annulated; lateral field with variable numbers Fergusobia tumifaciens (Currie, 1937) Chitw. of fine lines, usually about eight. Lip region and Chitw., 1950. slightly set off, lightly sclerotized. Stylet ADULT PARASITIC FEMALES (10): L = 866 tylenchoid. Anterior part of esophagus swol- (774-951) p.; W = 187 (165-207) ^ vulva = len, with valvelike structure in anterior part. 84 (77-88)%. Dorsal esophageal gland orifice approximately PARTHENOGENETIC FEMALES (20): L = 318 3 p, posterior to stylet knobs; short narrow duct (275-370) fi; a = 9.0 (6.8-11.0); b = 2.2 connects orifice with a single large ampulla; (1.8-2.6); c= 12.6 (9.6-14.9); stylet = 7 (5- from this, a wide sinus leads posteriorly to 8) p.; vulva = 82 (78-84)%. gland, narrowing for passage through isthmus. INFECTIVE STAGE FEMALES (10): L = 442 Lumen of esophagus is narrow to valvelike (417-489) p.- a =11.5 (10.0-12.6); b = 3.2 structure, broader posteriorly. Esophagus nar- (2.5-3.6); stylet = 8 (7-9) fl; vulva = 80 rows abruptly to a short isthmus which is en- (76-82)%. circled by nerve ring, then expands markedly FEMALES (20): L = 421 (370-492) ^ a = to contain large glands overlapping intestine. 11.6 (9.6-13.8); b = 3.1 (2.3-3.7); c = 11.3 Dorsal gland nucleus and nucleolus prominent. (9.7-13.6); stylet = 8 (6-9) ^. Excretory pore opposite beginning of intestine. DESCRIPTION OF MALE (Fig. 1A): Occurs in Gonad single, typically sphaerulariid; uterus gall. Ventral side of body concave when killed acts as extensive spermatheca, extending from by gentle heat. Cuticle finely striated; lateral vagina almost to base of esophageal glands; field with four lines. Lip region slightly set off, oviduct short, often with flexure; ovary with

A,B

Figure 1. Fergusobia curriei. A. Male; B. Fertilized infective stage female.

Figure 2. Fetgusobia curriei. A. Parthenogenetic female. B. Insect parasitic female.

Copyright © 2011, The Helminthological Society of Washington 44 • PROCEEDINGS OF THE HELMINTHOLOGICAL SOCIETY numerous ooeytes, often extending to nerve Biology and Life Cycle of ring; vulva inconspicuous, transverse, lips not Fergusobia curriei protruding. Anus an inconspicuous pore. Tail almost hemispherical. Numerous nuclei scat- The female nematode, parasitic within Fer- tered along length of intestine, also in hypo- gtisonina tillyardi, lives in the haemocoel of dermis and in tail region. adult female flies. No nematodes have been DESCRIPTION OF PARTHENOGENETIC FEMALE found in adult male flies, but all galls contain (Fig. 2A): Occurs in gall. Smaller than sexual nematodes. Eggs are laid in the body cavity generation. Ventral side convex when killed of the fly, and after hatching, the nematode by gentle heat. Cuticle finely striated; lateral larvae migrate into the oviduct of the fly. The field with four lines. Lip region slightly set off, fly deposits both insect eggs and nematode lightly sclerotized. Body begins to swell imme- larvae, separately, into Eucalyptus tissue diately posterior to lips. Stylet tylenchoid. where the nematodes quickly develop into Esophagus as in infective stage female but parthenogenetic females. Gall formation be- glands extend to almost 50% of body length. gins before the insect eggs hatch. The par- Excretory pore opposite beginning of intestine, thenogenetic nematodes soon lay eggs, and hemizonid anterior to pore. Intestine packed the resulting larvae develop into males while with granular material. Gonad single, extend- the maggots develop to the third instar. Eggs ing to region of nerve ring, sometimes flexed continue to be laid by the parthenogenetic females, and as the third instar maggots main older specimens; single ovarian cell, followed ture, some larval nematodes develop into by a large zone of maturation; uterus muscular, females. These females are thus produced late usually containing a single egg; vagina curves in the life of the third instar maggot. The toward vulva; vulval lips protrude in young nematodes mate, and shortly before pupation specimens. Anus a minute pore. Tail tapers of the maggot, up to seven (average 6) fer- to broadly rounded tip. Phasmids not seen. tilized infective-stage female nematodes pene- ADULT PARASITIC FEMALE (Fig. 2B): Occurs trate each fly larva. Penetration of the fly in body cavity of insect. Body large, swollen, maggots has not been observed. Whether both sausage-shaped, containing mostly reproduc- male and female maggots are penetrated by tive organs. Somatic setae not seen. Stylet the nematodes has not been determined. All tylenchoid. Esophagus degenerate. Cuticle late third instar maggots that were examined wrinkled, especially at anterior end; hypo- had nematodes within their body cavities, but dermis thick, with many large nuclei. Ovary nematodes were never found in adult male extensively developed, convoluted, occupies flies. During pupation, the parasites grow over 90% of body; ooeytes arranged about rapidly and are associated with the fat body central rachis; spermatheca with sperm, con- of the insect pupa, making the nematodes sisting of cylindrical tube lining wall of uterus; difficult to examine. The nematodes lay eggs shortly after emergence of the fly from the many eggs in uterus at same time; vulva trans- gall. verse, appearing as an enlarged, depressed slit. Anus not seen. Discussion FERGUSOBIINAE ( SPHAERULARIIDAE ): Diag- nosis emended. Containing four different Currie (1937) did not describe the infec- forms. (1) Adult Parasitic Female. Occurring tive stage female, and his drawing of the valve in the esophagus was difficult to interpret. in body cavity of insect. Body sausage-shaped. The valvelike structure in the esophagus is Uterus not everted. Oviparous. (2) Partheno- small and difficult to see, but use of the lacto- genetic Female. Not occurring in body cavity phenol-cottoii blue technique (Goodey, 1957) of insect, but in plant material. Oviparous. helped to clarify this. The structure appears (3) Infective Stage Female. Stylet present, as small cuticularized pieces in the walls of well developed, with knobs. Ovary with many the lumen of the esophagus; they are not cres- cells. Tail without supplements. (4) Male. centic in outline, and therefore, not criconem- Stylet present with knobs. Caudal alae present. atidlike as described by Siddiqi and Goodey Gubernaculum absent, spicules simple. (1963). Though the anterior part of the

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 45 esophagus is large and swollen as in the The life cycle of the nematode is unique in Criconematidae, the development of muscles that two female stages occur outside the insect in the metacorpus of Fergusobia is meager. in association with Eucalyptus. One female is The extensive overlapping of esophageal glands parthenogenetic, the other is the infective stage shows no affinity with the Criconematidae; female which is fertilized prior to penetration however it does show relationships with the of the insect maggot. The first eggs which the sphaerulariid genera. Furthermore, the infec- parthenogenetic females deposit develop into tive stage female was found for the first time males, and it is not until some time later, some and has the sphaerulariid type of gonad. The months in F. tillyardi galls, that the infective life cycle and insect parasitism also suggest stage females develop. that this genus is more correctly located in Currie's contention that the nematodes ini- the Sphaerulariidae. However, the possession tiate gall formation could not be confirmed. of a valvelike structure in the metacorpus is In every gall examined, both nematodes and not consistent with the diagnosis of the family, maggots were present, and in the early stages, but because we think this nematode has nematode development proceeded before the stronger relationships with the Sphaerulariidae fly's eggs hatched. It is conceivable that the than with other families it is placed there at nematodes contributed to gall formation, but this time. the possibility of a gall-inducing substance be- The question of the species of Fergusobia ing injected during oviposition by the female infesting Fergiisonina tillyardi is complex. fly or secreted by the maggot cannot be ex- The parthenogenetic females are considerably cluded. Although direct observations on feed- smaller than the measurements given by Cur- ing were not made, it seems possible that the rie, but our infective stage females are ap- parthenogenetic females may be true plant or proximately the same length as Currie's par- fungal parasites and that other stages (the thenogenetic females. This suggests that Cur- fertilized female and her larvae) are parasitic rie may have had both females, drew the in the insect. It is assumed that the partheno- parthenogenetic one, and measured the fer- genetic female must feed on something in the tilized one without recognizing the difference; gall to continue laying eggs, and that after or that Currie's parthenogenetic females were these eggs hatch, the larvae must also feed larger than ours, and in fact, we are dealing to grow into the adult stage. with a different species. Measurements of the The numbers of nematodes infecting differ- length of ten parthenogenetic females from ent species of flies varied. In F. tillyardi, six shoot tip galls on E. camaldulensis caused by nematodes (variation five to seven) were com- Fergiisonina lockharti averaged 357 (325- monly found in the body cavity of each fly. 402) fji and thus are intermediate between Two nematodes were found regularly in the those from F. tillyardi and Currie's material. body cavity of F. lockharti. Thus Currie's be- Our measurements of males and those of lief that flower-bud galling flies usually con- Currie agree closely, but males associated with tained smaller numbers of nematodes than F. lockharti are considerably longer at 481 shoot-galling flies has not been substantiated. (433-537) fji. In view of these variations in Summary length, we consider it best at the present time Fergusobia curriei (Johnston, 1938) Chris- to designate the nematode associated with F. tie, 1941, a parasite in the Eucalyptus gall fly, tillyardi as Fergusobia curriei. Fergiisonina tillyardi, from the red gum Eu- Though Fergusobia is similar to Heteroty- calyptus camaldulensis has been redescribed, lenchus Bovien, 1937, in having an alternation and new information on the life cycle is pre- of gametogenetic and parthenogenetic genera- sented. A missing stage, the infective stage tions, it differs in that the parthenogenetic female showing the correct sphaerulariid status generation is outside the insect where it may of this nematode, has been found. Studies on feed on plant or fungal tissue. Also, Fergusobia the valvelike structure in the esophagus show has morphological characters which separate it little similarity with the crescentic valve of the sufficiently from the other genera, and so it Criconematidae. The description is emended. is considered to be of separate subfamily rank. This paper supports the placement of Ferguso-

Copyright © 2011, The Helminthological Society of Washington 46 PROCEEDINGS OF THE HELMINTHOLOGICAL SOCIETY bia in the subfamily Fergusobiinae of the todes. (Revised by J. B. Goodey). Methuen, family Sphaerulariidae. London, 544 p. Johnston, T. H. 1938. A census of the free- Acknowledgments living and plant-parasitic nematodes recorded We thank Dr. D. H. Colless of CSIRO as occurring in Australia. Trans. Royal Soc. South Australia, 62: 149-167. Canberra for identifying the species of Fer- Nickle, W. R. 1965. On the status of Prothal- gusonina, and Mr. D. E. Symon of Waite Insti- lonema dubium Christie, 1938 (Nematoda: tute for identifying the species of Eucalyptus. Allantonematidae). Nematologica 11: 44. . 1967. On the classification of the insect Literature Cited parasitic nematodes of the Sphaerulariidae Christie, J. R. 1941. Parasites of invertebrates. Lubbock, 1861 (Tylenchoidea: Nematoda). In Introd. to Nematology, p. 246-266. Proc. Helm. Soc. Wash. 34: 72-94. Currie, G. A. 1937. Galls on Eucalyptus trees. Seinhorst, J. W. 1962. On the killing, fixation A new type of association between flies and and transferring to glycerine of nematodes. nematodes. Linn. Soc. N.S.W. Proc. 62: 147- Nematologica 8: 29-32. 174. Siddiqi, M. R., and J. B. Goodey. 1963. The Geraert, E. 1966. The systematic position of status of the genera and sub-families of the the families Tylenchuliclae and Criconemat- Criconematidae (Nematoda); with a com- idae. Nematologica 12: 362-368. ment on the position of Fergusobia. Nema- Goodey, J. B. 1957. Laboratory methods for tologica 9: 363-377. work with plant and soil nematodes. H.M.S.O. Wachek, F. 1955. Die entoparasitischen Tylen- 47 p. chiclen. Parasitolog. Schriftenreihe, 3: 1-119 Goodey, T. 1963. Soil and freshwater nema- Jena, Fischer.

Neoechinorhynchus constrictus sp. n., an Acanthocephalan from Texas Turtles

JOHN W. LITTLE AND SEWELL H. HOPKINS Department of Biology, Texas A & M University, College Station, Texas

It was assumed for many years that Neo- of the female and eggs, we recognized an echinorhynchus emydis (Leidy, 1851) was the undescribed species of Neoechinorhynchus, only Acanthocephalan present in North Amer- All 19 specimens were relaxed in cold, distilled ican turtles. Then Cable and Hopp (1954) water, fixed in AFA, stained with Alum described two additional species, N. chry- Cochineal Solution and cleared with Methyl semydis and N. pseudemydis. In the last few Salicylate. All measurements were made on years, two more have been described, N. whole mounts, in millimeters. There were ten emyditoides (Fisher, 1960) and N. stunkardi females and nine males; but only eight of the (Cable and Fisher, 1961). females, all mature, were measured. Twenty turtles, from in and near College Station, Texas were examined. Acanthocephala Neoechinorhynchtis constrictus n. sp. were collected from intestines of ten Pseudemys (Figs. 1-12) scripta elegans (Wied); all others, three Chely- DESCRIPTION: With the characteristics of dra serpentina (Linnaeus), three Trionyx mu- the genus Neoechinorhynchus. Trunk of fe- ticus (Le Sueur), two Sternotherus odoratus male to 23 mm long and 0.820 wide, males (Latreille) and two Terrapene Carolina (Lin- 19.2 long and 0.800 wide. Proboscis of female naeus) were negative. From two of the in- 0.127-0.196 long and 0.179-0.220 wide. Pro- fected hosts, on the basis of shape and size boscis of male 0.161-0.218 long and 0.161-

1 2

0.5mm "-" 0.2mm 0.5mm Figures 1-12. Neoechinorhynchus constrictus sp. n. Figures 1-4 were made with the aid of photomicro- graphs and 5-12 were made with the aid of a camera lucida. 1. Male, lateral view of posterior extremity. 2. Mature eggs. 3. Proboscis of holotype i'emale. 4. Posterior end of holotype female. 5-12. Posterior extremities of various females. 5-6. Immature females. 7. Female holotype. 8. Lateral view of another female. 9. Paratype, slightly rotated. 10-11. Lateral views of other mature females. 12. Ventral view of another mature female.

0.211 wide. Proboscis hooks in three rows of conical papilla 2 six hooks each, arranged quincunxially. In 2. Eggs almost round with conspicuous C- female, lateral hooks of anterior circlet pos- shaped vacuole .... emydis (Leidy, 1851) terior to others, 0.093-0.113 long; laterodorsal Eggs not round and without C-shaped and lateroventral hooks of that circlet 0.078- vacuole 3 0.092 long. Hooks of middle circlet similar 3. Posterior extremity deeply constricted .. and in line, measuring 0.046-0.069 long. Lat- constrictus (Little and Hopkins) eral hooks of basal circlet 0.027-0.044 long, Posterior extremity not deeply con- others 0.035-0.058. In male, lateral hooks of stricted 4 anterior circlet posterior to others, 0.078-0.131 4. Females up to 16 mm long long; laterodorsal and lateroventral hooks of .... chrysemijdis (Cable and Hopp, 1954) that circlet 0.064-0.097 long. Hooks of middle Females more than 16 mm long, up to circlet similar and in line, measuring 0.053- 39.2 mm 5 0.069 long. Lateral hooks of basal circlet 0.025 5. Eggs broadly spindle-shaped, up to to 0.046 long, others 0.041-0.062. Male repro- 0.054 mm long, middle shell mem- ductive system occupying 41—60% of trunk brane with uniformly distributed length; anterior testis 1.050-1.710 long, 0.270- tubelike structures 0.410 wide; posterior testis 1.150-1.950 long, ____ pseiidemydis (Cable and Hopp, 1954) 0,300-0.370 wide; cement gland 1.70-2.63 Eggs not broadly spindle-shaped, up to long, 0.280-0.370 wide; cement receptacle 0.030 mm long, no tubelike structures globular, posterior to cement gland. Mouth at end of middle membrane of uterine bell 0.800-0.920 from posterior end emyditoides (Fisher, 1960) of female; uterus exclusive of bell and selector apparatus 0.248-0.343 long, 0.046-0.121 wide; Discussion vagina 0.161-0.233 long, 0.041-0.069 wide. Neoechinorhynchus constrictus is distinct in Posterior end of female constricted about the possessing a deeply constricted posterior ex- level of vagina, reaching maximum constriction tremity. Although this worm resembles N. about the level of vagina sphincter, with gen- pseiidemydis more than the other members of ital pore ventral, in furrow. Embryonated eggs the genus, it differs from the latter by having in body cavity of mature female 0.030-0.037 smaller eggs (0.030-0.039 compared to 0.042- long, 0.007-0.012 wide; acanthor 0.030-0.034, 0.054) and a much smaller body trunk. The by 0.008-0.012; middle shell membrane with morphological variation in the posterior extrem- tubelike structures. ity of the female (Figs. 5—12) is probably due TYPE SPECIMENS: USNM Helm. Coll. No. to age. The immature ones (Figs. 5-6) have 62972 holotype female and eggs (two slides); longer and more narrowly constricted extrem- No. 62973 allotype male; No. 62974 paratypes ities than the mature worms. male and female. Male N. constrictus do not exhibit the same OTHER SPECIMENS: Laboratorio de Hel- visible external variations noticed in the fe- mintologia, Institute de Biologia, Universidad males. This relative stability is unlike some National Autonoma de Mexico. other members of the same genus. According TYPE LOCALITY: Brazos County, Texas. to Lynch (1936) referring to the fish parasite, TYPE HOST: Red ear turtle, Pseudemys N. veniistus, "The males of this species are scripta elegans. decidedly variable, so much so that at the two extremes of their range of variation they al- Key to the Species of Neoechinorhynchus most present the appearances of two distinct from North American Turtles, species." Based on Females A high degree of host specificity is apparent 1. Female apparatus sigmoid in shape and here as Acanthocephala were found only in posterior end with terminal conical P. s. elegans, yet several other species of turtles papilla were taken from the same area but contained stunkardi (Cable and Fisher, 1961) no worms. Acholonu (1967) also found that Female apparatus not sigmoid in shape only P. s. elegans out of a total of five different and posterior end without terminal species (77 specimens) of turtles examined

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 49 harbored Acanthocephala. N. emyditoides was oratory for providing us with specimens of the present in all ten specimens of P. s. elegans, described species of Neoechinorhynchus from several of which contained over 100 worms; the USNM collection. no other species of Acanthocephala was present other than the N. constrictiis in two hosts. Literature Cited It would seem possible that there still may Acholonu, A. D. 1967. Studies on the acantho- be unknown species of Acanthocephala from cephalan parasites of Louisiana turtles. Bull. turtles. Wildlife Disease Assoe. 3: 40. Cable, R. M., and F. M. Fisher, Jr. 1961. A Summary fifth species of Neoechinorhynchus (Acantho- cephala) in turtles. J. Parasit. 47: 666-668. Neoechinorhynchus constrictiis sp. n. is de- , and W. B. Hopp. 1954. Acantho- scribed from a Texas turtle, Pseudemys scripta cephalan parasites of the genus Neoechino- elegans. This new species is distinguished by rhynchus in North American turtles with the descriptions of two new species. J. Parasit. the females possessing a deeply constricted 40: 674-680. posterior extremity, small eggs, and small Fisher, F. M., Jr. 1960. On acanthocephala of trunk. A key is presented for the females of turtles, with the description of Neoechino- Neoechinorhynchus found in North American rhynchus emyditoides n. sp. J. Parasit. 46: turtles. Morphological variation in females is 257-266. probably clue to a difference in age. Leidy, J. 1851. Contributions to helminthology. Proc. Acad. Nat. Sci., Phila. 5: 205-209. Acknowledgments Lynch, J. E. 1936. New species of Neoechino- rhynchus from the western sucker, Catostomus We wish to express thanks to Dr. W. W. macrocheilns Girard. Tr. Am. Micr. Soc. 55: Becklund of the Beltsville Parasitological Lab- 21-43.

A Note on the Morphology of Cooperia punctata (Linstow, 1907) and Cooperia spatulata Baylis, 1938 MARTHA L. WALKER AND WILLARD W. BECKLUND Beltsville Parasitological Laboratory, Animal Disease and Parasite Research Division, Agricultural Research Service, USDA, Beltsville, Maryland

Until 1958, Cooperia spatulata Baylis, 1938, bution. Therefore, a comparison of the males a nematode parasite of cattle and sheep, had of these two species was made and the results not been reported outside Australia and the are presented as an aid in their identification. Federated Malay States (Baylis, 1938; Roberts, Thirty-nine specimens of C. spatulata from 1939; Secldon, 1947). In 1958 it was first re- Georgia and Florida cattle and 33 of C. punc- covered in the United States from cattle in tata from Georgia and Florida cattle and southern Georgia (Becklund, 1958). Since Georgia sheep were carefully studied and then, it has been reported from calves in measured. The specimens of C. punctata were Hawaii (Alicata, 1960), cattle in Florida especially chosen for their large size in order (Becklund, 1961a), sheep in Georgia (Beck- to determine if the maximum size of C. punc- lund, 1961b), and a lamb and steers in Missis- tata extended into the C. spatulata range. sippi (Knight, 1962). These recent records Linstow (in Schnycler, 1907) described C. show that this parasite has a wider distribution punctata as having a total body length of 5.3- than earlier believed. It is probable that C. 6 mm; our 33 specimens varied from 5.9-8.1 spatulata has been overlooked because of its mm, with an average of 7.1. Cooperia spat- similarity to C. punctata (Linstow, 1907) ulata was described as being 6.3-7.5 mm long Ransom, 1907, a common sheep and cattle (Baylis, 1938); our 39 specimens ranged from nematode with an almost cosmopolitan distri- 4.4-7.8 mm, with an average length of 6.4.

Figures 1-8. Spicules of Cooperia punctata (Figs. 1-4) and Cooperia spatulata (Figs. 5-8) from Georgia cattle. 1, 5. Ventral aspect. 2, 6. Dorsal aspect. 3, 7. Lateral aspect. 4, 8. Lateral aspect, removed from worm and flattened, a. Concavity, b. ventral flange. All drawings made with the aid of a camera lucida.

The average width, both of the head and of larger in C. spatulata than in C. punctata. the body at the esophagointestinal junction, These differences in the concavity and the did not differ by more than 2 p, between the ventral flange of the spicules are most im- two species. The average width of the body portant in distinguishing the two species. just anterior to the bursa was 116.4 ^ for Both C. punctata and C. spatulata are sim- C. punctata, 157.1 p. for C. spatulata. The ilar to C. africana, a species described by length of the dorsal ray of C. punctata ranged Monnig (1932) from three males collected from 105-139 p., with an average of 117.2; from an. eland, Taurotragus oryx, in Kenya. that of C. spatulata from 112-221 p., with an The writers wanted to include C. africana in average of 157.1. The variation and the over- this study, but were unable to borrow speci- lapping make it difficult to designate any of mens. Apparently, only one male of the type these measurements to separate the two spe- material still exists and it is damaged and not cies. Although in general the bursa of C. available on loan. spatulata is larger than that of C. punctata, close examination of the pattern of the rays Literature Cited and also of the structure of the genital cone revealed no apparent differential characters Alicata, J. E. 1960. Incidence of parasites in calves in Hawaii and the treatment of Coo- between the two species. peria punctata, with special reference to the Spicule length has been used to distinguish efficacy of Ruelene. Am. J. Vet. Res. 21: 410- C. spatulata and C. punctata. According to 415. Baylis, the range for C. spatulata was 230- Baylis, H. A. 1938. A new species of Cooperia 290 p,. In our 39 specimens, the spicule lengths (Nematocla) from cattle and sheep. Ann. and varied from 184-279 /j.. Cooperia punctata Mag. Nat. Hist. Ser. 11, 1: 68-73. was described as having a spicule length of Becklund, W. W. 1958. Cooperia spatulata re- 136-149 /j, (Linstow in Schnyder, 1907) and covered from cattle in the United States. J. Ransom (1911) reported a length of 120- Parasit. 44: 186. . 1961a. Helminth infections of healthy 150 p. However, when specimens which Ran- Florida cattle, with a note on Cooperia spat- som had identified as C. punctata were re- ulata. Proc. Helm. Soc. Wash. 28: 183-184. examined, spicule lengths up to 180 p, were 1961b. Helminthiasis of sheep in south- observed; and in our 33 selected large speci- ern Georgia. J. Am. Vet. Med. As. 139: 781- mens, the lengths were from 167—214 p.. This 784. overlap of the ranges of C. spatulata and C. Knight, R. A. 1962. Occurrence of the nema- punctata indicates that the spicule lengths todes, Trichostrongylus longi&picularis, Oster- cannot be vised as a specific character. How- tagia lyrata, and Cooperia spatulata in rumi- ever, although the spicules of the two are nants in Mississippi. Proc. Helm. Soc. Wash. 29: 145-147. rather similar in general conformation, they Monnig, H. O. 1932. New strongylid nema- can be quite readily used to differentiate the tocles of antelopes (preliminary notes). I. S. species. Afr. Vet. Med. As. 3: 171-175. The spicules of both C. punctata and C. Ransom, B. H. 1911. The nematodes parasitic spatulata are undivided, without branches or in the alimentary tract of cattle, sheep, and spurs. Each spicule has a concavity with a other ruminants. Bull. (127) Bureau Animal concentrically striated wall which opens on Indust., U. S. Dept. Agric., 132 p. the ventral surface. In C. punctata this con- Roberts, F. H. S. 1939. The occurrence and cavity is large with a distinct border which prevalence of gastrointestinal helminths in apparently healthy cattle in Queensland, Aus- projects laterally at an angle from the shaft of tralia. J. Comp. Path. Ther. 52: 160-165. the spicule. It is situated very slightly distal Schnyder, O. 1907. Eine neue Strongylusart. to the middle of the spicule (Figs. 1-4). In Centralbl. Bakteriol., 1 Abt., Orig. 43: 708- C. spatulata the concavity is small, indistinct, 709. and does not project appreciably from the Seddon, H. R. 1947. Host check list of hel- spicule. It is usually slightly more distal than minth and arthropod parasites present in do- that of C. punctata (Figs. 5-8). The spicules mesticated animals in Australia. Serv. Pub- of both species have a ventral flange distal to lication (Div. Vet. Hyg.) (2) Australia Dept. the concavity. This ventral flange is much Health. 41 p.

