蝶と蛾 Science 67（2）: 58-66, October 2016

Biology and chemical ecology of calleta（ ） in southern

Joseph H. Louwagie III and Richard S. Peigler Department of Biology, University of the Incarnate Word, 4301 Broadway, San Antonio, Texas 78209-6397 USAi

Abstract Field and lab observations were made on Eupackardia calleta, a large saturniid that in southern Texas. Leucophyllum langmaniae is newly reported as a hostplant in nature. An oviposition was found in nature comprising 17 eggs on 8 leaves. Samples of cocoons preyed upon by a mammal are illustrated. Three parasitoids are reported: the euplemid Anastatus from eggs, the tachinid Lespesia sp. near texana, and the chalcidid Conura maria from cocoons, the latter two for the first time. Predation on a third-instar larva by the vespid Polistes exclamans was observed. Phototactic behavior of males（ which are normally diurnal） and a multiple mating by a female are newly reported. Three new county records for Texas are cited. Biochemical analyses were done on unfed first-instar larvae, fed second-instar larvae, mature larvae, and adult , and the following compounds were found: 3,4-dihydroxyphenylalanine, dopamine, norepinephrine, hydroquinone, 2-（dimethylamino）ethylacetate, trehalose, and 3,5-dimethylphenol, some of which probably provide protection from predators. Trials on larvae were conducted in the lab and field with five species of ants（ Hymenoptera: Formicidae） to test for chemical protection against predation. It was observed that larvae have some chemical protection from ants, and because some of the above mentioned biogenic chemicals were found to be carried into the adult stage, it is proposed that moths of E. calleta may be Müllerian instead of Batesian mimics of Battus（ Papilionidae）.

Key words ants, biochemical defenses, Formicidae, Leucophyllum, mass spectrometer, , parasitoids, predators.

Introduction likely also serves as a model for E. calleta（ Waldbauer and Sternburg 1976）. and B. polydamas are common The large saturniid Eupackardia calleta（ Westwood） in Texas and southern Arizona where E. calleta occurs（ Collins （Figs 1, 2） ranges across southern Texas from the Coastal Bend 2007; d’Abrera 2016）. as far north as Cuero and Victoria in the east to the Big Bend and Trans-Pecos sections of the state in the west. It was long The brightly colored larvae of E. calleta（ Figs 6 , 7） are assumed that the distribution in eastern and western Texas was ostensibly aposematic, because they contain biogenic chemicals disjunct（ Tuskes et al. 1996）, but new records to the south and that repel predators（ Collins and Weast 1961; Deml and Dettner southwest of San Antonio reveal that the range is continuous. 1993, 1995, 1997）. Some of these biogenic compounds are The moth also ranges down to the Lower Rio Grande Valley in identified and discussed below in our results and discussion. The the southern tip of Texas and into Mexico. The primary hostplant purpose of this paper is to document new observations on the in southern Texas is cenizo（ , range, foodplants, parasitoids, predators, adult behavior, and the Scrophulariaceae）. Most of what is known about E. calleta in biochemical ecology of the larvae and adults. Texas was summarized by Collins and Weast（ 1961）, Ferguson （1972）, Miller（ 1976）, Tuskes et al.（ 1996）, and Peigler and Materials and methods Maldonado（ 2005）. We made five field trips to areas south of San Antonio to collect Because of their black ground color with red and white cocoons and larvae. Most of our research material of E. calleta markings, the adult moths of E. calleta are believed to be was collected on plantings of the hostplant in Devine（ Medina Batesian mimics of the papilionid Battus philenor（ Linnaeus）, County） and Pleasanton（ Atascosa County）. The main study like diurnal males of the saturniid promethea site in Devine consisted of an unpruned hedge of cenizo shrubs （Drury） and the butterflies Speyeria diana（ Cramer）, Limenitis more than 3 m tall and 40 m long, in a vacant lot adjacent to an arthemis astyanax（ Fabricius）, Linnaeus, Papilio old house. The males were attracted to captive females in San polyxenes asterius（ Stoll）, and the dark form of Antonio, Bexar County. The species has not previously been Linnaeus. In southern Texas, Battus polydamas（ Linnaeus） reported in literature from Atascosa, Bexar, and Medina counties. We reared larvae in cages using sprigs of cenizo inserted in i [email protected] bottles of water. Biology and chemical ecology of Eupackardia calleta（ Saturniidae） in southern Texas 59

