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John Janovy Publications Papers in the Biological Sciences

8-1993

Nubenocephalus nebraskensis n. gen., n. sp. (Apicomplexa: Actinocephalidae) from Adults of bipunctulata (: Zygoptera)

Richard E. Clopton Peru State University, [email protected]

Tamara J. Percival Cook Texas A&M University, [email protected]

John J. Janovy Jr. University of Nebraska - Lincoln, [email protected]

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Clopton, Richard E.; Percival Cook, Tamara J.; and Janovy, John J. Jr., "Nubenocephalus nebraskensis n. gen., n. sp. (Apicomplexa: Actinocephalidae) from Adults of Argia bipunctulata (Odonata: Zygoptera)" (1993). John Janovy Publications. 15. https://digitalcommons.unl.edu/bioscijanovy/15

This Article is brought to you for free and open access by the Papers in the Biological Sciences at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in John Janovy Publications by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. J. Parasitol.,79(4), 1993, p. 533-537 ? AmericanSociety of Parasitologists1993

NUBENOCEPHALUSNEBRASKENSIS N. GEN., N. SP. (APICOMPLEXA:ACTINOCEPHALIDAE) FROM ADULTS OF ARGIABIPUNCTULATA (ODONATA: ZYGOPTERA)

R. E. Clopton, T. J. Percival, and J. Janovy, Jr. School of BiologicalSciences, Universityof Nebraska,Lincoln, Nebraska 68588-0118

ABSTRACT: Nubenocephalusnebraskensis n. gen., n. sp. (Apicomplexa:Actinocephalidae) is described from trophozoites,sporonts, gamonts, and oocysts collectedfrom adultArgia bipunctulata (Odonata: Zygoptera). The new genus is distinguished from existing acanthosporinegenera by elongate dodecahedraloocysts, without equatorialfaces, that are hexagonalin equatorialcross section with equatorialand terminalspines. The epimerite is very broadly ovoid and truncatedposteriorly with equatoriallobiform tumidi that are posteriorlybilobate and do not form hooks, spines, or digitiform processes. The epimerite is borne on a long, slender stalk. The new speciesis also reportedfrom naiads ofArgia bipunctulataand adultsand naiadsofEnallagma civile(Odonata: Zygoptera).