The Synonymy of the Ruminant Parasites Nematodirus oiratianus Raevskaia, 1929 and Nematodirus lanceolatus Ault, 19441

KAY S. SAMSON USDA, Agricultural Research Service, Animal Disease and Parasite Research Division, Las Cruces, New Mexico

Nematodirus oiratianus Raevskaia, 1929 (see closer to the edge of the bursa than in N. Raevskaia, 1931) was first described from oiratianus and that a retrograde prong between Altai wapiti (Cervus canadensis asiaticus) and the spicules was lacking. Also, he was unable domestic sheep (Ovis aries) in Asiatic Russia. to find eggs as large as those described for It has since been recovered from other rumi- N. oiratianus. nant hosts in several localities in the USSR. The first report of N. lanceolatus from North The Index Catalog of Medical and Veterinary America was that of Gilmore and Allen (1960) Zoology lists the following host records: water who found it in pronghorn antelope in New buffalo (Bos bubalis), bezoar goat (Capra Mexico. They identified their specimens tenta- aegagrus), domestic goat (C. hircus), Siberian tively as N. lanceolatus because the measure- mountain goat (C. sibirica), roe deer (Capreo- ments of the spicules more nearly corresponded lus capreolns) and (C. pygargus bedfordi), to those described for that species than to European red deer (Cervus elaphus xanthop- N. oiratianus. This same tentative identifica- ygus), sika (C. nippon), Dzeren antelope tion was given to specimens from domestic (Gazella siibgutturosa), Siberian argali (Ovis sheep and Barbary sheep in New Mexico ammon), Armenian mouflon (O. ophion arme- (Allen, 1959 and unpublished data) and to niana), chamois (Rupicapra rupicapra cau- specimens from Colorado bighorn sheep (Pill- casica), and Saiga antelope (Saiga tatarica). more, 1961). Becklund and Senger (1967) The males of N. oiratianus can be distin- recorded N. lanceolatus from Montana big- guished by the characteristic shape of the horn. Apparently there is no published record terminal ends of their spicules. Each spicule of this species from cattle. divides and the resulting rod-shaped parts To clarify the apparent taxonomic confusion fuse distally to form a lancet-shaped structure, concerning N. oiratianus and N. lanceolatus, the point of which extends completely to the the author undertook the studies on morphol- posterior border of a surrounding membranous ogy described in this report. Since females expansion. The spicules illustrated by Raev- in the genus Nematodirus have few characters skaia (1931) had a retrograde prong at the of taxonomic value, this study involved males site of their junction. The females contained principally. Detailed comparisons were made unusually large eggs (0.255-0.272 mm long of 77 males from domestic sheep in New and 0.119-0.153 mm wide). Mexico, four from the same host in Argentina, Ault (1944) discovered nematodes similar that were supplied by Dr. C. N. Ault, and a to N. oiratianus in domestic sheep in Argen- total of 51 from Siberian mountain goats, tina, but because of differences he considered Siberian argali, domestic sheep, and domestic significant, Ault named his species N. lanceo- cattle in the USSR. The Russian specimens latus. Type specimens of N. oiratianus were were kindly supplied by Dr. S. N. Boev, Insti- not available to him for comparison, so his tute of Zoology, Academy of Sciences of the conclusions were based on the description Kazakh, Alma-Ata, USSR. given iii Travassos (1937). Ault observed in The structure of the distal ends of the spic- his specimens that the externo-dorsal rays were ules was carefully noted, as was the location of the externo-dorsal rays of the bursa. Other 1 A portion of a thesis submitted in partial fulfillment of the requirements for the degree of Master of Science, New characters examined were length of body, Mexico State University, Las Cruces. This work was carried out in cooperation with the New width of anterior end, length of esophagus, Mexico Agricultural Experiment Station and is a contri- width in front of bursa, length of spicules in- bution to the Western Regional Project W-35—Nematocle Parasites of Ruminants. cluding terminal ends, size of lateral lobes of

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 53 bursa, and length of dorsal rays. Comparisons mens examined in this study are considered were first made of all specimens from domestic minor and within the bounds of intraspecific sheep. Next, measurements of specimens from variation. Therefore, it is the opinion of the domestic sheep were compared with measure- author that N. lanceolatus Ault, 1944, should ments obtained from specimens from cattle and be placed in synonymy with N. oiratianiis wild ruminants in Siberia. The measurements Raevskaia, 1929. This opinion is supported by of all specimens obtained for this study were recent observations by Samson (1966) that then compared with the original measurements large eggs comparable in size with those in the given for N. oiratianiis by Raevskaia and those original description of N. oiratianiis are occa- given for N. lanceolatus by Ault. Some of the sionally associated with both natural and ex- data were treated by analysis of variance. perimental infections in domestic sheep in New A comparison of the spicules of the various Mexico. specimens revealed no important differences. Literature Cited The retrograde prong which Ault mentioned as being present in the illustration of IV. oira- Allen, R. W. 1959. The occurrence of Nema- tianiis was not discerned in any of the spicules. todirus lanceolatus in domestic sheep in the The externo-dorsal rays of the four speci- United States. J. Colo.-Wyo. Aeacl. Sci. 4 mens from domestic sheep in Argentina were (11): 53. situated somewhat closer to the edge of the Ault, C. N. 1944. Nematodes parasitos de los bursa than the rays in specimens from both bovinos y ovinos en la Argentina (Segunda nota). Rev. Med. Vet., Buenos Aires, 26: domestic and wild ruminants in Siberia and 497-509. domestic sheep in New Mexico, but this dif- Becklund, W. W., and C. M. Senger. 1967. ference alone is not considered sufficient to Parasites of Ovis canadensis canadensis in warrant placing the Argentina forms in a Montana, with a checklist of the internal and separate species. Total lengths of specimens external parasites of the Rocky Mountain big- varied considerably as might be expected, and horn sheep in North America. J. Parasit. 53: in some cases these differences were statis- 157-165. tically significant. There is, however, some Gilmore, R. E., and R. W. Allen. 1960. Hel- difficulty in measuring total lengths of males minth parasites of pronghorn antelope (Antilo- because they are invariably coiled. The great- capra americana) in New Mexico with new est difference in average total lengths was host records. Proc. Helm. Soc. Wash. 27: 69-73. between Siberian cattle (8.77 mm) and Si- Pillmore, R. E. 1961. General investigations of berian mountain goats (12.51 mm), with the diseases and parasites. Quar. Rept. Colo. Dept. specimens from other sources ranging between Game and Fish, Oct., p. 101-102. these two. It is of interest, though, that Raevskaia, Z. A. 1931. Zur Charakteristik der Raevskaia (1931) gives a maximum total Nematoden der Gattung Nematodinis, Ran- length of 16.5 mm and that a specimen from som 1907. (Versuch einer monographischen New Mexico domestic sheep measured only Bearbeitung). Ztschr. Infektionskr. Haustiere. 7.38 mm. In our collection of males from 40: 112-136. pronghorn antelope in New Mexico, there are Samson, K. S. 1966. Preliminary studies on the some specimens as short as 7.20 mm. Despite life history of Nematodirus oiratianiis, an intestinal parasite of ruminants. Program 42. this wide variation in total lengths of the Ann. Meet. Southwest, and Rocky Mountain worms, differences in lengths of spicules were Div. Am. As. Adv. Sci. p. 35-36. small and in no case significant, regardless of Travassos, L. P. 1937. Revisao da Familia host and geographical source. Trichostrongylidae Leiper, 1912. Monogr. The differences observed between the speci- Inst. Oswalclo Cruz. No. 1, 512 p.

Monogenean Parasites of Costa Rican Fishes. II. Proposal of Palombitrema heteroancistrium n. gen., n. sp.1

C. E. PRICE- AND W. A. BUSSING:!

Prior to the present investigation, only dissimilar anchors, one pair located ventrally four freshwater monogenetic trematodes were in haptor, the other dorsally. Rases of each known from Central America, all described anchor pair connected by a separate trans- from Costa Rican hosts. E. W. Price (1938) verse bar. Hooks 14 (7 pairs), greatly variable described two species of Cleidodiscus Mueller, in both size and shape. Cirrus provided with 1934 from the gills of Rhamdia rogersi (Regan), a basally articulated accessory piece, the latter the host taken from the San Pedro Montes de of two distinct pieces joined by connective Oca. Price (in press) has given reasons why tissue. Vagina opening near left body margin. these two parasites should be transferred to Vitellaria well developed, often arranged in the genus Urocleidus Mueller, 1934. patches. Intestinal crura apparently confluent In a study recently completed (Price and posteriorly. Bussing, 1967), two species of Cleidodiscus TYPE SPECIES: Palombitrema heteroancis- were removed from the gills of Astyanax trium. fasciatus (Cuvier) and subsequently described TYPE HOST: Astyanax fasciatus (Cuvier). as the first valid Cleidodiscus forms from TYPE LOCALITY: Guanacaste Province, Rio Central America. Montenegro; 23 km NW of Canas (on Inter- This paper is concerned with the descrip- american Highway), Costa Rica, Central tion and classification of Palombitrema, a new America. genus of Monogenea, recovered from the gills of Astyanax fasciatus (Cuvier) from Costa Rica. Palombitrema heteroancistrium n. sp. HOST AND LOCALITY: Asttjanux fasciatus Materials and Methods (Cuvier); Guanacaste Province, Rio Mon- Host specimens utilized in this study were tenegro, 23 km NW of Canas (on Inter- collected in Costa Rica, immediately frozen american Highway), Costa Rica, Central for several hours, and preserved in 3.5% for- America. malin prior to shipment to the United States. BODY REGION OCCUPIED BY PARASITE: Gill Gill trematodes were recovered and treated filaments. as prescribed by Price (1966). Appropriate SPECIMENS STUDIED: Ten. measurements and illustrations were made with TYPES: Holotype deposited in USNM Helm. the aid of a calibrated filar micrometer ocular Coll. (No. 62987), Washington, D. C. First and a camera lucida, respectively. All measure- paratype deposited in Museo de Zoologia (No. ments are expressed in microns; average mea- UCR-1), Departamento de Biologia, Univer- surements are given first, followed by minimum sidad de Costa Rica, Central America. Re- and maximum values enclosed in parentheses. maining paratypes in authors' collections. DESCRIPTION: A robust dactylogyrid of mod- Palombitrema n. gen. erate size, provided with a thin cuticle; length GENERIC DIAGNOSIS: Dactylogyridae, An- 502 (454-529). Greatest body width 100 cyrocephalinae. A form of moderate size pro- (88-105), usually near midlength. Two pairs vided with a thin cuticle. Two pairs of eye- of eyespots, members of posterior pair larger spots, members of posterior pair larger. Haptor and spaced farther apart than members of well set off from body proper. Two pairs of other pair. A few eyespot granules scattered in cephalic region. Peduncle short and stout, 1 This study was supported jointly by the Departamento de Biologia, Universidad de Costa Riea and by the Depart- setting haptor off well from body proper. Well ment of Biology, Augusta College. - Department of Biology, Augusta College, Augusta, developed anterior cephalic lobes; lateral ce- Georgia 30904. phalic lobes vestigial. Pharynx muscular, sub- 3 Departamento de Biologia, Universidad de Costa Rica, Central America. spherical in ventral view (whole mount: Fig. 1).

8 0. 04 mm

Figures 1-13. Camera lucida illustrations of Palombitrema heteroancistriwn n. gen., n. sp. 1. Entire worm (ventral view); 2. Ventral anchor; 3. Dorsal anchor; 4. Ventral bar; 5. Dorsal bar; 6-9. Hooks; 10. Vagina; 11. Seminal receptacle; 12. Cirrus; 13. Accessory piece. All illustrations except Figure 1 drawn to same scale.

Two pairs of dissimilar anchors. Ventral Discussion anchor without prominent roots and with a hollow base (Fig. 2); length 33 (30-34), This form was originally considered to be width of base 15 (14-17). Dorsal anchor a deviant species of Cleidodiscus Mueller, smaller, of a shape unusual for the Ancyro- 1934. Further study indicated that this para- cephalinae: the deep root, shaft, and point site possesses sufficient atypical features to form a solid structure, whereas the superficial prevent its inclusion in Cleidodiscus. root is thin and extremely flexible, appearing Palombitrema is morphologically close to to be a membranous appendage of the base- Cleidodiscus. The ventral bar and ventrally shaft region (Fig. 3); length 23 (21-25), located hooks (Nos. 1—5) are similar to those width of base 11 (10-12). Ventral bar of possessed by Cleidodiscus species. The major simple construction (Fig. 4); length 38 (36- features for differentiation of the two genera 40). Dorsal bar unusual for subfamily: main are: (1) the two-piece construction of the shaft is in form of a "V," the ends in form of accessory piece of Palombitrema is not present poorly developed sclerotized discs (Fig. 5); in any member of Cleidodiscus, (2) the dorsal length 36 (34-37). hooks of the latter genus are quite similar in Seven pairs of hooks, five pairs located both shape and size, whereas those of the ventrally on haptor, remaining two pairs former are grossly different both in morphology located dorsally. Ranges of both size and and in size, and (3) the large, irregularly shape of hooks constitute unusual features. shaped, and heavily sclerotized vagina present Pairs one to five (ventrally located) somewhat in this new form is lacking in all known species similar to each other (Fig. 9); dorsal members of Cleidodiscus. (pairs six and seven), however, are diverse in DERIVATION OF SCIENTIFIC NAME: The ge- comparison. Members of pair No. 6 each com- neric name is chosen to honor Professor Arturo posed of a base tapering uniformly to a sickle- Palombi of Napoli, Italy, in recognition of past shaped termination, exhibiting little or no shaft work performed on the Monogenea of Europe region (Fig. 8). Members of pair No. 7 rela- and also in personal appreciation of the senior tively large and very flexible (Fig. 6,7). Hook author for recent help and encouragement ex- lengths: No. 1, 13 (11-14); No. 2, 17 (16- tended by Professor Palombi. The species 18); No. 3, 19 (18-20); No. 4, 18 (17-19); name is derived from the Greek HETERO- ("dif- No. 5, 14 (13-15); No. 6, 17 (15-18); No. 7, ferent") and from the Greek ANCISTR- ("hook" 38 (37-40). or "spine"); this designation is chosen to indicate discrepancies in hook morphology and Cirrus arising from an expanded base, tapering to a tube of narrow diameter (Fig. 12); estimated length 42. Accessory piece artic- Summary ulated to cirrus base and complex in construction: composed of two distinct portions joined Ten specimens of a new genus of Mono- together by connective tissue (Fig. 13); over- genea, Palombitrema, were recovered from the all length 27 (25-30). Vagina heavily sclero- gills of a Costa Rican teleost, Asti/anax fasciatus tized (Fig. 10), opening ventrally in left body (Cuvier). half; vaginal tube opens into a thin-walled This parasite was originally considered to seminal receptacle (Fig. 11). Ovary pretes- be a deviant species of Cleidodiscus Mueller, ticular in position, compact, and somewhat 1934. Further study indicated that the form elongate; testis relatively small, subspherical presently studied was well differentiated from to elongate in outline. Prostatic reservoir Cleidodiscus, although the two genera are ap- single, folding back upon itself. parently related. Vitellaria well developed, composed of small The type species, Palombitrema hetero- granules of essentially uniform size and density; ancistrium, is described. tends to form lateral bands in two specimens, whereas there is a tendency toward formation Literature Cited of irregular patches in remaining specimens. In- Price, C. E. 1966. Urocleidus cavanaughi, a testinal crura apparently confluent posteriorly. new monogenetic trematode from the gills of

the keyhole cichlid, Aeqtiidens rnaroni (Stein- parasites of Costa Rican fishes. Part 1. De- dachner). Bull. Georgia Acacl. Sci. 24: 117- scriptions of two new species of Cleidodiscus 120. Mueller, 1934. Riv. Parass. 25: 81-86. . In press. Synonymy in the genus Price, E. W. 1938. The monogenetic trema- Cleidodiscus with the description of a new todes of Latin America. (In) Livro Jubilar species. Qtrly. J. Florida Acad. Sci. Prof. Travassos, Rio de Janeiro, Brazil. 3: —-, and W. A. Bussing. 1967. Monogenean 407-413.

A Soil Population Study of Ditylenchus dipsaci (Kuhii) Filipjev in an Alfalfa Field1

SHU-TEN TSENG, KEITH R. ALLRED, AND GERALD D. GRIFFIN-

Introduction Materials and Methods There have been several studies made on A field of 6-year-old Ranger alfalfa (Medi- the relationship of the environment to Dity- cago sativa L.) on Millville silt loam located lenchus dipsaci in the soil. Lewis and Mai near Smithfield, Utah, was selected for this (1960) found most specimens of this nematode study. The alfalfa was infected with the alfalfa species in the top 6 inches of fallowed soil. stem nematode, Ditylenchus dipsaci (Kiihn) This agrees with the observations of Wallace Filipjev. The experimental design consisted of (1962) who observed more D. dipsaci in the dividing the field into 16 sections in a ran- first 10 cm of soil than at lower depths in an domized square block, and marking off a infested oat field. Wallace further stated that the number of nematodes increased greatly square plot 5 by 5 meters in each section. The after a rain and decreased during the following distance between the centers of any two adja- dry period. Wallace (1961) also found that cent plots was 12 meters. horizontal as well as vertical movements of Soil samples were collected from each plot D. dipsaci were related to the moisture and at 2-week intervals, from 6 August 1965 to temperature gradients in the soil. Seinhorst 26 June 1966. Sampling depths were 0-10, (1956) found that heavy soils maintained a 10-20, 20-30, and 30-40 centimeters. Three larger population of nematodes than light soils, subsamples were taken from each plot at each and Sayre and Mountain (1962) reported a sampling date using a soil auger of 10 cm in high mortality of D. dipsaci in wet soils at diameter. These were thoroughly mixed, a 400 70 F. cc composite collected in a plastic container. The present investigation was initiated to Soil remaining from the subsamples was imme- study seasonal populations of D. dipsaci rela- diately replaced into the hole. During the tive to temperature and soil moisture condi- winter, a pick was used for breaking through tions in an alfalfa field. the frozen soil to facilitate the use of the soil auger. 1 Cooperative investigations of the Utah Agricultural Ex- periment Station, Logan, Utah and the Crops Research Soil moisture was determined on each sam- Division, Agricultural Research Service, USDA. Journal Paper No. 614 of the Utah State Agricultural Experiment pling date at the various sampling depths for Station. Part of a thesis submitted by the senior author in partial fulfillment of the requirements of the M.S. degree. four of the 16 plots. Soil moisture was deter- - Respectively former Graduate Assistant, Professor of mined by the gravimetric method, and ex- Plant Science, Utah State University, and Research Nema- tologist, Crops Research Division, Agricultural Research pressed as a per cent of field capacity. The Service, Crops Research Laboratory, Utah State University, Logan, Utah. field capacity of the soil was as follows:

Table 1. Results of the analysis of multiple regression between the nematode count and environmental factors. Variance Partial Constant in Sampling Source of Degrees of Mean ratio regression multiple depth variation freedom square (F) coefficient regression 0—10 cm soil moisture 1 631.1143 11.29** 0.34 52.28 season 1 1747.1661 31.27** —29.22 temperature deviation 1 1922.3260 34.40** -5.24 season X temperature dev. 1 460.2981 8.24** 1.85 model 4 1131.5568 error 19 55.8740 R- = 0.81 10-20 cm soil moisture 1 6.9889 3.34 0.06 2.38 season 1 10.2934 4.91* -1.76 temperature deviation 1 40.6180 19.38** -0,34 time lag 1 10.7239 5.12* 0.44 model 4 22.2856 error 19 2.0956 Ra = 0.69 20-30 cm season 1 4.4647 5.14* -1.62 5.80 temperature deviation 1 5.1363 5.92* -0.29 season X temperature dev. 1 0.9045 1.04 0.09 model 3 6.3795 error 20 0.8681 RS = 0.52 30-40 cm season 1 0.2139 1.88 -0.23 1.54 I 0.5433 4.26* 0 16 ( temperature deviation )-' 1 0.3459 2.71 0.01 time lag 1 0.9211 7.22* 0.47 model 4 0.8837 error 19 0.1276 R* = 0,59 * Significant at 1% level. * Significant at 5% level.

Sampling depth (cm) Field capacity (% the maximum nematode population at this 0-10 30.1 temperature); square of the temperature devia- 10-20 28.5 tion from 15 C (Temp. —15)2; variable in time 20-30 27.1 lag of 21 days, obtained by estimating the stem 30-40 29.0 nematode population 21 days before each sampling date (the life cycle for D. dipsaci Soil temperatures were taken each sampling at 15 C is 19-23 days, Yuksel, 1960); season elate at depths of 5, 15, 25, and 35 cm at times temperature deviation; and season times approximately 12:00 noon. (temperature deviation)-. The analysis was Soil samples were processed within 24 hours conducted by using all variables; then the after sampling by a sifting and gravity and variables were removed in turn until the R- Baermann funnel method and number of nema- value began to decrease appreciably (Table 1). todes determined. (R2 = coefficient of determination = multiple Since the study was made to determine rela- regression coefficient; this represents variation tionships between the environmental factors in nematode numbers that can be explained by and changes in number of stem nematodes in variation in independent variables of multiple the soil, and to compare predicted popula- regression.) tion values to the observed nematode population, a multiple regression analysis was used. Results The following items were considered as im- The largest number of alfalfa stem nema- portant variables: soil moisture at each sam- todes were recovered from the 0—10 cm layer pling depth; season, with the first 16 sampling of soil (Fig. 1). The nematode population dates set as one and the last eight dates as two; decreased as the sampling depth increased. deviation of soil temperature from 15 C (Temp. Two population cycles were observed during -15) (15 C was used because we recovered the sampling period (Fig. 1). In the upper

10 cm layer of soil, the mean number of stem The season times temperature deviation factor nematodes increased beginning the middle of showed a significant relationship with the stem August and seemed to peak in early September, nematode population for the 0-10 cm depth followed by a sharp decline during October due to the difference in direction of tempera- and November. The second mean population ture . change during the fall as compared to peak, which appeared in the middle of May, spring. was substantially lower than the fall peak. Figure 4 shows how the predicted popula- During the winter months the stem nematocle tion values for the 0-10 cm depth compared population was consistently low at all depths. with the actual nematode population. The pattern of change of the nematode population in the 10-20 cm depth was similar to Discussion that in the upper 10 cm layer of soil, but approached its peaks 2-3 weeks later. Populations of alfalfa stem nematode in Numbers of nematodes in the 20—30 and Millville silt loam, in the presence of alfalfa 30-40 cm depths also followed a similar cycle, plants, fluctuated greatly with seasonal but the totals were much lower. At the 20- changes. One population peak was observed 30 cm depth, a maximum of only 6 stem nema- between late August and early September in todes per sample were observed, while in the 1965, and the other peak occurred in the 30-40 cm depth less than 3 stem nematodes middle of May in 1966. For the 0-10 cm per 400 cc of soil were recovered. depth, the range per 400 cc of soil was from Soil moisture fluctuated most noticeably 50 nematodes in the fall to one stem nematode (Fig. 2) in the 0-10 cm depth; soil moisture in the winter. Most of the stem nematodes fluctuating between 35 and 85 per cent field were found in the upper 10 cm of soil, which capacity between early August and the middle agrees with Lewis and Mai (1960), and Wal- of October. At the lower depths during the lace (1962). The multiple regression study same period, soil moisture ranged from 45 showed that the observed population of nema- per cent to 80 per cent. todes compared favorably to the predicted Soil temperatures reflected seasonal changes population, showing a positive correlation be- in the weather, reaching a minimum range of tween those environmental variables considered 1—2 C in the winter, a maximum range of 20- in this study. 27 C in summer months. The 0-10 cm depth A positive correlation existed between soil had the widest fluctuations and range in tem- moisture values and numbers of stem nematode peratures (Fig. 3). during the summer. Wallace (1962) observed Soil moisture increased during late fall and an increase of D. dipsaci in the soil around oats remained high throughout the winter and early following increased rainfall, and a decrease spring months. During the winter months when the soil dried. when it was frozen, the soil in the 0-10 cm Peak populations of stem nematodes oc- depth was saturated. Soil moisture at all curred in early September, and again in late sampling depths decreased rapidly during late May and early June when the soil tempera- March and early April; however, during May ture was around 15 C. The greater the devia- and June there were large fluctuations that we tion from this temperature, the fewer nema- associated with rain and irrigation. todes in the soil. At depths below 10 cm the Nematode numbers at all depths were highly nematocle populations approached their peaks correlated with the temperature deviation from 2-3 weeks later than in the 0—10 cm layer, 15 C. Season was an important factor at the when temperatures were well below 15 C in 0-10, 10-20, and 20-30 cm depths, but had the fall and above 15 C in the spring at 0-10 little effect on the population at the 30—40 cm cm. This significant time lag might indicate depth. The time lag value was significant for that the nematodes migrated downward (Lewis the 10-20 and 30-40 cm depths. This indi- and Mai, 1950). Wallace (1961, 1962) stated cated that the stem nematode populations in that the horizontal as well as the vertical move- the lower soil depths approached the peaks ment of D. dipsaci was related to the tempera- significantly later than in the 0-10 cm depth. ture and moisture gradients in the soil.

O GO Depth (cm) O 0-10 u 40 u 10-20 O 20-30 30 30-40 L. aC9 20

•a o +-> 10 a E 0 631 12 10 7 5 5 2 14 11 Aug Scp Oct Nov Dec Jan Feb Mar Apr May Jun Sampling Date Figure 1. Population density ot Ditylenchus dipsaci in the Millville silt loam of an alfalfa field.

110

100

$ 80 'u oa 70 U s 60 o L 50 o 20-30 °~ 40 A---A 30-40

3O 631 12 10 7 5 5 2 14 11 Aug Sep Oct Nov Dec Jan Feb Mar Apr May Jun Sampling Date Figure 2. Moisture at different depths in an alfalfa field on Millville silt loam soil throughout the experimental period.

25 Depth (cm)

520

L 15 ZJ -»-> O a E 5

-5 631 12 10 7 5 5 2 14 11 Aug Sep Oct Nov Dec Jan Feb Mar Apr May Jun Sampling Date Figure 3. Temperature at different depths in an alfalfa field on Millville silt loam soil throughout the experimental period.

050 00

Observed U U A A Predicted O 030

0-20

C9 T3 O 10 a o z 0

-5 631 12 10 7 5 5 2 14 11 Aug Sep Oct Nov Dec Jan Feb Mar Apr May Jun Sampling Date Figure 4. A comparison of the observed and predicted population of Dictylenchus dipsaci in the 0-10 cm depth throughout the experimental period.

Summary indicate a nematode migration toward a more desirable temperature. A population of Ditylenchus dipsaci showed a seasonal fluctuation in Millville silt loam Literature Cited collected from an alfalfa field. Two population peaks were observed during the sampling Lewis, G. D., and W. F. Mai. 1960. Over- period from 6 August 1965 to 26 June 1966. wintering and migration of Ditylenchus dip- One population peak was observed between saci in organic soils of southern New York. late August and early September 1965, and Phytopathology 50: 341-343. Sayre, R. M., and W. B. Mountain. 1962. The the other peak occurred the middle of May bulb and stem nematode (Ditylenchus dipsaci) 1966. At 0—10 cm, where most of the nema- on onion in south western Ontario. Phyto- todes were found, the population ranged from pathology 52: 510-516. 50 nematodes per 400 cc of soil in the fall, to Seinhorst, J. W. 1956. Population studies on one nematode per 400 cc of soil in the winter. stem eelworms (Ditylenchus dipsaci}. Nema- These peak populations occurred when the soil tologica 1: 159-164. temperature was approximately 15 C, and the Wallace, H. R. 1961. The orientation of Dity- greater the deviation from this temperature, lenchus dipsaci to physical stimuli. Nemato- the smaller the nematode population. Nema- logica 6: 222-236. . 1962. Observations on the behavior of todes approached their peak at the 10-40 cm Ditylenchus dipsaci in soil. Nematologica 7: levels 2—3 weeks later than those at 0-10 cm, 91-101. when temperatures were well below 15 C in Yuksel, Hasan S. 1960. Observations on the the fall and above 15 C at the 0-10 cm level. life cycle of Ditylenchus dipsaci on onion This significant time lag at 10-40 cm might seedlings. Nematologica 5: 289-296.