To examine possible presence of several compounds that have once, and Crotch（ 1956） reported that a female of ricini been previously reported（ Deml and Dettner 1993） in E. calleta （Wm. Jones） emitted on three occasions 24 h apart, and cenizo leaves, we used an Advion® mass spectrometer of and mated each time. Morton（ 2009） recorded multiple matings the Single Stage Quadrupole model. For MS detection, the in females of , a saturniid that has diurnal ionizing method is ESI+ mode, and the sample injection method males and nocturnal females like E. calleta and S. pavonia. He was direct injection. It has a mass range of 10 m/z to 1,200 m/z, proposed various hypotheses for this polyandry, including and a scanning speed of 10,000 m/z units sec－1 and a mass increased fecundity and reduced predation. The latter was based resolution of 0.5-0.7 m/z units（ FWHM） at 1,000 m/z units sec－ on the hypothesis that because males of C. promethea are 1 over the entire acquisition range. The mass accuracy is ±0.1 mimics of toxic papilionids（ see Introduction above）, but m/z units over the entire acquisition range. For mass sensitivity, females are not, pairs mating during the day may afford the scan sensitivity is 100 pg Reserpine, and the SIM sensitivity additional protection to females from avian predators. However, is 10 pg Reserpine. although they are nocturnal, females of E. calleta also have the mimetic phenotype, so the reduced predation hypothesis Hemolymph samples from and moths were extracted probably would not apply to this species. using micropipettes after puncturing the integument. We tested six samples: unfed larva（ first instar）, 1 week old fed larva A captive pair of E. calleta reared from cocoons collected in （second instar）, mature larva hemolymph, mature larva scoli Devine mated in a cage on 30 March 2014. The female deposited （glandular secretions）, female moth hemolymph, and cenizo many eggs the next two nights in a paper bag, and those eggs leaf. We used a 10 μl sample（ sample + 40 μl de-ionized water） hatched 10 days later. She was placed in a cenizo shrub in the direct injection with a 70% methanol pump infusion. The cenizo front yard（ in northern San Antonio） of the second author after leaf preparation was slightly different than the larvae and moth nightfall on 1 April, and late on 2 April was observed to be samples. We placed one cenizo leaf into a mortar and pestle mating with another male that she had attracted. The next bowl, then added 2-3 drops of de-ionized water. The cenizo leaf morning she was gone, but the male was perched under a porch was crushed and the liquid from the leaf secretions mixed with light that was ca. 3 m from the shrub. It has long been accepted the water, and 10 μl of the sample were extracted. Once the data that males are diurnal and females are nocturnal, with only the were collected, we looked to see if there were certain chemicals latter coming to lights, but Howe（ 1963） illustrated a male he identified in E. calleta by Deml and Dettner（ 1993）. The collected at light in Cuero, DeWitt County, Texas, and our biogenic chemicals were tentatively assigned on the basis of observation verifies his report that males are occasionally molecular ions（ m/z） by using an on-line database called attracted to lights. We do not believe that males normally fly at MassBank, maintained by the National Bioscience Database night, and the above two instances of phototaxis possibly Center, Japan（ Horai et al. 2010）. occurred only because those males were very near lights when mating ceased. It is also possible that females occasionally fly to We ran some brief trials in the field and lab using five species of lights with males still attached to them, leaving the males at the ants to test the hypothesis that larvae possess some chemical lights when copulation ceases and then flying away to oviposit. protection. Larvae were placed beside entrances of ant nests in the field, and along with captive ants in petri dishes in the lab. We did not have the opportunity to take females of E. calleta to We were able to keep ants alive in plastic and glass containers field sites where high populations exist. However, in northern for a week or longer by providing small pieces of tissue paper San Antonio, males were attracted on the following dates and soaked with honey and water. The trials included offering first- times（ Central Daylight Time） in 2014 : 10 : 22 on 14 March; instar larvae（ 4 mm long） that had never fed, second-instar 9:34, 10:39, and 10:47 on 15 March; 9:52 on 16 March（ but larvae（ 8 - 9 mm long） that had fed for several days, mature windy after 10:00）; 13:00 on 21 March; 10:26 on 22 March; and larvae（ 65 mm long）, and mealworms（ Tenebrio molitor 9:30 on 29 March. The temperature was 7° Celsius on 29 March. Linnaeus）（ 12-15 mm long）. The latter were considered to be a Emergences from the cocoons occur around midnight, so that good control because they are palatable to a wide range of adults are ready for courtship behavior when the sun rises. In vertebrate pets in captivity（ birds, lizards, frogs, etc.）, and were southern Texas there are two distinct generations per year, with used as control subjects by Deml and Dettner（ 1993）, who flights in March and again in September to November, performed similar tests using European ants and larvae of E. apparently because the pupal stage is best suited to survive the calleta. Lengths of the ants used in our trials are given in Table 2. extremely hot and dry conditions of summers in southern Texas, as was proposed for this species in southern Arizona by Collins Results and discussion （2011）. In Texas, the generations overlap, with cocoons in Adult behavior. Multiple mating by females of E. calleta is captivity commonly holding over one to two years, and reported here for the first time. Tutt（ 1902） suggested that occasionally up to four years（ Prentiss 1976）. Cocoons remain females of pavonia（ Linnaeus） may mate more than on the hostplants a year or more after emergence, and we have 60 J. H. Louwagie and R. S. Peigler

