Early Stages and Natural History of Memphis Perenna Lankesteri

1 SDI Paper Template Version 1.6 Date 11.10.2012 2 Early stages and natural history of Memphis perenna lankesteri 3 butterfly (Nymphalidae, Charaxinae) from Costa Rica with the first 4 published female pictures 5 6 Jim Córdoba-Alfaro1*, Luis Ricardo Murillo-Hiller2 7 8 * (506)85597625, [email protected] 9 10 1Escuela de Ciencias Biológicas, Universidad Nacional de Costa Rica. 11 2 Centro de Acción Social e Investigación en Mariposas (CASIEM), 12 Escuela de Biología, Universidad de Costa Rica. 13 ______14 15 16 ABSTRACT The early stages of Memphis perenna lankesteri (Hall 1935) are described and illustrated in this paper, with the first published female pictures. The butterfly larvae were found in Croton niveus (Euphorbiaceae) in Mansiones de Montes de Oca (9o 56´N, 84o 01´W), San José, Costa Rica, with an immature stages duration of = 59 days (n=18). The larvae behavior, body shape larvae, host plant and female phenotype were discussed.

17 18 Keywords: Taxonomy, morphology, endemism, M. pithyusa, M. niedhoeferi, M. glauce, M. artacaena, M. xenocles, M. 19 arginussa, M. appias, Euphorbiaceae, Croton niveus. 20 21 22 23 1. INTRODUCTION 24 25 Memphis Hübner (1819) is composed of 62 species [1] throughout the Neotropic including Trinidad and Tobago Islands, 26 25 species of which occur in Costa Rica with a recent record of Memphis d. dia for this country [2]. 27 28 The natural history of this genus is poorly understood, only three species of Memphis have detailed descriptions of their 29 immature stages: two subspecies of M. moruus (Fabricius) (M. moruus boisduvali (Comstock) [3] and M. moruus stheno 30 (Prittwitz) [4]), M. pithyusa (R. Felder) [5], and M. appias [4]. In addition, 19 species have brief descriptions and/or 31 published illustrations of at least some stage of larval development [4, 6, 7, 8, 9]. 32 33 Knowledge of the immature stages in this group could help the understanding of phylogenetic relationships at the species 34 level as well as the possible intraspecific variation. 35 36 Memphis perenna Godman & Salvin (1884) show geographical disjunctions between the three subspecies [1]: M. p. 37 perenna Godman & Salvin (1884) is distributed from southern Mexico to El Salvador, M. p. lankesteri Hall (1935) is 38 endemic of Costa Rica [8] and M. p. austrina Comstock (1961) is found in Colombia and Venezuela [10]. 39 40 M. p. lankesteri is associated with premontane wet forest habitats, documented for Patarrá in Desamparados [8], San 41 Antonio in Escazú [11], and Mansiones in Montes de Oca [12], San José. It is present only from the end of the dry season 42 through the mid-rainy season [8], from 1000 to 2100 m. On sunny days, males can be seen feeding on dog excrement or 43 rotting fruit. The female of M. p. lankesteri was mentioned for Comstock (1961) however, there is not a detailed 44 description or illustrations of it. The goal of this manuscript is to describe the life cycle and illustrate the female of M. p. 45 lankesteri. 46