Septate gregarines have been reported from MATERIALSAND METHODS both odonate naiads and adults (e.g., Ellis, 1913, Adult damselflieswere isolated abdomen down in 1914; Devdhar and Deshpande, 1971; Abro, 5-ml cappedplastic test-tubes with 1 ml waterfor fecal Richardson and collection. Twenty-fourhours later each tube was ex- 1974, 1976, 1987; Janovy, 1990). amined for Shed Of the 274 described in Actino- gregarinegametocysts. gametocysts gregarine species were freedfrom feces, pipettedwith 10 jl of waterinto cephalidae, 36 species in 15 genera are reported individual wells of an HL-A tissue cultureplate (Lux from odonate hosts (Crawley, 1907; Watson, Scientific Corporation, Newbury Park, California), 1916; Levine, 1988; Richardson and Janovy, measured,and held for maturationand dehiscence. structureand dimensions were taken from 1990). Odonate have been reported Oocyst gregarines fresh preparationsof oocysts suspendedin water. Oo- from all 3 subfamilies of Actinocephalidae; how- cysts rotated freely, allowing elucidation of the full ever, North American species are known only 3-dimensionalstructure of the oocyst. Oocyst lengths, from 3 genera in 2 subfamilies: Geniorhynchus widths,and depthswere measured at theirwidest points and in and and are reportedin micrometers.Clopton et al. (1991, Actinocephalus Actinocephalinae demonstratedthe of in The third 1992) utility glycerin-basedoocyst Prismatospora Acanthosporinae. preparations.In the present study, however, oocysts subfamily, Menosporinae, is dominated by greg- collapsed in glycerinor Hoyer's medium-basedprep- arines reported from odonates (16 of 18 known arations,suggesting oocyst structuralinstability under species). The general paucity of the known North high osmotic tension. Aftercyst collection, were dissectedin saline without sucrose American fauna and the lack of known (Belton gregarine and Grundfest,1962) and examinedfor parasites.Six- North American menosporid species suggest the ty-nine of 119 damselfliesexamined between July and existence of substantial unknown fauna. September 1992 were infected. Measurementswere During a taxonomic survey of the gregarine taken on no more than 5 individualsper host. Widths of and deutomerites were taken at the parasites of odonates in the Salt Valley of south- protomerites widest points. eastern a distinct form was Nebraska, gregarine Measurementsare presented as range values fol- collected consistently from Argia bipunctulata lowed by means, standarddeviations, and samplesizes (Zygoptera: ). The secondary in parentheses.All measurementsare in micrometers. cuspidiform epimerite structure resembled that Levine's (1971) uniform terminology for Apicom- is used in this with a of Prismatospora evansi Ellis, 1914. a plexa paper single exception. However, "Sporont"is used to indicatea matureindividual that complete life cycle study revealed a unique group remainsattached to the host intestine, in contrastto a of morphological characters including a lobose, gamont(a matureindividual that is not attachedto the unhooked primary epimerite and spined, poly- host intestine and has formed an association) or to a hedral without faces. The trophozoite (an immature, vegetative individual that oocysts equatorial is attachedto the host for form of these structures the intestine).Terminology shapes unique prompted of planes is consistentwith that suggestedby the Sys- present proposal of a new genus and species of tematics Association Committee for Descriptive Bio- eugregarine. logical Terminology(Anonymous, 1962a, 1962b) and formsthe basis for derivationof terminologyfor shapes of solids used in this description. Received 13 November 1992; revised 16 February Drawingswere made with the aid of a cameralucida. 1993; accepted 6 March 1993. All observationswere made on a Wild binocularcom-

