Volume 62 Number 2 25 Aug 2008 ISSN 0024-0966 Journal of the Lepidopterists' Society Published quarterly by The Lepidopterists' Society THE LEPIDOPTERISTS’ SOCIETY Executive Council John H. Acorn, President John Lill, Vice President William E. Conner, Immediate Past President David D. Lavvrie, Secretary Andre V.L. Freitas, Vice President Kelly M. Richers, Treasurer Akito Kayvahara, Vice President Members at large: Kim Garwood Richard A. Anderson Kenn Kaufman John V. Calhoun Plarry Zirlin Amanda Roe Editorial Board John W. Rrovvn {Chair) Michael E. Toliver ( Member , at large ) Brian Scholtens ( Journal ) Lawrence F. Gall ( Memoirs ) 13 ale Clark {News) John A. Snyder {Website) H onorary Life Members of the Society Charles L. Remington (1966), E. G. Munroe (1973), Ian F. B. Common (1987), Lincoln P Brower (1990), Frederick H. Rindge (1997), Ronald W. 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Bliss, 28 DuPont Avenue, Piscataway, NJ 08854. Michelle DaCosta John H. Masters Michael G. Pogue The additional cost for members outside the U.S. is to cover mailing costs. Journal of The Lepidopterists’ Society (ISSN 0024-0966) is published quarterly by The Lepidopterists’ Society, % Los Angeles County Museum of Natural History, 900 Exposition Blvd., Los Angeles, CA 90007-4057. Periodicals postage paid at Los Angeles, CA and at additional mailing offices. POSTMASTER: Send address changes to The Lepidopterists’ Society, % Natural History Museum, 900 Exposition Blvd., Los Angeles, CA 90007-4057. Cover Illustration: Images from the study of Methona curvifascia immature stages at Garzacocha, Provincia Sucumbios Ecuador. Clockwise from top left: fifth instar, pupa, ventral adult and dorsal adult. J OURNAL The Lepidopterists’ OCIETY Volume 62 2008 Number 2 Journal of the Lepidopterists’ Society 61(2), 2007, 61-66 COMPARATIVE STUDIES ON THE IMMATURE STAGES AND DEVELOPMENTAL RIOLOGY OF FIVE ARGYNNIS SPP. (SURGENUS SPEYERIA ) (NYMPHALIDAE) FROM WASHINGTON David G. James Department of Entomology, Washington State University, Irrigated Agriculture Research and Extension Center, 24105 North Bunn Road, Prosser, Washington 99350; email: david_james@wsu.edu ABSTRACT. Comparative illustrations and notes on morphology and biology are provided on the immature stages of five Arg- ynnis spp. (A. ctjbele leto, A. coronis simaetha, A. zerene picta , A. egleis mcdunnoughi, A. hydaspe rhodope ) found in the Pacific Northwest. High quality images allowed separation of the five species in most of their immature stages. Sixth instars of all species possessed a fleshy, eversible osmeterium-like gland located ventrally between the head and first thoracic segment. Dormant first in- star larvae of all species exposed to summer-like conditions (25 ± 0.5° C and continuous illumination), 2. 0-2. 5 months after hatch- ing, did not feed and died within 6-9 days, indicating the larvae were in diapause. Overwintering oi first instars for ~ 80 days in dark- ness at 5 ± 0.5° C, 75 ± 5% r.h. resulted in minimal mortality. Subsequent exposure to summer-like conditions (25 ± 0.5° C and continuous illumination) resulted in breaking of dormancy and commencement of feeding in all species within 2—5 days. Durations of individual instars and complete post-larval feeding development durations were similar for A. coronis, A. zerene, A. egleis and A cybele (54.1—55.5 days from post-diapause first instars to adulthood). Development of A. hydaspe was significantly faster averaging 47 days. Larvae of all species readily accepted Viola adunca and V. glabella as host plants. Viola labradorica was also accepted by all instars of A. egleis, however, its acceptance was limited in the other species to later instars. Domesticated pansies ( Viola tricolor) were accepted by sixth instars of A. egleis, A. coronis and A. zerene, but only limited feeding occurred with sixth instar A. cybele. Additional key words: morphology, osmeterium, development, diapause, host plants, overwintering, instar durations Nine species of Greater Fritillary (Argynnis spp.) occur in Washington: A. cybele (F.), A. coronis (Behr), A. zerene (Boisduval), A. callippe (Boisduval), A. egleis (Behr), A. hesperis (Edwards), A. atlantis (Edwards), A. hydaspe (Boisduval) and A. mormonia (Boisduval) (Guppy and Shepard 2001, Pyle 2002, Warren 2005). Until recently, these species, along with till North American Greater Fritillaries were considered to belong to the genus Speyeria. However, recent morphological and molecular studies by Simonsen (2006) and Simonsen et al. (2006) showed that the North American species are better treated as members of the large, unified genus, Argynnis with Speyeria relegated to a sub-genus. The immature stages of Speyeria spp. are generally infrequently encountered and are thus poorly described or illustrated. Detailed morphological studies and descriptions of Argynnis eggs, larvae and pupae may yield important distinguishing characteristics that could be useful along with adult characteristics for resolving taxonomic issues with the many subspecies and putative subspecies that occur in this genus (Warren 2005). Similarly, the biology of immature Argynnis spp. is also imperfectly known and offers a field rich in potential for understanding mechanisms of diapause, defense, host plant relationships etc. This paper provides biological information on, and detailed illustrations of the immature stages of five Washington Argynnis spp., A. cybele leto (Behr), A. coronis simaetha dos Passos and Grey, A. zerene picta (McDunnough) , A. egleis mcdunnoughi (Guilder) and A. hydaspe rhodope (W. H. Edwards). Of the five, only the endangered Oregon coast subspecies of A. zerene (A. z. hippolyta (W. H. Edwards)) has received detailed attention to its immature stages within the past 25 years (McCorkle 1980, McCorkle and Hammond 1988). The early stages of eastern North American A. cybele and A. egleis from Nevada were described more than 125 years ago by W. H. Edwards (Edwards 1879, 1880). Descriptions and illustrations of late instars only, are available for A. cybele, A. coronis , A. zerene and A. hydaspe (Comstock and Dammers 1931, Dornfeld 1980, Scott 1986, Allen 1997, Guppy and Shepard 2001, Miller and Hammond 2003, Allen et al. 2005, Wagner 2005). Allen et al. (2005) described the late instar larva 62 Journal of the Lepidopterists’ Society of A. egleis but did not illustrate it. Aside from A. z. hippolyta , veiy little has been reported on the biology and ecology of the immature stages of these five species (Pyle 2002, Warren 2005). During August 2005 to April 2006, A. cybele, A. coronis, A. zerene, A. egleis and A. hydaspe were reared in the laboratory, for photography of all the immature stages (including each instar). Notes on coloration, patterning and dimensions of eggs, larvae and pupae were made. Observations were also made on aspects of biology such as overwintering mortality, diapause, host plant acceptance and developmental duration. Materials and Methods Gravid females of A. cybele leto (2), A. coronis simaetha (4), A. zerene picta (10), A. egleis mcdunnoughi (4) and A. hydaspe rhodope (4), were obtained during August 2005 from the Umatilla National Forest in SE Washington (A. c. leto , A. z. picta , A. e. mcdunnoughi) and Wenatchee National Forest on the eastern edge of the Cascade mountains (A. c. simaetha , A. h. rhodope). Females were placed in plastic buckets (31 cm deep, 28 cm diameter) with muslin- covered lids and held at 21-28° C under natural daylength. Butterflies were provided with potted violets ( Viola labradorica Schrank) or dessicating violet foliage (Viola adunca Site, V. labradorica, Viola glabella Nutt.) and paper toweling as opposition substrates and tissue pads soaked in sugar/water solution for nourishment. Butterflies oviposited freely under these conditions. Eggs were measured, photographed and left in the buckets to hatch. First instars were also measured and photographed and along with all violet foliage and paper toweling, were transferred to plastic boxes (30 x 23 x 10 cm) with muslin lids. The boxes were held at 20-28° C under natural daylength until September 3 when they were transferred to shaded outdoor conditions until October 31 (10-25° C). During this period, larvae were exposed to fine water-misting every 2-3 weeks. Larval diapause termination experiment. On October 31, five first instars of A. coronis , A. zerene, S. cybele and S. hydaspe were transferred to summer-like (25 ± 0.5° C, constant fluorescent illumination) conditions and placed on fresh, detached V adunca leaves laid upperside down on wet cotton wool in a muslin-covered plastic Petri-dish (13 cm diameter). Observations on behavior and mortality were made daily. On November 1, plastic boxes containing remaining dormant larvae were transferred to a dark constant temperature room set at 5 ± 0.5 C for overwintering. Relative humidity was maintained at 75 ± 5 %. Overwintering larvae were transferred to summer-like conditions (25 + 1° C, constant fluorescent illumination) during f 1-19 January. One group of 12 A. coronis larvae was transferred on January 4 to 15-21 ° C/ 9 hrs light. First and second instars were reared on detached Viola leaves placed upper surface down on moist cotton wool in muslin-covered plastic Petri dishes (13 cm diameter). Dried leaf debris was provided as shelter for the larvae. Third-sixth instars were reared in plastic boxes (25 x 15 x 6 cm) with muslin lids. Cut Viola spp. with stems in water was provided for food and shelter. Pre-pupal sixth instars were placed in larger boxes (30 x 23 x 10 cm) with a greater amount of foliage to provide pupation sites. All instars and pupae were photographed. Observations on larval morphology, coloration, behavior, development, host plant acceptance and mortality were made daily until pupation. Photographs were taken using a Canon EOS IDS Mark II, digital SLR camera mounted on a tripod. A Canon MP-E 65 mm L X - 5 X macro lens was used together with a Macro Twin Lite MT - 24 EX flash lighting system. Results Morphology of immature stages. Eggs, instars 1-6 and pupae of A. coronis simaetha, A. zerene picta, S. egleis mcdunnoughi, A. cybele leto and A. hydaspe rhodope are shown in Figs. 1-2. Eggs of all species were creamy white when freshly laid, turning orange or pinkish/tan/brown with development after 2-3 days. Egg dimensions were comparable between species, being 0. 