observed that both old and viable cocoons can be attached to West, Live Oak County, in December 2007（ Fig. 3）. There were stems that were ligatured by peduncles of cocoons, as noted by 17 eggs scattered within a few cm of each other, with 8 Miller（ 1976） and illustrated by Peigler and Maldonado（ 2005）. depositions on 8 leaves as follows: 3, 1, 2, 1, 2, 2, 3, 3. A few Larvae from the autumn brood are probably protected from days later 13 males of Anastatus（ Hymenoptera: Eupelmidae） parasitoids by being winter feeders. They grow slowly, surviving emerged from these, one parasitoid per egg. The remaining 4 freezing temperatures, and sometimes do not mature until eggs produced 2 larvae of E. calleta and 2 eggs remained January（ Tuskes et al. 1996）. According to Deml and Dettner unhatched. At this time, it is not possible to identify Anastatus to （1993）, larvae of E. calleta contain an unusually high amount of species level using only males, as noted by Burks（ 1967）. glycerol, a chemical that functions as an anti-freezing agent in Two flies of Lespesia sp., near but not, texana（ Webber） , but is more often found in diapausing pupae instead of （Diptera: Tachinidae: Exoristinae） were reared from a cocoon of larvae. E. calleta collected in Goliad, Goliad County, in January 2006 Hostplants. Although original stands of Leucophyllum by Robert A. Miranda and R. S. Peigler. The host cocoon was frutescens still grow in limestone and rocky fields across perfectly formed, but contained a black, shrunken larva of E. southern Texas（ Fig. 4）, most of those have been extirpated by calleta from which two maggots emerged and formed puparia. agriculture and ranching. All natural stands of cenizo that we Adult flies emerged indoors, a male on 12 January and a female found were on private ranches with fences, so were inaccessible on 18 January 2006. If the host cocoon had not been cut open, for field work or collecting. However, horticultural varieties of the maggots would have probably escaped through the exit valve this flowering shrub, such as L. frutescens ‘Green Cloud’ with before pupating at ground level. These flies were identified using greenish leaves and L. f. ‘White Cloud’ with white flowers, are the key of Sabrosky（ 1980）, plus the character he cited for commonly planted in the landscape（ yards, parks, plantings spiracles of the puparium. This apparently unnamed species of around businesses） in towns and cities throughout southern and Lespesia has also been recorded from the saturniids in central Texas, including all over the San Antonio area, which we Texas and Mexico and in Mexico（ Sabrosky 1980, believe has resulted in the expansion of the range of the moth to Peigler, 1996）. It was also possibly the species reported as the north, a point also noted by Collins and Weast（ 1961）. On “several parasitic flies” by Prentiss（ 1976） from a cocoon of E. the north side of the San Antonio metropolitan area, males of E. calleta collected in Corpus Christi, Texas. calleta are attracted to caged females occasionally, but not A cocoon of E. calleta was collected in early January 2013 in commonly, and we have never found cocoons in San Antonio. Devine, by R. S. Peigler and Gregory D. Muise, that produced Colder climate apparently limits the distribution of the moth 113 adults of both sexes of Conura maria（ Riley） farther north into the Edwards Plateau, so it would not be （Hymenoptera: Chalcididae） on 17-18 January 2013, after being expected in San Marcos, Austin, etc., although cenizo is also kept indoors. All emerged from a single hole chewed in planted in those cities. The fields containing wild cenizo the side of the host cocoon. We observed several cocoons at our undoubtedly support reservoir populations of E. calleta that are study site in Devine in 2014 with the same exit holes, indicating the sources of colonies on cultivated cenizo in towns