ARTHROPOD BIOLOGY

Biology and Behavior of Kapala (Hymenoptera: Eucharitidae)Attacking Ectatomma, Gnamptogenys, and Pachycondyla

(Formicidae: Ectatomminae and Ponerinae) in Chiapas, Mexico

GABRIELA PEREZ-LACHAUD,1 JOHN M. HERATY,2 ANDREW CARMICHAEL,2

AND JEAN-PAUL LACHAUD1, 3

Ann. Entomol. Soc. Am. 99(3): 567Ð576 (2006)

ABSTRACT Kapala (Hymenoptera: Eucharitidae) are among the most frequently collected chal-cidoid wasps from the Neotropics, but general information about their host plants, behavior, andlife history is scarce. A new species, Kapala izapa n. sp., parasitizing Ectatomma ruidum Roger(Hymenoptera: Formicidae), is described from Chiapas, Mexico, with a general account of its biology.Field observations on the behavior of adults of Kapala iridicolor (Cameron), which also attackE. ruidum, Gnamptogenys regularisMayr, Gnamptogenys sulcata (Fr. Smith), Gnamptogenys striatulaMayr, and Pachycondyla stigma (F.) (Hymenoptera: Formicidae), are included. Immature stages ofKapala izapan. sp. follow the general morphology and behavior described for other eucharitids knownto attack Ectatomminae and Ponerinae. More than one parasitoid can complete development on asingle host larva.K. iridicolor females were observed laying masses of eggs on undeveloped ßower budsofMelampodium divaricatum (L.C. Richard) D.C. (Asteraceae). Extraßoral nectaries of this plant arevisited by several insects, including predatory foraging worker ants ofE. ruidum. Dissection of recentlyemerged females of K. iridicolor yielded �4,500 fully developed eggs.

KEY WORDS Eucharitidae, Ponerinae, antÐhost association, Kapala, Ectatomma

Members of the family Eucharitidae (Hymenoptera:Chalcidoidea) are specialized parasitoids of ants andhave a highly modiÞed life cycle (Wheeler andWheeler 1937, Clausen 1941, Heraty and Darling 1984,Heraty 1994b). Instead of ovipositing directly ontotheir host ants, females deposit their eggs in differentlocations in or on plant tissue (Clausen 1940a, Heratyet al. 2004). Unlike most parasitic Hymenoptera, theactive Þrst instar, or planidium, is responsible for gain-ing access to the host ant larvae by using variousphoretic behaviors involving either attachment to anintermediate host that is ultimately taken directly tothe host larva or through direct attachment to foragingant workers and indirect access to the host larvaeduring trophallaxis or movement within the ant colony(Clausen 1940a, Heraty 2000). Once inside the antcolony, the planidium attaches to a mature host larvaand then waits until the host is ready to pupate beforecompleting its own development (Clausen 1941, John-son et al. 1986, Heraty 1994b).

Two of the three subfamilies of Eucharitidae areknown from the New World. The Oraseminae is

represented by the widespread Orasema Cameron.Nine of the 16 New World genera of Eucharitinaeoccur in Central America (Carletonia Heraty, Dilo-cantha Shipp, Isomerala Cameron, Kapala Cameron,Lirata Cameron, Lophyrocera Cameron, Obeza Her-aty, Pseudochalcura Ashmead, and PseudometageaAshmead). Except for Carletonia and Lophyrocera, allof these Central American genera are known to occurin Mexico. Although Kapala is the most commonlycollected genus of Eucharitinae in the Neotropics(Heraty 1995), only a few species can be accuratelyidentiÞed, and information dealing with their biologyand behavior is scarce (Clausen 1940a,b, 1941; Heratyand Darling 1984; Heraty 1994b, 2002; Howard et al.2001).Kapala are widespread throughout the Neotropical

region, with a few species extending north into thesouthern United States (Arizona, Texas, and Florida),and one disjunct species is found in central Africaand Madagascar (Heraty 1985, 2002, Heraty and Wool-ley 1993). Sixteen species currently are recognized,but as many as 60 may be eventually recognized (Her-aty 2002; A.C., and J.M.H., unpublished data). Severalspecies of Kapala have been reared from EctatommaFr. Smith and Gnamptogenys Roger (Ectatomminae)and from Odontomachus Latreille and PachycondylaFr. Smith (Ponerinae) (Wheeler and Wheeler 1937;Clausen 1941; Heraty 1994b, 2002; Lachaud and Perez-Lachaud 2001; Howard et al. 2001). The species de-

1 El Colegio de la Frontera Sur, Apdo. Postal 36, 30700 Tapachula,Chiapas, Mexico.

2 Department of Entomology, University of California, Riverside,CA 92521.

3 Centre de Recherches sur la Cognition Animale, CNRS-UMR5169, Universite Paul Sabatier, 118 route de Narbonne, 31062 Tou-louse Cedex 4, France.