Parasites of the Clapper Rail, Rallus longirostris Boddaert. I. The Current Status of the Genus Levinseniella with the Description of Levinseniella byrdi n. sp. (Trematoda: Microphallidae)1

RICHARD W. HEARD III Department of Zoology, University of Georgia, Athens, and Duke University Marine Laboratory, Beaufort, North Carolina

During the past 4 years, 192 clapper rails, Except for living material, worms used in this Rallus longirostris Boddaert, from the Atlantic study were killed in hot saline or hot (60 C) and Gulf Coasts of the United States have been 50% ethanol, then immediately placed in AFA examined for metazoan parasites. Specimens fixative. Material for whole mounts was of undescribed species of Levinseniella Stiles stained in Harris or Bullard's Hematoxylin and and Hassall, 1901 were found in the ceca, and mounted in piccolyte. Living worms were occasionally the rectum, of 59 of these birds. examined in 0.85% saline under a vaseline Infected birds carried from one to 80 worms. ringed coverslip. Both light and phase micros- Clapper rails were collected with a shotgun copy were used in studying living and stained or caught alive by hand during high tides. materials. All measurements are given in microns; the size range is given first followed 1 This study was supported in part by an appropriation from the Congress of the United States to the Southeastern by the average in parentheses. Measurements, Cooperative Wildlife Disease Study, School of Veterinary with the exception of the eggs, are based on Medicine, University of Georgia, Athens, with funds ad- ministered and research coordinated by the Bureau of Sport ten stained and mounted specimens which were Fisheries and Wildlife, Department of the Interior, through Contract No. 14-16-0008-676. killed in hot saline. Twenty eggs dissected

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 63 from specimens preserved in 70% ethanol were The generic affinity of L. camtshatica has also measured. been questioned by Belopolskaia (1963a). Historical Levinseniella byrdi n. sp. In reviewing the family Microphallidae, (Figs. 1-7) Belopolskaia (1963a) recognized the following DESCRIPTION: Small linguiform distomes 19 species of Levinseniella: L. brachysoma 1,180-1,720 (1,430) long and 323-494 (401) (Creplin, 1837) Stiles and Hassall, 1901; L. wide at broadest point (usually at level of propinqua Jiigerskiold, 1907; L. pellucida testes). Cuticular spines largest in region of Ja'gerskiold, 1907; L. howensis Johnston, 1917; oral sucker, extending posteriorly to level of L. cruzi Travassos, 1921; L. minuta Price, acetabulum. Oral sucker large, 152-243 (195) 1932; L. bucephalae (Yamaguti, 1935) Yama- wide and 124-209 (161) long, with a pair of guti, 1939; L. indica Lai, 1936; L. carcinidis lateral, glandular papillae. When oral sucker Rankin, 1939; L. charadriformis Young, 1949; is fully opened, these papillae are present on L. amnicolae Etges, 1953; L. gymnopocha prominent lateral protrusions. Well developed Coil, 1955; L. leptophalhis Coil, 1955; L. post-oral, muscular ring present. Acetabulum microovata Belopolskaia, 1958; L. fissicotyle with four papillae on inner margin, small, 95- Belopolskaia, 1958; L. somateriae Kulatach- 124 (105) in diameter; recessed in a shallow kova, 1958; L. tridigitata Deblock, Capron, cuticular pocket with contractile, circular and Biguet, 1958; L. camtshatica Morosov, margin. Prepharynx, 38-97 (76) long. Phar- 1960; and L. belopohkoi Chaun, 1962. Levin- ynx well developed, 62-70 (67) long and seniella polydactyla Deblock and Rose, 1962 48-67 (60) wide. Esophagus 164-296 (220) was not mentioned in this review. Since this long. Ceca lined with prominent epithelium, work, two additional species, L. ryjikovi Belo- relatively long, 456-696 (568), and extending polskaia, 1963b and L. carteretensis Coil and to lateral margins of testes. Testes equal, sym- Heard, 1966, have been described; and nine metrical, laterally elongate, 115-212 (155) by species have been synonymized or removed 67—125 (94). Seminal vesicle kidney shaped, from the genus. Deblock and Rose (1962) 115-235 (163) by 48-155 (72), lying just noted that L. somateriae more closely resem- anterior to acetabulum and slightly dextral to bled a member of the genus Microphallus longitudinal axis of body. Vas deferens greatly Ward, 1901, Microphallus claviformis (Brandes, reduced. Pars prostatica well developed, 72- 1888). Later (1964), they recognized L. 122 (93) by 22-28 (24), with numerous gland somateriae as a distinct species of Microphal- cells. Thin, nonmuscular membrane surround- lus, M. somateriae (Kulatachkova, 1958) n. ing retort-shaped seminal vesicle and pars comb. The same authors (1962) synonymized prostatica complex. Genital atrium sinistral, L. belopohkoi with L. tridigitata. Deblock at level of ventral sucker. Three dorso-laterally and Tran Van Ky (1966), after examining ad- directed male pockets located in lateral, glan- ditional material, synonymized L. tridigitata dular wall of genital atrium. Anterior pocket with L. brachysoma and L. belopohkoi with distinctly separate from other two; middle L. pellucida. In the same work, they re- pocket largest. Ductus ejaculatoris strongly described L. pellucida (three male pockets developed, discharging through a muscular, instead of four and no female pouch) and lobate, protrusible, male papilla 38-47 (41) L. propinqua, and synonymized the three long by 34-40 (36) wide, entering genital American species, L. carcinidis, L. leptophal- atrium anteriorly, mediad to male pockets. lus and L. gymnopocha, with L. propinqua. Each male pocket usually with a "hook" or Levinseniella minuta and L. charadriformis toothlike structure at its distal end. Genital have been removed from the genus to become pore a sinistral, slack, semicircular slit, lying Atriophallophorus minuta (Price, 1932), Deb- immediately lateral to acetabulum. Female lock and Rose, 1964 and Ascorhytis charadri- pouch absent. Metraterm thin-walled; extend- formis (Young, 1949), Ching, 1965, respec- ing from lateral wall of genital atrium to inter- tively. The incomplete and vague descriptions testicular region; entering atrium dorsally at of L. cruzi and L. howensis led Deblock and or slightly anterior to portion of atrial wall Rose (1962) to consider them incertae sedis. lying between anterior and two posterior male

Figures 1—6. Levinseniella byrdi n. sp. (Drawn with microprojector.) 1. Anterior end of specimen showing head papillae on lateral protrusions. 2. Ventral aspect of holotype with excretory system added freehand from living material; note head papillae and postoral muscular ring. 3. Terminal genitalia showing location, nature, and relative size of sclerotized male pockets, metraterm, and male papilla. 4. Section through pars prostatic complex (left), and seminal vesicle (right) showing the surrounding membrane. 5. Section through male papilla and anterior most male pocket. 6. Section through genital atrium showing opening of metraterm and associated cleft in lateral wall.

four male pockets (No. 61792) deposited in USNM Helm. Coll., Beltsville, Maryland. This species is named in honor of Dr. Elon E. Byrd. Comparisons and Discussion Levinseniella byrdi is most similar to L. pellucida, the only other described species of the genus with three male pockets and no female pouch. However, L. byrdi can be separated from L. pellucida and all the other species of Levinseniella by the spatial relationship of its male pockets (anterior pocket distinctly separated from the two posterior pockets ), its larger size, and the presence of a prominent postoral muscular ring and a pair of large glandular head papillae. The number and nature of the male pockets Figure 7. Photograph of genital atrium of L. have long been considered one of the major byrdi n. sp. showing the sclerotized "hook" struc- quantitative features in determining the spe- tures in each male pocket. cific identity of members of the genus Levin- seniella. Recently, however, Deblock and Tran Van Ky (1966) and Coil and Heard pockets. End of metraterm slightly dilated (1966) have reported that the number of with irregular, glandular lining. Weakly de- male pockets can vary in L. propinqua and veloped cleft in atrial wall just above opening L. carter etensis, respectively. However, in L. of metraterm. Small sphincter present at junc- byrdi, with but a single exception, the number tion of uterus and metraterm. Ovary 96-169 of male pockets was constant in the more than (137) by 72-96 (83), dextral to acetabulum 200 specimens thus far examined. A single and anterior to right testes. Ootype and asso- "abnormal" specimen possessed four male pock- ciated Mehlis' gland located between testes. ets (two smaller pockets replaced the posterior Laurer's canal and fertilization chamber pres- most one). ent. Vitellaria posterior to testes, acinose, seven Ching (1965) suggested that the presence to nine relatively large follicles (usually di- or absence of "hooks" in the male pockets may vided into two groups of from three to five be due to the kind of fixation used. This, how- follicles) on each side. Color of gravicl living ever, does not appear to hold for L. cartereten- worms, light yellow. Uterus posttesticular. sis and L. byrdi, since variations in the number Eggs brown, operculate, 19.2-21.6 (19.6) by and degree of development of "hooks" were 10.8—11.8 (11). Excretory pore subterminal. observed in living specimens. Whether these Bladder "V" to "U" shaped. Flame cell for- variations are intraspecific, or are due to the mula typically microphallicl, 2[(2+2) + (2+2)] degree of development, growth, age, or a = 16. combination of these factors has not been HOST: Rallus longirostris Bodclaert. determined. Information obtained from life LOCALITIES: Chatham County, Georgia history studies, currently being conducted on (type locality); Mobile County, Alabama; L. byrdi, may provide an answer to these Pinellas County, Florida; Indian River County, questions. Florida; Charleston County, South Carolina; The origin and function of the membranous, Carterette County, North Carolina; Accomac nonmuscular sac surrounding the seminal vesi- County, Virginia; Worcester County, Mary- cle and pars prostatica complex of L. bijrdi is land; Cape May County, New Jersey. uncertain. It may represent remnants of a SITE OF INFECTION: Ceca and rectum. vestigial cirrus pouch or it may be a second- HOLOTYPE (No. 61237), paratypes (Nos. arily derived structure. 61238, 61239) and "abnormal" specimen with An examination of the types or paratypes of

L. carcinidis, L. leptophallus, and L. gymno- pair of head papillae and a postoral muscular pocha., and specimens of L. propinqua from the ring may also be important characteristics in collections of Drs. Ching and Deblock, reveal any future generic revisions (Yamaguti 1967, close similarities among the four species. How- personal communication). ever, there are morphological differences in the At the present time, it is possible to recog- nature of the terminal genitalia in the speci- nize the following 18 species as belonging to mens examined which warrant further investi- the genus Levinseniella: L. brachysoma, L. gation. Therefore, until more American speci- propinqua, L. pellucida, L. howensis, L. cruzi, mens of the L. propinqua complex are L. bucephalae, L. indica, L. carcinidis, L. examined, I feel that these three forms should amnicolae, L. leptophallus, L. gymnopocha, be recognized. This view also is held by Dr. L. microovata, L. fissicotyle, L. camtshatica, Ching (1966, personal communication). With L. polydactyla, L. ryjikovi, L. carteretensis, reservations, L. howensis, L. cruzi, and L. and L. byrdi n. sp. camtshatica are retained in the genus pend- ing further study of original or additional Acknowledgments specimens. I In light of the recent redescriptions of L. The author wishes to express sincere appre- pellucida and L. propinqua by Deblock and ciation to Drs. Hilda Lei Ching, W. H. Coil, Tran Van Ky (1966), and the descriptions of S. Deblock, and Satyu Yamaguti for pertinent L. polydactyla and L. carteretensis, the only suggestions and observations. I would like to major characteristic consistently shared by all thank Mr. W. W. Becklund, Animal Disease the described species of Levinseniella is the and Parasite Research Center, Beltsville, Mary- presence of male pockets. As it now stands, land, for prompt loan of museum specimens the genus can be separated into four distinct vised in this study. Special thanks go to the morphological groups using the following com- Game and Fish Agencies of Alabama, Florida, bination of characteristics: (I) male pockets Georgia, Louisiana, Maryland, Mississippi, New few (i.e., four or less) and female pouch pres- Jersey, North Carolina, and Virginia for aid in ent (L. brachysoma, L. propinqua, L. carcin- procuring birds. Appreciation is extended to idis, L. gymnopocha, L. leptophallus, L. micro- the U.S. Fish and Wildlife Service and the ovata, and L. ryjikovi); (II) male pockets few Florida State Board of Health for use of lab- and female pouch absent (L. pellucida, L. oratory facilities. amnicolae, and L. byrdi n. sp.); (Ill) male Literature Cited pockets numerous (i.e., seven or more) and female pouch present (L. carteretensis); and Belopolskaia, M. M. 1963a. Family Microphal- (IV) male pockets numerous and female pouch lidae Travassos, 1920. In Skrjabin, K. L. absent or very greatly reduced (L. polydactyla). Trematodes of Animals and Man. Vol. 22: In the written descriptions of L. microovata 260-499. and L. ryjikovi, the presence of a female pouch . 1963b. Helminth fauna of shorebirds of the region of the Lower Amur during the is reported, but the number of male pockets in period of migration and nesting. Trudy both species is not given. The text figures for Gel'mint. Lab. Akad. Nauk SSSR. 13: 164- both species show three male pockets, there- 195. fore, they are tentatively included in group I. Ching, H. L. 1965. Systematic notes on some The descriptions for the remaining species, L. North American microphallid trematocles. howensis, L. cruzi, L. indica, L. bucephalae, Proc. Helm. Soc. Wash. 32: 140-148. L. fissicotyle, and L. camtshatica are too vague Coil, W. H., and R. W. Heard, III. 1966. Le- or incomplete to assign them to any of the vinseniella carteretensis sp. nov., a micro- above four groups. phallid trematode from the Wilson Plover, When additional species of Levinseniella are Charadrius ivilsonia. Proc. Helm. Soc. Wash. described and the incompletely described spe- 33: 54-56. Deblock, and F. Rose. 1962. Contribution a cies reexamined, the genus may warrant divi- la connaissance des Microphallidae Travassos sion into several genera based on the presence 1920 (Trematoda) des Oiseaux de France. or absence of a female pouch and the number VII. Description de Levinseniella polydactyla of male pockets. The presence or absence of a nov. sp. Vie et Millieu. Paris. 13: 773-783.

and 1964. Contribution a 1'etude parasites d'Anatides. Bull. Soc. Zool. France. des Microphallidae Travassos, 1920 (Trema- 89: 429-443. toda) des Oiseaux de France. XI. Descrip- , and Tran Van Ky. 1966. Contribution tion d'Anacetablitrerna surname n. gen., n. sp., a 1'etude des Microphallidae Travassos, 1920 de Maritrema macracetabulum n. sp. et de (Trematoda). Ann. Parasitol. humaine et Microphallus somateriae (Kulatsch.) n. comb., comparee. 41: 23-60.

Studies on Freshwater Larval Trematodes. Part XVI. Five New Species of Cercariae from Venezuela

PIR NASIR AND MARCOS TULIO DIAZ Laboratorio de Parasitologia, Depto. de Biologia, Escuela de Ciencias, Universidad de Oriente, Cumana, Venezuela

Uribe (1925) described two new species edge of the coverslip until a suitable pressure of nonvirgulate xiphidiocercariae, Cercaria rep- has been reached. tans and C. fausti, from Venezuela. Nasir All measurements are in millimeters. (1964; 1965) added two more species, C. baldai and C. cumanensis; Nasir and Acufia A. Xiphidiocercariae (1966; 1966a) described four additional spe- Cercaria marisa n. sp. cies, C. peculiaristylata, C. chacaracualensis, (Fig. 1, la) C. urceiis, and C. homocotylea. With the description of three new species, C. marisa, C. DESCRIPTION: Body spinose, without "fla- giiaraunemis, and C. allomacarapanensis, in gellets." Tail aspinose, without a fin fold, this paper, the number of nonvirgulate xiphid- subterminally attached; no caudal pockets, no iocercariae has been raised to eleven. The caudal depression. Stylet without a basal bulb gymnocephalic cercaria, C. pomacea, is the (Fig. la). Oral sucker considerably larger than third species of the gymnocephalous group of ventral sucker, both suckers without special larval trematodes described from the fresh- spines or papillae. Prepharynx and pharynx water snail, Pomacea glaiica, in Venezuela. present. Esophageal bifurcation extending up The other two species are: Cercaria sanlorezen- to anterior border of ventral sucker. Ceca not sis Nasir and Acuna, 1964, from an irrigation extending beyond anterolateral border of ven- canal, San Lorenzo, about 75 km east of tral sucker. Three pairs of penetration glands Cumana and C. macarapanensis Nasir and on each side of ventral sucker, extending Acuna, 1966, from an irrigation canal, San mostly posterior to it; anterior two pairs with Juan de Macarapana, about 15 km east of coarsely granular contents while posterior pair Universidad de Oriente, Cumana. Cercaria hyaline in nature; three penetration ducts on heteroglandula n. sp. is the first apharyngeate, each side of body. Excretory vesicle with two ocellate, distomate, brevifurcate furcocercaria lateral arms and a basal stem; arms of vesicle encountered in Venezuela. not extending to hyaline pair of penetration All observations were made on freshly glands; main excretory tubes arising terminally emerged cercariae. For the purpose of mea- from corresponding arms of vesicle and divid- surements, cercariae were mounted in 0.75% ing, at pharyngeal level, into anterior and saline; these mounts were irrigated by hot posterior lateral collecting excretory tubules. formalin (10%) applied on one side of the Flame cell formula, 2 [(3) + (3+3+3)] = 24. coverslip with the concomitant withdrawal of Measurements of 36 cercariae: body 0.162— liquid, by pieces of blotting paper, on other 0.197 by 0.095-0.129; tail 0.282-0.303 by

Figures 1-3. 1. Cercaria marina n. sp., ventral view. la. Stylet of C. marisa. 2. Cercaria guaraunensis n. sp., ventral view. 2a. Stylet C. guaraunensis. 3. Cercaria allomacarapanensis n. sp., ventral view. 3a. Stylet of C. allomacarapanensis, note the basal bulb.

0.027-0.037; prepharynx 0.002-0.004 long; Cercaria allomacarapanensis n. sp. pharynx 0.010-0.019 in diameter; esophagus (Fig. 3, 3a) 0.007-0.009 long; oral sucker 0.062-0.067 in diameter; ventral sucker 0.025-0.037 in diam- DESCRIPTION: Body spinose with six rows eter; stylet 0.045-0.057 long; width of stylet of "flagellets." Tail aspinose, without a fin at middle region 0.005-0.007. Development in fold, subterminally attached; no caudal depres- anteroposteriorly elongate, unbranched sporo- sion and no caudal pockets; oral sucker larger cysts of variable shape, measuring 0.137-0.260 than ventral sucker; both suckers without spe- by 0.112-0.180. cial spines or papillae. Stylet with a basal bulb HOST: Marisa cormiarietis (L.). (Fig. 3a). Prepharynx absent. Pharynx smaller LOCALITY: Poza Azul, about 90 km east of than ventral sucker. Esophagus poorly de- Cumana. veloped, represented by a longitudinal threadlike extension. Intestinal ceca well developed, Cercaria guaraunensis n. sp. not extending beyond equatorial level of ven- (Fig. 2, 2a) tral sucker. Three penetration glands on each side of body; anterior two pairs with finely DESCRIPTION: Body aspinose, possessing granular contents and situated about halfway eight rows of papillae with setae. Tail aspinose, along preacetabular extent, with a single duct without fin fold, subterminally attached; cau- opening in oral orifice on each side of body; dal depression and caudal pockets absent. posterior pair with coarsely granular contents Ventral sucker protrusible, smaller than oral and situated mostly posterior to ventral sucker. sucker; both suckers without special spines or Excretory vesicle V-shaped; main excretory papillae. Stylet without a basal bulb (Fig. 2a). tubes arising terminally and just anterior to Prepharynx absent. Pharynx anteroposteriorly anterior border of ventral sucker dividing into elongate. Esophagus relatively long, not ex- anterior and posterior lateral collecting excre- tending to ventral sucker. Intestinal ceca tory tubules. Flame cell formula 2 [ (2) + (2) ] considerably more dilated than esophagus, not — 8. Measurements of 36 cercariae: body extending past equatorial level of ventral 0.072-0.100 by 0.040-0.060; tail 0.077-0.100 sucker. Three pairs of penetration glands on by 0.012-0.019; oral sucker 0.014-0.030 in each side of ventral sucker, mostly anterior diameter; ventral sucker 0.012-0.017 in diam- to it; anteriormost pair with coarsely granular eter; pharynx 0.006-0.012 in diameter; stylet contents while posterior two pairs with finely including basal bulb 0.016-0.020 long; width granular contents; two penetration ducts on of stylet at shoulder 0.002-0.004; width at each side of body. Excretory vesicle U-shaped; shaft 0.001-0.003. Development in antero- main excretory tubes arising subterminally from posteriorly elongate, unbranched sporocysts corresponding arms of excretory vesicle and with considerable variations in contours, mea- dividing into anterior and posterior lateral col- suring 0.176-0.492 by 0.110-0.165. lecting excretory tubules about halfway along HOST: Marisa cormiarietis (L.) and Poma- esophagus. Flame cell formula 2 [(3+3) + cca glauca (L.). (3+3+3+3+3)]=42. Measurements of 36 LOCALITY: A pond near San Juan de Ma- cercariae: body 0.147-0.153 by 0.054-0.084; carapana, about 15 km south of Cumana. tail 0.273-0.340 by 0.027-0.034; oral sucker 0.025-0.039 in diameter; ventral sucker 0.018- B. Gymnocephalous Cercariae 0.025 in diameter; pharynx 0.015-0.025 by Cercaria pomacea n. sp. 0.009-0.013; esophagus 0.016-0.018 long; (Fig. 4, 4a) stylet 0.027-0.031 long; width of stylet at DESCRIPTION: Body and tail without spines, shoulder 0.005-0.006. Development in antero- papillae or flagellets. Tail without a fin fold, posteriorly elongate, unbranched sporocysts traversed by two pairs of longitudinal muscle with variable contours, measuring 0.131—0.282 bands, one ventral and other dorsal. No pig- by 0.075-0.112. men tecl eye spots. Both suckers almost iso- HOST: Marisa cormiarietis (L.). cliametric, without papillae or spines. Pre- LOCALITY: Los Guaraunos, about 160 km pharynx and pharynx present. Esophagus and east of Cumana. intestinal ceca absent. Cystogenous glands with

Figures 4, 5. 4. Cercaria pomacea n. sp., ventral view. 4a. A group o£ cystogenous gland cells of C. pomacea; note rodlike contents of cystogenous glands. 5. Cercaria heteroglandula n. sp., ventral view.

rodlike contents. Anterior margin of body bor- ciliated patches only in preacetabular region. dered with three pairs of apertures leading into Caudal excretory duct dividing into two lateral three pairs of ductlike extensions extending as branches in posterior half of tail. Flame cell far back as posterior border of oral sucker. formula 2 [(3+3+3) + (3+3+3)] = 36. Mea- Genital rudiments represented by two masses of surements: body 0.175-0.240 by 0.105-0.165; cells, one anterior and other posterior to ventral tail 0.200-0.273 by 0.032-0.045; oral sucker sucker. Pattern of excretory system shown in 0.042-0.057 in diameter; ventral sucker 0.040- Figure 4. Main excretory tubes, in preacetab- 0.052 in diameter; prepharynx 0.005-0.012 ular region, enclosing a varying number of long; pharynx 0.017—0.025 in diameter. De- refractile excretory granules of a double na- velopment in rediae having a muscular phar- ture. Secondary excretory tubules lined with ynx, a complete collar, a saccate gut and a pair

of posterior locomotor appendages. Measure- Measurements: body 0.187-0.242 by 0.062- ments of living rediae, chosen at random: body 0.085; tail stem 0.177-0.225 by 0.030-0.047; 0.405-0.770 by 0.132-0.264; pharynx 0.027- furcae 0.080-0.0140; anterior organ 0.070- 0.045 in diameter. 0.102 by 0.048-0.067; ventral sucker 0.020- HOST: Pomacea glauca (L.). 0.030 in diameter. Development in long LOCALITY: Los Bordones, en route to threadlike sporocysts. Puerto la Cruz, about 5 km west of Univer- HOST: Pomacea glauca (L.). sidad de Oriente, Cumana. LOCALITY: Los Guaraunos, about 200 km north of Cumana. C. Apharyngeate, brevifurcate, ocellate, distomate furcocercariae Discussion Cercaria heteroglandula n. sp. Nonvirgulate xiphidiocercariae, like the three species included in this paper, with three pairs (Fig. 5) of penetration glands, without fin folds on their DESCRIPTION: Body, tail stem and furcae tails and without eye spots are: Cercaria uniformly spinose. No "flagellets" on body or Camilla Faust, 1921, C. indicae X Sewell, 1922, tail stem. No special forward pointing spines C. indicae XVI Sewell, 1922, C. indicae XVIII at anterior end of body. About anterior half Sewell, 1922, C. indicae XIX Sewell, 1922, C. of anterior organ beset with an oral cap of indicae LIU Sewell, 1922, C. helvetica XII simple spines. Acetabular orifice beset with Dubois, 1929, C. cordiformes Wesenberg-Lund, a single row of acetabular spines. Tail stem 1934, C. tridena E. L. Miller, 1936, C. cerithidia without caudal bodies. Furcae provided with Porter, 1938, C. elizabethae Porter, 1938, C. a fin fold. A pair of pigmented eye spots mpingoensis Porter, 1938, C. globelaria Porter, present. Anterior organ divided into a larger 1938, C. hartebeestia Porter, 1938, C. nym- anterior thin-walled region and a narrow pos- phaea Porter, 1938, C. patentissima Porter, terior muscular region. Ventral sucker pro- 1938, C. spruitensis Porter, 1938, C. stonei Por- trusible. Esophagus divided into two stump- ter, 1938, C. spiralina Porter, 1938, C. um- like ceca, not extending to ventral sucker. holtuzana Porter, 1938, C. uvurensis Porter, Pharynx absent. Head gland consisting of five 1938, C. veta Porter, 1938, C. zuluana Porter, nuclei and finely granular contents. Six pairs 1938, C. pili Brooks, 1943, C. aciculata Fain, of penetration glands pre-, para-, and post- 1953, C. furtiva Fain, 1953, C. kunga Fain, acetabular. Bulk of penetration glands pos- 1953, C. liliputa Fain, 1953, C. schoetteri Fain, terior to ventral sucker. Anterior four pairs 1953, and C. triglandularis Probert, 1965. Of of penetration glands larger, with coarsely these 30 species of cercariae insofar as the num- granular contents, and including larger nuclei. ber of flame cells, that is 12 on each side of Posterior two pairs smaller, with finely granular body, is concerned C. Camilla, C. isipingoensis, contents and smaller nuclei. Two penetration C. spiralina, C. aciculata, C. furtiva, C. kimga, ducts, on each side, leading from penetration and C. liliputa are indistinguishable from C. glands with coarsely granular contents while mari-sa; however, none of these cercariae is only one duct emerges from penetration glands characterized with penetration glands having with finely granular contents. Penetration contents, i.e., coarsely granular in two anterior ducts opening on anterolateral border of an- pairs and hyaline nature of posterior pair, sim- terior organ and capped with six conical ilar to that of C. marisa and, therefore, this spines. Excretory vesicle bicornuate. Main character alone alludes to the separate entity excretory tubes, at anterolateral border of ven- of C. marisa. tral sucker, dividing into anterior and posterior From the standpoint of the nature of the lateral collecting excretory tubules. Caudal contents of the penetration glands (coarsely excretory duct, at caudal end of tail stem, granular in anterior pair and finely granular dividing into two furcal excretory branches in two posterior pairs) Cercaria helvetica XII, opening at tips of corresponding furcae. No C. cordiformes, C. cerithidia, C. mjmphaea, C. transverse excretory commissures. No ciliated furtiva, and C. schoetteri are very much like patches in main excretory tubes. Flame cell C. guaraunensis; on the contrary, C. gitaraun- formula, 2 [((1 + 1 + 1) + (1 + 1+(1))] = 12. ensis is marked with a total number of 42

Copyright © 2011, The Helminthological Society of Washington 72 PROCEEDINGS OF THE HELMINTHOLOG1CAL SOCIETY flame cells in contrast with a varying number Marshall, 1931, cercaria of Fasciola hepatica of flame cells met within other species. Linnaeus (1758) as described by Rees, 1932, Cercaria indicae XVI, C. isipingoensis, C. C. tubercidata Filippi (1854) as described by veta, and C. kunga are identical with C. allo- Wesenberg-Lund, 1934, cercaria of Sphaeridio- macarapanensis in the possession of two an- trema globuhis (Rudolphi, 1819) Odhner terior pairs of penetration glands with finely (1913) as described by Szidat (1937), Pro- granular contents and the posterior pair with bert, 1966, C. albinea Khan, 1960, C. den- coarsely granular contents but the number of sacutis Khan, 1960, C. Uanogorsensis Probert, flame cells, eight in all, in C. allomacara- 1965, C. cystogenata Probert, 1965, C. san- panensis conflicts with a different number of lorenzensis Nasir and Acuria, 1964 and C. flame cells found in the otherwise similar macarapanensis Nasir and Acufia, 1966. Cer- cercariae. caria pomacea can be easily separated from The nature of the contents of the penetra- these species in the possession of 36 flame cells tion glands and the number of flame cells have in all, where the number of flame cells is been taken into consideration as important known, and by the absence of esophagus and diagnostic characters for a clear cut separation intestinal ceca excepting C. complicata and of related species. In addition, Cercaria C. indicae XLI. In addition to these char- mama, C. guaraunensis, and C. allomacara- acters, C. pomacea differs in certain other panensis differ from the aforementioned 30 aspects which have been discussed below. species in the following aspects: shape and According to Dubois, 1929, the cystogenous size of stylet, pattern of digestive system, and glands of Cercaria helvetica XIX have granular in the arrangement of penetration glands in contents but in the redescription of the same relation to ventral sucker. To these may be cercaria as put forth by Wesenberg-Lund, added the geographical and host specificities; 1934, the inclusions of the cystogenous glands C. mama, C. guaraunensis, and C. alloma- are rod-shaped. Probably, Wesenberg-Lund carapanensis are parasites of Marisa cornuarie- was dealing with a different cercaria. In any tis and Pomacea glauca in contradistinction case, the ventral sucker of C. helvetica XIX is with other species which have been described considerably larger than its oral sucker in con- from different species of snails from Europe, trast with an almost isodiametric condition India, and USA. found in C. pomacea. The body of the cer- Cercaria peculmrishjlata Nasir and Acufia, caria of Psilotrema spiculigerum is spinose in 1966, parasite of Pomacea glauca from San the preacetabular region, there are 12 spines Juan de Macarapana, near Cumana, Venezuela, bordering the anterior margin of the oral sucker has three pairs of penetration glands like C. and the same sucker is also furnished with rows mama, C. guaraunensis, and C. allomacara- of spines whereas C. pomacea has a smooth panensis but is differentiated by the shape of cuticle and the corresponding spines are lack- the stylet; 18 flame cells on each side of body; ing. Cercaria sudanensis No. 3 presents spines and the uniformly granular contents of its on its body and tail, its oral orifice is bordered penetration glands. with spines and the oral sucker is provided In the literature on freshwater larval trem- with two cephalic glands in contradistinction atodes appear the following gymnocephalic with C. pomacea where there is a complete cercariae which like Cercaria pomacea are absence of spines and the cephalic glands in nonocellate, with cystogenous glands having the oral sucker are wanting. The body of the rodlike contents and without tin folds on their tails: C. pigmentosa Cawston (1919), Faust cercaria of Fasciola hepatica is covered with (1920), Porter (1920; 1938), syn. cercaria of rows of papillae and its ventral sucker is Fasciokt giganttca Cobbolcl (1856) as de- remarkably larger than oral sucker; in C. scribed by Porter, 1938, C. indicae XLI Sewell, pomacea the papillae are absent and its two 1922, cercaria of Psilotrema spiculigerum suckers are almost isodiametric. The presence (Muhling, 1898) Odhner (1913) as described of papillae in C. tubercidata is contrary with by Mathias (1924), Llewellyn, 1957, C. hel- the smooth cuticle of C. pomacea. In compari- vetica XIX Dubois, 1929, C. complicata Faust, son with C. pomacea the body of the cercaria 1930, C. sudanensis No. 3 Archibald and of Sphaeridiotrema globuhis is spinose.