* Tel.: (506+)85597625 E-mail address: [email protected]. 47 2. MATERIAL AND METHODS 48 49 A search for the butterfly larvae was carried out from 16 March 2008 to 25 September 2011 at Mansiones de Montes de 50 Oca (9o 56´N, 84o 01´W), San José, Costa Rica. The life zone of the locality presents a second growth habitat, defined as 51 Premontane Wet Forest [13]. Butterfly females were seen flying around the host plants and laying eggs between 1100 52 and 1300h. Due to the height of the host plants, eggs collection were impossible and only caterpillars were found. Each 53 collected larva was put into a plastic bag and fed with leaves of the host plant which were brought weekly from the 54 collecting site. The rearing of the larvae was carried out in the Insects Museum of the University of Costa Rica average 55 room temperature 24 C°. The adults obtained from this study and the empty pupae are deposited in the Córdoba-Alfaro 56 personal collection (CPCA). The photographs were taken with a Nikon Coolpix camera model 4500. 57 58 2. RESULTS 59 60 Host plant: Croton niveus Jacq. (Euphorbiaceae). An introduced tree from northern Central America, used in the coffee 61 plantations of Costa Rica for protecting it from wind [14]. 62 63 First instar: (Fig. 1: a) 2.0- 3.5 mm. Head capsule brown with six whitish tubercles and two white longitudinal lines on 64 frontal area, white frons, stemmata black. Body greenish brown with many small lateral whitish warts. 3-5 days to molt. 65 66 Second instar: (Fig. 1: b) 3.5- 6.5 mm. Front head black-brown with brown clypeus and whitish frons, lateral areas brown. 67 Many small warts white color surrounding the capsule and tree white lines beginning from antennal area crossing 68 longitudinal until the half of the capsule. Body green with a lateral line composed of small white warts surrounding the 69 spiracles. Spiracles black, the one on A8 bigger and more dorsally position than the rest. 6-7 days to molt. 70 71 Third instar: (Fig. 1: c, d & e) 6.5-9.5mm. Head capsule very similar to second instar but differs for the bearing of one 72 black head horn at each side of the epicranium which is composed by a crown of short spikes. Anteriorly to that structure 73 there is a conspicuous orange round wart. Body variable, green color with small white warts all over. Segment A2 can 74 present a transversal black and orange saddle that in some cases is connected with a black longitudinal line in lateral 75 position. Spiracles same than previous instar. 5 days to molt. 76 77 Fourth instar: (Fig. 1: f) 9.5-17.5 mm Head capsule very similar to third instar but the two black horns and six orange warts 78 are more prominent. Body brownish green spattered with white spots. The black “saddle” continues across second 79 abdominal segment lined posteriorly by a thin white streak. Caudal segments with longitudinal marginal blackish, line from 80 the A6-10. 4-7 days to molt. 81 82 Fifth instar: (Fig. 1: g, h, i & j) 17.5-28mm. Head capsule round shape, green, with three distinct black lines: one 83 beginning parallel to the adfrontal suture and the epicranial suture surrounding the epicranial horns and then descending 84 to the antennal base. The second is right on the adfrontal and epicranial suture. The third one is vertical in the middle of 85 the frons which can be from white to green color. Two epicranial black horns bearing numerous rounded projections. 86 Stemmata black. Green body with white spattered spots. Variable coloration: from the black “saddle” continuous dorsally 87 across A2 segment to discontinuous or absent. Lateral black longitudinal band continuous, discontinuous or absent. 88 Segments from A8 to A10 darker dorsally. 10-16 days to molt. 89 90 Pupa: (Fig. 2: a & b) Brown color with black cremaster. A white line surrounding dorsally the wigs area. Presents brown 91 markings on prothoracic legs, mesothoracic legs and mesothoracic wing cases. Abdomen incurved ventrally. A 92 transversal whitish band is present dorsally on the abdominal carina. A dark brown middle line is presented from protorax 93 to the abdominal carina. Duration: 25-27 days. 94 95 Female: (Fig. 3: b & d) Differentiated from male by presenting conspicuous blue spots on the sub medial forewings M3, M1 96 and R5 at the bifurcation level of R4 and R5. Prominent red orange spot on R1 cell in midway of the hind wing. 97 98 3. DISCUSSION 99 100 All instars of M. p. lankesteri larva show a spattered white warts body texture. This seems to have a protective function 101 through crypsis, since it highly resembles the trichomes covered leaves of the host plant. The larval body shape of M. p. 102 lankesteri is adapted to two different environmental scenarios according to its development: a) the exposed