and cities. that this parasitoid species attacked additional E. calleta. A Mature larvae and viable cocoons can be located on the second brood of Conura maria was reared from a cocoon of E. hostplants by defoliation on terminal branches（ J. A. McConnell, calleta collected in Three Rivers, Live Oak County, Texas, by R. personal communication）. S. Peigler and John A. McConnell in January 2016. The host In a landscape planting beside an office building in Devine, cocoon produced 45 wasps of both sexes that emerged 19-20 Medina County, we collected three cocoons on Leucophyllum February 2016. Conura maria oviposits into the host larva, but langmaniae ‘Lynn’s Legacy,’ which is a cultivar of a shrub native adults do not emerge until after the host forms its cocoon and to Chihuahua. There were also shrubs of L. frutescens growing pupates. It is recorded to attack most species of in at the site several meters from the shrubs of L. langmaniae. eastern North America（ Peigler 1996）. Larvae were also successfully reared from eggs in San Antonio Predators. At our study site in Devine, we found more than 20 on that cultivar. This is a new hostplant record for E. calleta, and cocoons that had been torn open by a predator（ Fig. 5）. These we note the possibility that the may utilize L. langmaniae were all near each other in one section of the large hedge of in nature in Chihuahua. cenizo bushes, indicating that the predator systematically Parasitoids. Compared to other large Saturniidae, E. calleta searched for and destroyed them. Possibly this predator was a appears to be attacked by very few species of parasitoids in species of the order Rodentia, or a Raccoon（ Procyon lotor Texas（ Collins and Weast 1961 ; Peigler 1996）, but we report Linnaeus）, or an Opossum（ Didelphia virginiana（ Kerr））, the here three records, two of which are new. Anastatus was latter a common marsupial in Texas. We doubt that a bird could previously reported to parasitize E. calleta by Peigler（ 1996）. A open cocoons in this manner. The attacked cocoons were a mix natural oviposition was located in a cenizo shrub in George of old and fresh ones, indicating that the predation events Biology and chemical ecology of Eupackardia calleta（ Saturniidae） in southern Texas 61 happened months（ or more than one year） apart, or that the Dettner found by using more sophisticated chemical analyses. predator could not distinguish between empty and viable We were not able to measure concentrations or percentages of cocoons. these compounds. Larvae of E. calleta do not have effective chemical protection The unfed larva spectrum（ Fig. 10） and the 1 week old, fed from paper wasps. On 7 April 2016 we observed a third-instar larva spectrum（ Fig. 11） were notably different. The fed larva larva on cenizo to be attacked and macerated by a female of had more peaks on its spectrum, as well as more intense peaks. Polistes exclamans Viereck（ Vespidae）. We netted the and Four compounds present in the fed larva were lacking in the pinned it for identification. unfed larva, three of which were also found in cenizo leaves. This suggests that the larvae derive some of the biogenic Biochemical analyses of caterpillars, moths, and hostplant. compounds directly from their hostplant. Moreover, six of the Figures 9 to 14 show the spectra for the various samples we seven compounds found in glandular secretions of mature larvae analyzed. We compared the molecular weights of the major were also found in the adult moth. Some of these compounds, peaks to compounds that Deml and Dettner（ 1993） identified in such as dopamine and norepinephrine, are known to affect the larvae of E. calleta. Although other compounds can have the nervous systems of vertebrates. The results are shown in Table 1. same molecular weights, we concluded that these peaks were highly likely to represent the same compounds that Deml and