9-1.0 mm in height and 0. 8-1.0 mm in diameter (Table 1). The eggs of A. zerene were generally the smallest (0.9 x 0.8 mm) and were noticeably more ovoid than eggs of the other species. Eggs of S. egleis and S. cybele were flattened basally, while the eggs of S. coronis and S. hydaspe were more cylindrical (Figure 1). First instars of all species measured approximatelyl.5 mm after hatching but increased to 1.75-2.0 mm after imbibing water droplets. Lengths of larvae at the beginning and end of each instar for the five species are shown in Table 1 . Generally, there was good correlation of instar sizes between species with an approximate doubling of size with each instar from one to three, a lessening of growth rate in instars 4 and 5, then an increase in the sixth instar. Mature larvae of A. cybele were largest at 45 mm with the other four species similarly sized, ca. 35-38 mm (Table 1). Coloration of unfed first instars varied with S. coronis and S. hydaspe generally lighter colored than the other three species (Figure 1). All species had black head capsules. In general the black tubercles running longitudinally down the body were darker and more prominent in first Volume 62, Number 2 63 instars of S. egleis and S. cybele than the other three species. Fine dark hairs arose from these tubercles and appeared in all species to secrete a small droplet ot fluid at the distal end. Second instars were characterized by the development of spines, replacing the hairs rising from the tubercles. Head capsules were black and the five species were generally similarly-colored in this instar. A light colored dorsal band was evident in A. cybele and all species had a lower lateral row of orange/tan-colored tubercles. Third instars showed greater spine development, particularly in A. hydaspe (Fig. 1) and the lower lateral row of orange tubercles was more prominent in all species. In addition, the upper lateral row of tubercles was also orange in this instar. This was particularly pronounced in A. coronis and A. zerene. Head capsules remained solid black except for A. hydaspe which showed limited orange/brown marking. All species except A. hydaspe had a prominent pale dorsal band in the third instar with a central darker colored intermittent stripe running through the center. The lower lateral row of orange tubercles was further developed in the fourth instar of all species. The upper lateral row was also strongly developed in A. cybele, A. coronis and A. zerene but virtually lacking in A. hydaspe and A. egleis. The pale dorsal band with intermittent central dark stripe was also further developed in fourth instar A. coronis, A. zerene and A. egleis, but absent in fourth instar A. cybele and A. hydaspe. Ground color of fourth instar A. coronis, A. zerene and A. egleis was gray/white rather than black as in A. cybele and A. hydaspe (Fig. 1). Head capsules of fourth instars were largely black with varying amounts of orange-brown on dorsal surfaces, most pronounced in A. cybele and A. hydaspe and least in A. coronis. Fifth instars were very similar to fourth instars in coloration, although A. coronis tended to be a little darker in this instar. The orange-brown dorsal markings on the head capsule were more pronounced in A. cybele, A. hydaspe and A. egleis in the fifth instar. Lateral and dorsal Hews of sixth instar larvae are shown in Fig. 2. The upper lateral row of orange tubercles was most developed in A. cybele and A. egleis, although they tended paler, almost white, in the latter species. In A. zerene the color of these tubercles was brown or black and blended in with background coloration (Fig. 2). The same was true for A. coronis except on the first two abdominal segments where the upper row of tubercles remained orange. The upper lateral tubercles on the first two abdominal segments of A. egleis sixth instars were also more vividly colored than the rest of this row. Dorsal coloration of A. zerene and A. coronis was palest contrasting with the black ground color of A. cybele and A. hydaspe. The dorsal ground color of A. egleis was intermediate lacking the distinctive gray/tan/white blotches of A. zerene and A. coronis (Fig. 2). The pale dorsal band containing an intermittent central dark stripe was still present in A. coronis and A. zerene but absent in the other species. Dorsal orange-brown coloration of the head capsule extending laterally was most developed in A. cybele and A. egleis, with only veiy minor orange coloration on the Table 1. Sizes (mm) of immature stages of five Argynnis spp. Egg dimensions are height x width. Larva] dimensions are lengths measured at commencement and end of each instar. Egg and larval data were obtained from examination of 2^4 individuals. Varia- tion was generally less than 0.1 mm. Pupae were measured from cremaster to tip of head (Mean ± SE) (number of pupae exam- ined in parentheses). S. coronis simaetha S. zerene picta S. egleis mcdunnoughi S. cybele leto S. hydaspe rhodope Egg 1.0 x 0.9 0.9 x 0.8 1.0 x 1.0 0.9 x 0.9 1.0 x 0.8 First instar 1.5 -3.0 1.5 -2.5 1.5 -3.0 1.5 -3.0 1.5 -3.0 Second instar 3.0 -6.0 2.5 - 4.0 3.0 - 5.0 3.0 - 5.0 3.0 -6.0 Third in star 6.0 - 10 4.0 -8.0 5.0 -8.0 5.0 -9.0 6.0 - 10 Fourth instar 10 - 15 8.0 - 13 8.0-13 9.0 - 15 10 - 15 Fifth instar 15-20 13-20 13-20 15 - 25 15 - 19 Sixth instar 20 - 35 20-38 20 - 35 25-45 19 - 37 Pupa 23.5 ± 0.2 23.6 ± 0.6 22.7 ± 0.2 28.5 ± 0.2 22.0 ± 0.9 (4) (10) (7) (4) (4) 64 Journal of the Lepidopterists’ Society S. coronis S. zerene S. egleis S. cybele S. hydaspe Fig. 1. Eggs and first four instars of A. coronis simaetha, A. zerene picta, A. egleis mcdunnoughi, A. cybele leto and A. hydaspe rhodope. Volume 62, Number 2 65 S. coronis S. zerene S. egleis S. cybele S. hydaspe Fig. 2. Fifth and sixth (ventral and dorsal views) instars and pupae of A. coron is simaetha, A. zerene picta, A. egleis mcdunnoughi, A. cybele leto and A lujclaspe rhodope. 66 Journal of the Lepidopterists’ Society head capsules of A. zerene and A. hydaspe. No orange coloration was seen on the all black head capsules of sixth instar A. coronis. Sixth instars of all species possessed a fleshy, eversible osmeterium located ventrally between the head and first thoracic segment (Fig. 3). A musky odor emanating from the organ was apparent when the larvae were handled roughly. F urther examination of A. egleis larvae showed that die organ was not present in first instars but appeared in the second instar and was present in all subsequent instars. Sixth instars were mature at maximum recorded lengths of 35 (A. egleis , A. coronis ), 37 (A. hydaspe ), 38 (A. zerene) or 45 mm (A. cybele ) (Table 1). The pupa of each species is illustrated in Fig. 2 and lengths are shown in Table 1. The pupae of A. coronis and A. zerene were most similarly colored (shades ol brown with variable black markings) although A. coronis pupae tended to be more darkly pigmented with more prominent black banding on the anterior edge of each abdominal segment. Wing venation in A. zerene pupae was generally highlighted in black (Fig. 2). The pupa of A. egleis was light brown with the least dark pigmentation of the five species. The pupa of A. hydaspe was similar to those of A. coronis and A. zerene although was generally more darkly pigmented. The pupa of A. cybele differed from the others by having a rougher texture and greater girth, as well as having greater length. It was similarly colored to the pupa of A. hydaspe. Oviposition, egg and pre-diapause biology. Oviposition by females of the five species occurred between 2 and 8 days after caging, resulting in ~50 eggs each for A. egleis , A. coronis and A. cybele. Approximately 30 eggs were obtained from A. hydaspe females and an estimated 250 from A. zerene females. Females of A. hydaspe and A. egleis took 2 days to oviposit after capture on August 1 and 3, respectively. Females of A. zerene took 6 days after capture on August 3 and A. coronis females oviposited 7 days after capture on July 30th. Females of A. cybele oviposited 8 days after capture on August 20. All females generally oviposited on dessieated Viola leaves and stems, paper toweling, and muslin lids. However, A. hydaspe and A. egleis initially (first 48 h of oviposition) only oviposited on potted V labradorica, ignoring dessieated foliage. In contrast, A. coronis oviposited only on dessieated leaves and stems despite the presence of potted V labradorica. Eggs of the five species took between 9 and 1 4 days to hatch under temperatures that fluctuated between 20-28° C (Table 2). First instars did not wander far from their egg shells but sought out refugia such as curled leaves before becoming quiescent. Examination of the larval cultures of all species on October 31 indicated substantial mortality of A. zerene larvae had occurred, leaving only an estimated 50 (from 250) larvae still alive. In contrast, very little (< 5 %) mortality was evident in the cultures of the other species. Larval diapause termination experiment. All dormant first instars exposed to summer-like conditions ~ 2. 0-2. 