533 534 THEJOURNAL OF PARASITOLOGY, VOL. 79, NO.4, AUGUST1993

poundmicroscope with 12.5 x wide fieldeyepieces and to depressed obvoid; protomerite length (LP) 91.3- American Optical-Spencer10x and 40 x objectives. 348.6 (223.1, 65.9?, 66), protomerite width (WP) Measurementsand color observationsin living spec- 124.5-489.7 (276.9, 82.6?, 66); truncated without imens were made over blue and daylight filters, re- constrictionat union with deutomerite,becoming cra- spectively. teriform anteriad in late trophozoites. Deutomerite DESCRIPTION broadly obvoid in very young trophozoites,narrowly obvoid in late trophozoites;deutomerite length (LD) Nubenocephalus n. gen. 141.1-1,651.7 (858.92, 321.4?, 66), deutomeritewidth Diagnosis: EugregarinidaLeger, 1900, sensu Le- (WD) 116.2-481.4 (257.1, 86.3+, 66); distended an- vine, Corliss,Cox, Deroux,Grain, Honigberg, Leedale, teriad at union with protomerite. Total length (TL) Loeblich, Lom, Lynn, Merinfeld, Page, Poljansky, 340.3-2,000.3 (1,082.0, 363.6?, 66); LP/TL 0.13-0.59 Sprague,Vavra, and Wallace, 1980; SeptatinaLankes- (0.22,0.08 ?, 66); LD/TL 0.41-0.87 (0.78, 0.08 ?, 66); ter, 1885, sensu Levine, Corliss, Cox, Deroux, Grain, LP/LD 0.14-1.41 (0.31, 0.22?, 66); WP/WD 0.32- Honigberg,Leedale, Loeblich, Lom, Lynn, Merinfeld, 1.44 (1.11, 0.20+, 66). Epimeriteborne on a narrow Page, Poljansky,Sprague, Vavra, and Wallace, 1980; basal stalk; length 20.0-54.0 (34.4, 8.6?, 54), width ActinocephalidaeLeger, 1892; AcanthsporinaeLeger, 10.0-16.0 (12.9, 1.7?, 54). Epimerite very broadly 1892. Oocystsdodecahedral, elongate, terminally trun- ovoid; length 22.0-46.0 (34.0, 5.5?, 54), width 30.0- cate, hexagonal in equatorial cross section, without 48.0 (37.0, 5.1 +, 54); truncateposteriad at union with equatorialfaces, with equatorialand terminal spines. epimerite stalk, with 6 equatorial lobiform tumidi; Epimerite broadly ovoid, truncated posteriad, with tumidi posteriorly bilobate and not forming hooks, broad, flexible, equatorial tumidi that do not form spines, or digitiformprocesses. Epimerite without vis- hooks, spines, or digitiformprocesses; borne on a long ible septum;obvious in young trophozoites,apparent slenderstalk. in some form in most trophozoite stages. Total epi- merite length 46.0-100.0 (68.4, 11.63+, 54). Nucleus Taxonomic summary narrowlyelliptoid to elliptoid;length 50.0-88.0 (69.1, Typespecies: Nubenocephalusnebraskensis n. sp. 9.3?, 52), width 20.0-42.0 (36.1, 5.2?, 52);placement Etymology: The genericname is given to mark the consistent, abaxial, and supraequatorial.Nuclear en- veiling nature of the crateriformprotomerite of the dosomes variablein shape and number.Fresh tropho- type species. zoiteswith endocytepartially granular when very young, becoming opaque with maturity,epicyte clear to light Remarks orange in color; width 8.0 (8.0, 0.0+, 66). Whitish Actinocephalidaehas been reviewedrecently as part under dissecting microscope and incandescent illu- of a largerwork (Levine, 1988). The new genus clearly mination, brown under compound microscope and is a memberof Acanthosporinaein that spinedoocysts daylightfilter. are unique to this subfamily. Oocyst structureis dis- Sporont(Fig. 3): Attached to host ventricularepi- tributed across the 20 genera that constitute Acan- thelium, solitary.Protomerite very broadlyobvoid to thosporinaeas follows: 4 genera with polyhedraloo- depressedobvoid; LP 157.7-597.6 (280.1, 110.1?, 54), cysts, 9 with biconical,4 with ellipsoidalor ovoidal, 2 WD 249.0-871.5 (434.7, 155.4+, 54); truncatedwith with unique asymmetricaloocysts, and 3 genera for constrictionat union with deutomerite;anterior mar- which the oocysts are unknown(Tschudovskaia, 1928; gins uncleft, expanded to form a broad, crateriform Devdhar and Amoji, 1978; Kori and Amoji, 1985; adhesive disk; apex distended to contact the host ep- Levine, 1988;Cokendolpher, 1991). Of the 4 described ithelium. Anteriormargins of protomeritein late spo- generapossessing polyhedral oocysts, the oocyst struc- ronts cleft or folded, becomingcorollate with 2 narrow ture of the new genusresembles only that of the mono- ventral lobes and 1 broad dorsal lobe. No epimerite typic Prismatospora.However, the new genus is dis- observed;transitional forms possess remnantsof epi- tinguishedby lack of equatorialoocyst faces. meritestalk. Deutomeriteobvoid to narrowlyobvoid; There has been no complete revision of Acantho- LD 506.3-2,672.6 (1,279.7, 458.2?, 54), WD 190.9- sporinae and Devdhar and Amoji (1978) considered 846.6 (391.6, 156.6+, 54);TL 713.8-3,237.0 (1,559.8, only epimeriteand oocyst structurein their review of 554.3 , 54); LP/TL 0.12-0.29 (0.18, 0.04 , 54); LD/ charactersdiagnostic among 12 member genera. Epi- TL 0.71-0.88 (0.82,0.04 ?, 54);LP/LD 0.14-0.41 (0.22, meritestructure varies greatly among genera and among 0.06+, 54); WP/WD 0.85-1.44 (1.14, 0.15?, 54). Nu- specieswithin a singlegenus (Baudoin, 1971;Hoshide, cleus elliptoid; length 48.0-190.9 (99.1, 36.6+, 22), 1977; Devdharand Amoji, 1978). The broad rangeof width 28.0-116.2 (56.4, 22.3?, 22); placement con- variationamong oocyst and epimeritestructure within sistent among individuals and developmental stages, Acanthosporinaehas led to genericdistinctions based abaxial and supraequatorialin deutomerite. Nuclear on cooccurrenceof oocystand epimerite structural types. endosomes variablein shape and number.Fresh spo- Althoughthe new genus is distinguishedupon oocyst ronts with granularendocyte, nonuniform and opaque morphologyalone, the epimerite structureof the new with clear epicyte; width 5.0-8.3 (7.0, 1.62?, 30). genusis unknownamong the 4 describedacanthospori- Whitishunder dissecting microscope and incandescent nid generapossessing polyhedral oocysts. light, translucentbrown to opaque black under com- pound microscopeand daylightfilter. Gamont(Fig. 4): Partiallyto enrobedin Nubenocephalus nebraskensis n. completely sp. host ventricularperitrophic membrane, solitary or in (Figs. 1-6) association.Protomerite transversely broadly elliptoid; Trophozoite(Figs. 1, 2): Attached to host ventric- LP 174.3-597.6 (351.4, 124.6?, 24), WP 439.9-871.5 ular epithelium,solitary. Protomerite broadly obvoid (564.4, 145.8?, 24); apex smooth, no evidence ofcra- CLOPTONET AL.-N. NEBRASKENSISN. GEN., N. SP. 535