first, second, 103 and third instars which rest on a frass chain at the tip of the host plant leaf and b) a roll up on the leaf apex where fourth 104 and fifth instar rest inside, both behaviors has been mentioned as normal for Memphis larvae [8, 10, 15].The first three 105 larval instars are cylindrical shape, although, the last two are more conical been the head capsule the wider area. When is 106 disturbed, the larvae retracts the head backward inside the shelter, making a hard shield. The various head warts then, 107 seem to be involved in a protective function, probably against parasitoid wasp or other predaceous insects such as bugs. 108 109 There is a marked similarity between the morphology of the larvae, pupae and adult of M. p. lankesteri with others species 110 of Memphis as M. pithyusa (R. Felder) [5], M. niedhoeferi (Rotger, Escalante & Coronado), M. glauce (Rober), M. 111 artacaena (Hewitson), M. xenocles (Westwood) [9], M. arginussa (Cordoba- Alfaro & Murillo-Hiller in prep.), and M. appias 112 (Hubner) [4]. 113 114 The larval behavior of M. p. lankesteri of building shelters on the host plant leaves resembles the ones of M. appias and 115 M. arginussa [4,16]. The last two species built the shelter perpendicular to the leaf and M. p. lankesteri build this shelter 116 either this way or at the apex of the leaf. According to Queiroz 2002 [16] and Dias et al. (2010) [4] this behavior is 117 important to be considered because might be and etological taxonomic character for grouping species. However, this may 118 not be right since other Charaxinae genera such as Anaea, Fountainea, and Consul also share this same behavior [15]. 119 120 There seems to be a tight relationship between M. p. lankesteri species group and their hostplants. This is because all the 121 proposed Memphis species in this group feed from Croton with scaled trichomes and share very similar pupal and larvae 122 morphological characteristics. Many Croton species with these distinctive trichomes are used as host plants by Costa 123 Rican Memphis. For example, C. schiedianus has been reported for M. artacaena and M. aulica. Also, C. megistocarpus 124 has been reported for M. aulica [9]. In the location where the immature stages of M. p. lankesteri were found, there is only 125 C. niveus, this is interesting because it is and introduced plant native from northern Central America. It would be valuable 126 to find which is the original locality host plant for further studies. 127 128 However, there are some species of Memphis such as M. aulica and M. forreri [9] that also feed from this group of species 129 of Croton, but, with very different early stages characteristics. Probably there is more than one species group 138 of 130 Memphis feeding from the plants genus Croton. 131 132 From the specimens studied, no seasonality is detected. Adults revised from Museo Nacional de Costa Rica, Instituto 133 Nacional de Biodiversidad, Museo de Insectos de la Universidad de Costa Rica and private collections shows low 134 abundance through all the year. In contrast, I. Chaon on DeVries (1987) [8] stated that this species is present from the 135 end of the dry season through the mid-rainy season in Patarrá, very close to the same locality of this study. 136 137 Hall (1935) [17] described Anaea lankesteri from two males collected in Cartago Costa Rica. However, the female was 138 described twenty six years later by Comstock (1961) [18] where he wrote that is hardly distinguishable from males except 139 in size and only traces of blue scales on R5 and R4. In this study we obtained nine females and all of them result to be 140 different than males presenting conspicuous blue spots on the sub medial anterior wings M3, M1 and R5 at the bifurcation 141 level of R4 and R5. Also, besides the Comstock description of the M. p. lankesteri female, no one else had either 142 described or illustrated it until now. The confusion may happen by misinterpretation of the only two Schaus and Barnes 143 females’ material he studied, with some other species of Memphis. 144 145 Specimens Studied 146 147 Museo Nacional de Costa Rica 148 149 (10♂) Costa Rica, Cartago, Coris, 1500 m, 25/VI/1980, G. Small; 12/VII/1980, I. Chacón; Heredia, Getsemani, 150 1400m, 5/IV/1981, R. Canet; Escazú, Bebedero, Pico Blanco, 2100 m, 15/II/1995, G. Vega; Cartago, La Unión, 151 Tres Ríos, La Carpintera, Campo Escuela Iztarú, 1870m, 2/VII/2006, 27/VII/2006, (2) 27/VI/2006, 27/IX/2006, 152 G. Vega; San José, Acosta, Palmichal, Río Negro, 1536 m, 29/VI/2010; (5♀) Cartago, Coris, 1/III/1980, I. 153 Chacón; Cartago, La Unión, Tres Ríos, La Carpintera, Campo Escuela Iztarú, 1700 m, 21/III/2007 (3), G. 154 Vega; San José, Escazú, Bebedero, Pico Blanco, 2100, III/97, G. Vega.