5 months after hatching, died within 6-9 days. All except one A. cybele larva remained quiescent during this period showing no sign of feeding. The single A. cybele larvae wandered a little but did not feed. Post-diapause biology. Examination of the larval cultures in early-mid January indicated good survival (>90%) of all species under 5° C conditions. Exposure of the larvae to summer-like conditions resulted in breaking of dormancy (using commencement ol feeding as the criterion) in all species within 2-5 days (Table 2). Developmental durations and overall mortalities for Fig. 3. Ventral gland of sixth instar A. coronis simaetha. Volume 62, Number 2 67 Table 2. Developmental durations (days) for eggs, larvae and pupae of five Argynnis spp. reared at 20-28° C (eggs) or 25 ± 0.5" C under continuous illumination (larvae and pupae). Pre-feeding durations represent the time between introduction of diapausing larvae into summer-like conditions and commencement of feeding. Instar duration data were obtained from first appearance of each instar among species cohorts. First-adult durations were calculated irom introduction of first instars to summer-like conditions to adult eclosion (Mean ± SE) (number of individuals completing development in parentheses). "Indicates significant difference from A. coronis simaetha A. zerene picta A. egleis mcdiinnoughi A. cybele leto A. hydaspe rhodope Egg Pre-feeding 14 13 14 12 9 period 25°C 3 5 2 4 3 First instar 9 9 8 9 9 Second instar 7 5 5 6 8 Third instar 8 6 5 6 3 Fourth instar 4 6 5 8 3 Fifth instar 4 6 6 5 3 Sixth instar 8 11 13 10 9 Pupa 12 13 12 13 12 First - Adult 54.2 + 0.7 55.0 ± 0.3 54.1 ± 0.2 55.5 ± 1.0 47.0 ± 1.0° % Survival (4) (10) (7) (4) (4) First-Adult 50 50 50 50 75 larvae during post-diapause development at 25 ± 0.5° C/ 24 h light are shown in Table 2. Due to limitations in host plant availability, starting cohort sizes for each species were necessarily small (4—30 larvae). Additional larval cohorts of A. egleis (3) and A. zerene (10) exposed to these conditions from January 1 1 instead of January 19, were slower to start feeding and took three weeks to reach the second instar instead of just over a week for the later group. An extra group of 12 A. coronis larvae exposed to 15-21 °C and short days (9 h) from January 4-19 failed to break dormancy and did not feed. Durations of individual instars and complete post-larval feeding development were similar for A. coronis , A. zerene, A. egleis and A. cybele (Table 2). From their introduction to 25 ± 0.5° C and continuous illumination on January 19, these four species averaged 54.1-55.5 days to reach adulthood. In contrast, development of A. hyclaspe was significantly faster than the other four species under the same conditions, averaging 47 days ( P < 0.05, Mann-Whitney Rank Sum Test) (Table 2). Fifty per cent survival was obtained for all species cohorts except A. hydaspe in which 3 of the 4 larvae reached adulthood (Table 2). Larvae of all species readily accepted V. adunca and V. glabella as host plants. Viola labradorica was also accepted by all instars of A. egleis. However, its acceptance was limited in the other species to later instars (5 and 6). First instars of A. coronis and A. zerene would not feed on V labradorica. Later instars of these two species led preferentially on V. adunca when supplied in combination with V labradorica. The sagebrush violet, V trinervata Howell, was provided to A. egleis third instars only and was not accepted as a host. Domesticated pansies ( Viola tricolor L.) were accepted by sixth instars ol A. egleis, A. coronis and A. zerene. Only limited feeding on pansy occurred with sixth instar A. cybele, and A. hydaspe was not evaluated. Mature larvae of all five species constructed 'leaf tents’ from strategically silked leaves for pupation, in which they spun a silk pad for cremaster attachment. This behavior was strongly entrenched, taking place even when insufficient space within the 'tent' was available for hanging’. The silken pads or cremaster attachments were insufficient in many cases with pre- pupae or pupae falling to the ground. Pupal development durations are provided in Table 2. The pupae of A. cybele were noticeably more active (wriggling at slightest provocation) than the pupae of the other species. Relative humidity of ~ 40% at 25 ± 0.5° C caused desiccation of many pupae. Noticeable protandry occurred only with A. zerene and A. cybele-, males of these species emerged - 4-5 days earlier than females. Discussion Comparative illustrations and notes on the morphology and biology of immature stages of five Argynnis spp. commonly found in the Pacific Northwest 68 Journal of the Lepidopterists’ Society are provided for the first time. High quality images allowed separation of the five species in most of their immature stages. Prior to this study, only the endangered, coastal A. zerene hippolyta had received detailed attention to its immature stages including aspects of biology (McCorkle 19 50%) occurred with S. coronis , S. zerene and S. cybele larvae held for 135 days under these conditions. In contrast, S. egleis larvae still showed little mortality after 135 days. This overwintering technique was a lot simpler than the small hollow wooden block technique reported by Mattoon et al. (1971). In the latter procedure blocks are soaked weekly to maintain high relative humidity. The key to good overwintering survival of first instar Argijnnis appears to be adequate moisture/humidity levels but requirements may differ between species. Diy ambient conditions during September-October may have caused the substantial mortality observed among A. zerene larvae, although the other species were not adversely affected. Post-diapause development rates of immature A. coronis , A. zerene , A. egleis and A. cybele , were very similar with each species taking about 54-55 days or 7. 7-7. 8 weeks to reach adulthood at 25° C. Although only three A. hijdospe larvae completed development, their mean duration of 47 days or 6.7 weeks from post- diapause first iustar to adult was significantly less than for the other four species. These developmental durations are similar to what has been reported previously lor Argynnis spp. For example, McCorkle and Hammond (1988) reported ‘most subspecies of A. coronis and A. zerene required six to seven weeks for males and seven to eight weeks for females’, when reared at 21-23° C. They also noted that A. cybele required a similar period of time. These authors did not mention A. hydaspe, but the duration recorded here for this species compares to the fastest rates reported in their paper for certain forms of A. atlantis , A. egleis and A. callippe. Although no systematic host acceptance or preference studies were conducted, it is likely as reported previously (Mattoon et al. 1971), that Argynnis spp. differ in their acceptance of different Viola spp. All five species readily accepted the two Pacific Northwest endemic Viola spp., adunca and glabella provided to them. Viola adunca is recorded as a natural host for four ol the five species, with A. coronis as the exception. Viola glabella is recorded as a natural host for A. hydaspe, A. cybele and A. zerene but not for A. egleis and A. coronis (Hammond 1983, Scott 1986, Pyle 2002, Warren 2005). In Washington, A. coronis is reported to feed mostly on V. trinermta (Pyle 2002, Warren 2005) but V. adunca and V. glabella also occur in many A. coroni.s -occupied habitats particularly on the eastern slopes of the Cascade mountains. This study suggests the larvae of this species would have no problem utilizing this host if they encounter it. Viola trinervata was not accepted by A. egleis as a host in this study and is unlikely to grow in habitats occupied by A. egleis. Viola glabella occurs in the Blue Mountain habitat from which females were collected for this study and could be utilized as a host, given its acceptability as a larval host recorded here. Viola labradorica is native to the north eastern USA, and varied considerably in its acceptability to Washington Argynnis. First instar A. coronis and A. zerene would not feed on it and later instars ol these species and A. cybele and A. hydaspe only accepted it when given no choice. In contrast, A. egleis accepted this host readily in all instars. Late in rearing it became necessary to supplement the diets of four species with pansies (V. tricolor). Argynnis egleis , A. coronis and A. zerene accepted this host readily, but A. cybele did not. Acknowledgements I thank Dave Nunnallee and Jon Pelham for their guidance and advice during these studies and also Dave McCorkle for in- formation on Argijnnis osmeteria. Literature Cited Allen, T. |. 1997. The butterflies of West Virginia and their caterpil- lars. University of Pittsburgh Press, Pittsburgh, 388 pp. — , J. P. Brock, & J. Glassberg. 2005. Caterpillars in the field and garden. Oxford University Press, New York. 232 pp. Comstock, J. A. & C. M. Dammers. 1931. Notes on the early stages of four Californian argynnids (Lepid.). Bull. S. Cal. Acad. Sci. 30(2): 40^14. Danks, H. V. 1987. Insect dormancy: An ecological perspective. Bio- logical Survey of Canada Monograph Series No. 1. Biological Sur- vey of Canada (Terrestial Arthropods) Ottawa. 439 pp. Dornfeld, E. J. 1980. The butterflies of Oregon. The Timber Press, Forest Grove. 276 pp. Edwards, W. H. 1879. Description of the preparatory stages of Arg- ynnis egleis, Bois. Can. Entomol. 11(9): 177-179. 1880. Description of the preparatoiy stages of Argynnis cybele , Fabr. Can. Entomol. 12(8): 141-145. Guppy, C. S. & J. H. Shepard. 2001. Butterflies of British Columbia. University of British Columbia Press, Vancouver. 414 pp. Hammond, P. C. 1983. The colonization of violets and Speyeria but- terflies on the ash-pumice fields deposited by Cascadian volca- noes. J. Res. Lepid.. 20(3): 179-191. Honda, K. & N. Hayashi. 1995. Chemical nature of larval osmeterial secretions of papilionid butterflies ( Luehdorfia and Papilio). In- sect Biochem. 20: 245-250. James, D. G. 1999. Larval development in Heteronymplia merope merope (Fabricius) (Lepidoptera: Nymphalidae). Austral. Ento- mol. 26(4): 97-102. Mattoon, S. O., R. D. Davis & O. D. Spencer. 1971. Rearing tech- niques for species of Speyeria (Nymphalidae). J. Lepid. Soc. 25(4): 247-256. McCorkle, D. V. 1980. Ecological Investigation Report: Oregon Sil- verspot butterfly ( Speyeria zerene hippolyta). United States For- est Service USDA, Pacific Northwest Region, Suislaw National Forest, Corvallis, Oregon. 