0013-8746/06/0567Ð0576$04.00/0 � 2006 Entomological Society of America

scribed herein is the Þrst available record of Kapalafrom cocoons of the Neotropical ant Ectatomma rui-dum Roger, collected in Chiapas, Mexico, and waspreviously regarded as K. sulcifacies (Cameron).K. sulcifacies was presumed to be a single highly vari-able species (Heraty and Woolley 1993); although itnow seems to be a complex of species (A.C., andJ.M.H., unpublished data). Howard et al. (2001) pro-vided information on cuticular hydrocarbons for boththe ant and the eucharitid wasp described herein (asK. sulcifacies), and behavioral interactions betweenthe two species within the ant nest. Further collectingshowed that different populations of E. ruidum areparasitized by Kapala izapa n. sp., Kapala iridicolor(Cameron), or by both species. An account of thegeneral biology and life history ofK. izapa parasitizingE. ruidum is given, the hosts and the host plant forK. iridicolor are identiÞed, and Þeld observations onadult behavior are noted.

Materials and Methods

Sampling Procedure and Study Sites. Several colo-nies of ants [Ectatomma ruidum, Gnamptogenys spp.,and Pachycondyla stigma (F.)] were sampled from acoffee plantation on the Experimental Station of theInstitutoNacionaldeInvestigacionesForestalesyAgro-pecuarias at Rosario Izapa, Tuxtla Chico municipality,Chiapas, Mexico (14�58�25� N, 92�09�19� W, 430 mabove sea level) between February 1997 and January1998, and from a secondary growth Þeld plot atCanton Leoncillos, Tapachula municipality, Chiapas(14�45�59� N, 92�24�12� W, 25 m above sea level) inAugust 2004. For each colony, the number of queens,males, workers, cocoons, and larvae were recorded.The cocoons were dissected and the number and stageof parasitoids (planidia, larvae, pupae, and adults)were recorded. Ant larvae were visually checked forany planidium or sign of attack (scar).Life History ofK. izapa. Because of the difÞculty of

rearing the parasitoids to adulthood, the life history ofthis eucharitid was reconstructed from observationsof individual parasitoids at different developmentalstages obtained during cocoon dissection. Twenty-three parasitized larvae of E. ruidum with attachedplanidia were obtained from cocoons. Each parasit-ized larva was placed in a small petri dish and kept ina climatized room (28�C � 2�C and 75% � 5% RHunder a photoperiod of 12:12 [L:D] h) to follow in-dividual development of the parasitoids. Another 28second- and third instars attached to the host wereobtained from dissections and kept under the sameconditions. Only pupae can be identiÞed to species.Because K. iridicolor and K. izapa both attack E. rui-dum, it is possible that larval stages could be attributedto either species; however, only K. izapa were rearedfrom colonies with larvae at Rosario Izapa, and we areconÞdent of the association. Adult wasps were cap-tured upon emergence from the host cocoons, placedin glass vials, and provided with a streak of honey anda piece of moist cotton. Vials were kept in the same

conditions and observed every day to determine sur-vivorship. Results are presented as mean � SD.LifeHistory ofK. iridicolor.Parasitoid behavior was

observed in the summer of 2002 at Canton Leoncillosin a Þeld plot of 500 m2 with a density of 1.3 E. ruidumnests per m2. During 2 wk from 14 July to 28 July 2002,K. iridicolor was observed in a Þeld plot dominatedby Melampodium divaricatum (L. C. Richard) D. C.(Asteraceae) and Lantana camara L. (Verbenaceae),and with several mango trees, Mangifera indica L.(Anacardiaceae). On 14 July, adults of K. iridicolorwere Þrst observed on M. divaricatum. Subsequently,daily observations where made at 8 a.m. and beforenoon. Additional observations were made in 2004, and,on this occasion, two colonies of E. ruidum were col-lected, and the cocoons dissected to conÞrm the as-sociation.Molecular Analysis. To conÞrm the identity ofK. iridicolor at the two study sites, the right legs orgaster of four adult females from the two sites andthree different hosts (Rosario Izapa: one femalereared fromG. regularis, 11-VIII-1997 and one femalereared from P. stigma, 18-VIII-1998; Canton Leon-cillos: one female collected in the Þeld 19-VII-2002,and one female reared from E. ruidum, 16-VIII-2004),were sequenced for the 28S-D2 ribosomal region fol-lowing the protocols in Heraty et al. (2004). Thesesequences were compared with other Kapala beingdone as a separate study (A.C. and J.M.H., unpub-lished data). All primary specimen vouchers are de-posited in the Entomology Research Museum at theUniversity of California, Riverside.Collections and Terms. Voucher specimens of the

oviposition host plant, ants, and parasitoids were de-posited in the botanical and entomological collectionsof El Colegio de la Frontera Sur at Tapachula, Chiapas,Mexico (ECO-TA). Additional specimens of Kapalawere deposited in the Entomology Research Museum,University of California, Riverside (UCR); the Na-tional Museum of Natural History, Washington, DC(USNM); and the Canadian National Collection ofInsects, Ottawa, Canada (CNCI). Terms for morphol-ogy follow Heraty (2002).