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FIGURES1-6. Nubenocephalusnebraskensis n. gen., n. sp. 1. Trophozoite,lateral view. 2. Primaryepimerite of trophozoite, lateral view. 3. Sporont, lateral view. 4. Mature gamont with disassociated nucleus, lateral view. 5. Oocyst, dorsal aspect. 6. Oocyst, lateral aspect. (Shallow depth of field and diffraction of lateral faces give the illusion of reduced equatorial width, reproduced here to facilitate diagnosis.) teriform margin observed. Deutomerite obvoid to nar- served. White underdissecting microscope and incan- rowly obvoid; LD 888.1-2,672.6 (1,590.8, 490.8?, 24), descent light, translucentto opaque black under com- WD 356.9-846.6 (525.67, 139.4+, 24); TL 1,062.4- pound microscopeand daylightfilter. 3,237.0 (1,942.2, 603.2 , 24); LP/TL 0.15-0.24 (0.18, Association: Syzygy late and ephemeral,latero-as- 0.03+, 24); LD/TL 0.76-0.85 (0.82, 0.03+, 24); LP/ sociative while caudallyenrobed in peritrophicmem- LD 0.17-0.31 (0.22, 0.04?, 24); WP/WD 0.98-1.28 brane. Sporont to gamont transformationmarked by (1.08, 0.09 , 24). Nucleus diffuseand indistinct.Fresh loss of anteriorcrateriform adhesive disk and an an- gamontswith opaqueendocyte, no distinct epicyte ob- teriad roundingof the protomerite,dissolution of the 536 THEJOURNAL OF PARASITOLOGY, VOL. 79, NO.4, AUGUST1993 nucleus, and a general shift from translucence to Remarks opaqueness.Syzygy begins as 2 adjacentsporonts fuse In 2 othergenera, the monotypicPrismatospora and caudally. Fusion line progressesanteriad to produce severalspecies within the Ancyrophora,transmutation similarin form and association.Association lengthto, of the primaryepimerite to a largercuspiform structure Associationfolds over but broaderthan, single sporont. has been reported (Ellis, 1914; Hoshide, 1977). All mem- on itself in roughlyequal thirds producing single known cases of epimeritetransmutation occur in spe- association mem- brane-boundgametocyst. Internal cies described or reported only from odonates. Ellis braneslost of- duringgamete production. Gametocysts (1914) suggestedthat this transformationmay serve to ten shed beforeloss of internalassociation membranes anchor the parasiteduring the violent displacements and developmentcontinues exogenously. Associations of the intestine associated with the rectal gill move- observed in the posteriorquarter of ventriculus.Ob- ments characteristicof immature how- that have not com- anisopterans; servationssuggest that associations ever, the same transmutationis observed in parasites and initiatedformation of the pletedsyzygy gametocyst of adultanisopterans and in adultand immaturezygop- membranedo not survive passagethrough the rectum. terans. Gametocyst: Opalescent and spherical; diameter 420.0-640.0 (532.6, 67.5?, 19);hyaline coat uniform, increasingdiameter to 920.0-1,270.0 (1,032.6, 107.8?, ACKNOWLEDGMENTS 19). Gametocystscollected and storedunder (ca. 22 C) This work was in Ashton C. water dehiscencebetween 60 and 72 hr. supported part by sporulateby Cuckler and U. S. Harkson awarded Oocyst(Figs. 5, 6): Oocystsdodecahedral; axial length fellowships 7.84 (7.84, 0?, 46), equatorialwidth 9.8 (9.80, 0?, to R.E.C. 46) (shallow depth of field and diffractionof lateral faces the illusion of an width of ca. 4.9 give apparent LITERATURECITED [Fig. 6]); elongate,terminally truncate; terminal width 1.47 (1.47, 0+, 46); hexagonalin