117 pp. & P. C. Hammond. 1988. Biology of Speyeria zerene hippolyta (Nymphalidae) in a marine-modified environment. J. Lepid Soc. 42(3): 184-192. 70 Journal of the Lepidopterists’ Society Miller, L. D. & P. C. Hammond. 2003. Lepidoptera of the Pacific Northwest: Caterpillars and Adults. USDA, Forest Health Tech- nology Enterprise Team, Washington, D.C. 324 pp. Pyle, R. M. 2002. The butterflies of Cascadia. Seattle Audubon So- ciety. 420 pp. Scott, J. A. 1986. The butterflies of North America. Stanford Univer- sity Press, Stanford. 583 pp. — & S. O. Mattoon. 1982. Early stages of Speyeria nokomis (Nymphalidae). J. Res. Lepid. 20(1): 12-15. SlMONSEN, T. J. 2006. Fritillary phylogeny, classification, and larval host plants: reconstructed mainly on the basis of male and female genitalic morphology (Lepidoptera: Nymphalidae: Argynnini). Biol. J. Linn. Soc. 89: 627-673. — , N. Wahlberg, A. V. Z. Brower & R. de Jong. 2006. Morphol- ogy, molecules and fritillaries: approaching a stable phylogeny for Argynnini (Lepidoptera: Nymphalidae). Insect Syst. Evol. 37: 405-418. Sims, S. R. 1984. Reproductive diapause in Speyeria (Lepidoptera: Nymphalidae). | Res. Lepid. 23(3): 211-216. Smedley, S. R., F. C. Schroeder, D. B. Weibel, J. Meinwald, K. A. LaFleur, J. A. Renwick, R. Rutoyvski & T. Eisner. 2002. May- olenes: Labile defensive lipids from the glandular hairs of a cater- pillar ( Pieris rapae). Proc. Natl. Acad. Sci. USA 99(10): 6822-6827. Wagner, D. L. 2005. Caterpillars of eastern North America. Prince- ton University Press, New Jersey. 512 pp. Warren, A. D. 2005. Butterflies of Oregon, their taxonomy, distribu- tion and biology. Lepidoptera of North America 6. Contributions of the C. P. Gillette Museum of Arthropod Diversity. Colorado State University, Fort Collins. 408 pp. Received for publication 27 Novemer 2006: revised and accepted 4 December 2007. Volume 62, Number 2 71 Jou rnal of the Lepidopterists’ Society 62(2), 2008, 71-79 HAWK MOTH FAUNA OF A NORTHERN ATLANTIC RAIN FOREST REMNANT (SPHINGIOAE) Jose Araujo Duarte Junior Programa de Pos-Graduagao em Ciencias Biologicas, Departamento de Sistematica e Ecologia, Universidade Federal da Parafba, 58059-900, Joao Pessoa, Parafba, Brazil; email: josejunior_lep@yahoo.com .br AND Clemens Schlindwein Departamento de Botanica, Universidade Federal de Pernambuco, Av. Prof. Moraes Rego, s/n, Cidade Universitaria, 50670-901, Recife, Pernambuco, Brazil; email: schlindw@ufpe.br ABSTRACT. We present results of a year-long faunistie survey of Sphingidae of the Brazilian northern Atlantic rain forest. The study was un- dertaken between August 2003 and July 2004, at the Private Nature Reserve (RPPN) Frei Caneca in the state of Pernambuco. Hawkmoths were captured using a 250-watt mercury-vapor light trap positioned against a white wall. We recorded 379 individuals of 50 species in 19 genera. The most abundant species were Erinmjis ello, E. elope , Neogene dynaeus and Protambulyx astygonus , which accounted for 44.2% of the collected individuals. More than one individual was recorded for all but eight species. Hawkmoths abundance was lowest in the months with intense rain- fall. The sphingid fauna of northeastern Brazil is compared with that of the Amazonian and southern Atlantic rain forest as well as with the ad- jacent caatinga, a tropical dry forest with abundant succulent plants. Species composition of Sphingidae of the northern Atlantic rain forest was most similar to that of the Amazonian forest. Additional key words: biodiversity, biogeography. Brazil, distribution, Pernambuco, South America, survey The Sphingidae (Bombycoidea) includes about 1200 species globally (Lemaire & Minet 1999) and ISO species in Brazil (Brown 1986). Sphingidae are cosmopolitan and show highest diversity in the tropics (Hodges 1971). In South America, surveys of Sphingidae are scarce and regional inventories are necessary to know their diversity and distribution and to elucidate their biogeographical relationships (Kitching & Cadiou 2000). The Brazilian Atlantic rain forest, which extends along the Atlantic coast between the states of Rio Grande do Norte and Rio Grande do Sul (Rizzini 1997), is currently highly fragmented, with only 5% of its original forest remaining (Ranta et al. 1998, Tabarelli et al. 2002). The first local survey of Sphingidae in the northern part of the Atlantic rain forest revealed 23 sjrecies (Duarte & Schlindwein 2005a). In an area of cerrac/o -like savannah vegetation of the Tabuleiro (tropical grassland with evergreen trees and shrubs), in the northeast Brazilian state of Parafba, Darrault & Schlindwein (2002) recorded 24 species of sphingids. In the caatinga, the hawkmoth fauna is poor, and only 14 species were recorded in an area of caatinga in Parafba (Gusmao & Creao-Duarte 2004) and 20 in Rio Grande do Norte (Duarte & Schlindwein 2005b). This contrasts to the high diversity of Sphingidae in the Amazon basin (Motta et al. 1998). The northern Atlantic rain forest is strongly influenced by the Amazonian biota (Prance 1982, Santos et al. 2007). The montane forests of northeast Brazil on the other hand form a refuge for several sjrecies of plants and animals, which, due to their cooler and more humid climate, differ from the arid caatinga that surrounds them (Andrade- Lima 1982). Several botanical studies in northeastern Brazil have revealed floristic disjunctions between the Amazonian forest and the north Atlantic rain forest (Andrade-Lima 1982). This is also true for many animal taxa (Bigarella et al. 1975, Coimbra-Filho & Camara 1996). In this study we determine sjDecies richness, abundance, and seasonality of Sphingidae of a preserved area of the Atlantic rain forest in Pernambuco, northeastern Brazil, and compare the sphingid fauna to that of the caatinga and Amazonian rain forest. Materials and Methods Study area. The study was carried out in the Reserva Particular do Patrimonio Natural RPPN Frei Caneca (Private Nature Reserve Frei Caneca) in the municipality of Jaqueira, Pernambuco, NE-Brazil. The study site is located at 8°42’41”S and 35°50’30”W at an altitude of 500-750 nr (Fig. 1). The reserve covers an area of 630.42 ha, with a mountainous relief and granite rocky outcrops. The climate is tropical, hot and humid, with a mean annual temperature of 22°C. There is a 4-5 month dry (less-humid) season between October and February and a rainy season between March and September (IBGE 1985). The mean annual rainfall. 72 Journal of the Lepidopterists’ Society Conventions Q Brazil CD Northeast Brazil • Natural Reserve Frei Caneca r 'mh * i— A i 1 }y//k jii f||| llx. N///r7V ^feGrande A Pli //// A^goas i y//s gp ZZZ/, r t/A Oapttic Scale ( Km ) Reference System SAD-69 Fig. 1. Geographical location of the study site RPPN Frei Caneca in the Atlantic rain forest of Pernambuco, Brazil. averaged over 47 years at Frei Caneca, which is 7 km from the reserve, is 1332 mm (unpublished meteorological data provided by Usina Colonia). Sampling of Sphingidae. The survey was undertaken between August 2003 and July 2004. A 250W mercury-vapor light source, positioned against a white wall of the Reserve Station inside the park facing the forest, was used to attract moths. Specimens were collected on two consecutive new moon nights per month from lS.OOh to 5.00h of the following day. Moths were killed by an injection of ethyl acetate in the posterior part of the thorax. Each specimen was then placed in an entomological envelope and prepared in the laboratory. Moths were identified using d'Abrera (1986) and Kitching & Cadiou (2000) and the reference collection at UFPE. Specimens were deposited in the Entomological Collection of the Federal University of Pernambuco (UFPE, Recife) and the Entomological Collection of the Department of Systematics and Ecology, Federal University of Paraiba (UFPB, Joao Pessoa). Three abundance criteria were established using Rabinowitz et al. (1986), based on the number of specimens collected per species: rare (1 to 2), common (3 to 19), and abundant (20 to 50). Bio-Estat 2.0 (Ayres et al. 2000) was used to calculate Pearsons correlation coefficients (Sokal & Rohlf 1996). The data were adjusted to lognormal distributions according to the model of Preston (1948), which groups the species into frequency classes of individuals on a logarithmic scale. The program “lognorm.bas” (Ludwing & Reynolds 1988) was used, according to the equation S(R)=S0e{“ R \ where S(R) is the estimated number of species in a given octave, R is the distance in relation 1,2,3,.. (Octaves), S0 is the estimated number of species in the modal octave, e is the natural logarithm base, and a an estimated constant calculated as a2 = l/(2.s)2 , where s is the standard deviation. Dates were compared with regional inventories of Sphingidae of the Amazonian rain forest, south Atlantic rain forest and eaatinga. Similarities were analyzed using NTSYS pc version 2. 1 Ot. Results Three hundred and seventy-nine individuals, representing 50 species in 19 genera were recorded, of which 15 species are new records for northeast Brazil (Table 1). The most abundant species were Erinnyis ello Linnaeus 1758, Erinnyis alope (Drury 1773), Neogene dynaeus (Hiibner [1825]) and Protambulyx astygonus (Boisduval [1875]), accounting together for 44.2% of the individuals recorded. Only 1 or 2 individuals were recorded for 17 species (Fig. 2). From October to December, the driest months of the study period (192 mm, 6.6% of total rainfall), 170 Volume 62, Number 2 73 70 , Fig. 2. Number of individuals of sphingid species recorded from August 2003 to July 2004 in the Atlantic rain forest of Frei Caneca, Pernambuco individuals were recorded, 45% of the total. In January, the month with the highest rainfall (554 mm, 19%), only 2 individuals were recorded, Adhemarius gannascus (Stoll 1790) and Protambidt/x astygonus (Boisduval [1875]). Other months with high rainfall, namely May, June and J uly (1120 mm, 392%) also showed low hawkmoth abundance (70 individuals, 18.5%). More than hall of the species (30 individuals, 58.8%) were recorded in March (Table. 