From the standpoint of the absence of the Literature Cited esophagus, intestinal ceca, spination of the Cawston, F. G. 1919. Further observations in body and tail and the almost isodiametric regard to South African cercariae. Mecl. J. condition of the suckers, Cercaria complicata South Africa, 14: 401-402. and C. indicae XLI are indistinguishable from Bearup, A. J. 1955. A schistosome larva from C. pomacea but C. complicata and C. indicae the marine snail Pyrazus australis as a cause XLI are markedly larger species and have of cercarial dermatitis in man. relatively larger suckers. . 1956. Life cycle of Auslrobilharzia ter- Cercaria sanlarenzensis Nasir and Acuna, rigalensis Johnston, 1917. Parasitology, 46: 1964, a parasite of Pomacea glauca and C. 470-479. Dubois, G. 1929. Les cercaires de la region de macarapanensis Nasir and Acuna, 1966, are Neuchatel. Bull. Soc. neuchat. Sci. Nat. 53: the other two gymnocephalic cercariae, from 1-177. the oriental part of Venezuela where the pres- Faust, E. C. 1920. A survey of Cawston's spe- ent investigation has been carried out, without cies of South African cercariae. Parasitology, eye spots, without fin folds, with cystogenous 12: 212-216. glands having rodlike contents and with six Llewellyn, C. 1957. The morphology, biology and apertures at the anterior end of the body, and, incidence of the larval digenea parasitic in therefore, are more closely related to C. certain freshwater mollusks. Ph.D. Thesis, pomacea; however, C. pomacea lacks an Univ. Wales. Mathias, P. 1924. Sur le cycle evolutif d'un esophagus and the intestinal ceca while C. trematode cle la famille des Psilostomidae sanlorenzensis and C. macarapanensis are pro- (Psilotrema spiculigerum Miihl.) CRAS 178: vided with these structures; moreover, each 1217-1219. of these two cercariae has 24 flame cells in Nasir, P. 1964. Studies on freshwater larval all in relation to C. pomacea which exhibits trematodes. Part V. A new polyadenous 36 flame cells in all. xiphidiocercaria, C. baldai, from Venezuela. Insofar as the identification of Cercaria Proc. Helm. Soc. Wash. 32: 28-30. heteroglandula is concerned, the cere-aria of . 1965. Studies on freshwater larval Aiistrobilharzia terrigalensis Johnston (1917) trematodes. Part I. A new species of a as described by Bearup (1955; 1956) is the microcotylous xiphidiocercaria, Cercaria cumanensis, from Venezuela. Proc. Helm. Soc. only other apharyngeate, brevifurcate, ocellate. Wash. 32: 103-105. distomate furcocercaria which is closely related Nasir, P., and C. A. Acuna. 1966. Studies on to C. heteroglandula in the possession of six freshwater larval trematodes. Part VIII. Two pairs of penetration glands with finely and new species of non-virgulate xiphidiocercariae coarsely granular contents, in the pattern of from Venezuela. Zool. Anz. 176: 291-296. the digestive system and in the number and . 1966a. Studies on freshwater larval arrangement of the flame cells. However, C. trematodes. Part XIII. Some new species of terrigalensis lacks a fin fold on furcae and the cercariae from Venezuela. Proc. Helm. Soc. Wash. 33: 96-100. anterior two pairs of its penetration glands Porter, A. 1920. The life history of the African have finely granular contents while the pos- sheep and cattle fluke, Fasciola gigantica. terior four pairs are coarsely granular in con- South Afr. J. Sci. 17: 126-130. trast with C. heteroglandula which is furnished . 1938. The larval trematodes found in with fin folds on its furcae and its two pos- certain South African molluska, with special terior pairs of penetration glands are finely reference to Schistosomiasis (Bilharziasis). granular in contradistinction with four anterior Publ. South Afr. Inst. Mecl. Res. 8: 1-492. pairs of coarsely granular contents. Moreover, Rees, G. 1932. An investigation into the occur- there is a multinucleate head gland in C. rence, structure and life histories of the heteroglandula whereas no such structure exists trematode parasites of four species of Lijm- naea, L. truncatula (Mull.), L. pereger in the cercaria of A. terrigalensis. These dif- (Mull.), L. palustris (Mull.), L. stagnalis ferences have been considered diagnostically (L.), and Htjdrobia jenkinsi (Smith) in important enough for the separation of C. Glamorgan and Monmouth. Proc. Zool. Soc. heteroglandula from the cercaria of A. ter- London, 1: 1-32. rigalensis. Szidat, L. 1937. tiber die entwicklungsgeschichte

von Sphaeridiotrema globulus Rucl. 1814 und Wesenberg-Lund, C. 1934. Contributions to die stelkmg der Psilostomiclae Odhner im nat- the development of the trematoda digenea. iirlichen system. I. Z. f. Parasitenk. 9: 529- Part II. The biology of the freshwater cer- 542. cariae in Danish freshwaters. K. Danske Uribc, C. 1925. Notes on Venezuelan xiphidio- Vidensk. Skri. Naturv. og math. Afd R9. 5: cercariae. J. Parasit. 11: 125-134. 1-223.

Further Studies on the Life History of Echinostoma lindoense Sandground and Bonne, 1940 (Trematoda: Echinostomatidae) with a Report of Its Occurrence in Brazil1

LIE KIAN JOE- G. W. Hooper Foundation, University of California Medical Center, San Francisco, California

This paper reports the finding of Echino- 24-27 C and fed red-leaf lettuce (Lactuca stoma lindoense in the larval stages in the sativa). Techniques for the study of the larval Brazilian snail, Biomphalaria glabrata (Say), and adult stages of the parasite were reported and supplements earlier studies on the life in detail in previous studies (Lie, 1963, 1965, history of this parasite. Previously it was 1966aandb). known to occur only in Southeast Asia. Sand- For comparison purposes, live specimens of ground and Bonne (1940) first described it the Malaysian snail Gyraulus convexhtsculus in villagers around Lake Lindoe in Central (Hutton) harboring larvae of E. lindoense were Celebes, Indonesia. Later it was found in air-shipped from Kuala Lumpur, Malaysia, to animals in Malaysia and probably also in man San Francisco. The life cycle of the Malaysian in Djakarta, Indonesia (Lie, 1964). Now it is parasite was subsequently established in B. reported in Brazilian snails and therefore seems glabrata as the first and second intermediate much more widely distributed. host and in California chicks as the final host. The description of the life cycle and measure- Materials and Methods ments presented in this paper refer to the Many B. glabrata, collected in Vila Cristina Brazilian strain of E. lindoense unless other- near Belo Horizonte, Brazil, and air-shipped to wise stated. All measurements are in microns. San Francisco for experimental use relating to other studies, contained echinostome meta- Results cercariae in the pericardial sac. In chicks and Larval stages in the snail host white mice these metacercariae developed into Eggs appeared in stools of chicks 10 days or adult worms identified as Echinostoma lin- more after infection (16 days or more in mice), doense. Eggs from the stools were used to in uncleaved condition, 104-116 by 64-74, infect laboratory-raised albino B. glabrata of yellow-brown, with thickening at nonoper- a strain obtained from the National Institutes culated end of shell. Eggs kept in distilled of Health, Bethesda, Maryland. Infected snails water in a petri dish at 28 C began to hatch were kept in clear plastic 2-gallon aquaria at in 10 days. Miracidia entered the snail host through the exposed parts while shedding the 1 Supported by the University of California International Center for Medical Research and Training (TJC ICMRT, epidermal plates. Complete penetration took Hooper Foundation, San Francisco School of Medicine) with Research Grant TW 00144 from the Office of Inter- about half an hour. Miracida fixed in hot 2% national Research, by Research Grant AI 07054, NIH, U. S. Public Health Service, and by the Rockefeller Foun- silver nitrate measured 67.9-90 by 33.1-47.4. dation Grant GA-MNS-6654. The structure of the apical papilla, eye spots, -Present address: Institute for Medical Research, Kuala Lumpur, Malaysia. and lateral processes and the number and

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 75 arrangement of epidermal plates were the same ture, and old and shriveled rediae. All rediae as described by Lie (1964). developing in the ventricular cavity produced Sporocysts developed in the ventricular cav- rediae only, not cercariae, even when rediae in ity of the heart, 340-396 by 170-188, some- the viscera had been producing cercariae for a what larger than those of E. lindoense in long time. Malaysia desribed by Lie (1964). They First cercariae released from the snail 21 started producing recliae 6 days after infection days after exposure or later. Measurements of or later. 25 cercariae fixed in hot water: body 332—474 Rediae released from sporocysts migrated by 130-185; tail 427-490 by 40-46; collar mainly to the ovotestis; some were found on 76-104 wide; oral sucker 44-52 wide; pre- the liver, in the albumen-gland region or in pharynx 10-32; pharynx 24-32 long by 20-24 the hemolymph spaces surrounding the lung wide; esophagus solid, consisting of eight cells, cavity; several remained in the ventricular 112-132; ceca solid, extending to level of cavity. Second-generation rediae were released excretory bladder. Thirty-seven collar spines 13 days after exposure or later. arranged as in adult. Patterns of integumen- Newly released rediae colorless, with con- tary papillae, cystogenous cell contents, and spicuous locomotor organs and continuous col- excretory system, including number and dis- lar, 339 long and 51 wide; pharynx 29 wide; tribution of flame cells, similar to those in cer- gut 105; collar 63 from anterior end; locomotor cariae of Malaysian E. lindoense. Tail, inade- organs 99 from posterior end. quately described in previous publications, Mature rediae containing rediae obtained with two dorsal, two ventral and two latero- 14—18 days after infection, orange with incon- ventral fin folds (Figs. 1 and 2). Paraesoph- spicuous collar, 990-2,000 long and 150-300 ageal gland cells present, their distribution wide; pharynx 42.5-60 wide; gut dark brown, similar to those in cercariae of Malaysian E. 250-660, 25-50% of body length; collar 68- lindoense, but often fewer visible. Body scales 133 from anterior end; locomotor organs 380- on dorsal surface almost reaching the posterior 650 from posterior end. Birth pore elevated, end while in Malaysian E. lindoense reaching dorsal, immediately posterior to collar. to acetabulum. Mature rediae producing cercariae; incon- Metacercarial cysts spherical, 155-175 in spicuous collar, 700-1,550 long and 130-275 diameter, with transparent outer cyst wall and wide; pharynx 37.5-52.5 wide; gut 175—360, opaque inner wall respectively 12 and 2 thick. 20-33% of body length; collar 55-182 from Encystment in pericardial sac and posterior anterior end; locomotor organs 220-560 from part of kidney in B, glabmta. Encystment also posterior end. Birth pore elevated, dorsal, im- possible in other freshwater snails. In snails meditely posterior to collar. harboring rediae, cysts also found in the viscera First-generation rediae produce rediae in and other tissues of the snail, and occasionally the early stages of infection. They also start also in their own rediae. Entry of cercariae producing cercarial embryos 18-24 days after into the pericardial sac through the urinary exposure. As cercarial embryos increase in orifice, kidney, and ciliated renopericardial number, redial production declines. By the duct. time mature cercariae are formed, most rediae Adult contain cercariae only. Second-generation rediae produce cercariae and an occasional third- Adult worms were found in chicks in the generation redia. Young, immature rediae were second half of the small intestine, ceca and found 70 days postexposure, a sufficiently long sometimes in the rectum, and in white mice period to make one suspect production of in the first half of the small intestine. Worms fourth-generation rediae. Old, shriveled rediae started producing eggs in chicks 10-14 days were found 26 days or more after exposure, after infection and in mice several days later. suggesting that rediae are short lived but new Measurements are based on 30 worms varying ones are constantly being formed. Rediae often in age from 22—55 days, all obtained from developed in the ventricular cavity of the heart. chicks. The redial population in the heart consists of Body elongate 12,420-17,200 by 1,400- various developmental stages: immature, ma- 2,230 with maximum width at midbody. Col-

sucker subterminal, 295-392 long and 280- 363 wide; prepharynx 30-68 long; pharynx 250-316 long and 200-268 wide; esophagus 780-1,185, bifurcating anterior to acetabulum; ceca extending almost to posterior end of body. Genital pore between acetabulum and bifurcation of esophagus, followed by shallow genital atrium. Testes tandem, with shallow indented margins; anterior testis 730-1,058 by 490-730; posterior testis 750-1,058 by 474-680. Cirrus sac 510-742 by 310-379, not extending beyond middle of acetabulum, containing seminal vesicle, pars prostatica, and unspined cirrus. Ovary more or less spherical, 316-505 in diameter. Mehlis' gland complex and distribution of vitelline gland cells the same as in E. lindoense (see Lie, 1964 and 1965). Malaysian E. lindoense Malaysian E. lindoense developed well in California chicks, attaining the same size as the Brazilian parasite, but often showing rather deeply lobed testes. Miracidia of the Malaysian strain readily penetrated B. glabrata snails and developed in the same way as the Brazilian strain, producing cercariae 21 days after exposure or later. Discussion The Brazilian larval and adult parasites closely resemble E. lindoense Sandground and Bonne from Southeast Asia. The only difference is the shape of the testes in the adult worm, i.e., they are superficially lobed in the Brazilian strain but rather deeply indented in the Asian specimens (see Sandground and Figures 1 and 2. Echinostonia lindoense (Camera Bonne, 1940; Lie, 1964). This difference is not lucida drawing, projected scales 50 /M). 1. Cercaria, absolute, however, as some Asian E. lindoense lateral view, showing tail fin folds. 2. Distal part have shallow-lobed testes and some Brazilian of cercarial tail, ventral view, showing ventrolateral specimens rather deep-lobed testes. fin folds. Sandground and Bonne (1940) were unable to infect local chicks with E. lindoense from the Celebes. I was also unable to infect lar well developed, 720-948 wide, with 37 Malaysian chicks (Lie, 1964) with the local collar spines, 65—92 long. Arrangement of E. lindoense, although the worm was once collar spines: five corner spines on each side, found in a chicken bought in the market in three oral and two aboral; six lateral spines Kuala Lumpur. California chicks, on the other on each side, in a single row; 15 dorsal spines, hand, are good hosts for both the Malaysian eight oral and seven aboral. Body scales and Brazilian parasites, which grow larger in distal to collar, covering anterior two-thirds them than in white mice. The average size of ventrally, anterior half laterally and anterior 14-day-old worms from mice is 4,600 by 771, sixth dorsally. Acetabulum 720-1,027 in diam- from California chicks 8,550 by 1,140. At- eter, situated in anterior fifth of body. Oral tempts to infect each of three Sprague-Dawley

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 77 rats with 100 metacercariae of the Brazilian generations. Certain aspects of its life cycle strain failed. not previously covered are described, and the Echinostoma lindoense also develops in the life cycle of the Brazilian strain is compared snail Biomphalaria straminea (Dunker), but with that of the Malaysian E. lindoense. Both the experimental infection rate is low. It varies strains developed well experimentally in B. from 0.5-3% upon exposure of each snail to glabrata and in California chicks. 20 miracidia, compared to 30-50% in B. glabrata. The first intermediate host of the para- Literature Cited site in Central Celebes is Gyraulus sarasinorum Lie, K. J. 1963. The life history of Echinostoma Boll, and in West Malaysia Gyraulus convex- malayanum Leiper, 1911. Trop. Geogr. Med. iusculus. In 1964 I reported that E. lindoense 15: 17-24. produces two redial generations in the snail, . 1964. Studies on Echinostomatidae but I now find at least three. Production of (Trematoda) in Malaya. VII. The life his- at least three redial generations has also been tory of Echinostoma lindoense Sanclground observed in other species of echinostomes, i.e., and Bonne, 1940. Trop. Geogr. Med. 16: 72-81. Echinostoma nudicaudatum Nasir, 1960; E. . 1965. Studies on Echinostomatidae audyi Lie and Umathevy, 1965; E. hystricosum (Trematoda) in Malaya. IX. The Mehlis' Lie and Umathevy, 1966; E. barbosai Lie and gland complex in echinostomes. I. Parasit. Basch, 1966; Echinoparyphium dunni Lie and 51: 789-792. Umathevy, 1965; Hypoderaeum conoideiim . 1966a. Studies on Echinostomatidae (Bloch, 1782); H. dingeri Lie, 1964; Isthmio- (Trematoda) in Malaya. XIII. Integumentary phora spiculator (Dujardin, 1845); and Pary- papillae in six species of echinostome cer- phostomum segregatum Dietz, 1909. cariae. J. Parasit. 52: 1041-1048. . 1966b. Studies on Echinostomatidae Summary (Trematoda) in Malaya. XIV. Body gland Echinostoma lindoense Sanclgrouncl and cells in cercariae of Echinostoma audyi Lie Bonne, 1940, a Southeast Asian parasite of and Umathevy and E. lindoense Sanclground and Bonne. J. Parasit. 52: 1049-1051. man and animals, is also found in Brazil. Its Sandground, J. H., and C. Bonne. 1940. Echi- first intermediate host is the snail Biomphalaria nostoma lindoensis n. sp., a new parasite of glabrata (Say). The worm also develops in man in the Celebes, with an account of its B. straminea (Dunker) under experimental life history and epidemiology. Am. J. Trop. conditions. There are at least three redial Med. 20: 511-535.

Revision of the Genus Anaplectus (Nematoda: Plectidae)

M. W. ALLEN AND E. M. NOFFSINGER Department of Nematology, University of California, Davis

Introduction cies, P. cirratus Bastian, 1865; P. longicaudatiis The genus Anaplectus was proposed by Biitschli, 1873; P. bland Hofmanner and Men- de Coninck and Schuurmans Stekhoven (1933) zel, 1914; P. schneideri de Man, 1880; and "for all those species, formerly reckoned to P. tubifer Cobb, 1914. Males of the first two Plectus Bastian, which possess a crown of four species do not possess preanal tubuli and re- cephalic setae and a set of preanal tubuli in the mained in Plectus. The last three species have male sex." Plectus granulosus Bastian, 1865 preanal tubuli and were placed in Anaplectus. was made the genotype. Prior to this The validity of the genus Anaplectus has time males were known for five other spe- been questioned by several authors. Schneider

(1939), Goodey (1951) and Goodey (1963) setae are relatively short and are sublateral in placed the genus as a synonym of Plectus position. Bastian, 1865. Chitwood and Chitwood (1937), The external openings of the amphids of Maggenti (1961), Brzeski (1963), and Killick Anaplectus are transverse and slitlike as indi- (1964) accepted Anaplectus. These differ- cated by Cobb (1914) in his description of ences of opinion resulted from the fact that Plectus tiibifer = A. tiibifer and by Killick the morphological characters upon which the (1964) for A. arenicola. When Bastian de- genus was based were not sound. Cuticularized scribed P. granulosus in 1865, no mention was preanal tubuli and four cephalic setae are pres- made of the lateral organs (amphids). How- ent in both Plectus and Anaplectus species. The ever, Biitschli in 1873 emphasized the fact that most recent generic diagnosis by Brzeski (1963) he was able to see lateral organs in all Plectus indicates that Anaplectus differs from Plectus species excepting P. granulosus. In 1876 and by the "widening prostom hexagonal in cross- 1884 de Man illustrated and described P. section." In this study of six species, additional granulosus as having circular amphids. This morphological characters of the genus have erroneous concept of the amphids of P. gran- been observed. These include the shape of ulosus has been perpetuated in the literature the amphid aperture, lip configuration, pres- to the present time excepting for the observa- ence of a collecting duct in the excretory sys- tions of Cobb and Killick. All of the standard tem, and the outstretched anterior testis. These reference works including de Coninck and characters are discussed in detail in the section Schuurmans Stekhoven (1933), Chitwood and on morphology and the redescription of the Chitwood (1937), Schneider (1939) and genotype. Goodey (1951, 1963) describe and illustrate A. granulosus as having circular amphids. In Morphology Goodey (1951, 1963) the nematode is de- Species of Anaplectus are typically cylin- scribed and figured as a typical representative drical and vary in length from 0.6 to 2.0 mm. of the genus Plectus and is figured as having Males and females are similar in appearance, circular amphids. Hirschmann in 1952 de- but males tend to taper less at the posterior scribed and figured P. submersus — A. sub- end. The cuticle is transversely striated from mersus as having circular amphids and more the lip region to near the terminus. Laterally recently Brzeski (1963) in his emended diag- there are two alae which begin near the nerve nosis of the genus Anaplectus stated, "the ring and extend nearly to the terminus. A amphids are circular or oval." He also de- cervical papilla is located between the alae scribes in the same paper A. magnus as having about one body width posterior to the nerve broadly oval amphids. These authors did not ring. Somatic setae occur only on the tails of see the amphids but described and illustrated females, the number varying from none to three Anaplectus to conform with the concepts prev- pairs. These setae are small and often obscure. alent in the literature. The lips are obscure in profile view; they All Anaplectus species have a large number may be set off by a constriction or continuous of conspicuous sublateral hypoclermal glands with the body contour. Most authors have which open to the surface of the cuticle in two indicated there are six lips; however, in face rows of pores, one on either side of the lateral views, the lip region is seen to consist of 12 sectors (Fig. 3b). There are six large sectors alae. The first of these glands is unpaired and which, conform to the lateral, subdorsal and the pore is located posterior to the amphid subventral lips, each bearing a single papilla. aperture. This pore might possibly have been The smaller sectors do not bear papillae. Sim- mistaken for the amphidial aperture by some ilar modification of the lip region has not been authors. In both sexes, there is always a papilla reported previously for Plectidae or in the associated with the first or second lateral hypo- Araeolaimida. The genus Plectus with which dermal pore. In most species it is dorsal to Anaplectus has been synonymized does not and near the first pore, but in one species this have this type of lip region. papilla is nearest the second lateral pore and it The four cephalic setae are situated two to may be dorsal, ventral, or anterior to the pore. five body annules posterior to the lips. These The laterodorsal series of glands extends to the

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 79 terminus, while the lateroventral series termi- closely appressed to the lateral chord. It ex- nates near the anal opening. tends to a point about 60 per cent of the body One species of Anaplectus is characterized length from the lip region. In one female, A. by having a dorsal and a ventral series of granulosus, 1.4 mm in length, the gland exten- hypoclermal glands with ducts opening to the sion terminated 0.85 mm from the head or 0.65 surface of the cuticle. These begin near the mm from the end of the esophagus. There are stoma and continue posteriorly to about the two subventral glands which appear to be con- level of the excretory pore. The dorsal and nected to the excretory system. These glands ventral series of pores appear to be similar are located just posterior to and on either side to the sublateral series and the glands appear of the ventral gland. Thus the excretory sys- to be located in the dorsal and ventral chords. tem in Anaplectus consists of the long cutic- The stoma of Anaplectus has been described ularized terminal excretory duct, a ventral cell by several authors. The cheilostom is globular (sinus cell) which is modified on one side to in shape and its walls, the cheilorhabdions, are form a lateral collecting canal presumably lo- strongly cuticularized and hexaradiate in cross cated in or near to the lateral chord, and two section. This is followed by the pro-meso- subventral glands. This system has many sim- metastom which is cylindrical, with less heav- ilarities to that in some Secernentea and sup- ily cuticularized walls which are roughly tri- ports previous suggestions (Chitwood and angular in cross section. Chitwood, 1938) that the lateral collecting The esophagus is nearly cylindrical, with its tubules in these systems could have arisen as diameter only slightly reduced posterior to the outgrowths of the ventral sinus cell (Fig. 2d—f). nerve ring. It is terminated by a large bulb The collecting tubule is not readily seen in with a strongly cuticularized denticulate valve. unstained specimens. If living specimens are The valve is similar to that illustrated by Mag- placed in very dilute aqueous solutions of genti (1961) for Plectus. The esophageal neutral red, the entire excretory system be- lumen has tuboid radii for a distance of about comes lightly stained and the posterior exten- one and one-half body diameters and then has sion of the ventral gland is easily seen when convergent radii. The esophagus is terminated the living specimens are viewed under oil by an elongate nearly cylindrical esophago- immersion. The collecting tubule was on the intestinal valve which is about one-half a body left side in all specimens examined. diameter in length. The lumen of the esophagointestinal valve is dorsoventrally compressed. Reproductive system The female reproductive system is similar Excretory system to that found in Plectus and consists of opposed The most obvious feature of the Anaplectus gonacls reflexed at the juncture of the ovary excretory system is the long cuticularized ex- and oviduct. The male reproductive system cretory duct (Fig. 1). Maggenti (1961) has consists of two opposed (diorchic) testes; an described the excretory system of Plectus in anterior outstretched testis and a posterior one detail as consisting of a cuticularly lined ter- that is reflexed near its center as indicated by minal duct which extends to a single large Cobb (1914). The anterior testis is connected uninucleate excretory cell. The odd looping of to the seminal vesicle by a duct (vas efferens) the terminal duct is largely within the tissue which lies along the side of the reflexed pos- of the excretory gland. Similarly Anaplectus terior testis. In all males that we have ex- has a terminal duct which loops ventrally amined there are a pair of uninucleate glands around the esophagus in a ventrally located associated with the vas deferens (Fig. 4f). excretory cell. This cell which partly sur- These glands appear to be connected to the rounds the esophagus is highly modified in vas deferens just anterior to the cloaca, and Anaplectus. On the right side there is a short may function as ejaculatory glands as in some posterior extension which connects to the Rhabditis species, Chitwood (1930). thicker central portion containing the nucleus. The male tail, spicules, gubernaculum, and On the left side, the gland has an elongated ex- cuticularized tuboid supplements provide the tension which extends posteriorly past the ter- best morphological characters for distinguish- minal esophageal bulb, and then enters or is ing the species of Anaplectus. The tuboid sup-

Copyright © 2011, The Helminthological Society of Washington 80 PROCEEDINGS OF THE HELMINTHOLOGICAL SOCIETY plements are large and conspicuous, and vary A single papilla present on the subdorsal, latin number from two to five. There is intra- eral, and subventral sectors. Four cephalic specific variation in the number of tuboid setae present. Stoma composed of two parts, supplements. Hirschmann (1952) concluded the globular cheilostom with hexaradiate walls, that intraspecific variation in the number of and the pro-meso-metastom with triradiate tuboid supplements was not likely to occur and parallel walls. Amphid openings transverse separated A. submersus from A. granulosus slits. Cervical papillae present. Esophagus partly on this basis. However, Brzeski (1963) consisting of corpus, isthmus and a terminal has pointed out that there may be variation in bulb with a denticulate valve. Esophageal the number of supplements amongst males in radii tuboid in anterior corpus. Esophago- populations of A. granulosus. This was also intestinal valve elongate; lumen dorsoventrally indicated by Cobb (1914) for P. tubifer. We flattened. Excretory duct cuticularized. Ven- have observed similar variations in other spe- tral excretory cell elongated posteriorly and cies. The tuboid supplements are connected forming a collecting tubule. Vulva about equa- to large uninucleate glands that are located in torial. Female gonads amphididelphic and re- the right side of the body. flexed. Tail conoid-cylindrical with three cau- There is a subventral and subdorsal series dal glands. Spinneret present, orifice usually of papillae on the male tail. The number and cuticularized. Male testes paired, opposed with position of these papillae are variable in both the posterior one reflexed. Males with two to series and minor differences are not useful five large, cuticularized preanal tubuli. Spic- specific characters. In order to minimize varia- ules arcuate, gubernaculum well developed tion, illustrations were prepared to show the with a caudal process and a bifurcate cuneus. right side of the tail. Subdorsal and subventral papillae present on The spicules are arcuate and vary in size male tail. amongst the species. Except for differences TYPE SPECIES: Anaplectus granulosus (Bas- in the size of the manubrium, the spicules are tian, 1865) de Coninck and Schuurmans Stek- similar in all of the known species. hoven, 1933. The gubernaculum is well developed and The genus Anaplectus differs from Plectus appears to consist of one piece. The corpus Bastian, 1865, in the shape of the stoma, the is a thin plate with lateral extensions that modified lip region with 12 sectors, the slitlike partly surround the spicules (Fig. 2i). Arising transverse amphid openings and the presence from the corpus there is a caudal projection of of a collecting duct in the excretory system, variable size which may or may not be asso- numerous lateral hypodermal glands and the ciated with paired caudal apophyses (Fig. 2i). outstretched anterior testis. There is sometimes a small posteriorly directed process at the distal end of the gubernaculum. Anaplectus granulosus (Bastian, 1865) The well developed cuneus projects between de Coninck and Schuurmans Stekhoven, the spicules and is bifurcate at the distal end 1933 (Fig. 1 and Fig. 2, a-i) (Fig. 2h). There are three caudal glands which open at the terminus, and except for one spe- SYNONYMS: Plectus granulosus Bastian, cies, the spinneret is strongly cuticularized. Trans. Linn. Soc. Lond. 25. 1865: 120; Plectus bland Hofmanner and Menzel, Zool. Anz. 44. Systematics 1914: 82; Plectus schneideri de Man, Tijclschr. ned. dierk. Ver. 5. 1880: 48; Plectus tubifer GENUS: Anaplectus de Coninck and Schuur- Cobb, Trans. Am. Microsc. Soc. 33. 1914: 87; mans Stekhoven, 1933. Syn. Marinoplectus Anaplectus arenicola Killick, N. Z. J. Sci. 7. Kreis, 1963. 1964: 65 (New Syn.); Marinoplectus tetra- DIAGNOSIS (Emended): Cuticle with trans- papillatus Kreis, Zoology Iceland 2. 1963: 13 verse striae and lateral alae. Lateral hypo- (New Syn.). dermal glands in two sublateral rows, sometimes a partial dorsal and ventral series present. Dimensions Lip region set off or continuous with body con- FEMALES (84): L = 0.7-1.5 mm; a = 20- tour. Lips complex consisting of 12 sectors. 39; b = 4-6; c = 12-23; V = i<>--'->46-57!»-~-;

Figure 1. Anaplectus gramilosus male.