equatorialcross sec- ABRO,A. 1974. The gregarineinfection in different tion, with 6 equatorialspines; length 3.92 (3.92, 0+, species of Odonatafrom the same habitat.Zoolo- 46); 1 spine at each equatorialangle and 12 terminal gica Scripta3: 111-120. spines;length 2.94 (2.94, 0?, 46); 1 spine at each ter- 1976. The mode of gregarineinfection in minal angle. Each oocyst has an equatoriallylocated Zygoptera(Odonata). Zoologica Scripta 5: 265- sphericalresiduum; diameter 1.25 (1.25, 0+, 46). Oo- 275. cysts very uniform in size and shape. 1987. Gregarineinfection of Zygopterain diverse habitats.Odonatologica 16: 119-128. ANONYMOUS.1962a. Systematics Association Com- Taxonomic summary mittee for DescriptiveBiological Terminology. II. Type host: Argia bipunctulata(Zygoptera: Coena- Terminologyof simple symmetricalplane shapes grionidae). (Chart 1). Taxon 11: 145-155. Host records:Argia bipunctulatanaiads and adults; 1962b. SystematicsAssociation Committee Enallagma civile (Zygoptera:coenagrionidae) naiads for Descriptive BiologicalTerminology. IIa. Ter- and adults. minology of simple symmetrical plane shapes Type locality: Bowling Lake (Section 2, Township (Chart la). Taxon 11: 245-247. 10 North, Range6 East),Lancaster County, Nebraska. BAUDOIN,J. 1971. Etude comparee de quelques gre- Specimens deposited: Two (1 trophozoite, 1 spo- garinesAcanthosporinae. Journal of Protozoology ront) type slides have been deposited in the United 18: 654-660. StatesNational MuseumHelminthological Collection, BELTON,P., AND H. GRUNDFEST. 1962. Potassium Biosystematic Parasitology Laboratory, Beltsville, activation and K spikes in muscle fibers of the Maryland.The trophozoite (USNM Helm. Coll. No. mealworm larva (Tenebrio molitor). American 82671) is the holotype and clearlydemonstrates char- Journalof Physiology203: 588-594. acteristic primary epimerite structure;however, the CLOPTON, R. E., T. J. PERCIVAL, AND J. JANOVY, JR. sporont(USNM Helm. Coll. No. 82672) clearlydem- 1991. Gregarinaniphandrodes n. sp. (Apicom- onstratesthe more typical maturebody shape and sec- plexa:Eugregarinorida) from adult Tenebriomol- ondaryanterior crateriform margin and shouldbe con- itor(L.) with oocyst descriptionsof othergregarine sideredin any revision. parasitesof the yellow mealworm.Journal of Pro- Infection site: Trophozoites were observed in the tozoology 38: 472-483. anterior2/3 of the ventriculus,attached to the epithe- , AND . 1992. Gregarinacuneata lium and outside the peritrophicmembrane. Sporonts n. sp. (Apicomplexa:Eugregarinida) described from were observed in the lower 1/2 of the ventriculus,at- adults of the southerncorn rootworm,Diabrotica tached to the epithelium and outside the peritrophic undecimpunctatahowardi (Coleoptera:Chryso- membrane.Gamonts were observedin the lower /3 of melidae). Journalof Protozoology39: 417-420. the ventriculus, caudally enrobed in the peritrophic COKENDOLPHER,J. C. 1991. Cosmetophilus vonones, membrane. n. g., n. sp. (Apicomplexa:Actinocephalidae) in Etymology: The specificname nebraskensisis given the harvestman Vonones sayi (Arachnida:Cos- to mark the type locality and to mark the 100th an- metidae).Journal of Protozoology38: 461-464. niversaryof parasitologyresearch and curriculumat CRAWLEY,H. 1907. The polycystid gregarines of the the University of Nebraska. United States (thirdcontribution). Proceedings of CLOPTONET AL.-N. NEBRASKENSISN. GEN., N. SP. 537