1). The male to female sex ratio was 4:1, with 80% of captured moths being males. For almost half the species (26/50) only males were recorded. In the north Atlantic rain forest, species richness is lower in the rainy months. Pearson's correlation coefficient between rainfall 2003/2004 anti abundance was -0.6813, and between rainfall 2003/2004 and species richness was -0.6741, both negative and significant (< 0,05). The species accumulation curve (Fig. 4), shows that by the fifth month of collecting 70.5% of the total number of sphingid species had been recorded and after the tenth month the number of species did not increase. The estimated number of species in the study area was 52 (Fig. 5). DfSCUSSION The sphingid fauna of the north Atlantic rain forest shows a much higher species richness and abundance than that found in another survey, undertaken in the Atlantic rain forest. Nature Reserve of Gurjau in Pernambuco, using the same methodology and the same sampling effort, in which only 23 species were recorded (Duarte & Schlindwein 2005a). Of these, only Pachylia syces , Protambnlyx eurycles, P. goeldii and Manduca brasilensis were not recorded in the present study. The Nature Reserve, Frei Caneca is only 150 km distant f rom the reserve Gurjau and is characterized by a little disturbed rain forest in a good level of conservation. The latter area is heavily fragmented, close to the metropolitan area of Recife City, surrounded and strongly impacted by sugar cane monocultures. The high number of additions to the list of the northeast Brazilian sphingid fauna (15 species) shows that the sphingid fauna of this area is still poorly known. No surveys are available for any other northeast Brazilian states (Bahia, Sergipe, Alagoas, Ceara, Piaui and Maranhao). These woidd most likely further increase the number of species in this region. Our study corrects earlier statements made about the apparently restricted distributions of some species, such as Xylophones anubus , Amphimoea walked and Madoryx plutonius, which are now known to be more widespread in the neotropical region than previously thought. The sphingid fauna of the Atlantic rain forest of Pernambuco shows no strong seasonal pattern, but we found a significant negative correlation between rainfall and both species richness and abundance, such that the rainiest months showed the lowest species richness and abundance. In Costa Rica adult sphingids were almost absent at the end of the diy season, abundant in the rainy season but scarce in October, the most humid month of the year. Abundance of Sphingidae was associated with the presence of leaves on their larval host-plants (Haber & Frankie 1989). In the south Brazilian state of Parana, which is outside the tropics, sphingid abundance was correlated with temperature and abundance drastically diminished in winter (Marinoni et al. 1999). The sphingid fauna is also highly seasonal in the dry northeast Brazilian caatinga, where greatest abundance is in March at the beginning of the rainy season, when larval host-plants provide their leaves (Duarte & Schlindwein 2005b). Similar results were found in Mexico, where 77% of sphingids were recorded only in the rainy season (Gomez-Nucamendi et al. 2000). 74 Journal of the Lepidopterists’ Society Table 1. Species of Sphingidae recorded in the Atlantic rain forest of Frei Caneca, Pernambuco, Brazil, from August 2003 to July 2004. The abundance categories follow Rabinowitz et al. (1986). Taxon Month Rare Common Abundant Male Female Total Macroglossinae MACROGLOSSINI Xylophones cf. amadis (Stoll, 1782) Aug X 1 0 1 Xylophones anubus (Cramer, 1777) ° Feb- May, Jun X 9 1 10 Xylophones pluto (Fabricius, 1777) Mar, Nov X 3 0 3 Xylophones t. tersa (Linnaeus, 1771) Apr, May, Aug, Nov, Dec X 5 2 7 Xylophones cliiron (Drury, 1773) ° Mar, Sep, Oct Dec X 6 0 6 Xylophones libya (Druce, 1878) Sep X 1 0 1 Xylophones loelia (Druce,1878) Fev, Mar-Jun, Oct X 8 1 9 Xylophones t. thyelia (Linnaeus, 1758) ° Mar, Oct, Dec X 3 0 3 DILOPHONOTINI Aleuron iphis (Walker, 1856) ° May X 1 0 1 Callionima falcifera (Gehlen,1943) ° Mar, Dec X 4 0 4 Callionima g risescens elegans (Gehlen,1935) Feb X 1 1 2 Callionima inuus (Rotlisch. & Jordan, 1903) * Mar, May, Jun, Oct, Dec X 4 2 6 Callionima nomius (Walker, 1856) ° Feb-Apr, May, Dec X 8 1 9 Callionima parce (Fabricius, 1775) Feb, Mar, May, Aug, Nov X 4 1 5 Erinnyis a. alope (Drury, 1773) Feb-Apr, Sep-Dec X 50 11 61 Erinnyis crameri (Schaus, 1898) Aug X 1 0 1 Erinnyis e. ello (Linnaeus, 1758) Mar-Jun, Sep-Dec X 37 26 63 Erinnyis lassauxii (Boisduval, 1859) Mar, May X 4 0 4 Erinnyis o. obscura (Fabricius, 1775) Mar X 2 0 2 Enyo 1. lugubris (Linnaeus, 1771) Sep X 1 0 i Enyo ocypete (Linnaeus, 1758) May X 2 0 2 Hemeroplanes triptolemus (Cramer, 1779) Feb, May, Jun, Sep, Nov, Dec X 9 3 12 Isognathus allamandae Clark, 1920 May, Jun, Oct, Dec X 7 0 7 Isognathus c. caricae (Linnaeus, 1758) Feb-May, Oct, Dec X 7 0 7 Isognathus leachii (Swainson, 1823)° Feb -Jun, Dec X 7 2 9 Isognathus menechus (Boisduval, [1875]) Feb-May, Sep, Dec X 8 2 10 Isognathus swainsonii (Felder & Felder, 1862)° Mar.Oct, Nov X 8 0 8 Madoryx plutonius (Hiibner, [1819])° Oct X 1 0 1 P achylia ficus (Linnaeus, 1758) May, Oct, Nov X 2 3 5 Pachylioides resumens (Walker, 1856) ° May X 2 1 3 Pseudosphinx tetrio (Linnaeus, 1771) Feb, Mar, Sep-Dec X 7 2 9 PHILAMPELINI Eumorpha anchemolus (Cramer, 1779) Nov X i 0 1 Eumorpha f. fasciatus (Sulzer, 1776) Apr, May X 2 0 2 Eumorpha l. labruscae (Linnaeus, 1758) May X 0 2 2 Eumorpha obliquus (Rothseh & Jord, 1903) ° Mar, Oct, Dec X 3 0 3 Eumorpha v. vitis (Linnaeus, 1758) Mar, Apr X 3 0 3 Volume 62, Number 2 75 Table 1. Continued. Taxon Month Rare Common Abundant Male Female Total Smerinthinae AMBULYCINI Adheinarius gannascus (Stoll, 1790) Jan -May, Dec x 6 1 7 Protambulyx strigilis (Linnaeus, 1771) May, Nov, Dec x 5 0 5 Protambulyx astygonus (Boisduval, [1875]) Jan-Mar, Oct-Dec x 18 1 19 Sphinginae SPHINGIN1 Agnus cingulatus (Fabricius, 1775) Jun, Sep x 2 1 3 Amphimoea walkeri (Boisduval, [1875]) ° Mar, Jun x 2 0 2 Cocytius antaeus (01X117,1773) Mar- May, Oct x 4 2 6 Neogene dynaeus (Hiibner, [1825]) Feb-Jun, Aug-Dec x 20 4 24 Manduca cf. clarki (Rothsch. & Jordan, 1916) Mar x 1 0 1 Manduca contractu (Butler, 1S75) * Feb, Mar, May, Jun x 12 2 14 Manduca diffissa tropical is (Rothsch. & Jord, 1903) Mar, Sep., Oct x 4 0 4 Manduca h. Hannibal (Cramer, 1779) May, Nov x 1 1 2 Manduca florestan (Stoll, 1782) ° Mar, Apr X 2 0 2 Manduca r. nistica (Fabricius, 1775) Feb, Sep, Dec x 4 1 5 Manduca sexta paphus (Cramer, 1779) Mar X 1 1 2 Total 304 75 379 “New occurrence in northeastern Brazil 76 Journal of the Lepidopterists’ Society Table 2. Presence of sphingid species in the northern Atlantic rain forest (this study), Amazonian rain forest (Motta et al. 1998; Motta & Andreazze 2002), southern Atlantic rain forest (Laroca & Mielke 1975, Marinoni et al. 1999) and caatinga (Duarte et al. 2001, Gusmao & Creao-Duarte 2004, Duarte & Schlindwein 2005b). Taxon N -Atlantic Rainforest Amazonian Rainforest S -Atlantic Rainforest Caatinga Adhemarius gagarini (Zikan, 1935) X Adhemarius gannascus (Stoll, 1790) X X X Adhemarius palmeri (Boisduval, 1875) X X Aellopos ceculus (Cramer, 1777) X Agrius cingulatus (Fabricius, 1775) X X X X Aleuron chloroptera (Perty, 1834) X Aleuron iphis (Walker, 1856) X X Aleuron n. neglectum (Rothschild & Jordan, 1903) X Amphimoea walkeri (Boisduval, [1875]) X X Callionima falcifera (Gehlen, 1943) X Callionima grisescens elegans (Gehlen, 1935) X X Callionima inuus (Rothschild & Jordan, 1903) X X X Callionima nomius (Walker, 1856) X X X Callionima p.pan (Cramer, 1779) X Callionima parce (Fabricius, 1775) X X X X Cocytius antaeus (Drury, 1773) X X X X Cocytius beelzebuth (Boisduval, 1875) X Cocytius duponchel (Poey, 1832) X X Cocytius lucifer (Rothschild & Jordan, 1903) X Enyo g.gorgon (Cramer, 1777) X X Enyo l. lugubris (Linnaeus, 1771) X X X X Enyo ocypete (Linnaeus, 1758) X X X Erinnyis a. alope (Drury, 1773) X X X X Erinnyis crameri (Schaus, 1898) X X X Erinnyis e. ello (Linnaeus, 1758) X X X X Erinnyis lassauxii (Boisduval, 1859) X X X X Erinnyis o. obscura (Fabricius, 1775) X X X X Erinnyis oenotnis (Stoll, 1780) X X Eumorpha anchemolus (Cramer, 1779) X X X Eumorplia capronnieri (Boisduval, 1875) X Eurnorpha eacus (Cramer, 1780) X Eumorpha f. fasciatus (Sulzer, 1776) X X X Eumorpha 1. labruscae (Linnaeus, 1758) X X X X Eumorpha obliquus (Rothschild & Jordan, 1903) X X X Eumorpha phorbas (Cramer, 1775) X Eumorpha v. vitis (Linnaeus, 1758) X X X X Eupyrrhoglossum venustum (Rothschild & Jordan, 1903) X Hemeroplanes triptolemus (Cramer, 1779) X X X Hyles euphorbiarum (Guerin & Percheron, 1835) X Isognathus allamandae Clark, 1920 X X Isognathus australis Clark, 1917 X Isognathus c. caricae (Linnaeus, 1758) X X Isognathus excelsior Boisduval, 1875 X Isognathus leachii (Swainson, 1823) X X Isognathus m. mossi Clark, 1917 X Volume 62, Number 2 7 / Table 2. Continued. N-Atlantic Amazonian S-Atlantic Taxon Rainforest Rainforest Rainforest Caatinga Isognathus menechus (Boisduval, [1875]) Isognatlius rimosus (Grote, 1865) Isognathus scyron (Stoll, 1780) Isognathus swainsonii (Felder & Felder, 1862) Isognathus zebra Clark, 1923 X X X X X X X Madoryx plutonius (Hiibner, [1819]) Manduca brasilensis Jordan, 1911 Manduca brunalba (Clark, 1929) X X X X X X Manduca cf. clarki (Rothschild & Jordan, 1916) Manduca lucetius (Cramer, 1780) X X X Manduca contracta (Butler, 1875) Manduca d. dalica (Kirby, 1877) X X Manduca diffissa tropicalis (Roth. & Jordan, 1903) X X X Manduca florestan (Stoll, 1782) X X X Manduca h. hannibal (Cramer, 1779) X X X Manduca I. lefeburei (Guerin, 1844) Manduca p. pcllenia (Herrich-Schaeffer, 1854) X X X X Manduca r. rustica (Fabricius, 1775) Manduca sexta paphus (Cramer, 1779) Neococytius cluentius (Cramer, 1775) X X X X