Results

Kapala izapa Carmichael n. sp.(Figs. 1, 2, 5Ð19)

Type Material. Holotype male, MEXICO: Chiapas,Rosario Izapa, IV.1997, G. Perez and J.-P. Lachaud, exEctatomma ruidum. Deposited in USNM collection(on permanent loan from UCR). Paratypes: same data(Þve females, three males, UCR; Þve females, twomales, USNM; three females, one male, ECO-TA);COSTA RICA: Heredia, La Selva, 3 km S Pto Viejo,10�26�N84�01�W,H.A.Hespenheide, 19-IV-1986(twomales), 28-VI-1986 (three males), 29-III-1987 (onemale), 19-VII-1982 (one male), 2-IV-1987 (one male)(allUCLA); same location,butT.W.Sherry, 25-X-1978

568 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 99, no. 3

(two males, UCR); Alajeula, 5Ð10 mi W La Fortuna,300 m, J. Helava, 25-IX-1972 (one male, CNCI).Etymology. An arbitrary combination of letters

based on the type locality, Rosario Izapa.Diagnosis. K. izapa can be placed within theK. sulcifacies complex by the following characters:Þnely carinate face with a ßat superclypeal area(Fig. 6); high proÞle of the mesonotum (Figs. 1 and 8);and abrupt elevation of the posterior margin of thescutellar disc above the scutellar spines (Figs. 1, 5, and8). K. izapa can be distinguished from K. sulcifacies(Nicaraguan type material) by differences in the an-

tennae: ßagellar branches of males 1.7Ð1.8 times heightof head (Figs. 1 and 2; compared with 1.2Ð1.3 times inK. sulcifacies, Fig. 3, arrow), ßagellum of females 1.2Ð1.3 times height of head (Fig. 8; compared with 0.95Ð1.05 in K. sulcifacies); longer; frenal spines archingboth laterally and dorsally (Figs. 1 and 5; comparedwith relatively level and straight inK. sulcifacies); andfrenal spines are entirely brown (some females) orapically brown and black basally (entirely black in K.sulcifacies). K. izapa can be distinguished from otherundescribed members of the complex located in Pan-ama and Nicaragua by having the ßagellar segments

Fig. 1–7. 1Ð2, K. izapa (male): 1, habitus (lateral); 2, antenna. 3, K. sulcifacies, paralectotype male habitus (lateral); 4,Kapala sp. near sulcifacies (Trinidad), male habitus (lateral). 5Ð7, K. izapa (male): 5, habitus (dorsal); 6, head (frontal); 7,mesosoma (lateral).

May 2006 PEREZ-LACHAUD ET AL.: BIOLOGY AND BEHAVIOR OF Kapala 569

light brown with a slightly lighter scape, comparedwith a uniformly dark brown or black antenna, and arelatively smooth mesepimeron (Fig. 7), comparedwith a mesepimeron with distinct horizontal carinaethat cross the femoral depression. An undescribedspecies from Trinidad (�K. sulcifacies in Heraty andWoolley 1993) also has the long antennal rami (Fig. 4),but these have a higher mesonotum (1.2Ð1.3 timeshead height compared with �1.0 times inK. izapa;Fig.4, arrow). The frenal spines are robust and linear inlateral view in the Trinidad specimens.Male. Length 5Ð5.3 mm. Head, mesosoma, coxae,

and petiole black with a dark green sheen; frenalspines black basally, becoming brown apically; eyesgray; legs entirely yellow; gaster dark brown; antenna,mandibles and wing venation light brown; wings hy-aline.

Head subtriangular, 1.7Ð1.8 times as broad as high(Fig. 6); interocellar distance 1.2Ð1.3 times headheight. Eye height 0.5Ð0.6 times head height, eyesbare. Face, excluding supraclypeal area, uniformlyÞnely carinate, carinae transverse ventrally and ver-tical adjacent to scrobes (Fig. 6), face with Þne semi-appressed setae, setae becoming denser above toruli.Scrobal depression without striae, but some specimenswith a weak median carina. Clypeus smooth and bare,laterally demarked by deep sulci, anteclypeus distinctand linear; supraclypeal area bare and weakly rugose,almost smooth, swollen medially and not demarked

laterally or ventrally. Malar space 1.2Ð1.3 times widthof clypeus. Maxillary palpi with three segments andlabial palpi two segmented; labrum with seven elon-gate digits. Antenna with 12 segments (Fig. 2); scape0.26Ð0.32 times head height, 2.6Ð3 times as long asbroad; ßagellomeres 1Ð9 with single long branch al-ternating in origin along midline, branch of basal ßag-ellomere 1.7Ð1.8 times head height, branch of ßag-ellomere 9 1.4Ð1.6 times head height, ßagellomere 10as long as branch of preceding ßagellomere; in proÞle,longest branch exceeds height of mesonotum by atleast one-third of its length (Fig. 1).

Mesosoma narrower than head (Fig. 5) and 1Ð1.1times as long as tall (Fig. 1). Dorsal aspect of me-soscutum, axilla, and scutellar disc roughly level.Mesonotum, axilla, scutellar disc, and posterior verti-cal face of scutellar disc densely setose. Midlobe ofmesoscutum transversely carinate and 1Ð1.1 times astall as head height; mesoscutum in frontal viewbroadly rounded dorsally, in lateral view, anterior faceof midlobe of mesoscutum broadly rounded (Fig. 1);carinae on lateral lobe of mesosoma more pronounceddorsally. Axillae transverse with strong parallel cari-nae; scutellar disc medially with carinae converging tomidline; axillular carina deep and parallel to dorsum,axillula with carinae becoming weaker posteriorly.Frenal spines 1.1Ð1.2 times longer than rest of themesosoma and 14Ð16 times as long as wide; spinesbowed laterally and dorsally and gradually tapering

Fig. 8–11. K. izapa: 8, female habitus (lateral); 9, female habitus (dorsal); 10, unfed planidium attached to ant larva; 11,fed Þrst instar from prepupa.