Fig. 1. Eupackardia calleta, female, Devine, Texas, photographed on Leucophyllum langmaniae. （ Specimen in collection of Stefan Naumann, Berlin）. Fig. 2. Eupackardia calleta, male, Beeville, Texas. Fig. 3. Eggs of E. calleta on cenizo, found in George West, Texas. Fig. 4. Natural stands of cenizo（ Leucophyllum frutescens） along Highway 59, southwest of Beeville, Texas. Fig. 5. Cocoons from Devine, Texas, that had been torn open by vertebrate predator（ s）. Fig. 6. Mature larva of E. calleta, Devine, Texas. Fig. 7. Mature larva of E. calleta, Goliad, Texas, showing droplets of glandular secretion on several scoli, indicated by arrows. Fig. 8. Entrance to nest of red harvester ants（ Pogonomyrmex barbatus） in Devine, Texas. 62 J. H. Louwagie and R. S. Peigler

Figs 9-11. Graphs showing peaks of chemicals in: 9 cenizo leaves, 10 unfed larva, 11 fed larva. Numbers on peaks refer to numbers assigned to compounds in Table 1. Biology and chemical ecology of Eupackardia calleta（ Saturniidae） in southern Texas 63

Figs 12-14. Graphs showing peaks of chemicals in: 12 mature larva hemolymph（ HL）, 13 mature larva glandular secretion （GS）, 14 adult moth. Numbers on peaks refer to numbers assigned to compounds in Table 1. 64 J. H. Louwagie and R. S. Peigler

Table 1 Major Peaks unfed 1 week mature mature adult cenizo Compounds found in samples （m/z） larva fed larva larva HL larva GS moth HL leaves 1 3,5-dimethylphenol 107, 122 + + + + 2 hydroquinone 109, 110 + 3 3,4-dihydroxyphenylalanine 123, 197 + + + + 4 dopamine 124, 153 + + + + + +

5 norepinephrine 139, 151, 169 + + + + + +

6 trehalose 155, 163, 342 + + + + + 7 2-（dimethylamino）ethylacetate 131 + + + + +=compound present in sample; HL=hemolymph; GS=glandular secretion, from scoli

Table 2. Ants（ Formicidae） used in predation trials Scientific name Common name Length Source Pogonomyrmex barbatus（ F. Smith） red harvester ant 6-7 mm Devine, Texas Crematogaster hespera Buren acrobat ant 4 mm San Antonio, Texas Pseudomyrmex gracilis（ Fabricius） elongate twig ant 8-10 mm San Antonio, Texas Solenopsis invicta Buren red imported fire ant 3-5 mm San Antonio, Texas Neoponera villosa（ Fabricius） hairy panther ant 15 mm Devine, Texas