X X X X X X Neogene dynaeus (Hiibner, [1827]-[1831]) Nyceryx c. continue (Walker, 1856) Orecta I. lycidas (Boisduval, 1875) On/ba kadeni (Shaufuss, 1870) Pachylia darceta Druce, 1881 X X X X X X P achylia ficus (Linnaeus, 1758) Pachylia syces (Hiibner, [1819]) Pachylioides resumens (Walker, 1856) Perigonia lusca lusca (Fabricius, 1777) Perigonia pallida Rothschild & Jordan, 1903 X X X X X X X X X Perigonia pittieri Lichy, 1962 Phryxus caicus (Cramer, 1777) Protambulyx astygonus (Boisduval, [1875]) Protambulyx eurycles (Roth. & Jordan, 1903) X X X X X Protambulyx goeldii (Roth. & Jordan, 1903) X Protambulyx strigilis (Linnaeus, 1771) Pseudosphinx tetrio (Linnaeus, 1771) Xylophones aglaor (Boisduval, 1875) Xylophones anubus (Cramer, 1777) Xylophones ceratomioides (Grote & Robinson, 1867) X X X X X X X X X X X X X Xylophones cf. amadis (Stoll, 1782) Xylophones chiron (Drury, 1773) Xylophones indistincta Closs, 1915 Xylophones isaon (Boisduval, 1875) Xylophones libya (Druce, 1878) X X X X X X X X Xylophones loelia (Druce, 1878) Xylophones pluto (Fabricius, 1777) Xylophones porcus continentalis Rothschild & Jordan, 1903 Xylophones s. schausi (Rothschild, 1894) X X X X X X X X Xylophones t. tersa (Linnaeus, 1771) X X X X Xylophones t. thyelia (Linnaeus, 1758) Xylophones titana (Druce, 1878) Xylophones tyndarus (Boisduval, 1875) Xylophones xylobotes (Burmeister, 1878) X X X X X X Total 54 71 63 26 Species in common with the N-Atlantic Rain Forest 42 (59 %) 32 (58%) 20 (77%) 78 Journal of the Lepidopterists’ Society saaszsza Species Richness - - - - Prec Pluriannual (1983-2003) ■ Abundance -Precipitation during study penod Fig. 3. Abundance and richness of sphingids, mean annual precipitation and precipitation during the study period in the Atlantic rain forest of Frei Caneca from August 2003 to July 2004. Months Fig. 4. Species accumulation curve of Sphingidae during the study, from August 2003 to July 2004, in the Atlantic Rain For- est of Frei Caneca. ■ Observed Adjusted Fig. 5. Distribution of species recorded in the Atlantic rain forest of Frei Caneca by abundance class (octaves), adjusted to a lognormal curve. In the north Atlantic rain forest, the vegetation is not deciduous and host-plants have leaves throughout the year, a factor that probably exercises great influence on the sphingid abundance. In Para (Amazonian region), Moss (1920) noted that periods of heavy rainfall reduced the abundance of hawlcmoths, because they dislodged larvae, mainly those newly-emerged from the egg- Callionima grisescens elegans is a sphingid subspecies endemic to northeast Brazil (Schreiber 1978), occurring abundantly in the caatinga (Duarte & Schlindwein 2005b) and tropical montane forest in Paraiba (Gusmao & Creao-Duarte 2004). In the Atlantic rain forest, only two specimens were recorded, suggesting that it is a resident species of the caatinga. The high male to female ratio in onr study follows a pattern similar to the 10:1 male to female capture ratio reported in Costa Rica by Janzen (1983). The expected male to female proportion is 1:1 (Kitching & Cadiou 2000). It appears that sampling of sphingids with light traps distorts this ratio. Janzen (1983) supposed that the two sexes could have a physiological susceptibly to light, with males showing a high mobility using light sources as reference points in finding females. Comparison of the present short-term survey of the sphingid fauna of the northern Atlantic rain forest with those conducted in the southern Atlantic rain forest (Laroca & Mielke 1975, Marinoni et al. 1999), caatinga (Gusmao & Creao-duarte 2004, Duarte & Schlindwein 2005b) and the Amazonian forest (Motta et al. 1998; Motta & Andreazze 2002) showed that 78% (42) of the species recorded in the northeastern Atlantic rain forest also occur in the Amazonian forest, 59% (32) in the southern Atlantic rain forest, and 37% (20) in the caatinga. Protambulyx astygonus , Callionima falcif era, Pachylia syces , Manduca contractu and Xylophones libtya were recorded only in the northeastern Atlantic rain forest (Table 2). Nevertheless, these comparisons have to be treated with care, because the compiled species lists of all regions are results of short-term su rveys. The present study shows that the sphingid fauna of the north Atlantic rain forest is most similar to that of the Amazonian forest when compared to other regions. Several authors have demonstrated a close faunistic and floristie relationship between these two forests (Andrade-Lima 1982, Bigarella et al. 1975, Vanzolini 1970, Haffer 1982, Santos et al. 2007), which today are separated by about 1500 km. The xeromorphic caatinga forest, that occupies the gap between the rain forests, is characterized by a highly endemic flora (Queiroz et al. 2006). The sphingid fauna of the caatinga, however, is impoverished and formed almost exclusively from Volume 62, Number 2 79 elements of the Atlantic and the Amazonian rain forests. Acknowledgements We thank IB AM A for the license to work at the RPPN Frei Caneca. Andre Mauricio Melo Santos, Reisla Oliveira, Fernando Zanella helped with statistical support. Olaf H. H. Mielke (Fed- eral University of Parana, Curitiba), Catarina Motta (National Research Institute of Amazonas, INPA, Manaus) and I J. Kitch- ing (The Natural History Museum, London), kindly confirmed identification of specimens. This study was supported by grants from CAPES to J.A.D and CNPq to C.S. Literature Cited Andrade-Lima, D. 1982. Present-day forest refuges in northeastern Brazil. Pp. 245-251. In G.T. Prance (ed.). Biology diversification in the tropics, Columbia University Press. New York. 714pp. Ayres, M., Jr., D. L. Ayres, &. A. S. Santos. 2000. BioEstat 2.0: apli- cagoes estatisticas nas areas de ciencias biologicas e medicas. So- ciedade Civil Mamiraua e CNPq, Brasilia. 272pp. Bigarella, | .J., D. Andrade-Lima, & P. J. Riehs. 1975. Consider- agoes a respeito das mudangas paleoambientais na distribuigao de algumas especies vegetais e animais no Brasil. Anais da Academia Brasileira de Ciencias 47: 411- 464. Brown, K. S., fr. 1986. Diversity of Brazilian Lepidoptera: History of study, methods for measurement, and use as indicator for genetic, specific and system richness. Pp. 221-253. In C. E. Bicudo & N. A. Menezes (eds.). Biodiversity in Brazil: a first approach. Sao Paulo. 326pp. Coimbra-Filho, A. F, & Camara, I. G. 1996. Os limites originais do bioma Mata Atlantica na Regiao Nordeste do Brasil. Fundagao brasileira para a Conservagao da Natureza. Rio de Janeiro. 86pp. D'Abrera, B. 1986. Sphingidae Munch. Hawk moths of the world. Faringdon, Oxon, United Kingdom, E.W. Classey. 225pp. Darrault, R. O. & C. Schlindwein. 2002. Esfingideos (Lepidoptera, Sphingidae) no Tabuleiro Paraibano, Nordeste do Brasil. Abundaneia, riqueza e relagao com plantas esfingofilas. Revista Brasileira de Zoologia 19 (2): 429-443. Duarte, A. J., Jr., C. S. Motta & A. Varella-Freire. 2001. Sphingi- dae (Lepidoptera) da Estagao Ecologica do Serido, Serra Negra do Norte, Rio Grande do Norte, Brasil. Entomologia & Vectores 8(3): 341-347. Jr. & C. Schlindwein. 2005a. Riqueza, abundaneia e sazonal- idade de Sphingidae (Lepidoptera, Heterocera) num fragmento de Mata Atlantica de Pernambuco, Brasil. Revista Brasileira de Zoologia 22(3): 662-666. ., Jr. &.C. Schlindwein. 2005b. The highly seasonal hawkmoth fauna (Lepidoptera, Sphingidae) of the caatinga of northeast Brazil: a case study in the state of Rio Grande do Norte. Journal of the Lepidopterists1 Society 59(4): 212-218. G6mez-Nucamendi, O. L., R. W. Jones & A. Moron-Rios. 2000. The Sphingidae (Heterocera) of the "El Ocote" Reserve, Chiapas, Mexico. Journal of the Lepidopterists1 Society .53(4): 153—158. Gusmao, M. A. B. & A. J. CreAo-Duarte. 2004. Diversidade e analise faum'stica de Sphingidae (Lepidoptera) em area de brejo e caatinga no Estado da Paraiba, Brasil. Revista Brasileira de Zo- ologia 21(3): 491-498. Haffer, J. 1982. General aspects of the Refuge Theory. Pp. 6-24. In G. T. Prance (ed.). Biological diversification in the tropics. Co- lumbia University Press, New York. 714pp. Haber, W. A. & G. W. Frankie. 1989. A tropical hawk-moth commu- nity: Costa Rican dry forest Sphingidae. Biotropica 21(2):155— 172. Hodges, R. W. 1971. Sphingidae. The moths of America north of Mexico, volume 21. E. W. Classey and R.B.D. Publications, Lon- don. 158pp. IBGE. 1985. Atlas nacional do Brasil. IBGE, Rio de Janeiro. Janzen, D. H. 1983. Costa Rica natural history. Univ. Chicago Press, Chicago. 816pp. Kitching, I. J. & J.-M. Cadiou. 2000. Hawkmoths of the world: an annotated and illustrated revisionary checklist (Lepidoptera: Sphingidae). Cornell University Press, Ithaca, New York. 227pp. Laroca, S. & O. H. IT Mielke. 1975. Ensaios sobre ecologia de co- munidades em Sphingidae na Serra do Mar, Parana, Brasil. (Lep- idoptera). Revista Brasileira de Biologia 35(1): 1-19. Lemaire, C. & | Minet. 1999. The Bombycoidea and their relatives. Pp. 321-353. In De Gruyter (ed.). Lepidoptera: moths and but- terflies. 1. Evolution, systematics, and biogeography. Part 35. Handbook of zoology. IV, Berlin, New York. 576pp. Ludwing, J. A. & J. F. Reynolds. 1988. Statistical ecology. New York: John Wiley, 337pp. Marinoni, R. C„ R. R. C. Dutra & O. Id. H. Mielke. 1999. Levan- tamento da fauna entomologica no Estado do Parana. IV. Sphin- gidae (Lepidoptera). Diversidade alfa e estrutura de comunidade. Revista Brasileira de Zoologia 16(2): 223-240. Moss, A. M, 1920. Sphingidae of Para. Brazil. Early stages, food plants, habits, etc. Novitates. Zoological 27: 334-357. Motta, C. S., F. J. Aguilera-Peralta & R. Andreazze. 1998. Aspec- tos da Esfingofauna (Lepidoptera: Sphingidae), em area de terra firme, no Estado do Amazonas, Brasil. Acta Amazonica 28(1): 75-92. . & R. Andreazze. 