Figure 2. A. granulosus: a, male head; b, female tail; c, variation in tuboid supplements; d, e, f, left lateral, ventral, right lateral views of excretory system; g, spicule, gubernaculum and first tubule; h, gubernaculum cross section through cuneus; i, gubernaculum cross section through caudal projection and caudal apophysis. A. varicaudatus: j, male head; k, female tail; i, male tail; m, spicule, gubernaculum and first tubuli.

Stoma* = 15-30 p.; Amphid* = 5-11 p- Ex. between first preanal tubuli and cloacal open- Pore* = 69-146 p; T/ABD** = 1.6-2.7. ing. A coelomocyte located anterior to caudal MALES (75): L = 0.7-1.2 mm; a = 21-39; glands. Spinneret cuticularized. b = 4.0-6.4; c = 11-19; T = 47-84; Stoma = FEMALE: Similar to male. Lateral papillae 15-28 p, Amphid = 5-11 p- Ex. Pore = 73- near and dorsal to first lateral pore. Lips of 132 p.; T/ABD = 1.4-2; 1st Tuboicl Supple- vulva protruding in fixed specimens. Two ment = 14-27 p.; Spicules = 35-50 p.; Guber- lateral alae, marked by three lines. Ovaries naculum = 8-15 p. reflexed. Usually not more than one egg in MALE (Neotype): L = 1.0 mm; a = 35; uteri. Rectal glands present. Approximately b = 5.4; c = 15.3; T = 73; Stoma = 22.0 p.; 162 sublateral hypodermal pores on each side Amphid = 7.5 p.; Ex. Pore = 103 p.- T/ABD = of body. Tail conoid and curved. Three cau- 1.8; 1st Tuboid Suppl. = 17,5 p.; Spicule = dal glands and associated coelomocyte present. 40.0 p.; Gub. = 10.5 p.. Three pairs of small setae near terminus. Spin- Lip region set off by constriction, not neret cuticularized. striated. Cephalic setae arising at level of NEOTYPE: Male collected 11 December third annule. Amphids located eight annules 1959, by Mr. F. C. Peacock. Catalogue num- from lip region, and anterior to middle of ber 838, University of California Nematode stoma. First lateral pore 2.7 p, posterior to Survey Collection, Davis. amphid opening. A papilla adjacent and dor- TOPOTYPES: Two females and one male sal to the first lateral pore. Annules in cervical same data as neotype. region 0.07-1.6 p, wide. Cervical papillae in TYPE HABITAT: Soil around stubble. lateral field one body width posterior to nerve TYPE LOCALITY: Broadmoor, England. ring. Lateral field consisting of two alae be- Bastian's original description of A. gran- ginning posterior to stoma and extending almost ulosus was based upon females. A male from to terminus. The lateral field appears as three the type locality is designated as a neotype lines. A sublateral series of about 160 hypo- because the morphological features of males dermal glands. provide a better basis for species recognition The stoma consists of a globular cheilostom than do those of females. and the subcylindrical pro-meso-metastom. The pro-meso and metarhabdions enclosed in Diagnosis esophageal tissue. Corpus and isthmus not A. granulosus differs from A. submersus, A. well differentiated, nerve ring appears to en- similis n. sp., A. varicaudatus n. sp., and circle the posterior end of corpus. Esophago- A. porosus n. sp. in having the lip region intestinal valve elongate, one-half body width set off by constriction. From A. magnus it in length. Excretory pore opening one-half differs in size, the shape of the spicules and body width posterior to nerve ring. the absence of a distal projection on the Three heavily cuticularized preanal tubuli; gubernaculurn. the first opening 13 p., the second 37 p., and Paratype females of A. arenicola Killick, the third 85 p., anterior to the cloacal opening. 1964 could not be distinguished from females First tubuli less than one-half the length of of A. granulosus. The presence of the slitlike the spicules. Spicules arcuate, manubrium ex- amphidial opening of A. arenicola was given as panded, wider than calmus and blade. Guber- a differentiating character but as is indicated naculum with a dorsal projection from corpus. in the generic diagnosis this is characteristic Paired caudal apophysis present (Fig. 2i). of all Anaplectiis species. Marinoplectits tetra- Cuneus projecting between spicules and bi- papillatus Kreis, 1963 was described as a furcate at distal end (Fig. 2h). Margins of marine species from collections made in Ice- corpus partially enclosing spicules, serving as land. The figures and description indicate that crura (Fig. 2i). Subventral papillae; four on the single male and female are Anaplectus the tail and three preanal. Six subdorsal papil- granulosus despite the author's statement that lae extending to midway of the spicules. A a valvular apparatus was not present in the large median papilla located about midway esophageal bulb. Collections made by Dr. J. Klingler from the type locality of Plectus * Distance from anterior end. ** Ratio of tail length to anal body diameter. bland Hofmanner and Menzel, 1914 contained

Figure 3. A. subrnersus: a, male head; b, face view; c, male tail; d, variation in tuboid supplements; e, female tail; f, spicule, gubernaculum and first tuboid supplement. A. magnus: g, female tail; h, female head; i, spicule, gubernaculum and first tuboid supplement; j, male tail.

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 85 many specimens of A. granulosus and we extending almost to terminus. Length of believe that this species is a synonym of A. esophago-intestinal valve about one-third body granulosus as indicated by other authors. width. Excretory pore opening adjacent to Commonly the males of A. granulosus have nerve ring. Four heavily cuticularized tuboid three preanal tuboid supplements, but occa- supplements, first 9 p., second 35 /x, third 91 p., sional specimens are encountered with two or fourth 158 p. anterior to cloacal opening. First four supplements or rarely with none. tubuli about two-thirds the length of the spic- Specimens of this species have been collected ules. Spicules with manubrium not exceeding in the United States from California, Florida, width of the adjacent part of spicule. Guber- Hawaii, Kansas, Maryland, Minnesota, New naculum with a dorsal projection from the cor- Mexico, New York, South Carolina, South pus and a small projection at the distal end. No Dakota, Utah, Washington, and Wisconsin. paired caudal apophysis. The dorsal projection Also from Austria, England, Galapagos Islands, of the cuneus is bifurcate and sharply pointed. Germany, India, Ireland, Italy, Lichtenstein, Six subventral papillae on the tail, and three Netherlands, Republic of South Africa, Sweden, preanal papillae which extend anterior to the Switzerland, and Yugoslavia. second tubuli. Seven subdorsal papillae on the tail. A median papilla is located halfway be- Anaplectus submersus (Hirschmann, 1952) tween the cloacal opening and the first tubuli. Maggenti, 1961 (Fig. 3, a-f) Spinneret cuticularized. SYNONYM: Plectus submersus Hirschmann, FEMALE: Similar to males. Lateral papillae Zool. Jb., Jena, 81. 1952: 313-407. near and dorsal to first lateral pore. Cephalic setae fourth to sixth annules lip region. Lips Dimensions of vulva protruding only slightly in fixed specimens. Two lateral alae marked by three lines. FEMALES (32): L = 1.0-1.7 mm; a = 23- Ovaries reflexed. Rectal glands present. Num- 41; b = 4.5-6; c = 12-23; V = 11--M44-568--°; ber of lateral hypodermal pores variable (110- Stoma = 20-34 p.- Amphid = 8-13 ^ Ex. Pore 186). Tail conoid, curved. Three caudal = 114-186 (j,- T/ABD = 2-3. glands and associated coelornocyte present in MALES (30): L = 0.8-1.9 mm; a = 26-44; tail. Two pair of small setae near tail ter- b = 4-6; c = 13-20; T = 51-69; Stoma = 19- minus. Spinneret cuticularized. 33 p, Amphid = 7-13 ^ Ex. Pore = 100-178 p.; T/ABD = 1.7-2.6; 1st Tuboid Suppl. = 21- Diagnosis 33 p.- Spicules = 34-51 ^ Cub. = 10-17 p. Specimens of A. submersus were not avail- A. submersus can be distinguished from able from the type locality and the following other species of Anaplectus by the continuous description is made of a male collected in lip region, the size of the manubrium, absence Wageningen, The Netherlands. of paired caudal apophysis and the large tuboid MALE (Wageningen): L = 1.9 mm; a = supplement. This species resembles A. similis 36; b = 6; c = 20; T = 62; Stoma = 30 p.; but differs in lacking paired caudal apophysis Amphid = 8 p.; Ex. Pore = 168 ^ T/ABD = 2; and in having a longer first tuboid supplement. 1st Tuboid Suppl. = 30 p,; Spicule = 47 p,; Thirty-eight males of A. submersus were ex- Gub. = 16 p.. amined; 24 had four tuboid supplements, 13 Lip region continuous, no striations. Base had three, and one had five. As in other spe- of cephalic setae at fourth annule. Amphidial cies in the genus, identification cannot be based opening located anterior to middle of stoma, solely upon the number of tubuli in the male. at about the eighth annule. First lateral pore Distinguishing morphological differences are 3 p, posterior to amphidial opening. A lateral spicules, gubernaculum and supplement size. papilla is located 1 p, posterior and dorsal to Females of A. submersus and A. similis differ first lateral pore. Width of cervical annules in size, length of tail, and shape of the lip range from 1.1 to 1.5 p.. Cervical papilla two- region. thirds body width posterior to the nerve ring. Specimens of this species have been exam- Lateral field consists of two alae and three ined from Canada, Japan, The Netherlands, lines, beginning anterior to the nerve ring and California, Colorado, and Utah.

Figure 4. A. siniilis: a, female head; b, female tail; c, male tail; d, variation in tuboid supplements; c, spicule, gubernaculum and first tuboid supplement; f, gland associated with vas deferens.

Anaplectns siniilis n. sp. Lip region not set off by a constriction, no (Fig. 4, a-f) striations. Base of cephalic setae at about the Dimensions fourth annule. Amphidial opening anterior to FEMALES (44) : L = 1.2-1.6 mm; a = 26- middle of stoma, at the seventh annule. First 48; b = 5-7; c= 13-18; V = 7~ lateral pore 2.5 p, posterior to amphidial open- Stoma = 26-35 ^ Amphid = 7-13 ^; Ex. Pore ing. A lateral papilla located dorsal and adja- = 124-172 p.- T/ABD = 1.6-2.7. cent to first lateral pore. Width of annules in MALES (19): L = 1.1-1.7 mm; a = 25-37; b the cervical region from 1.0—1.5 p.. Cervical = 5-7; c = 12-17; T = 56-68; Stoma = 26-37 papilla about two-thirds body width posterior p.; Amphid = 8-11 p.- Ex. Pore = 132-168 p.; to nerve ring. Lateral field composed of two T/ABD = 1.9-2.3; 1st Tuboid Suppl. = 19- alae and three lines, beginning at level of nerve 26 p.; Spicules = 44-53 ^ Gub. = 10-14 p.. ring and extending almost to terminus. Pos- MALE (Holotype) : L — 1.4 mm; a = 35; terior bulb about one-half the width of the b = 5.4; c = 16.6; T = 64; Stoma = 34 p.; Am- body. Esophago-intestinal valve one-half body phid = 8 p.; Ex. Pore = 154 ^ T/ABD = 2; width in length. Excretory pore at level of 1st Tuboid Suppl. = 20 p.; Spicule = 48 ,t.- nerve ring. Four heavily euticularizecl preanal Gub. = 12 p.. tuboid supplements, first located 15 p. anterior

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 87 to cloacal opening, second 46 p., third 114 p., the adjacent part of the spicule, and a shorter and the fourth 174 //,. First tubuli about one- first tuboid supplement which is about one-half half the length of the spicules. Spicules and the length of the spicules. gubernaculum similar to A. gmnulosus but with Males of A. similis usually have four tuboid smaller manubrium. Five subventral papillae supplements. In the populations examined, 19 on the tail and three preanal. Eight subdorsal of the males had four tubuli and three had five. papillae extending to the level of the second Specimens of this species were also found at tubuli. A large preanal median papilla located Wisconsin Rapids, Wisconsin, in soil around midway between the cloacal opening and first the roots of Jack Pine. tuboid supplement. FEMALE: Similar to male. Vulva protrudes Anaplectus porosus n. sp. slightly in fixed specimens. Approximately 142 (Fig. 5, a-f) lateral pores on each side of body. Lip region Dimensions not set off by constriction, no striations. Base FEMALES (2): L = 1.6 mm; a = 34-35; b = of cervical setae at fourth annule. Amphidial 5.7; c = 22-24; V = i^53-5411-12; Stoma = opening anterior to middle of stoma at seventh 23-25 p.; Amphid = 8-9 p- Ex. Pore = 160- annule. First lateral pore opens 2.5 ^ posterior 164 p.; T/ABD = 2.3-2.5. to amphidial opening. Lateral papilla located MALES (5): L = 1.5-1.7 mm; a = 30-39; dorsal and adjacent to first lateral pore. An- b = 5-6; c = 20-24; T = 54-68; Stoma = 26- nule width in cervical region from 1.0-1.5 p.. 30 p; Amphid = 7-8 p.; Ex. Pore = 140-160 p.- Cervical papilla about one-half body width T/ABD = 1.8-2.1; 1st Tuboid Suppl. = 37-42 posterior to nerve ring. Lateral field consists p.; Spicules = 46-52 p- Gub. = 11-13 p.. of two alae and three lines, beginning anterior MALE (Holotype): L = 1.5 mm; a = 30; b to nerve ring and extending almost to terminus. = 5.6; c = 20; T = 54; Stoma = 26 p- Amphid Oval posterior bulb about one-half the body = 7 ^; Ex. Pore = 140 M; T/ABD = 1.9; 1st width. Esophago-intestinal valve about one- Tuboid Suppl. = 38 p.; Spicule = 47 p.; Gub. half body width in length. Excretory pore = 12 p.. adjacent to nerve ring. Tail conoid and elon- Lip region continuous with body contour. gate. Two small setae on the right and one Base of cephalic setae at fourth annule. Am- on the left side near terminus. Spinneret phidial opening at the seventh annule, anterior cuticularizecl. to middle of stoma. First lateral pore 3.5 p HOLOTYPE: Male collected 19 July 1957, posterior to amphidial opening. A lateral pa- by Mr. Gerald Thorne. Catalogue number 839, pilla 9.5 p, posterior to first lateral pore. Cer- University of California Nematocle Survey Col- vical annule width from 1.0-1.5 p,. Cervical lection, Davis. papillae about one-half body width posterior PARATYPE: Forty-three females and 20 to nerve ring. Lateral field consists of two males, same data as holotype distributed as alae and three lines, beginning anterior to follows: 42 females and 19 males, University nerve ring and extending almost to terminus. of California, Davis; one female and one male Posterior bulb about two-thirds body width. deposited USDA Nematode Collection, Nema- Esophago-intestinal valve less than one-half tology Investigations, Beltsville, Maryland; one body width in length. Excretory pore adja- female and one male Wageningen, The Nether- cent to nerve ring. A dorsal series of 12 hypo- lands. dermal pores beginning one-fourth body width TYPE HABITAT: Soil around the roots of posterior to nerve ring, extending to lip region. Maple. Ventrally a series of eight hypodermal pores TYPE LOCALITY: Wasau, Wisconsin. beginning anterior to excretory pore and extending to lip region. About 200 lateral hypo- Diagnosis dermal pores on either side of body. Three A. similis can be distinguished from A. gran- heavily cuticularized preanal tuboid supple- idosiis by the shape of the lip region and ments, first 10 p anterior to cloacal opening, the number of tuboid supplements. It differs second 33 p., third 88 /JL. First tuboid supple- from A. submersus in having paired caudal ment three-fourths as long as spicules (38/47). apophysis, manubrium exceeding the width of Gubernaculum with a long, narrow dorsal and

Figure 5. A. porosus: a, male head; b, anterior end showing dorsal, lateral and ventral series of hypodermal glands; c, male tail; d, male with three tuboid supplements; e, spicule, gubernaculum and first tuboid supplement; f, end of female tail. a small distal projection from corpus. Paired the excretory pore and extending to the lip caudal apophyses present. Cuneus projecting region. Lip region not set off by constriction. between the spicules, the distal ends bifurcate, Base of cephalic setae at fourth annule. Am- long and sharply pointed. Five subventral phidial opening at seventh annule and located caudal papillae; three preanal papillae extend- anterior to middle of stoma. First lateral pore ing about to level of the second tuboid supple- 3 p, posterior to amphidial opening. A lateral ment; one adanal papilla. Six subdorsal pa- papilla located 8 ^ posterior to first lateral pore. pillae extending anteriorly to level of cloacal Width of cervical annules from 1.3 to 1.5 /<,. opening. One lateral papilla located just an- Cervical papillae located about one-half body terior to tail terminus. No median papilla width posterior to nerve ring. Lateral field between cloacal opening and first tuboid sup- consists of two alae and three lines, beginning plement. Spinneret cuticularized. anterior to nerve ring and extending posterior FEMALE: Similar to male. Vulva not pro- almost to terminus. Esophageal bulb about truding in fixed specimens. About 194 lateral two-thirds body width. Esophago-intestinal hypodermal pores on each side of the body. valve about one-third body width in length. Thirteen dorsal hypodermal pores beginning Excretory pore adjacent to nerve ring. Two about one-half body width posterior to nerve small setae near terminus. Spinneret cutic- ring and extending to lip region. A series of ularized. nine or ten ventral pores beginning anterior to HOLOTYPE: Male collected 14 October 1962,

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 89 by Dr. D. J. Raski. Catalogue number 840, T/ABD = 1.3-1.8; 1st Tuboid Suppl. = 15-17 University of California Nematode Survey Col- p, Spicules = 35-48 ^ Gub. = 7-12 M. lection, Davis. MALE (Holotype): L = 0.76 mm; a = 29; PARATYPES: Two females and four males, b = 5; c = 18; T = 78; Stoma = 26 ^ Amphid same data as holotype, distributed as follows: = 5^; Ex. Pore = 94/i; T/ABD = 1.4; 1st two females and three males, University of Tuboid Suppl. = 15 p.; Spicule = 40 p; Gub. = California, Davis; one male USDA Nematode V Collection, Nematology Investigations, Belts- Lip region not set off by constriction, not ville, Maryland. striated. Cephalic setae short and robust, base TYPE HABITAT: Soil around the roots of at second annule. Amphidial opening located Pinus sp. at posterior end of cheilostom, about at third TYPE LOCALITY': Tangmarg, Kashmir, India. annule. First lateral pore 2 ^ posterior to On trail halfway between Tangmarg and Gul- amphidial opening. A lateral papilla located marg, elevation about 6,500 feet. dorsal and adjacent to first lateral pore. Width of cervical annules from 1.0-1.5 p.. Cervical Diagnosis papillae located about one-third body width A. porosus can be distinguished from other posterior to nerve ring. Lateral field with two species by the presence of an anterior series alae and three lines. Esophageal bulb about of dorsal and ventral hypodermal pores. The two-thirds body width. Length of esophago- number of glands in the dorsal and ventral intestinal valve about one-half body width. Ex- series is variable amongst the five males in the cretory pore opening adjacent to nerve ring. collection from India. The dorsal series varied Three cuticularized tuboid supplements, first from 10 to 14 in number and the ventral series 11 p,, second 28 p., third 56 ^ anterior to cloacal from eight to 11. Specimens from two other opening. First tubuli about one-third the localities, Scotland and Australia, show some length of the spicules. Manubrium set off deviation in the number and position of these from the rest of the spicules. Gubernaculum pores. The ventral series has one to three pores with a dorsal projection from corpus. Four posterior to the excretory pore. In addition the subventral papillae on the tail, two preanal position of the papilla associated with the first papillae located at anterior ends of first and lateral pore differed in these populations. In second tubuli. Five subdorsal papillae on the the Scotland and Australian specimens the tail. A median papilla located halfway between papilla was near to the first lateral pore rather cloacal opening and first tubuli. Spinneret not than the second as in the India specimens. cuticularized. Coelomocyte located anterior to These variations are considered to be within glands. the probable morphological variation of the FEMALE: Similar to male. Vulva protrudes species. in fixed specimens. About 98 lateral hypo- The five males from India also vary in the dermal pores on each side of the body. Lip number of preanal tubuli. Three of the males region not set off by a constriction. Cephalic have three tubuli and two have four. Two setae short and robust, arising at second annule. males from Scotland each have four tubuli. Amphidial opening at second annule, about The collection from Australia consisted of two level with posterior part of cheilostom. First females. lateral pore 2.5 p. posterior to amphidial opening. Lateral papilla adjacent and dorsal to Anaplectus varicaudatus n. sp. first lateral pore. Width of cervical annules (Fig. 2, j-m) from 1.0 to 1.8 ^. Cervical papilla about one- Dimensions third body width posterior to nerve ring. FEMALES (8): L = 0.8-0.9 mm; a = 25- Lateral field width of two alae and three lines. 33; b = 5-7; c = 16-20; V = ^--'^S-SS14--0; Esophageal bulb about one-half as wide as Stoma = 22-26 p.; Amphid = 4-7 p.; Ex. Pore body. Esophago-intestinal valve length slightly = 81-102 p, T/ABD = 1.8-2.5. less than one-half body width. Excretory pore MALES (8): L = 0.7-1.2 mm; a = 25-32; opening adjacent to nerve ring. Spinneret not b = 4-6; c = 17-22; T = 42-78; Stoma = 21- cuticularized. Two pair of small setae near 31 /*; Amphid = 4-7 p.; Ex. Pore = 86-125 p.; terminus.

HOLOTYPE: Male collected 19 December not seen. First tuboid supplement less than 1957, by Dr. R. P. Esser. Catalogue number one-half the length of spicules. Manubrium 841, University of California Nematode Survey of spicules not exceeding width of calmus. Collection, Davis. Series of nine subventral papillae, five on the PARATYPES: Seven females and four males, tail and four preanal papillae extending almost same data as holotype, distributed as follows: to second tuboid supplement. Five subdorsal six females and three males, University of Cali- papillae on the tail. A large median papilla fornia, Davis; one female and one male, USDA located one-half way between the cloacal open- Nematode Collection, Nematology Investiga- ing and the first tuboid supplement. Spinneret tions, Beltsville, Maryland. cuticularized. TYPE HABITAT: Soil around roots of Zoysia FEMALE (Allotype): Similar to male. Vulva grass. not protruding in fixed specimen. Lip region TYPE LOCALITY: Monticello, Florida. set off by a deep constriction. Base of cephalic setae at third annule. Amphidial opening lo- Diagnosis cated at sixth annule. First lateral pore 17 /;, A. varicaudatus differs from other species of posterior to amphidial opening. A lateral Anaplectus in lacking a cuticularized spinneret. papilla is located 5.5 ^ posterior to amphidial The amphidial openings are more anterior than opening and anterior to first lateral pore. in other species and the first tuboid supple- Width of cervical annules range from 1.0-1.5 ^. ment is less than half the length of the spicules. Excretory pore opening adjacent to nerve ring. Specimens of this species were also collected Posterior bulb width about one-half body in soil from Kansas and South Carolina. width. Three setae on tail. Spinneret cutic- Anaplectus magnus Brzeski, 1963 ularized. HOLOTYPE: Male collected 1961, by M. (Fig. 3, g-j) Stradowski. Catalogue number 442, University Dimensions of California Nematode Survey Collection, FEMALE (Allotype): L = 2.0 mm; a = 39; Davis. Deposited at Davis by M. Brzeski. b = 7; c = 19; V = i«471(S; Stoma = 30 /x; Am- ALLOTYPE: Female, same data as holotype. phid = 16 ^; Ex. Pore = 149 p.; T/ABD = 2. Catalogue number 443, University of Califor- MALE (Holotype): L = 1.9 mm; a = 58; nia Nematode Survey Collection, Davis. b = 7; c = 26; T = 72; Stoma = 30 p.; Amphid TYPE HABITAT: Soil around psammon. = 13 p.; Ex. Pore = 143 ^ T/ABD = 1.5; 1st TYPE LOCALITY: Wolin Island, Poland. Tuboid Suppl. = 20 /x; Spicule = 49 p.; Gub. = Diagnosis 15 p.. A. magnus differs from A. gmnulosus by the MALE (Holotype): Lip region set off by shape of the gubernaculum, the reduced diam- a deep constriction. Base of cephalic setae at eter of the manubrium and the number of third annule. Amphidial opening at eighth tuboid supplements. The set off lip region annule. First lateral pore 18 p, posterior to distinguishes A. magnus from other species in amphidial opening. A large lateral papilla is the genus. located 6.5 ^ posterior to amphidial opening, A. magnus is known only from the type almost one-half the distance between first lat- specimens. In the description of the species eral pore and amphid aperture. Width of Brzeski (1963) indicated that the amphid cervical annules from 0.8-2.0 p,. Excretory apertures were oval. However, examination of pore opening adjacent to nerve ring. Cervical the types indicates that the large circular papilla located one-third body width posterior papilla posterior to the slitlike amphid aper- to nerve ring. Posterior bulb more than one- ture was illustrated as the amphid opening. third body width. Esophago-intestinal valve In the original description the vulva was indi- about one-third body width. Lateral field with cated to be at 39%. Measurements of the same two alae and three lines. Two cuticularized female specimen shows the vulva to be at 47%. tuboid supplements located anterior to cloacal opening, the first 26 ^ and the second 46 /jt. Summary Gubernaculum with a dorsal and distal projec- Anaplectus granulosus is redescribed from tion. Paired caudal apophysis present. Cuneus a neotype collected from the type locality.