the Academy of Natural Sciences of Philadelphia KORI,S. S., AND S. D. AMOJI. 1985. Tetrameridion- 59: 220-228. ospinispora karnataki, new genus new species: A DEVDHAR,M. J., AND S. D. AMOJI. 1978. On a new new cephaline gregarine from the , actinocephalid gregarine, Contospora opalnia gen. Agriocnemis sp. Acta Protozoologica 24:139-146. nov., sp. nov. from the gut of an arachnid Opalnia LEVINE,N. D. 1971. Uniform terminology for the sp. Archiv fur Protistenkunde 120: 182-189. protozoan subphylum Apicomplexa. Journal of ,AND S. DESHPANDE.1971. On a new gregarine Protozoology 18: 352-355. Menospora nonacantha, n. sp., from the odonate 1988. The protozoan phylum Apicomplexa, Agriocnemis sp. (fam. Coenagriidae) from Dhar- Vol. I. Chemical Rubber Company Press, Boca war, India. Indian Zoologist 2: 41-54. Raton, Florida, 203 p. ELLIS, M. 1913. A descriptive list of the cephaline RICHARDSON,S., AND J. JANOVY,JR. 1990. Actino- gregarines of the New World. Transactions of the cephalus carrilynnae n. sp. (Apicomplexa: Eugreg- American Microscopical Society 32: 259-296. arinorida) from the blue damselfly, Enallagma ci- .1914. An acanthosporid gregarine from North vile (Hagen). Journal of Protozoology 37: 567-570. American dragonfly nymphs. Transactions of the TSCHUDOVSKAIA,I. 1928. Uber einige Parasiten aus American Microscopical Society 33: 215-222. dem Darmkanal der Sciara-larven. Archiv fir HOSHIDE, K. 1977. Notes on the gregarines in Japan Protistenkunde 60: 287-304. 8: Three new species of Eugregarina from Odo- WATSON, M. E. 1916. Studies on gregarines. Illinois nata. Bulletin of the Faculty of Education, Ya- Biological Monographs 2(3): 1-258. maguchi University 27: 93-125.