2002. Sphingidae (Lepidoptera) de Querari, Sao Gabriel da Cachoeira, Amazonas, Brasil. Entomologia & Ve- tores 9(3): 329-337. Prance, G. T. 1982. Forest refuges: evidences from woody an- giosperms. Pp. 137-159. In G. T. Prance (ed.). Biological diversi- fication in the tropics. Columbia University Press, New York. 714pp. Preston, F. W. 1948. The commonness and rarity of species. Ecology 29: 254-283. Queiroz, L. R, A. A. CoNCEigAo & A. M. Giulietti. 2006. Nordeste semi-arido: caracterizagao geral e lista das fanerogamas. Pp. 15-364. In A. M. Giulietti, A. A. Conceigao & L. P. Queiroz (eds.), Diversidade e caracterizagao das fanerogamas do semi- arido brasileiro. Recife, Associagao Plantas do Nordeste, Vol. 1. 488pp. Rabinowitz, D., S. Cairns & T. Dillon. 1986. Seven forms of rarity and their frequency in the flora of the British Isles. Pp. 205-217. In M. E. Soule (ed.). Conservation biology: The science of scarcity and diversity. Sinauer, Sunderland, Massachusetts. 584pp. Ranta, P, T. Blom, J. Niemela., E. Joensuu & M. Siitonen. 1998. The fragment rain forest of Brazil: size, shape and distribution of forest fragment. Biodiversity & Conservation 7(3): 385-403. Rizzini, C. T. 1997. Tratado de fitogeografia do Brasil. Ambito cultural Edigao Ltda, Rio de Janeiro, Brasil. 747pp. Santos, A. M. M., D. R. Cavalcanti., J. M. S. Cardoso & M. Tabarelli. 2007. Biogeographical relationships among tropical forests in north-eastern Brazil. Journal of Biogeography 34: 437^46. Schreiber, II 1978. Dispersal centres of Sphingidae (Lepidoptera) in the neotropical region. Biogeographica. 10: 195pp. Sokal, R. R. & F. J. Rohlf. 1996. Biometry. Freeman & Company, New York. 887pp. Tabarelli, M., J. F. Marins & J. M. C. Silva. 2002. La biodiversidad brasilefia. Investigation y Ciencia 308: 42^9. Vanzolini, P. E. 1970. Zoologia sistematica, Geografia e a origem das especies. Universidade. S. Paulo. Instituto Geografico. Series teses e Monografias 3: 1-56. Received for publication 29 September 2005; revised and accepted 20 December 2007. 80 Journal of the Lepidopterists’ Society Journal of the Lepidopterists’ Society 62(2), 2008, 80-83 NOTES ON PAPILIO MACHAON ALIASKA (PAPILIONIDAE) POPULATIONS NEAR FAIRBANKS, AK Shannon M. Murphy Department of Ecology and Evolutionary Biology, Corson Hall, Cornell University, Ithaca, NY 14853, USA; Present Address: Department of Biological Sciences, The George Washington University, Lisner Hall, Suite 340, 2023 G Street, NW, Washington, DC 20052, USA; email: smurph@gwu.edu ABSTRACT. I present results from a mark-recapture study ot Papilio machaon aliaska swallowtail butterflies from four sites near Fairbanks, Alaska. The sites were alpine-tundra hilltops and butterflies were caught throughout the month of June in 2000-2003. Only males (n=569) were marked and released while females (n=31) were kept for other experiments. Adult males tended to fly earlier in the season than did females and also were found flying earlier in the day than females. About one sixth of the males that were marked were later recaught and some were caught multiple times (up to six times for one male). Most males were recaught within four days of their initial catch date, but a few were caught many days later. Thus, these data indicate that some males may live for up to two to three weeks under natural field conditions. The research pre- sented here support the claim that P. m. aliaska is a hilltopping swallowtail butterfly. Additional key words: flight behavior, flight times, mark-recapture. Swallowtail butterflies from the Papilio machaon group use plants of the Apiaeeae as their primary hosts (Feeny et al. 1983; Sperling 1987; Thompson 1995; Wiklund 1981). Apart from occasional use of plants in the family Rutaceae, an ancestral host family for the genus Papilio (Sperling 1987), P. machaon swallowtails have rarely incorporated non-apiaceous plants into their diet. In Alaska and northwestern Canada, Papilio machaon aliaska Scud, oviposits and feeds not only on the local apiaceous host, Cnidium cnidiifolium (Turcz.) Schischk., but also on Artemisia arctica Less, and Petasites frigidus (L.) Franeh. (Scott 1986) in the Asteraceae. This host-range expansion by P. m. aliaska appears to represent an intermediate step towards a complete host shift. Previous work has demonstrated that shared chemical cues in ancestral and novel host plants may have provided the opportunity for the establishment of the host expansion onto the two novel host species (Murphy & Feeny 2006). However, these host plants are not equal in terms of larval survival in the field (Murphy 2004) or the laboratory (Murphy 2007a). In the absence of predators, P. m. aliaska larvae survive best on the ancestral host plant, C. cnidiifolium, but in the presence of predators, larval survival is greater on the novel host plants. In the field, the novel host plants seem to offer larvae enemy-free space that is not found on the ancestral host plant simply because of their different local environments. Predators are common in the ancestral host plants environment and larval mortality on C. cnidiifolium can be very high in the field; enemy-free space on the novel host plants may be the selective pressure maintaining the host expansion, possibly driving the incipient host shift to completion. Despite the environmental differences and physical distance between the locations where the larval host jflants can be found, P. m. aliaska is thought to be a typical hilltopping swallowtail butterfly (cf. Lederhouse 1982; Shields 1967). Hilltopping is a widespread behavior in butterflies and has been documented in at least five Lepidoptera families, including Papilionidae (Shields 1967). When males and virgin females emerge from their pupae, they fly towards a local topographic prominence (Pe'er et al. 2004), which may be quite minor in appearance (Baughman & Murphy 1988), and congregate at the summit. Hilltopping behavior may be an effective method for finding mates in low-density species (Scott 1968) or in species that do not mate on or near their larval host plants (Rutowski 1991). Males tend to establish territories (or perches’ sensu Scott 1974) and exhibit aggressive behaviors towards other males as well as other butterfly species (Lederhouse 1982). Virgin females, or females that mate multiple times in some species, also summit the hilltop, mate with the males, and then return to lower elevations to search for oviposition sites (Shields 1967, but see Pe’er (2004) for a discussion of whether this downhill movement is active or passive). Thus, on these hilltops, males tend to be numerically more common than are females since females only summit long enough to mate while males defend their territories and wait for new mates at the top of the hill (Alcock 1985; Shields 1967). Lederhouse (1982) found that for the black swallowtail butterfly, Papilio polyxenes, early-emerging males were more likely to defend a preferred territory, and these preferred territories were visited more frequently by females. Here I present data that I gathered when I was collecting P. m. aliaska individuals in the field, including a mark-recapture study on P. m. aliaska males. The goal of this research is to investigate the flight behaviors of P. m. aliaska butterflies in the field. In addition to learning Volume 62, Number 2 81 more about peak flight time and longevity under natural conditions, I aimed to determine if my observations of P. m. aliaska flight behavior near Fairbanks, AK are consistent with patterns associated with other hilltopping butterflies (e.g. female rarity and males that either remain or return to a hilltop regularly lor several days). Materials and Methods With help from many field assistants, I collected P. m. aliaska individuals from four sites in Alaska. The sites were alpine-tundra hilltops (domes) near Fairbanks, AK: Ester Dome (64°52’N, 148°4’W, ~720m), Murphy Dome (64°57’N, 148°21'W, ~890m), Wickersham Dome (65° 13’ N, 148°3’ W, ~977m) and along the Pinnel Mountain trail southwest of Table Mountain (65°25’ N, 145°57’ W, ~l,200m). The two closest sites. Ester and Muqihy domes, are about 18 km apart while the two sites that are farthest from each other are about 120 km apart (Ester and Pinnel Mountain). These four sites have populations of the host plants Artemisia arctica and Petasites frigidus and vegetation characteristic of open tundra. Ester and Muiphy Domes are characterized by low birch and willow scrub ( Betula , Salix spp.) with a few small spruces ( Picea ) as well as dwarf scrub ( Andromeda , Anemone, Carex , Empetrum, Epilobium, Ledum, Lupinus, Pedicularis, Petasites, Pyrola, Salix, Vaccinium, Valariana). The Pinnel Mountain trail and Wickersham Dome are more open, without any trees on the tops of the domes, and the terrain is covered by the dwarf scrub described above. In 2000 I was the only person in the field collecting butterflies. In 2001 and 2002, however, I had a field assistant so the number of butterflies caught reflects the efforts of two people. During these two field seasons I would often drop my assistant off at one dome and then I would travel to another dome. We were thus sampling two sites per day, each with the effort of a single person. In 2003 I had two field assistants, but this year we all sampled a single site together. We spread out and were able to sample each site more extensively. During each field season, we began searching for butterflies by May 25 and continued searching for flying adults until early [uly. All butterflies that were caught were marked and numbered (see Carter & Feeny 1985) and during the 2001, 2002 and 2003 field seasons the time of day that the butterflies were caught was also recorded. Females were kept for experiments. Most males were released at the end of the day although some were kept overnight so that we could mate them with the females. The males that were kept were released within a day or two and always at the same field site. Sampling effort varied by site; the sites that were closer to Fairbanks (Ester Dome and Murphy Dome) were sampled more frequently than the sites that were more distant (Pinnel Mountain and Wickersham Dome). Ester Dome was sampled a