570 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 99, no. 3

apically (Figs. 1 and 5); spines longitudinally carinate,lateral margins in dorsal view looking serrate.Propodeal disc mostly smooth with a strong borderingcarina; propodeal furrow and metepimeron areolatewith strong carinae; callus mostly smooth and pilose.Posterodorsal corner of prepectus triangular; prepec-tus and dorsolateral half of pronotum pilose, pronotumventrolaterally bare with areolate sculpture (Fig. 7).Mesepisternum and upper mesepimeron mostly smoothand shining with few weak carinae and rugae ven-trally, bare except for sparsely setose posterodorsalcorner of mesepimeron (Fig. 7); femoral depressionmore or less parallel-sided and crenulate. Hind coxasmooth with a single row of setae; fore coxa with twoweak carinae and scattered setae; fore femur Þve timesas long as broad; hind femur 6.5Ð7 times as broad aslong;basitarsus as longasother fourcombined.Forew-ing 2.5Ð2.7 times as long as broad, pilose and lackingwing fringe; marginal vein eight times longer thanstigmal vein; postmarginal vein at most 0.3 times aslong as the stigmal vein. Marginal vein of hind wing0.6Ð0.7 times length of hind wing; hind wing withfringe.

Petiole with Þne, parallel carinae, petiole 7Ð9 timesas long as broad, 2.5Ð2.7 times as long as hind coxae.First gastral tergite 1.1Ð1.2 times as long as the petiole;tergal scar present and linear. Phallobase parallel sid-ed; aedeagus ßat and tonguelike; paramere with threeto Þve stout setae and paramere exceeding the digitus;digitus discoidal with three to four marginal spines.Female. Length 4.9Ð5.2 mm. As in male, except for

the following: scutellar spines black or brown basally;antennal ßagellum weakly serrate with eight ßagellarsegments, terminal clava with three fused segmentsand slightly expanded (Fig. 8); scape and pedicel yel-low; ßagellum light brown; frenal spine tapering to asingle ßattened point (Fig. 9); hind femur 6Ð6.5 timesas long as broad; petiole � 4 times as long as wide, 2times length of hind coxa; gaster bulbous; hypopygiumwith two tufts of stout setae.Planidium (Figs. 10, 11, 13–17). The morphology

and setal pattern are typical of most Eucharitinae:pleurostomal spine present; four pairs of placoid sen-silla on cranium, second pair from anterior marginlarge and with two narrow lines of weakness leadingto anterior sensillum and to lateral margin of cranium,dorsal cranial spines absent; hatchet-shaped scleritepresent; pleurostoma extending posteriorly as an elon-gate triangular process; ventral transverse process ofcranium Þngerlike and separated medially; tergopleu-ral line (tp) separating pleural and dorsal tergitespresent but faint on tergites (T) IIÐVII; TI and II fuseddorsally, with two pairs of small setae ventrally andtwo pairs of placoid sensilla dorsally; TIII with threepairs, dorsally with placoid sensilla; TVI with stoutlateral spine (Fig. 13); TVII with minute seta on ven-tral margin; TIX entire and with long lateral process;caudal cerci as long as TXI�XII and stiff.

The planidium of this species is very similar tothe planidium of a Kapala sp. (sulcifacies complex)(Heraty and Darling 1984), except for the ventralsetae on TI and II, faint tergopleural line, and presence

of two pairs of anterior sensilla on the cranium. Theplanidium burrows partially into the host, and the siteof entry becomes melanized (Fig. 14). Most planidiawere found burrowed into the sixth and seventh ab-dominal segments of the host larva (Fig. 15, blackdots), but some planidia found on the larval thorax andanterior abdomen were lightly attached (dotted cir-cles) and had not invoked melanization by the host,which suggests that feeding had not taken place.Third Instar (Fig. 18). This instar is virtually iden-

tical to the description of the same stage by Clausen(1940b) (his Fig. 7) for Chalcura deprivata (Walker),with the exception that only Þve pairs of spiracles areapparent as opposed to eight pairs in Chalcura. Thelarva of Schizaspidia convergens (Walker), a memberof the same clade of large poneromorph parasites, issimilar but has a prominent dorsal expansion of theÞrst thoracic segment (Clausen 1940b) (his Fig. 8).Dorsally the integument of K. izapa is covered byminute papillae (Fig. 18). A minute pair of weaklybidentate mandibles is located ventrally on the headcapsule, and the Þrst instar exuvium (and presumablythe second instar exuvium) remains attached ventrallyto the basal abdominal segments (Fig. 18, pl-ex).Pupa (Fig. 19). The pupa resembles the adult, with

the elongate scutellar spines readily apparent. Fea-tures that are shared with the pupa of Chalcura andSchizaspidia (J.M.H., unpublished data) include thepronounced conical projections from each of thethree ocellae (Fig. 19, ocellar horns), and the bulbousprojections from the lateral margins of the Þrst ab-dominal segment, and dorsally and laterally on thesecond segment (Fig. 19, abpr). These projections arenot associated with any features found on the adult,and their function is unknown. The larval exuviumremains attached to the terminal segment of the pupa.Pupation occurs within the host cocoon.