A. ma gnus is redescribed from the type speci- freilebenden Nematoden. Nova Acta Leop. mens and an emended description of A. sub- Carol. 36: 1-144. mersus is presented. A. similis n. sp., A. vari- Chitwood, B. G. 1930. Studies on some physio- caudatus n. sp., and A. porosus n. sp. are de- logical functions and morphological characters scribed. The generic diagnosis of Anaplectus of Rhabditis (Rhabditidae, Nematodes). I. Morph. and Physiol. 49: 251-275. is emended to include new information on the , and M. B. Chitwood. 1937. An in- excretory system, lip region and amphids. A. troduction to nematology, Sec. 1, Pt. 1. arenicola and Marinoplectus tetrapapillatus are Monumental Press, Baltimore, Mel. 53 pp. synonymized with A. granulosus. Cobb, N. A. 1914. The North American freeliving freshwater nematocles. Trans. Am. Key to Males Microsc. Soc. 33: 69-134. 1. Cuticularized spinneret present 2 dc Coninck, L. A. and J. H. Schuurmans Stek- Cuticularizecl spinneret absent hoven. 1933. The freeliving marine nemas A. varicaudatus n. sp. of the Belgian Coast. II. With general re- 2. Dorsal and ventral hypodermal glands marks on the structure and the system of present A. porosus n. sp. nemas. Mem. Mus. Hist. Nat. Belg. 58: 1- Dorsal and ventral hypodermal glands 163. Goodey, J. B. 1963. Soil and freshwater nema- absent 3 todes. (Revision of T. Coodey, 1951.) Me- 3. Lip region set off by constriction 4 tliuen, London. 544 pp. Lip region not set off by constriction 5 Goodey, T. 1951. Soil and freshwater nema- 4. Manubrium expanded, larger than tocles. Methuen, London. 390 pp. blade, gubernaculum without distal Hirschmann, H. 1952. Die Nematoden der Was- projection, tuboid supplements usu- sergrenze Mittelfrankischer Gewasser. Zool. ally three A. granulosus Jb. 81: 313-407. Manubrium not expanded, gubernac- Hofmanner, B., and R. Menzel. 1914. Neue ulum with a prominent distal projec- Arten freilebender Nematoden aus cler Schweiz. Zool. Anz. 44: 80-91. tion, tuboid supplements two Killick, J. L. 1964. A new species of the genus A. magnus Anaplectus de Coninck and Schuurmans Stek- 5. First tuboid supplement small, one-half hoven, 1933 (Nematoda: Plectidae). N. Z. or less length of spicule. Paired cau- J. Sci. 7: 165-168. dal apophysis present _„ A. similis n. sp. Kreis, H. A. 1963. Marine nematoda. Zoology First tuboid supplement large exceed- Iceland 2: 1-68. ing one-half length of spicule. Paired Maggenti, A. R. 1961. Revision of the genus caudal apophysis absent.... A. submersus Plectus (Nematoda: Plectidae). Proe. Helm. Soc. Wash. 28: 139-166. Literature Cited de Man, J. G. 1880. Die einheimischen, frei in Bastian, H. C. 1865. II. Monograph on the der reinen Erde und im siissen Wasser lebende Anguillulidae, or free Nematoids, marine, land Nematoden. Tijdschr. Ned. Dierk. Ver. 5: and freshwater; with descriptions of 100 new 1-104. species. Trans. Linn. Soc. Loncl. 25: 73-184. . 1884. Die frei in der reinen Erde und Brzeski, M. 1963. Review of the nematocle im siissen Wasser lebenden Nematoden der genus Anaplectus de Coninck, Sell. Sth. Niederliindisclien Fauna. Eine systematische— (Nematoda, Plectidae). Bull. Acad. Pol. Sci. faunistische Monographic. Leiden. 206 pp. Cl. II Ser. Sci. Biol. 11: 35-38. Micoletzky, H. 1922. Die freilebenden Erd- . 1963. Contribution to the knowledge Nematoden. Arch. Naturg Abt. A. 87: 1- of the Polish species of the genera Plectus 650. Bastian and Anaplectus de Coninck and Sch. Schneider, W. 1939. Wurmcr oder Vermes. II. Stekh. (Nematoda, Plectidae). Fragm. Faun. Fadenwurmer oder Nematoclen. I. Freilebencle 11: 21-30. und pflanzenparasitische Nematoclen. Tierwelt Biitschli, O. 1873. Beitriige zur Kenntnis cler Deutschlands (Dahl) 36: 1-260.

Observations on Diphyllobothrium sebago Plerocercoids in the Fish Hosts1

MARVIN C. MEYER AND ROLF VIK-

Plerocercoids of Diphyllobothrium Cobbolcl Lakes drainage. The perch were examined constitute a normal part of the larval tape- during the summers of 1960 and 1963. The worm fauna of both anadromous and fresh- smelts were taken from 28 April through 7 water Salmonidae and related fishes. Working May 1961 at the mouth of the Kennebago in Maine, Meyer and Vik (1961), Meyer and River as the fish ascended the river from Robinson (1963) reported that plerocercoids Mooselookmeguntic Lake on the spawning run. of Diphyllobothrium sebago (Ward) occur in Only fresh fish were used. As soon as landlocked salmon, Salmo solar Linnaeus; possible after capture they were subjected to brook trout, Salvelinus fontinalis (Mitchill); routine examination. The salmonids were allo- and landlocked American smelt, Osmerus mor- cated a reference number and measured for dax (Mitchill). The plerocercoid is the most total length; sex was observed, and scales were commonly encountered stage in the worm's life removed for age determination; the perch were cycle, yet it is the least studied stage. Some not examined in detail. The smelts, 450 of of the diphyllobothrians infecting salmonids which were examined during each of 10 con- normally mature in gulls, as does D. sebago. secutive days, were assigned a reference num- In elucidating the life cycle of D. sebago, ber, weighed, and sexed; 50 were digested Meyer and Vik (1963) have shown that the individually and the remaining 400 were di- life cycle is typically diphyllobothrian, involv- gested in lots of 50. The viscera of salmonids ing copepods and fish as intermediate hosts. and perch were removed and digested, apart The final host, normally the herring gull Lams from the flesh, to determine the distribution of argentatus Pontoppidan, becomes infected the larvae. Terms common to ichthyologists through eating fish or fish offal containing are used in the aging of teleosts. Fish in their viable plerocercoids. This study includes fur- first year of life are termed "0" group; those ther data of a quantitative nature on the larvae at the end of their 1st year and just entering in the fish. their 2nd year of life are called "I" group; while those caught toward the middle of their 2nd Materials and Methods year are known as "I+" individuals; the fish From June 1959 through 1963, 97 salmonids, of the other ages are similarly defined. The 96 yellow perch [Perca flavescens (Mitchill)], term "incidence of infection" is used to mean and 4,500 smelt were digested by the peptic- the ratio of the number of infected fish to the HCl digest method for Diphijllobothriiim se- total number of fish examined (in some in- bago plerocercoids. Except for three trout and stances, this ratio is expressed as a percentage); an occasional salmon digested during the sum- the term "intensity of infection" is used to mean mers, the salmon were obtained as casualties of the number of parasites per infected fish. stripping for artificial propagation during each of the Novembers. The fish were from Observations and Discussion Rangeley Lake and Mooselookmeguntic Lake, Location of the plerocercoids except for 11 salmon from Sebago Lake. Since there is continuous water passage between The plerocercoids of D. sebago occur in Rangeley Lake and Mooselookmeguntic Lake, salmon and trout beneath the serosal layer of fish are free to migrate between these lakes the stomach and caeca, especially between the and the other larger lakes in the Rangeley folds formed by the shorter arm of the stomach and the intestinal arm from the stomach; the 1 Department of Zoology, University of Maine, Orono, plerocercoids also penetrate the kidneys, pan- Maine. Supported by research grant (AI-02759) from the National Institutes of Health, U. S. Public Health Service. creas, spleen, liver, swim bladder, heart, and - Now at the Zoological Museum, University of Oslo, Oslo 5, Norway. gonads of both sexes; the plerocercoids also

Figure 1. Anterior portion of the alimentary canal of S. salar, viewed from the right. (The broken line separates the esophagus from the stomach, and the solid line marks the level of the pyloric valve. Most of the pyloric caeca are displaced ventrally to show the relationship between the stomach and the anterior region of the intestine.) occur free in the coelom, immediately beneath individual fish of a population and among fish the serosal layer of the body wall; and they in different lakes, it is true that within the occur in the flesh. While no part of the fish limits of our study the incidence of infection is always free of plerocercoids, the greatest and the intensity of infection are associated concentration of larvae occurs between the with age. The 11+ fish were less than 30 cm; arms of the stomach and the intimately asso- with one exception, the III+ constituted the ciated pylorie caeca (Fig. 1). 30-34 cm group; the majority of the 35-39 cm The larvae are often encysted in great num- category were IV+, and the 40-44 cm group bers, but they may also be free in the tissues was predominantly V+. Beginning with V+, with little or no evidence of host-tissue reac- the length:age relationship is much obscured; tion. Pimplelike, raised areas were observed while the oldest fish were VII+, the longest under the skin of a 25 cm trout; while each (69 cm) was VI+. raised area contained a larva, no exudate or Based upon fish length classes of 5 cm, both inflammation of any kind was present and the incidence of infection and the mean in- there was no evidence of encystment. In an- tensity of infection with D. sebago plerocer- other single case, the anterior end of a larva coids increase with the length of the salmon was seen projecting several mm through the (Table 1 and Fig. 2). If an analysis of the body wall of a salmon. Nothing can be said 94 salmon is made on the basis of two length about the location of the plerocercoids in smelt, classes, 27-42 cm and 43-69 cm, groups ap- since larval recovery was limited to digestion proximately equal in number, the findings are of the complete fish. even more striking. In the smaller size category, the great majority of fish are V+ and Relationship between the length younger; the incidence of infection is 64% and of salmon and occurrence the mean intensity of infection is 16. In the of the plerocercoids 43-69 cm category, including fish which are Fish size is, generally speaking, a function predominantly V-f- and older, the incidence of of age. Although precise relationships between infection is 86% and the mean intensity of body length and age vary somewhat among infection is 31. Previous workers (Hobmaier,

XIII fc INCIDENCE OF INFECTION -%

MEAN INTENSITY OF INFECTION XXV

A LENGTH CLASSES-CM Figure 2. Incidence of infection and mean intensity of infection of S. salar by D, sebago plerocercoids. A, Analysis based upon fish length classes of 5 cm; B, Analysis of same fish based upon two length classes (27 to 42 and 43 to 69 cm). The number of hosts in each group is indicated by the Roman number adjacent to it.

1927; Petruschewsky, 1931; Kuhlow, 1955; The three trout examined (11+ and III+), Vik, 1957; Tallqvist, 1962; Romanov, 1964) 35-37 cm in total length, were infected. Eighty have also reported that large fish have more plerocercoids were recovered: 65 from the vis- Diphijllobothrium plerocercoids than small fish cera, and 15 from the flesh. Among the 4,500 of the same species. smelts, 209 larvae were recovered, 4.64 per

Table 1. Incidence of infection and mean intensity of infection of Salmo salar by Diphyllobothrium sebago plerocercoids. A, Analysis based upon fish length classes of 5 cm; B, analysis of same fish based upon two length classes (27—42 and 43—69 cm).

Total Number of fish Incidence Nu: 1 f ms Mean intensity of infection of infection (cm) Examined Positive (%) Viscera Flesh Total ( Range ) "A" <30 2 1 50 3 0 3 3 30-34 6 o 33 12 8 20 10 (1-19) 35-39 25 17 68 232 28 260 15 ( 1-66 ) 40-44 25 20 80 286 44 330 17 ( 1-55 ) 45-49 18 14 78 287 23 310 22 (1-86) 50-54 13 12 92 351 27 378 32 (6-109) 55-59 3 3 100 411 7 418 139 (51-312) >59 2 o 100 59 1 60 30 (11-49) Total 94 71 78* 1,641 138 1,779 25* (1-312) "B" 27-42 45 29 64 409 50 459 16 (1-66) 43-69 49 42 86 1,232 88 1,320 31 (1-312) * Weighted mean.

100 fish. In the 500 digested individually (50 Acknowledgments each day for 10 consecutive days), the inci- Grateful appreciation is due certain person- dence of infection was 28/500 (6%) and the nel of the Fisheries Division, State of Maine mean intensity of infection was 30/28, or 1.07 Department of Inland Fisheries and Game: per infected fish. The 96 perch were negative W. Harry Everhart and Stanley J. Linscott, for plerocercoids. Chief of Fisheries and Superintendent of the We believe that the food of the salmon is Hatchery Division, respectively, for providing significant in relation to the transfer of the field laboratory quarters; Arlo Copp, through plerocercoicl. As a matter of fact, our attention whose cooperation most of the salmon were was first focused on the smelt as a host in the obtained; and Charles F. Ritzi, Raymond A. transmission cycle of D. sebago, when plero- DeSandre, and Richard B. Anderson for aging cercoid-infected smelt were found in the stom- the scales of the salmonids. We are indebted ach of salmon during the summer of 1960 by to Harold E. Young for setting up the experi- one of us (RV). ment involving the digestion of the smelts. Cooper (1940) reported that smelts consti- Special thanks are also due John E. Watson, tute one of the most important forage fishes for whose critical reading of the manuscript con- landlocked salmon in Maine. In a stomach tributed to its improvement. analysis of salmon from Mooselookmeguntic Lake, the source of all the smelts and most of Summary the salmon examined by us, Cooper found that 85% of the volume consisted of smelts, and In an examination of 4,693 freshwater fishes, less than 1% consisted of aquatic insects. In comprising 94 landlocked salmon, Salmo salar; 40 salmon stomachs from three lakes in the three brook trout, Salvelinus fontinalis; 4,500 Rangeley Lakes region, he found 49 smelts, landlocked American smelt, Osmenis mordax; one sucker, one lake chub, and five fish di- and 96 yellow perch, Perca flavescens, for gested beyond recognition. plerocercoids of Diphyllobothrium sebago, only

Copyright © 2011, The Helminthological Society of Washington 96 • PROCEEDINGS OF THE HELMINTHOLOGICAL SOCIETY the salmon, trout, and smelt were found in- scription and occurrence of Diphijllobothrium fected. In the salmon, ranging from 27 cm sebago (Ward, 1910). J. Parasit. 49: 969- through 69 cm and 11+ through VII+ years, 973. the incidence of infection and the mean inten- , and R. Vik. 1961. Sparganum sebago, sity of infection increased with host size. A incidence and location in host fishes. J. Para- sit. 47 (4, Sect. 2): 56. 59 cm, VII+ salmon harbored 312 larvae; , and . 1963. The life cycle of 309 in the viscera and three in the flesh. Diphijllobothrium sebago (Ward, 1910). J. Among the 4,500 smelt, 209 plerocercoids were Parasit. 49: 962-968. recovered. Petruschewsky, G. K. 1931. Uber die Verbrei- tung der Plerocercoide von Diphijllobothrium Literature Cited latwn in den Fischen der Newabucht. Zool. Cooper, G. P. 1940. A biological survey of the Anz. 94: 139-147. Rangeley Lakes, with special reference to the Romanov, I. V. 1964. Study of foci of diphyl- trout and salmon. Maine Dept. Inland Fish. lobothriasis in the Gorkov region. Med. Para- Game, Augusta, Fish Survey Kept. No. 3, zitol. Parazitarn. Bolenzni. Moscow 33: 289- 182 pp. 294. [Russian: English summary, p. 294.] Hobmaier, M. 1927. Wie kommt die Infektion Tallqvist, Johanna. 1962. Occurrence of plerocercoids of the fish tapeworm (Diphijlloboth- der Raubfische mit clem Plerocerkoid von rium latum) in perch and ruff from Lake Dibothriocephalus latus zustande? Zbl. Bakt., lisvesi and Lake Niinivesi (SE Finland). 2 Abt. 72: 268-273. Acta Soc. Fauna Flora Fenn. 77: 1-10. Kuhlow, F. 1955. Untersuchungen iiber die Vik, R. 1957. Studies of the helminth fauna of Entwieklung des Breiten Bandwurmes (Di- Norway. I. Taxonomy and ecology of Diphijl- phi/llobothrium latiim). Zeit. Tropenmed. lobothrium norvegicum n. sp. and the plero- Parasit. 6: 213-225. cercoid of Diphijllobothrium latum (L.). Nytt Meyer, M. C., and E. S. Robinson. 1963. De- Mag. Zool. 5: 25-93.

Three New Species of Actinolaimidae (Nematoda: Dorylaimoidea) from India

M. SlIAMIM JAIRAJPURI1

Two new species belonging to the genus Carcharolaimus masoodi- n. sp. Carcharolaimus Thorne, 1939 and one to the (Fig. 1, A-H) genus Actinolaimus Cobb, 1913 were found in FEMALES (15): L = 1.70 mm (1.57-1.85 soil samples collected in Uttar Pradesh, India mm); a = 51 (44-55); b = 4.6 (4.3-5.0); from the districts Pilibhit, Moradabad, and c = 84 (70-88); V = °51» («-1()48-547-1(J). Saharanpur, respectively. This appears to be HOLOTYPE (female): L = 1.75 mm; a = 44; the first record of Actinolaimidae from India. All the observations and measurements were b = 4.5; c = 83; V = i«5410. taken on the specimens killed and fixed in JUVENILES (4th Stage) (5): L = 1.51-1.64 mm; a = 46-48; b = 4.7-5,3; c = 70-76. hot 4% formalin and mounted in dehydrated glycerine. Description Body straight, tapering very slightly towards 1 Section of Nematology, Department of Zoology, Aligarh Muslim University, Aligarh, U.P., India. extremities. Cuticle smooth. Lateral chords - Named after Mr. Masood T. Zaidi, Assistant Engineer who collected the soil samples containing these worms. distinct, about a quarter of body width near

.A,C — D-H sou —.8

Figure 1, A-H. Car char olaimus masoodi n. sp. A. Head end. B, Anterior sexual branch. C, Head end showing glandular organs and amphids. D, Cardia region. E, Anterior part of basal expanded portion of esophagus. F, Vulva. G, H, Tail ends.

20JU , A r 30U , D,E,CH SOU

Figure 2, A-H. Car char olainnm mujtabai n. sp. A, Head end. B, Anterior sexual branch. C, Head end showing glandular organs and amphid. D, Cardia region. E, Anterior part of basal expanded portion of esophagus. F, Vulva. G, H, Tail ends.

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 99 middle and provided with 100-125 glandular 1939 from which it differs in having a slender organs which are conspicuous towards tail end. body (a = 37 in C. teres), smooth cuticle, Each glandular organ connected to a lateral wider lip region (one-fourth as wide as base pore in its centre. There are 22-28 glandular of esophagus in C. teres), longitudinal vulva organs in the esophageal region and only one (transverse in C. teres) and a shorter tail (c = on tail. Lip region about two-thirds as wide 50 in females of C. teres). as body at base of esophagus. Lips large, angular; labial papillae visible. No denticles Carcharolaimus mujtabai* n. sp. in. the labial basket. Amphids cuplike, their (Fig. 2, A-H) apertures 7-8 p, wide, situated 8-10 p. from FEMALE: L = 1.52 mm; a = 31; b = 3.5; anterior end. c = 64; V = "4811. Spear 11-13 p., cylindrical; aperture 6-7 p.. HOLOTYPE (female): L = 1.60 mm; a = 34; Guiding ring 10-12 p, from anterior end, single. b = 4.0; c = 73; V = M810. Spear extension 20-25 p., simple. Anterior JUVENILES (2): L = 0.98-1.08 mm; a = 25- slender part of esophagus forms an oval swell- 30; b = 3.7-4.0; c = 60. ing before joining the basal expanded portion through a distinct constriction. Basal expanded Description portion of esophagus 56-64% of the total Body plump, straight or slightly curved, only esophageal length; its width more than half slightly tapering towards extremities. Cuticle of corresponding body and one-twelfth to one- smooth. Lateral chords very distinct, about a fourteenth of its own length. Esophageal lu- quarter body width wide near middle and men prominent, cuticularized, 3-5 p, wide. provided with very conspicuous 120—126 Opening of dorsal esophageal gland 173—181 glandular organs, each connected to a lateral p, or 43-48% of the esophageal length from pore in its centre. There are 28-34 glandular anterior end. A pair of subventral glands have organs in the esophageal region, 27-30 be- orifices 140-150 p. behind the dorsal esoph- tween base of esophagus and vulva and 61-64 ageal gland opening. Nerve ring 94-105 ^ between vulva and anus and only one on tail. from anterior end. Base of esophagus sur- Lip region about two-thirds of body width rounded by a muscular pad; cardia hemi- at base of esophagus. Lips very prominent and spheroid to spathulate. large, angular; labial papillae visible. Amphids Vulva a longitudinal slit. Vagina 11-14 p. shallow cuplike, their apertures 8-10 ^ wide, or one-third to one-fourth across the body. situated 8-10 p, from anterior end. Each gonad branch has a uterus about one Spear 20-21 p,, cylindrical; aperture 12-13 body width long, an oviduct two to three body p,. Guiding ring 17-18 p. from anterior end, widths long and an ovary with a single flexure single. Spear extension 27-30 ^, simple. An- and eight to 12 oocytes. Sperms not present terior slender part of esophagus same as in in the uteri. Prerectum 33-51 p., one and one- C. masoodi except that the swelling before the half to two anal body widths long. Rectum constriction is spherical. Basal expanded por- 20-24 p, about one anal body width long. Tail tion of esophagus 60-62% of the total esoph- 20-24 p., rounded. ageal length; its width about half of corre- MALE: Not found. sponding body and one-twelfth to one-sixteenth HABITAT: Soil around roots of grasses (un- of its own length. Esophageal lumen strongly identified), 3 km from Pilibhit Railway Station cuticularized, 4—7 p, wide. Opening of dorsal on Pilibhit-Bareilly Road near River Bridge, esophageal gland 173-182 ^ or 41-44% of District Pilibhit, U.P., India. the esophageal length from anterior end. A TYPE SPECIMENS: Collected by Mr. Masood pair of subventral glands have orifices 156 p. T. Zaidi in March 1967; holotype mounted on behind the dorsal esophageal gland opening. slide MS]/Carcharolaimus masoodi/1; 14 para- Nerve ring 112—114 p, from anterior end. Base types and five juveniles on slides MSJ/Car- of esophagus surrounded by a muscular pad; charolaimus masoodi/2-4; deposited with the cardia rounded. Zoology Museum of Aligarh Muslim University. DIFFERENTIAL DIAGNOSIS: Carcharolaimus :l Named after Mr. Syed Hasan Mujtaba Baqri from whose garden the soil samples containing the worms were masoodi n. sp., comes close to C. teres Thorne, collected.

SOU ,pr,

Figure 3, A-G. Actinolaimus armatus n. sp. A, Head end. B, Cardia region. C, Vulva region. D, Anterior sexual branch. E, Amphid. F, Anterior part of basal expanded portion of esophagus. G, Tail end.

Vulva a longitudinal slit. Vagina 12-15 ^ c = avg 84 masoodi n. sp. or one-fourth to one-fifth across the body. Lip region one-fourth of body width at Uterus in each sexual branch about one body base of esophagus; vulva transverse; width long, oviduct two to three body widths c = 50 teres Thorne, 1939 long and ovary has a single flexure and eight SPECIES INQUIRENDA: Carcharolaimus rotun- to 10 oocytes. A weak sphincter present be- dicauda (de Man, 1880) Thorne, 1939 is trans- tween uterus and oviduct. Sperms not present ferred to species inquirenda because its de- in the uteri. Prerectum 21—27 /JL, less than one scription is inadequate to distinguish it from anal body width long. Rectum 28-30 /m, as other members of this genus. long as one anal body width. Tail 21-24 p., rounded. Actinolaimus armatus n. sp. MALE: Not found. (Fig. 3, A-G) HABITAT: Soil around roots of White Mul- berry, Moms alba Linn., from the garden of FEMALES (6): L = 2.05 mm (1.92-2.13 Mr. Syed Hasan Mujtaba Baqri, Said Nagli, mm); a = 61 (57-65); b = 4.3 (4.0-4.6); District Moradabad, U.P., India. c = 21 (19-24); V = 852S (7-1050-568-

134 ft, from anterior end. Base of esophagus elaboratus (Cobb, 1906) Thorne and Swanger, surrounded by a muscular pad; cardia conoid. 1936; A. cinctus (Cobb unpubl.) Thorne, 1939 Vulva a transverse slit. Vagina 10-12 ^ or and A. zealandicus Clark, 1963. It differs from one-fourth to one-fifth across the body, pro- all these three species in having a slender body vided with distinct cuticularized pieces. Uterus (a = 36 in A. elaboratus-, 32 in A. cinctus and in each sexual branch about two to three body 38-50 in A. zealandicus}, and cuticularized widths long, oviduct is equally long, thin, and pieces in the vagina. It further differs from tubular but expands before joining the ovary. A. ekiboratus in having a longer prerectum; Ovary with a single flexure, contains 10-12 from A. cinctus in having a differently shaped oocytes. Sperms not present in the uteri. lip region and nature of the vestibule and from Prerectum 100-148 /*., five to eight anal body A. zealandicus in the shape of the lip region. widths long. Rectum 32-35 ^, one and one- half anal body widths long. Tail 100-110 /,„ Summary conoid, filiform with acute tip, about five anal Three new species of Actinolaimidae (Nema- body widths long. Two minute caudal pores toda: Dorylaimoidea), two belonging to the visible. genus Carcharolaimus Thorne, 1939 and one to MALE: Not found. the genus Actinolaimus Cobb, 1913, were col- HABITAT: Soil around roots of plum, Primus lected in Uttar Pradesh, India. A key to the communis Huds. from L. R. Brothers Nurseries, species of Carcharolaimus is provided. C. ro- Saharanpur, U.P., India. tundicaiida is transferred to species inquirenda. TYPE SPECIMENS: Collected by the author in October 1962; holotype, one paratype and Literature Cited a juvenile mounted on slide MS]/Actinolaimus Clark, W. C. 1963. New species of dorylaimoid armatus/1; five paratypes and two juveniles on nematodcs belonging to the genera Ptingentus Thorne and Swanger, Actinolaimus Cobb, and slide MSJ/Actinolaimus armatus/2; deposited Donjlaimellus Cobb. N. Z. Jl. Sci. 6: 565- in the Zoology Museum of Aligarh Muslim 576. University. Thorne, G. 1939. A monograph of the nema- DIFFERENTIAL DIAGNOSIS: Actinolaimus ar- todes of the superfamily Dorylaimoidea. matus n. sp., is most closely related to A. Capita Zool. 8: 1-261.

Susceptibility of Gerbils and Young White Rats to Simultaneous Infection with Trichostrongylus axei and Trichostrongylus colubriformis

K. C. KATES AND D. E. THOMPSON1

Lelancl (1961, 1963a, b) reported that Mon- young white rats "were very susceptible" to golian gerbils could be infected with Tricho- infection with T. colubriformis, a common strongylus axei, a common nematode parasite intestinal nematode parasite of ruminants. We in the stomach of ruminants and equines, and (1967) reported that Mongolian gerbils coukl Williams and Palmer (1964) reported that be infected with both nematode species simul- taneously, and that this host—parasite system 1 This \vork was done while the authors were employed by the Division of Veterinary Research, Bureau of Veter- showed promise as a primary anthelmintic inary Medicine, Food and Drug Administration, Depart- ment of Health, Education, and Welfare, Beltsville, Mary- screen. Because of the possible utility of such land. Present address: Beltsville Parasitological Laboratory, host—parasite systems in anthelmintic research Animal Disease and Parasite Research Division, ARS, USDA, Beltsville, Maryland. and for other purposes, three experiments were

Table 1. Experiment I. Trichostrongylus spp.1 recovered at necropsy from gerbils and young white rats (modified Wistar strain). Larval inoculation 975 ± 36 SE/animal; culture 58.