Trials with ants in field and lab. When three unfed larvae were placed near the entrance, they were all carried off within 4 minutes, and dropped more than 15 Ants（ Hymenoptera: Formicidae） are usually the most important cm from the nest entrance. In another nest a few meters away, an insects in ecosystems, frequently preying on caterpillars unfed larva was picked up by an ant within 10 seconds and （Hölldobler and Wilson 1990）. However, some larvae of carried away from the entrance for a distance of 38 cm. A Lepidoptera have chemical defenses to protect them from ants. second-instar（ fed） larva was dragged by two ants and dropped Collins（ 2013） reported two instances in California in which 8 cm away from the entrance, and then left. ants were killed when attacking caterpillars of Saturniidae. Crematogaster hespera. Using specimens from San Antonio, we We made anecdotal observations with caterpillars of E. calleta placed five ants of this species in a petri dish and fed them honey by testing predation in the field and lab using five species of ants water. On 21 October 2014 we offered a mealworm, a first-instar （Table 2）. All five species can inflict a painful sting if handled unfed, and a second-instar fed larva of E. calleta. None of these by humans. When kept in petri dishes, all five species of ants were attacked, although the ants are aggressive in nature and will moved toward the sides of the dishes closest to a window, so sting humans if handled. were observed to be phototactic. Pseudomyrmex gracilis is an arboreal ant that we collected in Pogonomyrmex barbatus, a nest（ Fig. 8） was located within 2 m San Antonio. On 20 October we placed two ants of this species of the cenizo hedge in Devine. On 12 October 2014 we placed in a petri dish and fed them honey water. On 21 October we two first-instar larvae that had never fed near the entrance to the offered a mealworm, a first-instar unfed, and a second-instar fed nest, and within two minutes these were taken down into the larva of E. calleta. None of these were attacked, although the hole of the nest. Two second-instar larvae（ which had obviously ants are aggressive in nature and have a painful sting. We have fed） were not attacked by the ants, and crawled away. Ants from observed these ants carrying nymphs of Cicadellidae this colony were captured and brought to the lab. In a petri dish （Homoptera） in nature, probably as prey. with five ants, we offered a mealworm, which was immediately attacked. We also offered unfed first-instar larvae and fed Neoponera villosa. On 12 October 2014 we collected five of second-instar larvae, but these were not attacked by the ants. these large, black ants foraging on the ground at our field site in Devine. The next day we placed them into a petri dish and In April 2015 we repeated these tests in the field in Devine. Biology and chemical ecology of Eupackardia calleta（ Saturniidae） in southern Texas 65