155 Authors collections

156 (5♂) Costa Rica, San José, Montes de Oca, Mansiones, 13/IX/2008, 9/III/2010, 4/IX/2011, 1/VI/2012, J. 157 Córdoba-Alfaro; Cartago, La Unión, Tres Ríos, Campo Escuela Iztarú, 1400 m, 4/VIII/1999, R. Murillo-Hiller; 158 (1♀) Alajuela, Grecia, IX/2011, Ricardo Murillo-Hiller.

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161 162 Bred specimen

163 164 (9♂), Costa Rica, San José, Montes de Oca, San Rafael, Mansiones (9o 56´ 42´´N, 84o 01´ 08´´W). 165 18/IX/2010, 25/IX/2011 (4), 1/X/2011, 10/VII/2012 (2), 6/X/2012. (9♀) 6/XI/2010 (2), 4/IX/2011, 11/IX/2011, 166 25/1X/2011, 23/X/2011, 29/IV/2012, 23/VI/2012, 10/VIII/2012. Col: Jim Córdoba-Alfaro. 167 168 169 170 ACKNOWLEDGEMENTS 171 172 Thanks to Enrique Lorenzoni by literature contribution. Also, to Hugo Aguilar for facilities at the Museo de Insectos of the 173 Universidad de Costa Rica. To Germán Vega and Cecilia Pineda for allowing access to the Museo Nacional de Costa 174 Rica and to the Escuela de Ciencias Agrarias de la Universidad Nacional de Costa Rica for images facilities. Finally, to 175 Adrea Gonzalez Karlsson for correcting an earlier draft of the manuscript. 176 177 178 COMPETING INTERESTS 179 180 Authors have declared that no competing interests exist. 181 182 AUTHORS’ CONTRIBUTIONS 183 184 Author # 1 & Author # 2 established the research goal, collected the data and did the field work. Manuscript writing, 185 specimens breeding, literature researching and image and photographic work were done together. 186 187 188 REFERENCES 189 190 1. Lamas, G. (2004) Nymphalidae. Charaxinae. Tribe Anaeini. In: Lamas, G. (Ed.) (2004) Checklist: Part 4A. 191 Hesperioidea-Papilionoidea. In: Heppner, J.B. (Ed.) Atlas of Neotropical Lepidoptera. Association for Tropical 192 Lepidoptera/Scientific Publishers, Gainesville, 428 pp. 193 194 2. Córdoba-Alfaro, J. & L. R. Murillo-Hiller. (2012). The first record of the butterfly Memphis d. dia (Lepidoptera: 195 Nymphalidae, Charaxinae) in Costa Rica. Rev. peru. biol. 19(3): 327-328. 196 197 3. Muyshondt, A. (1975a) Notes on the life cycle and natural history of butterflies of El Salvador. V.Anaea (Memphis) 198 morvus boisduvali (Nymphalidae). Journal of the Lepidopterists’ Society, 29, 32–39. 199 200 4. Dias, F. M. S., Casagrande, M.M. & Mielke, O.H.H. (2010). Biology and external morphology of immature stages 201 of Memphis appias (Hübner) (Lepidoptera: Nymphalidae: Charaxinae). Zootaxa. 2668: 21–32. 202 203 5. Muyshondt, A. (1975b). Notes on the life cycle and natural history of butterflies of El Salvador. VI.Anaea 204 (Memphis) pithyusa (Nymphalidae). Journal of the Lepidopterists’ Society, 29, 168–176. 205 206 6. Müller, W. (1886) Südamerikanische Nymphalidenraupen. Versuch eines natürlichen Systems der Nymphaliden. 207 Zoologischen Jahrbücher, 1, 417–678. 208 209 7. Rydon, A.H.B. (1971) The Systematics of the Charaxidae (Lep: Nymphaloidea). Entomologist's Record and 210 Journal of Variation, 83, 219–233, 311–316, 336–341, 384–388. 