Gerbils Rats 4M 4F All 4M 5F All GROUP A. 14 days postinfection T. axei 11 (3-32)2 15 (5-32) 13 ( 3-32 ) 0 ( 0-0 ) 0.2 (0-1) 0.1 (0-1) T. cohtbriformi.t 136 (40-202) 138 ( 74-266 ) 137 (40-266) 4 (1-8) 2 (0-5) 3 (1-8) Both species 147 (44-232) 153 (125-271) 150 (44-271) 4 (1-8) 2 (0-5) 3 (0-8)

GROUP B. 28 days postinfection''1 T. axei 43 (11-106) 19 (1-46) 31 (1-106) 0 0 0 T. colubriformiH 90 (25-165) 39 (7-76) 64 (7-165) 0 0 0 Both species 133 (36-230) 58 (15-122) 95 (15-230) 0 0 0 Composite EPG 410 10 210 0 0 0

] Nematodes in 4th or early 5th stage at 14 days PI, feces negative for eggs; 99+% were mature at 28 days PI. - Average, followed by range in parentheses. :l Ten rats in this group. An added 21-day rat group was also negative for parasites. conducted to compare the susceptibility of search Division, Agricultural Research Center, gerbils and young white rats to simultaneous Beltsville, Maryland, and the S-.B/P strain infection with both T. axei and T. colubri- (Sprague-Dawley rats crossed five times with formis. Also, we wished to confirm, if possible, NIH black rats by S. Poiley, then inbred) the unique report of Williams and Palmer obtained from the National Cancer Institute, (1964) that young white rats are satisfactory NIH, Bethesda, Maryland. Because Williams hosts for T. colubriformis, and Palmer (1964) inoculated young white rats weighing 50 g (strain not specified) with Materials and Methods T. colubriformis larvae, we inoculated our rats The gerbils (Meriones unguiculatus) used in with larvae when their average weight was these experiments were either purchased from 50 g (range 40-60 g) and when they were Tumblebrook Farm, Brant Lake, New York, 27 to 34 clays old. or obtained from a small laboratory breeding Three experiments (Tables 1-3) were con- colony. At the time of larval inoculation the ducted to compare the susceptibility of gerbils gerbils averaged about 5 months of age and and young white rats of both sexes to simul- about 65 g (range 50-75 g) in weight. taneous infection with the two aforementioned Two strains of young white rats were used, species of Trichostrongylus. Each experiment namely, a modified Wistar strain obtained usually consisted of two groups of gerbils and from a colony maintained for over 30 years rats; one group was necropsied for quantifica- at the Dairy Branch, Animal Husbandry Re- tion of the infections on the 14th day post-

Table 2. Experiment II. Trichostrongylus spp.1 recovered at necropsy from gerbils and young white rats (S-B/P strain). Larval inoculation 1,000 ±11 SE/animal; culture 59a.

Gerbils Rats 5M 5F All 5M 5F All GROUP A. 14 clays postinfection T. axei 361 (255-598)-' 203 (115-328) 282 (115-598) 7 (0-34) 5 (0-12) 6 ( 0-34 ) T. colubriformis 65 (9-140) 42 (16-101) 53 ( 9-140 ) 12 (0-29) 12 ( 0-30 ) 12 ( 0-30 ) Both species 426 (332-738) 245 (131-368) 335 (131-738) 19 ( 0-63 ) 17 (0-30) 18 ( 0-63 ) GROUP B. 28 clays postinfection''1 T. axei 177 (131-220) 252 (186-306) 214 (131-306) 0 0 co o T. colubriformis 36 (0-77) 33 (10-96) 35 ( 0-96 ) 0 0 Both species 213 (151-283) 285 (201-402) 249 (151-402) 0 0 Composite EPG 639 390 514 0 0 0

1 Nematodes in 4th or early 5th stage at 14 clays PI, feces negative for eggs; 99+% were mature at 28 days PI. - Average, followed by range in parentheses. 3 Only 4M and 4F gerbils in this group.

Table 3. Experiment III.1 Trichostrongylus spp.2 recovered at necropsy from gerbils and young white rats (S..B/P strain). Larval inoculation 1,260 ± 16 SE/animal; culture 59b.

Gerbils Rats 4M 4F All 5M 5F All GHOUP A. 14 days postinfection T. axei 202 (160-282)-' 96 (28-165) 149 (28-282) 4 (1-11) 4 (0-12) 4 (0-12) T. colubriformis 117 (79-180) 88 (48-152) 103 (48-180) 29 (7-59) 13 (7-23) 21 (7-59) Both species 319 (240-462) 184 (76-317) 252 (76-462) 33 (8-64) 17 ( 7-35 ) 25 (7-64) GROUP B. 28 days postinfection T. axei 231 (190-271) 221 ( 143-332 ) 226 (143-332) 0 0 T. cohibriformifi 64 (6-122) 80 (21-109) 72 (6-122) 0 0

Both species 295 (196-390) 301 (234-412) 298 (196-412) oo o 0 0 Composite EPG 1,236 1,851 1,543 0 0 0

1 This experiment was a replication of Exp. II, but another larval suspension from the same culture was employed. - Nematodes in 4th or early 5th stage at 14 days PI, feces negative for eggs; 99+% were mature at 28 days PI. :! Average, followed by range in parentheses. infection (the time of necropsy reported by Results and Discussion Williams and Palmer, 1964), and the other The results obtained are summarized in group was necropsied 28 days postinfection Tables 1-4. Substantial infections, similar to when the bulk of the parasite populations are those previously reported by vis (1967), were usually mature. In experiment I only (Table established on the average in both male and 1), a third group of infected rats was included female gerbils in all experiments, although an and these were necropsied 21 days postinfec- occasional gerbil had minimal infection of one tion for the purpose of obtaining worm reten- or the other species of Trichostrongylus; one tion data for the interval between 14 and 28 male gerbil in Group B, Experiment II, had days postinfection. Each group of gerbils and no T. colubriformis at necropsy. The average rats in all three experiments consisted of sub- percentages of the larval inocula recovered groups of four or five animals of each sex; these from all gerbils (Table 4) at 14 days post- subgroups were maintained in separate small infection were 15.3, 33.5, and 20, and at 28 animal cages. The animals were given a stan- days postinfection were 9.8, 24.9, and 23.6, dard, pelleted feed and water ad lib. for experiments I to III, respectively. Similar The procedures used for culture of Tricho- percentages of worm recoveries from young strongylus spp. larvae from feces of infected white rats were 0.3, 1.8, and 2 at 14 days lambs, for larval inoculations, for necropsy of postinfection; zero in experiment I at 21 days the animals, for recovery and enumeration of postinfection; and zero in all three experiments the worm populations, etc. were identical with at 28 clays postinfection. Some of the rats had those previously reported by us (1967). no parasites 14 days postinfection, and no Larval cultures were prepared from feces of parasites at all were recovered from a total of lambs with substantial mixed infections of T. 40 inoculated rats 21-28 days postinfection. axei and T. colubriformis only. The percentage Furthermore, the few nematodes recovered of infective larvae of each species in the larval from some rats at 14 days postinfection were suspensions was not determined prior to animal all immature, as were all the nematodes re- inoculation, because of difficulties in quantita- covered from all gerbils necropsied at 14 days tively differentiating infective larvae of these postinfection; no eggs were seen in the feces two closely related species. The larval closes of any of the animals at this time. per animal in experiments I to III were 975, The data on worm recoveries from gerbils 1,000, and 1,260, respectively. and rats showed no consistent differences in Prior to necropsy of the animals, 24-hour sex susceptibility for either host. The overall composite fecal samples were collected from average percentage of larval inocula recovered each sexed subgroup of gerbils and rats, and from both male and female gerbils in three the numbers of trichostrongyle eggs per gram experiments was 19.4 (Table 4), and there of feces were determined by the zinc sulfate was no significant differences in the small direct centrifugal flotation method. numbers of immature parasites recovered from

Table 4. Summary of average per cent recovery of larval inocula of Trichostrongyhis spp. at necropsy from gerbils and young white rats in three experiments.

Per cent of inocula recovered at necropsy Gerbils Rats Male Female All Male Female All

Experiment I ( Gerbils vs. Wistar Rats )l Group A: 14 days PI 15 15,5 15.3 0.4 0.2 0,3 Group B: 28 days PI 13.6 6 9.8 0 0 0 Experiment II ( Gerbils vs. S.-.B/P Rats ) Group A: 14 days PI 42.6 24,5 33.5 1.9 1.7 1.8 Group B: 28 days PI 21.3 28.5 24.9 0 0 0 Experiment III (Gerbils vs. S.-.B/P Rats)- Group A: 14 days PI 25.3 14.9 20 2.6 1.3 2 Group B: 28 days PI 23.4 23.8 23.6 0 0 0 Avg at 14 days PI — All experiments 27.6 18.3 22.9 1.6 1.1 1.4 Avg at 28 days PI — All experiments 19.4 19.4 19.4 0 0 0 Avg 14 + 28 day PI data — All experiments 23.5 18.8 21.1

1 A similar group of infected rats necropsied at 21 days postinfection (PI) were negative for parasites. - This experiment was essentially a replication of Experiment II. male and female rats at 14 days postinfection. have become better adapted to rat passage Much greater variation occurred in the fecal than a strain used directly from ovines. egg counts of the gerbil groups at 28 days It was also reported by Williams and Palmer postinfection than in the necropsy worm counts (1964) that the T. colubriformis recovered in the three experiments. The composite egg from young rats were adults 14 days postinfec- counts varied from 10 EPG in the female sub- tion, and that young male rats were more sus- group in experiment I to 1,851 EPG in the ceptible to infection than young female rats. female subgroup in experiment III. However, We were not able to confirm either of these in experiments II and III (Tables 2, 3) the observations. All specimens we recovered at EPG counts of the male and female subgroups 14 days postinfection from gerbils and rats in each experiment were relatively consistent, of both T. colubriformis and T. axei were im- 639 and 390 EPG in experiment II and 1,236 mature although some were in the early fifth and 1,851 EPG in experiment III. stage, and no significant differences were noted That substantial numbers of T. colubriformis in the susceptibility of male and female rats to larvae were present in our larval suspensions infection. is shown by average recoveries of 137, 53, and 103 T. colubriformis per gerbil at 14 days post- Summary infection, and 64, 35, and 72 per gerbil at 28 Young white rats of two strains were almost days postinfection. The apparent success re- completely refractory to simultaneous infection ported by Williams and Palmer (1964) in with Trichostrongylus axei and T. colubri- inducing significant infections in young white formis, whereas Mongolian gerbils under iden- rats with T. colubriformis (no detailed data tical conditions were readily infected with both were reported), and our failure to do so, is species. No significant differences by sex were difficult to explain. Some possible reasons for noted in the susceptibility of either host to these different results are as follows: (1) The infection with these nematodes. rat strain used by the authors above was more susceptible to infection than our strains. (2) Literature Cited They employed pure cultures of T. colubriformis, whereas we used mixed cultures of two Kates, K. C., and D. E. Thompson. 1967. Ac- species. (3) Their source of eggs for culture tivity of three anthelmintics against mixed infections of two Trichostrongylus species in of infective larvae of T. colubriformis was from gerbils, sheep, and goats. Proc. Helm. Soc. a rabbit adapted strain, which had been serially Wash. 34: 228-236. passaged 50 times in this host, and thus may Leland, S. E. 1961. Some aspects of experimcn-

tal infection of the Mongolian gerbil (Meri- ostrongylus axei in the Mongolian gerbil as a ones unguiculatus) with Trichostrongylus axei. screening system for anthelmintics of domestic J. Parasit. 47 (4, sec. 2): 19. animals. J. Parasit. 49 (5, sec. 2): 15-16. . 1963a. Studies on Trichostrongylus axei Williams, G. A. H., and B. H. Palmer. 1964. (Cobbold, 1879). VIII. Some quantitative Trichostrongylus colubriformis (a nematode aspects of experimental infection of the Mon- parasite of sheep and other ruminants) as a golian gerbil (Meriones unguiculatus). J. test organism in screening for sheep anthel- Parasit. 49: 617-622. mintics in the laboratory. Nature 203: 1399- . 1963b. Preliminary evaluation of Trich- 1400.

A Technique for Isolating and Maintaining Cultures of Meloidogyne

JACQUELINE R. SHEPPERSON AND WILLARD C. JORDAN Department of Biology, Winston-Salem State College, Winston-Salem, North Carolina 27102

Various species of Meloidogyne cause root puncturing aerial organs (stems and leaf mid- galling on a variety of cultivated plants (Chit- ribs) of susceptible plants with glass capillary woocl, 1949). In nature, the endoparasitic tubes containing a known number of Me- female worms deposit eggs in roots or soil loidogyne incognita larvae or eggs. In the and the larvae upon hatching penetrate rootlets stem or leaf, the nematodes mature and pro- to feed and mature. Under optimum condi- duce offspring that have not been exposed to tions, the life cycle is completed in 33 days contamination with other soil borne nematode (Peacock, 1959). Linford (1941) demon- species. This technique is a simple, inexpen- strated that root-knot nematodes can develop sive method for maintaining an abundant sup- and reproduce in leaves and stems of tomato ply of Meloidogyne incognita in the laboratory. plants, Coleus, Begonia, etc. Under the influ- By utilizing glass capillary tubes for inocula- ence of successive generations of parasites, tions, single egg sacs can be isolated to estab- massive galls containing nematodes in all lish and perpetuate pure nematode cultures. developmental stages develop on roots as well as aerial parts of the hosts. Materials and Methods Krusberg (1963) reviewed host responses to The inocula consisted of viable larvae or root nematodes and indicated that many as- ruptured egg sacs suspended in sterile water. pects of host—nematode interactions require Egg masses were removed from the surface of further investigations. He pointed out that the infected tomato roots with forceps, placed in lack of sufficient specimens frequently impeded hanging drops and incubated at 27 C for 3 the progress of physiological studies. Plant days. Hanging drop preparations were kept tissue cultures have been utilized to propagate in moist chambers to prevent desiccation. The some species of tissue invading nematodes. In hatched larvae were suspended in water on most studies on Meloidogyne spp., investigators shallow depression slides. After about 30 min, have employed larvae or eggs from roots grow- larvae tend to congregate at the center of the ing in infested soil. Powell and Moore (1961) concavity. Finely drawn capillary tubes (1 mm described a technique for cultivating Me- in diameter and 50 mm in length) were used to loidogyne incognita acrtta which involved the pick up the concentrated larval suspensions inoculation of larvae into leaves with a hypo- from the depression slide preparations. Using dermic syringe. 40 X magnification, the larvae drawn into each The technique described herein involves capillary tube were counted. The inocula used

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 107 contained 1, 25, or 50 larvae. Glass capillary The life cycle of M. incognita in Begonia tubes containing macerated egg masses were leaves and stems and in the tomato stems used to inoculate some host plants. paralleled that in the tomato roots. The first Capillary tubes containing larvae or eggs visible swellings appeared in about 7 to 10 were inserted into stems and leaf midribs of days. Inoculum containing only one larva did Begonia, young tomato, peanut, and potato not initiate infection. All susceptible plant plants. Beginning at the base of stems, wounds organs that were inoculated with macerated were made 1 inch apart ascending the stem suspensions of egg sacs or 50 larvae showed until three capillary tubes had been so placed. swellings at the site of inoculation by the 8th The tubes were placed in a downward direc- day; inoculum with 25 larvae produced en- tion in the stems at a 45° angle to the organ largements by day 10. so that the larvae or eggs could descend into Microscopic examinations of infected tissues the plant tissues. Single capillary tubes were from roots, stems and leaves showed nematodes inserted into midribs of individual leaves of in similar developmental stages. In 30-day-old the four host plants. Roots of six tomato plants infected materials, most female worms had were exposed to larvae in order to compare begun to secrete egg masses. Adult male worm development in aerial and underground worms were occasionally observed in the organs. tissues. All inoculated plants were grown under As development progressed, some egg greenhouse conditions. Observations were masses protruded through the epidermis. The made for at least 2 weeks on all plants. Some egg sacs appeared on the surface as small plants were sacrificed and examined at weekly dark elevations. These external sacs were intervals; others were examined and studied easily removed with forceps without injury for 3 months. Microscopic examinations of to the egg-producing females or the host fresh and fixed sections were made period- plants. Eggs in external sacs did not hatch ically. For the demonstration of nematodes in until they were removed and placed in water. roots, stems, and leaves, sections were pre- Eggs embedded in the cortex or pith hatched pared and stained according to the methods and larvae migrated to other sites in the stem of McBeth et al. (1941). or leaf. Several generations of nematodes were Results and Discussion produced in a host plant. Thus such cultiva- tions circumvent frequent transfer of worms. Results of this study indicate that the stems With host sacrifice or removal of infected of tomato and Begonia plants are ideal ma- leaves, nematodes embedded deep in stem or terials for maintaining uncontaminated Me- leaf tissue serve as a source of uncontaminated loidogyne populations in the laboratory. The material. The worms can be dissected from the growth of worms in midribs of Begonia ribs tissues and used in various types of studies. affords an easily accessible plant part for Meloidogyne development. By utilizing glass Summary capillary tubes, the number of nematodes con- Pure cultures of the root-knot nematode, tained in an inoculum can be readily ascer- Meloidogyne incognita, can be established in tained. The capillary tubes also mark the site aerial organs of Begonia and tomato plants. of inoculation. The midribs of Begonia leaves, tomato, and By inoculating stems with glass capillary tubes Begonia stems supported growth and matura- containing sterile eggs or larvae, several generation of Meloidogyne incognita. Worms did not tions of the nematodes are produced in a single thrive in tomato leaves, nor in leaves and stems host plant. Worms that develop in aerial of potato and peanut plants. Apparently, the organs are not exposed to soil-borne contam- leaves were irreparably damaged during initia- inants. Egg sacs that protrude through the tion of infections. In most cases, these leaves stem epidermis are easily accessible and can withered and died in the early stages of the be removed from the plant with forceps. experiment. Extensive mechanical damage did Midribs of Begonia plants support growth of not occur in the larger midribs of Begonia nematodes and produce a copious supply of leaves that were inoculated. material for various investigations.

Literature Cited McBeth, C. W., A. L. Taylor, and A. L. Smith. Chitwood, B. G. 1949. "Root-Knot Ncma- 1941. Note on staining nematodes in root tocles"—Part I. A revision of the genus Me- tissue. Proc. Helm. Soc. Wash. 8: 26. loidogyne Goekli, 1887. Proc. Helm. Soc. Peacock, F C. 1959. The development of a Wash. 16: 90-104. technique for studying the host parasite rela- Krusberg, L. R. 1963. Host response to nema- tionship of the root knot nematode Meloi- tocle infection. Annu. Rev. Phytopathol. 1: clogyne incognita under controlled conditions. 219-240. Nematologica 4: 43-55. Linford, M. B. 1941. Parasitism of the root- Powell, N. T., and E. L. Moore. 1961. A knot nematode in leaves and stems. Phyto- technique for inoculating leaves with root- pathology 31: 634-648. knot nematodes. Phytopathology 51: 201-202.

A Peripheral Nervous System in Nematoda with a Discussion of its Functional and Phylogenetic Significance

N. A. CROLL* AND A. R. MAGGENTI Department of Nematology, University of California, Davis

Previously, a technique has not been recog- tonic media because of the influx of water nized for demonstrating a peripheral nervous (Croll and Viglierchio, in press). The rapid system in nematodes. This has probably been exchange of water resulting from the osmotic due to the inadequate specificity of the stains differences of the immersion media, has been tried, or the inability of the stains to penetrate employed to flush dilute AgNO8 through the the cuticle. Using the new physiological stain- nematode cuticle. ing technique described below, a peripheral Living T. californicum were transferred from nervous system of longitudinal and transverse seawater to 10 per cent NaNOs for five min- nerves has been observed in a nematode for utes, after which there was a considerable the first time and is here described as it occurs reduction in size. They were then placed in in Thoracostoma californicum (Steiner and 0.5 per cent AgNO;! for 15 seconds, and finally Albin, 1933) Weiser, 1953. transferred to distilled water. Because of prior In describing the peripheral nervous system exosmosis in NaNO3, when placed in hypotonic we have adopted a terminology consistent with AgNO3, endosmosis occurred causing an ex- that standardized for invertebrate nervous sys- pansion of the worm and dilute silver was tems by Bullock and Horridge (1965) and that flushed into the tissues. The uptake of silver utilized by Chitwood and Chitwood (1950). was further enhanced when the nematodes were placed in distilled water. Upon immer- Materials and Methods sion in dilute silver, the nematodes became Thoracostoma californicum were collected moribund and activity stopped after a few from the holdfasts of Laminaria digitata and minutes in distilled water. After illumination Egregria laevigata at lowtide from Dillon the peripheral nervous system of nematodes Beach, California. Specimens were stored at processed in this way became apparent through 5 C iv\d seawater which was changed the deposition of colloidal silver. at weekly intervals. This technique, depending as it does on the When placed in hypertonic solutions, T. osmotic properties of the media has been found californicum becomes reduced in size through successful only for living worms and is not exosmosis, and expands if transferred to hypo- satisfactory for dead or fixed specimens. Small cylindrical portions of the stained worm about * Fulbright Scholar: Department of Zoology, Imperial College, London. 0.3 mm long were excised and then cut longi-

Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 35, NUMBER 1, JANUARY 1968 109 tudinally on one side, opened, flattened, fixed are connected by transverse mtra-neural com- in 2 per cent formalin and mounted on a glass missures (Fig. 3A). slide. These together with totomounts were the LONGITUDINAL PERIPHERAL NERVES: Dorsal basis of the observations and descriptions re- peripheral nerve. Anteriorly the dorsal periph- ported here. The peripheral nervous system eral nerve (dpn) consists of two elements also remained visible in glycerin and persisted (Fig. 2B), the peripheral dorso-dorsals (ddpn). even after drying. These longitudinal nerves unite approximately 1 mm from the anterior extremity to form Morphology of the Peripheral a single longitudinal dorsal peripheral nerve Nervous System (dpn), (Fig. 1C). In this manner the nerve BASIC PATTERN: Along the entire length of extends along the remaining length of the T. californicum there is a peripheral system nematode. consisting of a latticework of nerves. This The peripheral dorso-dorsals are connected latticework appears to form a completely uni- by transverse intra-neural dorso-dorsal com- fied system with apparent cytoplasmic con- missures (ddc). tinuity between all the nerve fibers, architec- LATERAL PERIPHERAL NERVES: Anteriorly turally consistent with the orthogon plan. The the lateral peripheral nerves (Ipii) may be bilaterally symmetrical system is composed of single but splits to form paired nerves, the transverse and longitudinal elements associated dorso-lateral peripheral nerve (dlpn) and the with the outermost region of the hypodermis. ventro-lateral peripheral nerve (vlpn), (Fig. The latticework is further complicated by the IB). The split becomes definite approximately presence of local elaborations of nerve fibers 1 mm behind the anterior extremity. These or plexuses. The above described pattern (Fig. longitudinal nerves are latero-lateral commis- 1A) is common to all specimens of T. cali- sures (lie) which occur at regular intervals fornicum examined, within the common plan, along the length of the body (Fig. 3A). however, there is a degree of individuality. VENTRAL PERIPHERAL NERVE: The ventral The differences in the basic transverse peripheral nerve (vpn) consists of two longi- nerves are often correlated with the distribu- tudinal ventro-ventral peripheral nerves (vvpii) tion of somatic setae. All of the setae, includ- which are joined by transverse intra-neural ing the cephalic setae, are connected to the ventro-ventral commissures (we), (Fig. 1A peripheral nervous system. This is most appar- and 2C). The ventro-ventral nerves widen ent in the plexuses where the increase in the around the vulval region and are elaborated density of neural anastomses is correlated with into a vulval plexus (Fig. 2C). an increase in the density of setae. When the TRANSVERSE COMMISSURES FORM AND FRE- setae are not directly over a major element QUENCY: The frequency of the inter- and of the peripheral latticework, a separate minor intra-neural commissures is not constant, even neural connection appears to join the scolopoid though the peripheral neural net is continuous body (Maggenti, 1964) to a major element in all directions. All inter-neural commissures (Fig. 2C). When the setae are situated occur at relatively constant intervals of 25— directly over a major element they appear to 35 p. They normally consist of single nerves connect directly with it by way of the scolopoid that directly link two longitudinal peripheral body (Fig. ID). However, there may be a nerves. Anteriorly, however, they frequently connection between the scolopoid and a major inscribe a posteriorly directed curve (Fig. 2A). peripheral nerve which is not apparent with These nerves may also fuse or be incomplete, the light microscope. The detailed irregu- looping back onto the nerve of origin without larities and asymmetries of the major periph- completing the transverse inter-neural com- eral nerves may be seen in Figures 1 and 2. missure (Fig. 2C). The different series of longitudinal periph- The latero-lateral intra-neural commissures eral nerves are connected to one another by occur at fairly constant intervals but are transverse inter-neural commissures: dorso- slightly less frequent than the inter-neural lateral commissures (die) and the latero- commissures. The ratio between these intra- ventral commissures (Ivc). The paired sub- and inter-neural commissures is 1:1 or 2 divisions of each longitudinal peripheral nerve (Fig. IB).

Anterior to the dichotomy of the dorsal atoda is based on: (1) the definite connection peripheral nerve, the ratio of the intra-neural of setae with the peripheral latticework; (2) dorso-dorsal commissures to the inter-neural the ability to stain with silver; (3) the simi- commissures is approximately 1:3 or 4 (Fig. larity between the staining reaction of the 2B). setal filaments, scolopoid body and peripheral The frequency of ventro-ventral intra-neural fibers; (4) the connection of the scolopoid commissures shows a pronounced localized body with the central nervous system, and (5) variation in specific regions of the body, this the cephalization of the peripheral network variability was not present in the other intra- and its elaboration into plexuses both in re- neural commissures. Anteriorly there is a ratio gions where sensory stimuli could be of special of approximately 1:1 with the inter-neural significance. These observations are further commissures (Fig. 2A); over the rest of the supported by the comparative morphology of body (excepting the vulval and anal regions) invertebrate nervous systems; the size and the ratio of intras to inters is approximately nature of the fibers; and the fact that this 1:4 to 6 (Fig. 1A). system does not correlate with any other non- RADIALLY SYMMETRICAL PLEXUSES: At the nervous systems previously described in nem- anterior extremity there is a cephalic plexus atodes or other invertebrates. consisting of a whorl of nerves of almost equal The discovery of the peripheral nervous sys- mesh, supplying the cephalic setae in the form tem requires that our understanding of the of a corona around the cephalic capsule (Fig. central nervous system be reviewed. It is im- 2A). portant that existing knowledge be used to Posterior to the cephalic plexus and extend- integrate the two systems into one functional ing for approximately 1 mm, in adult worms, whole. In doing so it is necessary to compro- there is a concentration of nerves here desig- mise or refute earlier assumptions, not only of nated as the cervical plexus. This region is the nervous system but of the morphological demarcated anteriorly by the cephalic plexus configuration of the hypodermis. and posteriorly by the split in the dorsal Chitwood (1950) reported that the genital peripheral nerve and the fusion of the lateral papillae connect with the central nervous sys- peripheral nerves (Fig. 2A, 2B). tem by way of the ventral nerve, ventro-lateral ZYGOMORPHIC PLEXUSES: Approximately 0.4 nerves and the caudal nerve through the lum- mm anterior and posterior to the vulva and bar ganglia. Maggenti (1964) concurred in within the ventro-ventral peripheral nerves, these observations for T. californicum. In the there is an increase in the density of trans- anterior end of T. californicum Hope (1964) verse intra-neural ventro-ventral commissures described pseudocoelomic chords containing (Fig. 2C). Coincident with this increase in large numbers of axonic fibers. The numbers density of the intra-neural commissures, there of axonic fibers within these chords closely is a divergence of the ventro-ventral peripheral correlates with the number of cervical setae. nerves to encompass the vulva. Maximum den- Therefore, the somatic setae and papillae have sity of intra-neural commissures is reached at several pathways whereby impulses can be about 150 ^ from the vulva; this area is further transmitted either directly to the circum- complexed by the addition of minor longitu- esophageal commissure or through transverse dinal nerves forming a dense network here commissures to motor centers. designated as the vulval plexus. The peripheral neural net is considered here Discussion to be sensory rather than motor. The neural The conclusion that we have observed and net is a continuous meshwork from anterior to described a peripheral nervous system in Nem- posterior, linking throughout the somatic setae

Figure 1. Thoracostoma californicum stained with silver. A. Flattened integument anterior to vulva split along dorsal line showing the relationships of all of the peripheral nerves. B. Lateral peripheral nerve showing intra- and inter-neural commissures. C. Dorsal peripheral nerve with details of incomplete inter-neural commissures and anastomoses. D. Scolopoid hody in relation to inter-neural commissures and its connection to the ventral peripheral nerve (oil immersion).

\ \

100 ji

Figure 2. Peripheral nervous system of Thoracostoma californicum, camera lucida illustrations. A. Ventral view, cephalic and cervical plexus. B. Dorsal view, dichotomy of dorsal peripheral nerve. C. Ventral view, vulval plexus.