allowed them to settle and adjust to their new surroundings for a detectable on human hands. Deml and Dettner（ 1993） listed the few hours. In our first trial, we placed an unfed first-instar larva sugar trehalose as one of the components of glandular secretions into the petri dish. Within 1 minute, three ants began to attack in E. calleta. Collins（ 2007） hypothesized that such sugars the larva. One of the ants picked up the larva with its mandibles might be attractive to ants, and therefore the ants might protect and carried it around, while two of the other ants continually the caterpillars from attack by predators and parasitoids. We crawled over and interacted with it. This continued this for about agree with Collins that this should be tested with field 4 minutes, and then the two ants began to interact less and less observations, but we have not observed larvae of E. calleta to be with the larva, and the ant that was holding it released it. We tended by ants. Deml and Dettner conceded that the hemolymph removed the unfed larva and waited several minutes, and then of caterpillars of E. calleta is attractive to ants, based on their placed a second unfed larva into the petri dish, and this time the own trials, but they hypothesized that trehalose opens ants made no contact at all. The five ants also avoided a second- biochemical pathways for storing toxins, thus conferring an instar larva（ fed） and a mealworm that were subsequently added increased chemical protection from avian predators. to the petri dish. Although Waldbauer and Sternburg（ 1976） considered Solenopsis invicta is an introduced, invasive species found Eupackardia calleta to be a Batesian mimic of Battus, we commonly in all locations where we collected E. calleta. In a propose that adults of E. calleta should be considered Müllerian mound that we slightly disturbed, we added first-instar unfed and mimics of Battus, because our experiments showed that they second-instar fed larvae of E. calleta on 21 October 2014. The also contain defense chemicals. Most of the chemicals we found ants immediately interacted with the unfed larva, and several in larvae were also found in the adult moths, as shown in our ants carried it around until it was carried down into the mound Table 1. 3-4 minutes later. The ants also interacted with the second-instar larva, biting it several times. However, after 1-2 minutes they Acknowledgments began to avoid it and no longer picked it up in their mandibles. Weeks later a mature larva of E. calleta was placed on a fire ant Cynthia E. Franklin（ Feik School of Pharmacy） was maximally mound on 29 November, and the ants immediately attacked it. helpful in providing training, data, and figures using the mass After two minutes we removed the larva and knocked off spectrometer. We are grateful to Nicholas A. Leed and Robert N. numerous ants. It sustained several stings, visible hours later as Garner（ both University of the Incarnate Word） for assistance in dark spots on the integument. It secreted a lot of dark fluid our biochemical analysis and presentation. The ants were （“saliva”） from its mouth over the next 30 hours, and ejected six identified by Robert A. Johnson（ Arizona State University）. The pellets of frass during the next 10 hours, but was not able to manuscript was critically reviewed by Michael M. Collins, who cling to a stem nor feed. On the third day it was able to attach suggested useful improvements. One anonymous reviewer for Lepidoptera Science itself to a stem of cenizo and commenced feeding, so was provided valuable corrections and transferred to a cage indoors. It fed normally on numerous leaves improvements. Assistance in field collecting was provided by over the next 10 days, but failed to grow, and eventually died on Michael John Baker, John A. McConnell, Robert A. Miranda, 12 December. Following the comments by Deml and Dettner and Gregory D. Muise. The color figures were prepared by Adela （1993）, the dark spots on the integument and the oral secretion M. Gott. Much of this study was conducted for an undergraduate were probably melanin, produced when tyrosine is catalyzed by research project by Joseph Louwagie. Voucher specimens of Polistes tyrosinase. This reaction is a normal defense against predation parasitoids, ants, cocoons torn open by a predator, wasp, L. langmaniae that the uses to protect its integument from its own and moths reared from cocoons collected on have toxic glandular secretions. It is reasonable to assume that this been deposited in the insect collections of Texas A&M mature larva would have easily survived attacks from one or two University and Clemson University. foraging ants, a more likely scenario in nature, because it lived and fed for almost two weeks after suffering multiple stings. References d’Abrera, B., 2016. Butterflies of the Neotropical region, part 1: Conclusions Papilionidae & Pieridae. New and revised edition. 273 pp. Hill House Publishers, Melbourne & London. Although our trials with ants in the field and lab are anecdotal, Burks, D. B., 1967. The North American species of Anastatus our observations collectively indicate that larvae of all stages Motschulsky（ Hymenoptera: Eupelmidae）. Trans. Amer. have some protection from ants by biogenic chemicals, even Entomol. Soc. 93: 423-431, pls. 1-2. before they have fed. Larvae that have fed apparently derive Collins, M. M., 2007. Moth catcher: an evolutionist’s journey some of these chemicals directly from the hostplant, and they are through canyon and pass. 176 pp., 14 pls. University of Nevada able to synthesize additional chemicals after feeding. Mature Press, Reno. larvae emit a strong odor when handled, which is clearly Collins, M. M., 2011. Cocoons: reflections on their unappreciated 66 J. H. Louwagie and R. S. Peigler