211 212 8. De Vries, P. J. (1987) The butterflies of Costa Rica and Their Natural History: Papilionidae, Pieridae, 213 Nymphalidae. Princeton University Press. Chicago. USA. 120-127 p. 214 215 9. Janzen, D.H. & Hallwachs, W. (2012) Dynamic database for an inventory of the macrocaterpillar fauna, and its 216 food plants and parasitoids, of Area de Conservacion Guanacaste (ACG), northwestern Costa Rica. Available 217 from http://janzen.sas.upenn.edu (accessed May 22, 2012). 218 219 10. Neild, A. (1996). The butterflies of Venezuela Part 1: Nymphalidae I (Limenitidinae, Apaturinae, Charaxinae). 220 Meridian Publications. Greenwich, London. 144p. 221 222 11. Montero-Moreno, J. R. (2003). A new locality for the Endemic Charaxine Memphis lankesteri in Costa Rica. News 223 of the Lepidopterists’s Society. 45:1 224 225 12. Córdoba-Alfaro, J. (2011). Diversidad de Mariposas (Lepidoptera: Papilionidae, Pieridae, Nymphalidae) en 226 Mansiones de Montes de Oca, San José, Costa Rica. Brenesia. 75-76: 121-123. 227 228 13. Bolaños, R., V. Watson & J. Tosi. (2005). Mapa ecológico de Costa Rica (zonas de vida) según el sistema de 229 clasificación de zonas de vida del mundo de L. R. Holdridge, escala 1:750 000. Centro Científico Tropical. San 230 José, Costa Rica. 231 232 14. Estrada, A & J. E. Sánchez. (2012). Árboles y arbustos de importancia para las aves del Valle Central de Costa 233 Rica. EINBio. Santo Domingo de Heredia. Costa Rica. 153 p. 234 235 15. Miller, J.C., D, Janzen & W. Hallwachs. (2006). Portraits from the Tropical Forest of Costa Rica. The Belknap 236 Press of Harvard University Press. Cambridge, Massachusetts, USA. 264 p. 237 238 16. Quiroz, J.M. (2002) Host plant use among closely related Anaea butterfly species (Lepidoptera: Nymphalidae: 239 Charaxinae). Brazilian Journal of Biology, 62, 657–663. 240 241 17. Hall. (1935). Anea lankesteri. Entomologist, London, Vol. 68, p 224. 242 243 18. Comstock, W.P. (1961). Butterflies of the American Tropics. The genus Anaea Lepidoptera Nymphalidae. A study 244 of the species heretofore included in the genera Anaea, Coenophlebia, Hypna, Polygrapha, Protogonius, 245 Siderone and Zaretis. American Museum of Natural History, New York, 214 pp. 246 247 248 Legend

249 Fig. 1: Memphis perenna lankesteri early stages: a= first instar, b= second instar, c, d= third instar, e= third instar head 250 capsule, f= fourth instar, g, h, i= fifth instar polymorphism, j= fifth instar head capsule. 251 252 Fig. 2: Memphis perenna lankesteri pupa and prepupa: a & b=pupa, c= prepupa. 253

254 Fig. 3: Adult of Memphis perenna lankesteri (male: Costa Rica, San José, Montes de Oca, San Rafael, Mansiones, 255 1/X/2011 rec. J. Córdoba-Alfaro; female: Alajuela, Grecia, IX/2011, Rec. R. Murillo-Hiller): a=male dorsal, b=female 256 dorsal, c=male ventral, d=female ventral. 257