Ivc —

vvpn vpn we vvpn

Figure 3. A. Nomenclatorial diagram: dpn, dorsal peripheral nerve; ddpn, dorso-dor.sal peripheral nerve; ddc, dorso-dorsal commissure; die, dorso-lateral commissure; Ipn, lateral peripheral nerve; dlpn, dorso-lateral peripheral nerve; Ivc, latero-ventral commissure; vpn, ventral peripheral nerve; vvpn, veil- tro-ventral peripheral nerve; we, ventro-ventral commissure. B. "Epithelial Cells" (redrawn after Retzius (1906)). C. Schematic hypodermal cell plan (redrawn after Filipjev (1924)).

and papillae. In T. californicum the density tern offers a limited means for communicating of the pattern is directly correlated with the the environment to the nematode. In the density of the setae. absence of free nerve endings or a peripheral The only apparent connection of the network neural network under the cuticle, the localiza- with the underlying central nervous system is tion of sensory organs and the consequent lack through the scolopoid bodies. If the numbers of sensory receptors over the general body and position of setae are mapped for the surface would leave areas insensitive to stim- general body, it is apparent that the setal pat- ulation. Therefore, it may be that the network

Copyright © 2011, The Helminthological Society of Washington 114 PROCEEDINGS OF THE HELMINTHOLOGICAL SOCIETY acts to coordinate impulses from seta to seta In addition, the staining reactions resemble the and thence to the central nervous system. Such epithelial cell walls described by Retzius a neural network would then compensate for (1906), who also employed a silver technique. the paucity of sensory organs. This is not to Our findings lead us to believe that what imply that the peripheral network may not also Retzius in 1906 referred to and illustrated as function in its own capacity, that is, responsive the epithelial cell walls of Oncliolaimii.s vul- to special external stimuli not necessarily re- garis (Fig. 3B) and Enoplus communis were lated to that which will stimulate setal con- in reality elements of the peripheral nervous nections. In which case the message relayed system. The similarity between his figures and to the central nervous system from setal stim- the peripheral nervous system of T. califor- ulation would be entirely different from that nicitm as described here is unmistakable. relayed by the peripheral nervous system Retzius' misinterpretation in nematocles may per se. This latter assumption correlates well have been influenced by his earlier descrip- with the increased density of setae in the an- tions of epithelial cells in other lower inverte- terior region, at the vulva and in the caudal brates (1902). region of males. Concentrations of setae in The misinterpretation of Retzius has implica- the localized areas where sensory stimuli could tions on the understanding of basic nematode be of special significance would seem to indi- morphology. The distribution of hypodermal cate that setal nerves relay specific information. cells described by Filipjev (1924) perpetuated The peripheral nervous system and the cen- the error (Fig. 3C). The classic illustration of tral nervous system both follow an orthogon Retzius (1906) with its original designation plan and there is a slight measure of similarity has appeared in many major compendia on between the two systems. In both, the longi- nematodes (Chitwood and Chitwood, 1950; tudinal fibers are closely associated with dorsal DeConinck, 1965; Hyman, 1951 and Kreis, lateral and ventral lines. Anteriorly, the cen- 1934). tral nervous system does not clearly follow the A peripheral nervous system of the type sagittal and frontal planes, neither does the described above occurs in both platyhelminths peripheral nervous system. The cervical plexus and priapulids, but not in rotifers, gastrotrichs, of the peripheral nervous system ends posterior and kinorhynchs, although extensive studies of to the nerve ring and is a more radially sym- these groups have been made (Bullock and metrical basket than the plan in the remainder Horridge, 1965). This fact, together with of the body. This change being associated Clark's (1963) reassessment of the phylo- with setal distribution. Anteriorly modifica- genetic significance of a coelom and pseudo- tions are, for the most part, with the number, coelom may again open the question of the position and cephalization of sensory organs. evolutionary relations of these groups to It seems significant that alterations in both nematocles and the validity of the phylum systems are associated with corresponding Aschelminthes. changes in the general pattern of sensory organs. For example, the vulval plexus with Summary the increase in setal density is correlated with A new physiological technique using silver the vulval ganglia in the central nervous sys- nitrate has been employed to demonstrate a tem; the same sort of manifestation occurs in hitherto unknown peripheral nervous system in the caudal region. Circumstantially these ob- the marine nematode Thoracostoma calif or- servations are accepted as further support for nicum. This system is described and a no- the connection of the peripheral nervous sys- menclature proposed for the elements of the tem to the central nervous system through the orthogon plan. The functional significance sensory organs. and importance of this system to Aschelminthes It is not known whether the peripheral is discussed. nerves are covered by glial cells but the sim- Literature Cited ilarity in staining of the scolopoid bodies and Bullock, T. H., and G. A. Horridge. 1965. nerves possibly suggest a continuous neuri- Structure and function in the nervous system lemma unlike the unmyelinatecl axons reported of invertebrates. Vol. I and II, pp. 1719. for Haemonchus contortns larvae (Ross, 1967). W. H. Freeman and Co., San Francisco.

Chitwood, B. G., and M. B. Chitwood. 1950. morphology of Thoracostoma californicum An introduction to nematology. Baltimore Steiner and Albin, 1933. Ph.D. thesis, Uni- Monumental Publishing Co., Sec. 1, Anatomy. versity of California, Davis. Rev. Ed. 213 pp. Hyman, L. H. 1951. The invertebrates. Vol. Clark, R. B. 1963. The evolution of the celom Ill: Acanthocephala, Aschelminthes and En- and metameric segmentation. The lower nieta- toprocta—the Pseudocoelomate Bilateria. Mc- zoa, comparative biology and phylogeny. Graw-Hill, New York, Toronto, London, 572 Edited by Dougherty et al., Univ. of Calif. pp. Press, pp. 91-107. Kreis, H. A. 1934. Oncholaiminae Filipjev 1916 Croll, N. A., and D. R. Viglierchio. 1968. eine monographische studie. Capita. Zool., Osmoregulation in a marine nematode. In 4(5): 1-270. press. Maggenti, A. R. 1964. Morphology of somatic DeConinck, L. 1965. Traite de zoologie anat- setae: Thoracostoma californicum (Nemata: omic, systematique, biologic. Published under Enoplidae). Proc. Helm. Soc. Wash. 31(2): Pierre P. Grasse. Nemathelminthes (Nema- 159-166. todes) Tome IV Sascicole II. Masson et Cie Ret/ius, G. 1902. Weiteres zur Kcnntniss der Editeurs Libraires dc L'Academie de Med- Sinneszellen der Evertebraten. Biol. Unter- ecine, pp. 732. such. n.F. 10: 25-33. Filipjev, I. N. 1924. Uber das /ellmosiak in . 1906. Zur Kenntniss der Hautschicht der epidermis von Paroncholaimus zernovi. der Nematoden. Biol. Untersuch. n.F. 13: Zool. Anz., 61(11/12): 268-277. 101-106. Hope, W. D. 1964. A taxonomic review of the Ross, Mignon M. R. 1967. Modified cilia in genus Thoracostoma Marion, 1870 (Nemafz: sensory organs of juvenile stages of a parasitic Lcptosomatidae) and a study of the histologic nematode. Science 156(3781): 1494-1495.

Presentation 1967 Anniversary Award of The Helminthological Society of Washington 429th Meeting 11 October 1967

Gerard Dikmans, whom we honor at this According to his own account, he went to meeting with The Anniversary Award of the Louisiana in 1920, fully intending to be a Helminthologieal Society of Washington, is bacteriologist and pathologist. However, the recognized as one of the outstanding veterinary late Drs. W. H. Dalrymple and B. H. Ransom parasitologists of the United States. persuaded him to undertake some parasi- He was born at Bolsward, Friesland, Nether- tological work for the Louisiana Agricultural lands (1885), and came to this country in Experiment Station. He became intensely in- 1905. He holds a B.A. degree from Washing- terested in parasitology and never left this ton College, Tennessee (1914); B.S.A. from field of research. the University of Tennessee (1917); D.V.M. From 1922 to 1924, Dr. Dikmans was Assist- from Michigan State College (1920); M.S. ant Parasitologist at the University of Minne- from the University of Minnesota (1927); and sota. He was appointed Associate Parasitologist Ph.D. from Georgetown University (1931). at the Puerto Rico Agricultural Experiment Sta- From 1914 to 1917, Dr. Dikmans was assist- tion (USDA), Mayaguez, Puerto Rico, in 1924 ant in bacteriology, University of Tennessee, and remained in Puerto Rico until 1926. At and from 1920 to 1922, he was Assistant Pro- this time, he was transferred to the Zoological fessor of Veterinary Science and Assistant Division of the Bureau of Animal Industry and Veterinarian at the Louisiana State Agricultural was stationed at Jeanerette, Louisiana, until Experiment Station, Baton Rouge. 1929. He was then moved to Washington,

senior author, of eight. Eighty-three of the papers were written on the subject of worm parasites of ruminants, 12 on trichomoniasis of bovines, ten on anaplasmosis, ten on general parasitology, seven on the worm parasites of rodents, two on anthelmintics, two on nasal granuloma, and 18 on miscellaneous parasites. He was responsible for the original descriptions of 26 new species of nematode parasites: 15 from rodents and lagomorphs, 10 from ruminants, and one from the opossum. On July 11, 1949, Dr. Dikmans received the annual Twelfth International Veterinary Congress Prize at the meeting of the American Veterinary Medical Association held in Detroit, Michigan, in recognition of his life's work which, according to President Hurt of that Association, had done so much to keep the study of parasitology abreast of the more spectacular branches of medical science. Presi- dent Hurt also stated that the honor marked Dr. Gerard Dikmans Dr. Dikmans as one of America's foremost veterinarians whose accomplishments rank with D. C., and remained there and at Beltsville, those of L. G. Neumann, A. Railliet, Cooper Maryland, until his retirement on May 31, Curtice, B. H. Ransom, Maurice C. Hall, and 1953. In October 1957, Dr. Dikmans became such celebrities as James Law and W. H. associated with the State Animal Disease Diag- Dalrymple, who foresaw the significance of nostic Laboratory, Kissimmee, Florida, where metazoal pathology in animal production. he remained about a year. He now lives in Dr. Dikmans was a member of the Com- Ionia, Michigan. mittee on Parasitology of the American Vet- Although Dr. Dikmans was interested in all erinary Medical Association from its inception parasites of all animals, it was necessary for in 1939-40 through 1947 and served as its him to confine his principal interest to one Chairman in 1945. He held the office of group, because the field of parasitology takes Recording Secretary of the Helminthological in far more territory than any one man can Society of Washington and was an ex-officio cover. His publications show that his chief member of the editorial committee of its interest was centered in the parasites of rumi- Proceedings during the year 1934-35. On nants, both domestic and wild. He was inter- July 17, 1936, he helped the Society establish ested in these parasites as causative agents of the Brayton H. Ransom Memorial Trust Fund. disease, but he did not neglect other phases Dr. Dikmans was President of the Helmintho- of the subject, such as taxonomy and morphol- logical Society of Washington during the year ogy, because he felt that he had to know the 1937-38. In 1943-44, he served on the com- parasites he encountered in order to learn how mittee appointed to revise the Society's con- to deal with them. The helminth parasites of stitution. He is a life member of the Society. cattle, sheep, goats, and deer, and anaplasmosis To honor him for his accomplishments in and bovine trichmoniasis, have claimed the helminthological research and in appreciation major portion of his time. of his contributions to and interest in the During the 36-year period from 1921 to Society, The Helminthological Society of Wash- 1957, Dr. Dikmans was the author or coauthor ington is privileged to present to Gerard Dik- of 144 papers on parasitological subjects. He mans this Anniversary Award.— (Committee: was the sole author of 117 of these papers, the Lund, Sadun, Becklund. Presentation: John senior author of 19, and the coauthor, but not S. Andrews.)

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Bruce, J. I., Jr. • , Boisvenue, R. Jr •'••-,••' Go6d.;J.xM., Jr. ,, > i- "/•-^"' '^Cairns, E. J. • /'- '->-•'_, Buchheit, J. R. - / , Cable, R.M. : ^ Hechler, :Helen Carol ..; A -v Ford.B. R. ,. ^ M Burke, J.C. : ' Ehrenford, F. A. , Herhch,H. ^;^J Frandsen, J. C. i. < . Duxbury, R. E. . Ferris, Virginia R. - , Herman, C.-"M. ,-. . x ; "'. ' - " Isenstein, R. S. Erickson, D. G. ': : Shumard, R. F. ' -J. , Hernandez, X.- - ' s •'>' '-•'. J Rogers, W. A. Habermann, R. T. - Thorson, R.-E. -. r Hill, C. H. Wellborn, T. L., Jr. Hope, W. D. .Iowa , •, : '. • _--- vl ; .' Hblbrook, A. A; " \^ . Alaska •.-.• . o ; • -:.. • Kenworthy, F. T. . / Blankespoor, H. D. - , Huff C^ G. ' "~- •. ' ' ; V -Tx{./," Rausch,R. ;,i lancicome, D. R. Ellis, C. J. Humphrey, Judith M. V ' !. •--•' £chuits,'L. M. :,: '•-> ; (. Martin, L. K. / \ Epstein, A. H; - v \- Jachowsld, L. A., Jr. , , • . './.-' - Arizona ' •) • -' '\ ', OKvier,L.J. / • Goss.'R. C. . •'-'. ;• i IK Jacobs, L. . I.';-, • -' ' "Anderson, G. A. , ^ Price, D. L. ' , CGreve, J.H. : - 'Jahnes, W. G. ; >/":'''<;. Nigh,E. L,Jr. ', -• • Redington, B. C. '" , Hsu,H. F. -*ates,:K.C.' j -->.'• ''-'/:^'' r Reynolds, H. W. Sadun,E. H. ~^ Norton, D- C. :'., Knight, R. A. ^. ".;.'/-- Vy WfikeS, S. N. :-;.•• , / Slie,RitaB. , i, Uhner, M. J. . x( ' ,, '"•' Krusberg, L. R. v_- '0 • Arkansas ."--V.- •< .-••"-.. Florida •.:•:,-• •v.-i>-:- Zimmermann, W. J. : ' Lawless, D. K, K ' < / , Slack, D. A. Christie, J. R. - v 1 Kansas • ;_': . \ -^ ;V? Leek, R. G. , . -'.>;-v '• "•• : (\ 'California i •- ..'.' ••;-..• ; .. : ••"-. Ellis, H. ;-::/> •••..- •.:;.,- ,' \Ackert, I.E. . •* Lesser, E.'- " '\'- ''••;•' -'; v- -•':^,tLund,E. EJ , x? ." . ' ; ', -'-.' 'Cavenessi- F;E. ; • r /Overman, Amegda J. / ,Hansen,,M;,F. "/' ,: 'Lunde,1 M. N, 1 • ••., '.'-' .(-... • • <"•:. ;' $>waa, F.L.. : •• -';' / . v. Perry;.. V. G.- -.Kentucky- ' :.,' j.. • •-; •' LuttermoserrG. W. 'r- ' • ;' x^''' : VHart, Mary Ann /. Premvati, Mrs. Chapman, R. A.- / - , ; | Malakatis, G. M. i/ ";> ' "v . •• , j'v , . Hart, W. iH, • i> • '•• • ~ Rhoades, H. L. , , . v .Eversmeyer, H. E. '.-. -' Marchbank, Dprothy F. ; ••A; ..:V / Hasbrouck, E. R. '; •Short, -R.iB. : Whittaker, F. H. -* McIntosh,,A. " .'-•.•^y Heyneman, D. Smart, G. G., Jr, .Louisiana ._ j -/ McLoughlin, D. KT 7 i? ••'',;,';. • Holdeman, O. L;," "., .Tarjan, A. G. : ^ , Acholonu, , A. -D. , • .;- .. Morris, J. A. ': '•. ...-'. r/V'\, S. M. '•" ^ Taylor, A. L. :, ':.-,. :>: { ,Birchfield, W. 'jLg >Murphy, D. G. ^ ..--,'' ' vV-,;; "•;'-' iiamberti, .F.; -1 ' ,- /; '.Georgia :; '— v ' •'.;:.' ./"• Hollis, J.P,,Jr. / i Nickle 'W. R.> '-••'• ; ;; : • -N •' .;. . , Lear B. ' • ' - ': '""-• '•( ^Bizzell, W. E. ;'. ' 'Lee,'S.H. -'>',->'-. •,Otto,C.',F.^ ,' L-' '"--':-'" •><•" v. .'•-<;-,'}\;(/ )'-,^' Lembrightj'H. 'W. -^'' " CSordia, H. , ~7i—': Lumsdeh, R. D. l\vs!v Phillips, B. P. . ; : , ':•'•- ;/" v v'' Lie'Kian-Joe Gross, H. F.' -^-: Marfin^W. J. ;; Pipkin, A. C. - .- .'. ' •' ' j • f. ',••'. • -A Ldos. G. • A - " T ' ' Denton, J.\F. .^"^ '<;;-, '- .Newsom, L. D. ; '-"' ; Pofley, Judith A/ ,:- "v >A'-V J-;Mkggenti,A. R;' ' *-; . ;v* ' iGoodchild* C. G. ';' Sogandares-JBernal, F. • ' Poole, J; iB. ' ,; -_" .-r^ , . "'? * Mankau, H. .t , Heard, Ri W., Jr. Warren, L. G. .: ^Powers, 'K.G. "' 1 " '.?' • 'jiff "'<:-•; :•*•'. : '•'. 'j'McBetil, C. W. ' • ''"...... Kagan, I. G. -.'~- 'Maine !X -- • ^>- "-\-; Qtngley , -Diane / (, -.-0 .;.", > , McGuire, -W. C. Minton, N. .-A. ••• •', .''' • /'; :' Meyer, M. C-' .'"'/'-'• /'.-'•': >Richards,C. S. V /c\ ;to Mizelle, T. D. ',/'-• Pripe.C.E. . - '.- .'Maryland ' '<<4 -. • -i.. Rpby, T. O. '. ;v.^"'-- /^Nalihas, F;M. '•- - Ruehle J. L. •• / '- - /Abrarn'y/J. B. ' • ^ !'•• ^ '/•• y' Reimanowski,''R. D/ \"~*- /• /. ^ ; ; ; i Noffsihger, Ella Mae Stewart, T.vB. _V •--. Anastos, G. -'"-••'.' Vr"' -''V' ' ;Rose,'J,;E.-. > ••7-- ,'-,^ - • ^ / ~ jPaxman, G. A. ./ .: Anderson, ;R. L / ">v / Rozeboom,"L.'-E. x- " ' -: v: '•' /"?' 'pbinar^-G. O.; Jr. - ^HawaliV3P' •' •' '>-''-/;' •". Andrews, J. S. i. , -' 'Sarles, M; P^ , > --, I1'? ' <-; \;''- ;.,,-':•'••'•• '.•Alabam; Raskia ' . ;'•-..-D;. ;J". Li/. ; Alicata, J. E. ; - *, Anthony, D. W. Sawyer T.iK. S ' ,/ -^-:r. ;.''\: :'i'yRossan,:RrN,.-. ;:•' • ApvWlJ' V-- '.*:.>,> Bang.F. B. . ;\ . ' s; _ > • iRothman, A. H. - Cheng, ;T. C., , -•-•r-' Beckfund, W. W. ' JS VA [;:-, '-..,\, S.A. > '- -Goto.'S. '/ .:,v,v,.- -'- - ••von Brand, TV ; 5, / Schiller, E. L. " r v -r, '« ,Steefe,A.' E. : ;, ' Idaho .. •' >' •(-' ;>.'•' Buhrer, Edna M. ij ' - Schneider, C. M. . ;::'~y Van Gundy, S. D. DaUiinore, C. E. , " ' Cannon; L. X '•.'- T" Scott,J. A: •• '• , '?"''4' . '.'. '- ' \, D. R. > Illinois Castillo, Jessica M- Segal, Dorothy B. .; /I--; ,, '•'. |X'. . •- 'Voge, Marietta '-, . •Bauman, P. *M. Chitwood, May B. . ;- \ "v. jj- -Wagner, E. D/- •'-, ' • . Dunagan, T. T. , 'Colglazier, M. L. i V. Shiroishi,-:Tsugiye / x "K i^';- /Edwards, D. I. i i-./ DeWitt, W. B. :'• "Solomon, G. B. \' '" f,';V' '>•-"• ,'•.'-.••;/< Conol Zone -^ . Garoian, G. ' -' - - Diamond, 'L'.\S., :: i Spindler, L.:A. ,.,,,:'.:: • , Walton, B.C. , ;; '• Huizinga, H. W. . • ", Doran, D. J.', U . Sprague, V. Kruidenier,.-F. J. , Doss, 'Mildred A. & '-'\ Stirewalt, Margaret A- i : fc^ i X S, ••''•''. Ballard *N.~'-. • '' / ' • s' , Levine, 'N. D. '' *' ' Douvres, F. W. •> .': . • • Tarshis.1 B.(" :/-..• :'X \\. .Ai. ' /'•] ' * •" ?X ; ! ; .""'•.'• '/•' ;''• *•'' ••. Olsen O. 'W. .- ' '!• 1.'- McCrae, RiC.v \i T. LDropkin. ;, V. H._ 3^;i "; \ - y^ •- v Schmidt, G. D. Durbin, C..G.X \' Cv v Stabler, R. Mi . ""i Pillai, J.K. ' Endo,B.Y. :. '-" ;-;;*, Tromb'a, F. G, ' , /'• : Connecticut /• -.../ " ••. -.',- Schneider, M.'.D, Elll E. U. . • ; •: • i-L, : .''/. ' Penner, L. H. f", Siddiqui, I. A. ;•.: ' i Fair, Marion M; ; ' " -'i- Yegors, H. H. V*'f^ --• ./ -Delaware ' v ..,-;, , Singer, I. Feldinesser, J. "".j . '(. Wehr, E. E. • V "I-,7 • '.'!?««« ' TIJT 'C» -' ' .' ' 1 --: - Fielding, M.J. Taj?lor,uD.:P. " >/ , JP erguson*' jvi* • o. - •'.Weinstein.'^?.'?.''1! ,'••'•"• >t' '"%, ]" Dittrietibf Columbia >•-.__ -Thomas, L. J. - .Foster j A. ,O. ;..\ , Wiest, L. M., Jr. ,,:•'- :, '< ' "V'^' \ -" ,Baisden,--L. A. ; Todd, K. S.,:jr. , Friedman, -W. - v . ' ( ''• '• Williamson, F. S. LI ) ;. ';.. 'iBetz, D. O., Jr. , '.. Ubelaker,'J. E; •; '.-•• Goldberg. A. Wiseman, H. G. - , (. v £,&[ Indiana , .-. - M./-V .' ,-'•'• j.r ;Gpiaen,-A..k -;,/'/':

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^'EABD: , iliGHAHD W., IH... Parasites ''oftiie Clapper Ilail, Rallus lohgirostris. Boddaert./ ; y I( The (Current /Status/ of tiie Genus Lfeviweifalla* yn\h &e Description of ' ' ; s&niella byrdi^a. sp. (Trematoda: 62 ; HECHLER/ HEL£N GABOL. Postembrybnic Develophient and Reproduction in DfpZo- .'•';' scopter corotiata (Nerrptoda: Rhabditidae ^ — -_-_ T_i-4-j_.i...^.;7__-_._.l_.._ _ _.._._.___i.i ' ' ' ' ' ' ' ••• • ' .' x- " ' • • ,/ ' "• • -v ' ;' .- ' •'••. - > •• i,f, / HOLLOWAY, HARRY L., JR., AND -}EFTREY W. BIJSR. Helicon/ietra antarcticoe sp. ripv. ^ ; froin Antarctic Coastal Fishes ^-^^.-.^..-^v... 30 V In'' Memoitia' ' m ' tfj) <• "Ehree New Species of Actinolalirnidae (Neriiatoda: Dory- laimoidea ) from India ._^_!.._.i;...—'—.—._-rj——. .-_._..—_; ____ L _____ -r—.-.l..l.....______96

;:l •'/ ;l-(r rKATEs,;'iC, p.^ 'Ai«),D.;E. 'THOMpppN. Suceptibility ,of Gerbfls arid Youhg White Rats to - , ..' : i Simultaneous Infection with axei and Tnchostrongylus colubrifoiinis 'W^r>( - / (; *,' LIE, KIAN JOE. Further Stiijdiesôhlthe Life History ipf Echinostoma lindoense Sand- -, '4> "'';.-. •• u ground arid -Boiihe^ 1940 XTreiâtodaîEchindstomaydiae) with # Report^pf ][tSi ' ' . \e >in (Brazil ' ..H.^t.~~ — 1^.^_1. —;^ — .. J-u.,.^. — 1 — ; —i____.^L.. Jr ____ .„_.:..„. '\74/1 • .Neoec/M^M>rhynchus corW«ru)tt

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'.:.'/<•' tipirif n.;sp. ( Nematoda: ilopllaimidae) -frbm Forest (/'^:!'] L '/ Nurseries in Jap'an ._„.._., _____ .„•_ •::r$8'': -.-v ; >'"-,.-::•-'. •;••.. v-: .. . :•'-, 4 ••? ^ . •• ; : y f ; tMlEYER, MARVIN C., ;ANJ? ROLF VEK. jQbseryafclons on Diphyltobdthrium ^sebago :Plero- ' ' ' .'i' '! •* .cercoids din the ;Fish :iiHosts ..^... - - . ., ,^ , - . .; 'A MARCOS .Tuiao/DiAZ. ^ Studies on Freshwatisr LarvaKTremittodes. :Earfc -/v,2CWii' Five .New ISpecies of: , Cercariae. from Venezuela' .;_-.._-,_,-,._.4.^i._™_c...i...^ •;:67/:fy -; •lf- .-/;• r- 'V-lj ' -'.' V ":. V- .; '-' ': • •' ' l-y' |A i— i •• •' ' ".'" • ^ ' /' V\. '' '\,'' • '. •''• . -.'•'. """'..i' <' V n>Presentatiori-^-19i87 Anniversaryi Award of The H^lminthologieal Society of 'Washington, ' -429th Meeting vKL 'vOctdber 1967' ^i:.,..^..^^...r.^.^:...r^...^ ____4 .: ______ify£a$+±- IIS, . - " "" '' . ' ' ' ' ""i ' ' "'1 ' PRICE, C. *E.,/Aj>JD A. BUSSING. Monogenean Parasites of Costa Rican iFishes. ,11. ' , Proposal of J54 '' .,'; '•'; '•;';'' '•';*];: 'SAMSON, KAY^S; , The Synonyiny'of the Ruminant 'Parasitej s Nematodirus biratianus ,".'j -} ' r:' x '/Baevskaia, 1929 and Nematodirus:JanceQlatus Ault, '1944 • ^,.-.-.L_,-..^_i;^.^..._... ;- 52 K •. 1 , , . ... • '' ; .V v '' S&EPEERSON, JACQUEUKE :R,, AND G. JORDAN.' , A Technique for Isolating arid ' A: Maintaining Cultures of ' •.'• •; r :,--' - .' " •' •']

\.y. . _ ; /; N -'I..'' , • :. I : . ^ AOl^J-^VJTSENGj .U£*\J- LJM.*t), SHU-TEN AVJUjJLAAi '•••»••, ' KEITH R. ALLBED/ AND GERALD D. GRIFJTN.; \ Soil ^Populatiori Study (Kiihn) Filipjev iri an Alfalfa ^Field ....._..^.._.:^C1.1^1.. f^^V^vv.'^ •^'-'••/ P^ tDityienchus dipsaci ;••'>'-«7i'-.; ., /i/\-'n :' -:/l':r"5V:"'u iil^^VV^liER^MARTkAV.'!.., ANDI JWniARD W. B^ckLUNp. A Note on the Morphology of ''", . .,'•''•'• V.'; . -!'.'it/'; > . ,>n;- _;..'_ .'.j. fl. _i_t'_!-/Ti pooperia,punctata ;(Linstpw, -1907/) and iCooperia ppatttlata Raylis, 1938 ;.-.^..L!"_!_• J 49::.-; ^;i , -,'-.•' /fT>N-, •-',' . . R.'iBARKER. Hatchpg Response .of Hypsoperine ctiersoni •'.' - i ;l- to Ganarygrass Root Bmariations .^^ WiHiTTAK^R, FRED H., AND LOR?N G, Hux. ' Proteocepholus cJwlogosteri sp. 'n. (Ces- '•'« ?lj|'> .^^4, , V ( toda: Proteocephalidae) from theV Spring Cave'fish^ Chojogoster , j'TToo (jpiSCes;/Afnblyopsidae) of v^^r^li ~: •':..,. .1 v'-ii:.-''-^

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