Life History of Kapala izapa

Development and InstarDuration.The reconstruc-tion of the life history of K. izapa was determinedthrough dissection of cocoons and direct observations.The Þrst instar of K. izapa (Figs. 10 and 13) measures0.13 mm in length and is Þrmly attached externally tothe cuticle of last instar of E. ruidum. The earlierinstars of the host seem not to be attacked. A circularscar indicates the attachment of a planidium to thesurface of the host (Figs. 14 and 16). Typically, oneparasitoid per host is observed, but up to three cancomplete development on one host. In these cases, theadult parasitoids are much smaller than average. In asample of 96 parasitized hosts (belonging to severalnests), 84 host larvae (87.5%) had one planidium, 10had two planidia (10.4%), and only two host larvae hadthree planidia (2.1%). In this sample, planidia werepreferentially attached to the ventral region of thehost abdomen (43.7%) or just posterior to the head ofthe host larvae (31.2%) (Fig. 15). Of the remainingplanidia, 14.6% attached to the ventral side of thethorax, 7.3% anchored into the dorsal region of theabdomen, and 3.1% were attached at the anal region.

May 2006 PEREZ-LACHAUD ET AL.: BIOLOGY AND BEHAVIOR OF Kapala 571

Planidiawerenotobservedon thedorsal regionsof thethorax or the head of the host larva.

Like other Eucharitinae, planidia remain attachedexternally to the ant larva or prepupa within the co-coon (Clausen 1928, 1940b; Heraty 1994b) and do notburrow into the cuticle as has been observed inOraseminae or Gollumiellinae (Heraty et al. 2004).Only after the ant larva Þnishes weaving its silk cocoonand begins pupation does the planidium begin to feedand develop. As the planidium feeds, it swells (Fig. 14)and remains attached to the host prepupa at the siteof its initial attachment. When fully fed, the planidiameasure 0.67 � 0.02 mm in length (n� 5) (Figs. 11 and17). When the host is ready to pupate, the engorgedÞrst instar detaches and begins a wandering phase,during which it migrates over the host surface. Thiswandering phase coincides precisely with the larva-to-pupa molting of the host. After the host has Þnishedshedding its larval exuviae, the wandering Þrst instarresumes feeding on the ventral region of the thoraxjust under the last pair of legs of the newly trans-formed ant pupae. All (n � 28) second instars werefound in this position on ant pupa. The Þrst and pre-sumably second instar exuviae remain attached to theventral thoracic region of the parasitoid larva (Fig. 18,pl-ex). Larvae from all castes of E. ruidum (females,males, and workers) were parasitized by K. izapa.

Only four cases (n� 23) could be followed from theplanidium stage to the wandering phase due to thehigh mortality observed when parasitized host larvaewere experimentally removed from their cocoons.The mean duration of feeding by Þrst instars was8.0 � 0.8 d (n� 4). None of the Þrst instars observedwandering on artiÞcially “naked” ant pupae (n � 15)were able to reattach themselves to the host.

After relocation to the thoracic region of the hostpupa, the parasitoid larva continues feeding until theentire host is consumed and only the integument isleft. All the larval exuviae remain attached to the lastinstar, and inK. izapa there are three instars. The thirdinstar remains with its mouthparts inserted in theintegument of the completely drained ant pupa afterfeeding is completed. Once the larva has Þnished feed-ing on the host, metamorphosis begins. There is noexpulsion of the meconium at this stage. The pupatransforms inside the host cocoon. The last sign of thecomplete transformation of the larva into a pupa is theformation of the characteristic frenal spines.

Duration of the pupal stage was 13.33 � 3.06 d (n�4) for females and 7.00 � 1.41 d (n � 3) for malesunder laboratory conditions. Pupation and emergenceof adults take place within the host nest. Adult waspsemerge inside the host cocoon, cast off the pupalexuviae, expel the meconium and cut an opening atthe apical end of the host cocoon with their mandibles.Adults must leave the nest to mate and lay eggs. Fe-males of K. izapa are proovigenic. Although pairs ofvirgin adults were put together in the laboratory, themating behavior was not observed.Adult Longevity. Males of K. izapa kept in labora-

tory conditions and provided with diluted honey andwater survived 4.6 � 1.0 d (n � 7) and females 4.0 �

1.4 d (n � 10). Although they were provided honey,adults were not observed feeding; however, they wereobserved taking water from cotton saturated with wa-ter.Interaction inside Nests. Howard et al. (2001) pro-

vided an account of the interaction between adults ofK. izapa (reported as K. sulcifacies) and adult workerants inside the ant nest. Brießy, adult parasitoidsemerging in artiÞcial nests in laboratory conditionswere not attacked while initially searching for the nestexit hole. If the K. izapa became trapped in the arti-Þcial nest, the worker ants would eventually killthem. The cuticular hydrocarbon proÞle of recentlyemerged adult wasps partially mimics that of the hostbrood (Howard et al. 2001). This chemical resem-blance may explain the tolerance toward eucharitidsand may allow the parasitoids to exit by their ownmeans or by being transported by ant workers. Thissituation is similar to that found betweenOrasema sp.(�O. xanthopusCameron; see Heraty et al. 1993) andits ant host, Solenopsis wagneri Santschi (�S. invictaBuren) (Myrmicinae) (Vander Meer et al. 1989).