natural history. News Lepid. Soc. 53: 39-43. Sonnenschein, London; Friedländer & Sohn, Berlin. Collins, M. M., 2013. On the finding of dead ants attached to Waldbauer, G. P. and J. G. Sternburg, 1976. Saturniid moths as saturniid caterpillars: evidence of successful deterrent mimics: an alternative interpretation of attempts to demonstrate chemistry? J. Lepid. Soc. 67: 62-63. mimetic advantage in nature. Evolution 29: 650-658. Collins, M. M. and R. D. Weast, 1961. Wild silk moths of the United States: Saturniinae. 138 pp. Collins Radio Co., Cedar Rapids, 摘 要 Iowa. Crotch, W. J. B., 1956. A silkmoth rearer’s handbook. The Amateur テキサス州南部におけるカレッタシロスジサン Entomologist 12: 165 pp., 26 pls. Eupackaridia calleta（ヤママユガ科）の生活史および化学生 Deml, R. and K. Dettner, 1993. Biogenic amines and phenolics 態（J. H. Louwagie III・ R. S. Peigler） characterize the defensive secretion of saturniid caterpillars テキサス州南部産のカレッタシロスジサン Eupackardia （Lepidoptera: Saturniidae）: a comparative study. J. Comp. calleta（ Westwood）を野外及び室内で観察した．ゴマノハ Physiol, B, 163: 123-132. グサ科レウコフィルム属の一種 Leucophyllum langmaniae Deml, R. and K. Dettner, 1995. Effects of emperor moth larval を野外における新たな寄主植物として記録することができ secretions, hemolymph, and components on micoorganisms and predators. Entomol. Experim. Applic. 76: 287-293. た．メス成虫は上記のレウコフィルム属の一種の 8 枚の葉 Deml, R. and K. Dettner, 1997. Chemical defence of emperor moths に 17 個の卵を産卵した．蛹の繭は脊椎動物によって捕食 and tussock moths（ Lepidoptera: Saturniidae, Lymantriidae）. されることが示された．また，3 種の捕食寄生者が見つかっ Entomol. Gen. 21: 225-251. た．ナガコバチ科 Anastatus 属は本種の卵から，ヤドリバ Ferguson, D. C., 1972. : Saturniidae（ part 2）, In R. B. エ科の Lespesia sp.（ nr. L. texana）とアシブトコバチ科の Dominick et al.（ eds.）. The moths of America north of Mexico. Conura maria は本種の繭から出てくるのが確認された．後 fasc. 20.2B: 155-275, pls. 12-22. E. W. Classey Ltd., London. 2 者は初記録である．また 3 齢幼虫が Polistes exclamans ア Hölldobler, B. and E. O. Wilson, 1990. The ants. 732 pp. Harvard シナガバチによって捕食されるのが観察された．通常昼行 University Press, Cambridge, Massachusetts. 性である本種のオスの走光性も確認され，メスの多数回交 Horai, H. et al. 2010. MassBank: a public repository for sharing 尾も確認された．この事実は初記録である．テキサス州内 mass spectral data for life sciences. J. Mass Spectr. 45: 703- の 3 郡から新たに本種が確認された．1 齢，2 齢，終齢幼 714. DOI: 10.1002/jms.1777 虫および成虫について生化学分析を行い以下の化合物を検 Howe, W. H., 1963. Our butterflies and moths. 208 pp. True Color 出した． Publishing Co., North Kansas City, Missouri. Miller, T. A., 1976. Observations of Eupackardia calleta in southern 3,4-dihydroxyphenylalanine, Texas（ Saturniidae）. J. Lepid. Soc. 30: 127-130. dopamine, Morton, E. S., 2009. The function of multiple mating by female promethea moths, Callosamia promethea（ Drury） norepinephrine, （Lepidoptera: Saturniidae）. Amer. Midland Nat. 162: 7-18. hydroquinone, Peigler, R. S., 1996. Catalog of parasitoids of Saturniidae of the world. J. Res. Lepid. 33: 1-121. 2-（dimethylamino） ethylacetate, Peigler, R. S. and M. Maldonado, 2005. Uses of cocoons of trehalose, Eupackardia calleta and Rothschildia cincta（ Lepidoptera: Saturniidae） by Yaqui Indians in Arizona and Mexico. Nachr. 3,5-dimethylphenol Entomol. Ver. Apollo, N.F. 26: 111-119. これらのうちのいくつかの化合物はおそらく捕食者から身 Prentiss, J. B., 1976. Time variations of pupal stage of Eupackardia を守るための防御に使用されていると考えられた．本種幼 calleta（ Saturniidae）. J. Lepid. Soc. 30: 187. 虫に対する 5 種のアリによる捕食実験を室内と野外におい Sabrosky, C. W., 1980. A revised key to the Nearctic species of て行って，捕食に対する化学防御の有無を確認した．その Lespesia（ Diptera: Tachinidae）. Ann. Entomol. Soc. Amer. 73: 結果幼虫はアリに対して化学防御をしており，これらの化 63-73. Tuskes, P. M., J. P. Tuttle and M. M. Collins, 1996. The wild silk 合物が成虫からも見出されていることから，推定して，カ moths of North America: a natural history of the Saturniidae of レッタシロスジサンはアオジャコウアゲハ属 Battus に the United States and Canada. 250 pp. Cornell University ベーツ擬態をしているのではなく，ミュラー擬態している Press, Ithaca, New York. 可能性が示唆された． Tutt, J. W., 1902. A natural history of the British Lepidoptera: a ［文責：坂巻祥孝／Yositaka Sakamaki］ text-book for students and collectors, 3. 558 pp. Swan