Kapala iridicolor (Cameron)

Collections and host associations. MEXICO: Chia-pas, Rosario Izapa (RI), 11-VIII-1997, ex nest ofGnamptogenys regularisMayr (one female UCR); RI,18-VIII-1998, ex nest of Pachycondyla stigma (F.) (onefemale UCR); RI, 7-XI-1997, ex nest ofGnamptogenyssulcata (F. Smith) (one female, one male, UCR);12-VIII-1998, ex nest of Gnamptogenys striatula Mayr(one female UCR); Chiapas, Canton Leoncillos (CL),19-VII-2002, collected while ovipositing on Melampo-dium divaricatum (one female, one male UCR); Can-ton Leoncillos, 5-VII-2004, collected on Panicum sp.(one female UCR); 16-VII-2004, collected on M. di-varicatum (one female, one male, UCR); 16-VIII-2004,exEctatomma ruidumnest #2 (one female, two males,UCR); 16-VIII-2004, ex.E. ruidumnest #1 (two males,two females, UCR).Molecular Diagnosis. Because of the different host

associations at the two localities, we were interestedin the genetic similarity of the two populations totest for potential morphologically indistinct (cryptic)species. All four K. iridicolor from the two localitieswere identical for 637 bp of 28S-D2 (GenBank no.DQ004748). These differ by one base from a morpho-logically similar population of K. iridicolor fromColumbia (position 516: AG; GenBank no.AY671874; Heraty et al. 2004), and by no 3 bp fromother populations belonging to the K. iridicolorcomplex (GenBank nos. AY671842, AY671847,AY671853Ð54, AY671856Ð57, AY671859, AY671861Ð63, AY671866Ð67, AY671871Ð72, AY671876, AY671886;Heraty et al. 2004). The 28S-D2 forK. iridicolor differsfrom K. izapa collected in Costa Rica (GenBank no.AY671893 refered as K. nr sulcifacies #6; Heraty et al.2004) by a total of 10 bp, which includes a single basepair insertion, eight transitions, and one tranversion(A.C., and J.M.H., unpublished data). Thus, theMexican K. iridicolor are identical for 28S-D2, despite

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the different host associations. The minimal number ofbase differences between geographically distant pop-ulations of K. iridicolor needs to be evaluated, but itlikely is not enough to support these as being differentspecies (Heraty 2004). However, the differences be-tweenK. izapa andK. iridicolor are more than enoughto support these as two distinct species (Heraty 2004).Mating and Predation.A mating pair ofK. iridicolor

was observed on unidentiÞed vegetation �20 cmabove the entrance hole of a nest of E. ruidum at theCanton Leoncillos site. Because the beginning of thesequence was not observed, evidence of courtship isnot available. Immediately after mating, the maletook ßight and the female remained on the vegetation,preening herself for �10 min before ßying away.Males of this species also were observed hovering overant nests or resting on the vegetation around and overthe ant nests. On one occasion, a robber ßy (Diptera:Asilidae) that was perched on the iron stakes used tosignal the entrance of an ant nest was observed eatinga K. iridicolor male.Oviposition Behavior. At the time of emergence,

the abdomen of the adult female is completely dis-tended, with each ovary comprised of 18Ð20 ovarioles;recently emerged females had an average of 4,578 �623 fully developed eggs (n� 5). Eggs obtained from

dissections of gravid females are white with a smoothchorion (Fig. 12). Their overall length is 0.28 � 0.01mm (n � 8), including a translucent white, narrowcaudal stalk which is slightly longer than the egg body(stalk, 0.15 mm; egg body, 0.13 mm).

Two females of K. iridicolor were observed; ovipo-sition took place directly on the involucral bracts ofundeveloped ßower buds ofM. divaricatum that were�1.2 m above the ground. Although there were sometaller plants (1.6Ð1.7 m), females wasps were onlyobserved on plants �1.4 m in height and were onßower buds �1.2 m above the ground. Higher ßowerswere examined for signs of oviposition but no eggswere found. Females walked on ßower buds with thetips of their antennae touching the surface, raisedtheir abdomen, and laid eggs in masses directly be-tween the bracts or on the surface of the ßoral buds.

Other plants in ßower were present in the Þeld plot(e.g., L. camara), but females were observed ovipos-iting only on M. divaricatum and one female was col-lected on a grass. While ovipositing, females wereoriented head down on the unopened ßower buds,with oviposition lasting from 1 to 2 min. Upon com-pletion, females would either reorient themselves onthe same ßower bud or ßy to another plant. Occa-sionally, the same female would oviposit on the same

Fig. 12–19. K. izapa: 12, egg; 13,planidium,dorsal andventral view(tp, tergopleural line); 14,planidiumpartiallyburrowedinto host cuticle; 15, location of attachment of planidia to host larvae (n� 96 host larvae, 110 planidia; black dots, burrowedwith melanization); 16, host Ectatomma with attached planidium (pl); 17, fed planidium from host prepupa; 18, third instar(pl-ex, planidium exuvium); 19, pupa (abpr, abdominal projection).

May 2006 PEREZ-LACHAUD ET AL.: BIOLOGY AND BEHAVIOR OF Kapala 573

involucral bract just below the Þrst mass of eggs. Thenumber of eggs present in a mass varied from 11 to 242(n� 9; median � 52; ranked sizes, 11, 18, 22, 33, 52, 67,88, 102, and 242). The largest value of 242 may cor-respond to two closely placed egg masses laid by thesame female.Interaction with Worker Ants. Workers of E. rui-dum foraged actively on M. divaricatum. Some work-ers collected the liquid produced by extraßoral nec-taries located on the axils of the veins on the abaxialside of leaves and transported it back to the nest as adrop between their mandibles. Other workers wereobserved standing still in a classic ambush predatoryposition (Schatz and Wcislo 1999) on branches andon the stems of leaves and ßower buds. Planidia werenot directly observed in the Þeld, and their exactmeans of entering the nest remains uncertain. Otherspecies of Kapala have been observed attaching toinanimate objects (insect pins) or other eucharitids(e.g., planidia are occasionally found attached exter-nally to museum specimens; J.M.H., unpublisheddata). Most likely, the planidia attach externally toforaging workers. Eventual tending of the ant broodby foragers probably results in the transfer of planidiato the host larva.Transport of Parasitized Cocoons by E. ruidum

Workers. Worker ants were observed transportingwhat seemed to be empty cocoons far away from theircolonies. This behavior contrasts with the recycling ofempty cocoons that are normally provided to lateinstars by worker ants to initiate cocoon weaving(workers chew empty cocoons and provide smallpieces to larvae). Three cocoons that had been trans-ported this way were collected and taken to the lab-oratory. All of the cocoons contained the character-istic exuvia and meconium (see above) of eucharitidparasitoids and the remains of the parasitized ant pu-pae. One cocoon also contained a small, dead K. iridi-color female that had failed to emerge. Another femaleparasitoid had successfully emerged from this cocoon.

Discussion

Host information, whether for plant oviposition orant host, is known for only 29 of the 53 recognizedgenera of Eucharitidae (Heraty 1994b, 2002). Accu-rate host associations (parasitoids reared directly fromhosts) are known for only a few species (e.g., Wheeler1907; Clausen 1923, 1928, 1941; Ayre 1962; Johnson etal. 1986; Heraty and Barber 1990; Heraty 1994a, 2000,2002; Lachaud et al. 1998; Lachaud and Perez-Lachaud2001).

In this study, we showed that two species ofKapalaparasitize E. ruidum and conÞrm our previous reportof K. izapa (formerly reported as K. sulcifacies) usingE. ruidum as a host in the southern part of Mexico. K.iridicolor also attacks Gnamptogenys sulcata, G. stria-tula, G. regularis, and P. stigma at Rosario Izapa. Thisconstitutes the Þrst report of hosts for K. iridicolor.Other species of Kapala, most of them undescribed,are known to be associated with medium to largeponeromorph ants: Pachycondyla crassinoda (La-

treille) (Myers 1931),Odontomachus insularisGuerin-Meneville (referred to asO. haematodes insularis pal-lens; Clausen 1941), Odontomachus bauri Emery,Odontomachus hastatus (F.), P. harpax (F.) (Heraty2002), Odontomachus brunneus (Patton), Odontoma-chus laticeps Roger, and Odontomachus opaciventrisForel (Lachaud and Perez-Lachaud 2001). Only onespecies,Kapala floridana (Ashmead), was reported toparasitize Myrmicinae, Pogonomyrmex badius (La-treille) (Wheeler 1907), but this association, basedonly on the opinion of Ashmead (Wheeler 1907, p. 17),is probably incorrect.

The life history and morphology of the immaturesof K. izapa do not differ greatly from those Þrst de-scribed by Wheeler (1907) for other eucharitids.Planidia were found attached externally posterior tothe mouthparts and laterally on the mesosoma butwith most located on the ventral or lateral regions ofthe abdomen. The attachment of planidia by K. izapais similar to that of another eucharitine, Pseudo-chalcura gibbosa (Provancher), that parasitizes Cam-ponotus (Heraty and Barber 1990).

As for other eucharitids, the Þrst instar of K. izapacompletes its development on the host mature larvaÐprepupa within the cocoon, is cast-off with the hostÕslarval exuvium, and relocates itself to the mid-sectionof the pupa to resume feeding. By initially attaching tothe larval stage of the host, feeding on the prepupa,and completing development on the pupa, K. izapacan be classiÞed as a koinobiont, larvalÐpupal, ecto-parasitoid of E. ruidum. Only a few hymenopteranectoparasitoids can be considered as koinobionts,which include ichneumonids (Adelognathinae, Try-phoninae, and polysphinctine Pimplinae), some roga-dine braconids, most dryinids, and a few pompilids(Gauld and Hanson 1995). K. izapa is a solitary para-sitoid but two and three individuals can exceptionallydevelop on a single host pupa, similar to P. gibbosa(Heraty and Barber 1990) and toDilocantha lachaudiiHeraty (Perez-Lachaud et al. 2006).

Plant hosts are known for only four species ofKapala and include species from seven differentfamilies: Asteraceae, Geraniaceae, Euphorbiaceae,Boraginaceae, Malvaceae, Amaranthaceae, and an as-clepiadlike cactus (Table 1). As far as oviposition isconcerned, there seems to be little uniformity in habiteven among species of a genus; females seem to beattracted to the physical qualities of certain types ofplant structures and not so much to a speciÞc planttaxon (Clausen 1940a, Heraty 1994b). In our study,K. iridicolor females were observed laying eggs di-rectly on the surface of unopened ßower buds ofM. divaricatum, without making any puncture,whereas it was reported previously ovipositing intothe ßower buds of Cordia macrostachya (Jacquin),as was also K. sulcifacies (Heraty and Darling 1984,Heraty and Woolley 1993). K. iridicolor is the Þrsteucharitid known to deposit their eggs both on thesurface and inside ßower buds of different plant spe-cies. The native range ofM. divaricatum is from south-ern United States and Mexico, throughout CentralAmerica (Guatemala, Belize, Honduras, El Salvador,

574 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 99, no. 3

Costa Rica, and Nicaragua), and into Colombia andBrazil. Its distribution seems to follow that of theK. sulcifacies complex and of theK. iridicolor complex(see Fig. 6 from Heraty and Woolley 1993), but this isthe Þrst association of M. divaricatum with Kapala.

Some of the tropical Eucharitidae, likely those thathave overlapping generations throughout the year,may change their oviposition plants with the seasons(Clausen 1940a). At the Canton Leoncillos site, adultsof K. iridicolor and ßowering M. divaricatum plantsare present in the Þeld during the Þrst part of the rainyseason (from July to September; J.-P.L., and G.P.-L.,unpublished data). At Rosario Izapa, 35 km fromCanton Leoncillos, M. divaricatum is absent, and theplants used for oviposition by K. izapa and byK. iridicolor were not identiÞed. More observationsshould be done throughout the year and at this andother sites, to determine the existence of alternativehost plants for these eucharitid species in the Soco-nusco region.

It is generally assumed that the adults of the ma-jority of species do not feed; yet, females of Kapalafurcata (F.) of Panama were observed upon foliageheavily infested with aphids, apparently feeding uponhoneydew (the plant species was different from thatused for oviposition; Clausen 1941), and females ofOrasema valgius (Walker) were observed feedingupon the nectar of its host plant,Exocarpos sp. (Heraty2000). It seems thus likely that both male and femaleeucharitids gather some nutritive resources fromplants and reports do exist of plants being visited byeucharitids. For example, plants of Hyptis capitata L.(Lamiaceae), which possess extraßoral glands on theteeth of leaves, are visited by Eucharitidae (Mexzonand Chinchilla 1999). However, these authors did notdirectly observe feeding or oviposition on this plantspecies.M. divaricatum also possess extraßoral glands,and several insects are attracted to these nectaries,including a single record of a nongravid K. iridicolor

female. It is, however, clear that the association be-tween K. iridicolor and E. ruidum is based on theforaging behavior of worker ants which do gathernutritive substances directly from Melampodiumplants and also prey on insects attracted to this foodsource. Presumably, sugars produced by M. divarica-tum (Bohlmann and Le Van 1977, Huther et al. 1999),could act as a strong attractant to bring potential anthosts to the ßowers and facilitate planidia waiting forphoretic transport to the ant colony, as also was sug-gested for Orasema valgius (Heraty 2000).

Finally, Heraty and Woolley (1993) suggested thatthe morphological variation within Kapala may beresolved as supporting few polymorphic species or atremendous array of new species. Molecular studiesare supporting the latter hypothesis, with minor mor-phological variation distinguishing distinct lineages(A.C., and J.M.H., unpublished data). Informationabout plant or within-nest biology for this and otherspecies as well as oviposition habits and ant hoststhroughout their geographical range will prove invalu-able in further resolving the limits and relationships ofthis species complex.

Acknowledgments

We thank Jurgen Pohlan and Annie Damon (El Colegio dela Frontera Sur) for identifying the host plant. Don CiprianoGonzalezand familykindly letusworkat their garden-mangoplantation. Thanks are also due to Lupita Nieto for theSEM photo, Jose Antonio Lopez for technical support, andTim Batchelor (University of Nottingham) and two anony-mous referees for useful comments on a previous versionof this article. The research was supported by grant fromSEP-CONACyT, Mexico (project number 44417).

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Table 1. Host plant records for Kapala

Eucharitidspecies

Host-plant FamilyPlant structure used for

ovipositionCountry Reference

Kapala sp. Cordia sp. Boraginaceae Into undeveloped ßowerbuds

Trinidad Heraty and Darling 1984

Kapala sp. Tragia volubilis L. Euphorbiacea Random, completely exposedon the undersides ofleaves

Cuba Clausen 1940a

K. furcata (F.) Mikania micrantha (L.)Kunth.

Asteraceae In ßower buds Panama Clausen, 1940a

Undetermined sp. Amaranthaceae PanamaK. iridicolor

(Cameron)Cordia macrostachya

(Jacquin) Roemer andSchultes

Boraginaceae Into closed ßower buds Trinidad Heraty and Woolley1993

Gossypium hirsutum L. Malvaceae NicaraguaCordia cana Boraginaceae Costa RicaMelampodium divaricatum Asteraceae On closed ßower buds Mexico This study

K. sulcifacies(Cameron)

C. macrostachya Boraginaceae Into closed ßower buds Trinidad Heraty and Woolley1993

Gossypium hirsutum L. Malvaceae NicaraguaUndetermined sp. Asclepiad-like cactus Nicaragua

K. terminalisAshmead

Tragia volubilis L. Euphorbiacea On upper surface of leaves Cuba Clausen 1940aCasearia spinescens

(SW.) Griseb.Flacourtiaceae Cuba

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Received 13 July 2005; accepted 14 January 2006.

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