total of 25 days (5 days in 2000, 6 days in 2001, 8 days in 2002 and 6 days in 2003). Murphy Dome was sampled a total of 18 days (4 days in 2000, 4 days in 2001, 6 days in 2002 and 4 days in 2003). Pinnel Mountain was sampled a total of 6 days (3 days in 2000, 2 days in 2001 and 1 day in 2003). Wickersham Dome was sampled a total of 1 1 days (4 days in 2000, 4 days in 2001, 1 day in 2002 and 2 days in Table 1. Number of male and female P in. aliaska individuals collected at each field site during each year of the study. Site Year Ester Dome Murphy Dome Pinnel Mtn Wickersham Dome Totals Females 2000 1 2 4 7 2001 4 2 3 1 10 2002 2003 6 2 1 5 2 12 Totals 11 5 5 10 31 Males 2000 14 62 26 23 125 2001 26 22 27 49 124 2002 53 71 34 158 2003 32 76 7 47 162 Totals 125 231 60 153 569 82 Journal of the Lepidopterists’ Society 2003). The sites were visited more frequently than the number of days given above, but only days in which butterflies were actually caught are counted in the tallies. Results and Discussion Males were plentiful and easy to find and catch. During the four years of data presented here, we caught 569 males (Table 1). Males were often observed circling an object (a bush, rock or piece of debris) as well as other males that approached. Females were more difficult to find. Over the four years of data presented here, we collected only 31 females (Table l). I do not think that this reflects a skewed sex ratio as I have reared the progeny of both wild-caught and lab-reared females and the sex ratio of their offspring has never been significantly different from 50:50 (S. Murphy, unpublished data). Rather, the difference in the number of males and females caught probably represents a difference in their behaviors; indeed female rarity is common at hilltop sites in other hilltopping butterflies (Shields 1967). My observations of how rare females are on the hilltops is consistent with the notion that females only stay on top of the domes long enough to mate and then they fly downhill towards larval host plant sites. Once my field assistants and I had caught all of the males that were present on a dome upon our arrival, new males were observed flving up from lower elevations and they began to occupy the newly unoccupied perches. Given the difficulty in accessing some of the field sites, we were not able to sample every site eveiy day. Hence, any females that arrived on days that we were not present were able to mate and fly away without our having ever encountered them. However, any males that arrived at a site on a day that we were not Time of Day (Hour in Military Time) Fig. 1. Female (black bars) and male (gray bars) Papilio machaon aliaska butterflies that were caught during each hour of the day given as a percentage of the total caught. The data is given for all four field sites combined, but only for butterflies caught during the 2001-2003 field seasons (Females n=24; Males n=444). 80 60 40 20 11* a [| ILUIJl 13 17 Day in June 25 29 Fig. 2. Number of female (black bars) and male (gray bars) Papilio machaon aliaska butterflies that were caught during the month of June for all years (2000-2003). present were likely caught the next time we visited that site given their propensity to remain on the hilltops. For these reasons, I feel that my data are a rather accurate representation of the number of males that were present at these dome sites, but that the number of females has been significantly underestimated simply because their behavior makes them more difficult to catch when we could not be present at eveiy dome site everyday. I was able to find both males and females at each of the four sites tlescribed in the methods section above (Table 1). Males tend to be caught earlier in the day than females (Fig. 1), although none was ever caught before 9:00 hr. Their peak flight time was between 1 1:00-12:00 hr while the majority of females was caught slightly later, between 13:00-14:00 hr, but these two distributions for flight time did not differ significantly (P > 0.1, Wilcoxon signed-rank test on ranks). Females were never caught before 10:00 hr. Females also were caught a few days later than the first males were caught (Fig. 2). The earliest females were caught on June 9 while the latest females were caught on June 24. The earliest male was caught on June 1 while the latest male was caught on June 26. Nearly 17% of the males (n=96) were recaught at the same field site during a subsequent visit (Fig. 3); males were never found to have traveled between sites, which is not surprising given the significant distances between them. The majority of these males were only recaught once, but a few were caught several times. One male was recaught six times in the same location on a dome, which I interpret to mean that he was occupying the same perch or territory for several days. Most males were recaught within four days of their initial catch date. A few, however, were caught many days later. This gives us some insight as to how long males can live in the field. For instance, at least one male lived a minimum of 18 days in the field. Volume 62, Number 2 S3 100 80 s 60 s 40 20 0 1 A r i i i , — , — — 2 3 4 5 6 Number of Times Recaught Number of Days Before Last Recatch Fig. 3. Many male Popilio machaon aliaska butterflies (n=96) were caught, marked and released and then re-caught during this study (2000-2003). Of these males that were re- caught, many were caught multiple times. A) The number of times that the marked males were re-caught. B) The number of days that passed between the first time a male butterfly was caught and the last time he was caught. Although not directly tested, my observations are consistent with the idea that males tend to establish territories at the top of the dome that they then occupy, as evidenced by the number of males that were recaught on the domes along with personal observations of their behavior before they were caught. Males tend to emerge earlier in the season than do females and also fly earlier in the day than females. Finally, while males are commonly found on the hilltops, females are rarer. Together, these data support the claim that P. in. aliaska is a hilltopping swallowtail butterfly. Acknowledgements I thank L. Hough, S. Rothman, R Bennett and ]. Goodman for assistance in the field and P. Feeny as well as three anony- mous reviewers for helpful comments that greatly improved this manuscript. This work was supported by the Andrew W. Mellon Foundation, American Museum of Natural History Theodore Roosevelt Memorial Fund, Edna Bailey Sussman Fund, Ex- plorer's Club Exploration Fund, Sigma Xi, Cornell University Department of Ecology and Evolutionary Biology, a NSF Doc- toral Dissertation Improvement Grant awarded to S.M.M. (DEB-0104560) and a NSF research grant awarded to P. Feeny (IBN-9986250). Literature Cited Alcock, ]. 1985. Hilltopping in the nymphalid butterfly Chlosyne Cal- iforniai (Lepidoptera). American Midland Naturalist 113: 69-75. Baughman, }. F., & D. D. Murphy. 1988. What constitutes a hill to a hilltopping butterfly? American Midland Naturalist 120: 441-443. Carter, M., & P. Feeny. 1985. Techniques for maintaining a culture of the black swallowtail butterfly, Papilio polyxenes asterius Stoll (Papilionidae). Journal of the Lepidopterists1 Society 39 : 125-133. Feeny, R, L. Rosenberry. & M. Carter. 1983. Chemical aspects of oviposition behavior in butterflies. Pp. 27-76, In S. Ahmad (ed.). Herbivorous insects: host-seeking behavior and mechanisms. Ac- ademic Press, New York. Lederhouse, R. C. 1982. Territorial defense and lek behavior of the black swallowtail butterfly Papilio polyxenes. Behavioral Ecology and Sociobiolog)' 10: 109-118. Murphy, S. M. 2004. Enemy-free space maintains swallowtail butter- fly host shift. Proceedings of the National Academy of Sciences 101: 18048-18052. . 2007a. The effect of host plant on larval survivorship of the Alaskan swallowtail butterfly ( Papilio machaon aliaska). Ento- mologia Experimentalis et Applicata 122: 109-115. . 2007b. Inconsistent use of host plants by the Alaskan swallowtail butterfly: Adult preference experiments suggest labile oviposi- tion strategy. Ecological Entomology 32 : 143-152. & P Feeny. 2006. Chemical facilitation of a naturally occurring host shift by Papilio machaon butterflies (Papilionidae). Ecologi- cal Monographs 76: 399—114. Pe'er, G., D. Saltz, H. II. Thulke, & U. Motro. 2004. Response to topography in a hilltopping butterfly and implications for model- ling nonrandom dispersal. Animal Behavior 68: 825-839. Rutowski, R. L. 1991. The evolution of male mate-locating behavior in butterflies. American Naturalist 138: 1121-1139. Scott, |. A. 1968. Hilltopping as a mating mechanism to aid the sur- vival of low density species. Journal of Research on the Lepi- doptera 7: 191-204. . 1974. Mate-locating behavior of butterflies. American Midland Naturalist 91: 103-117. . 1986. The butterflies of North America: a natural history and field guide. Stanford University Press, Palo Alto, CA. Shields, O. 1967. Hilltopping — an ecological study of summit con- gregation behavior of butterflies on a southern California hill. Journal of Research on the Lepidoptera 6: 69-178. Sperling, F. A. H. 1987. Evolution of the Papilio machaon species group in western Canada (Lepidoptera: Papilionidae). Quaes- tiones Entomologicae 23: 198-315. Thompson, J. N. 1988. Variation in preference and specificity in monophagous and oligophagous swallowtail butterflies. Evolution 42: 118-128. - — 1995. The origins of host shifts in swallowtail butterflies versus other insects. Pp. 195-203. In J. M. Scriber, Y. Tsubaki, & R. C. Lederhouse (eds.). Swallowtail butterflies: their ecology and evo- lutionary biology. Scientific Publishers, Gainesville, FL. WlKLUND, C. 1981. Generalist vs. specialist oviposition behaviour in Papilio machaon (Lepidoptera) and functional aspects on the hi- erarchy of oviposition preferences. Oikos 36: 163-170. Received for publication 29 May 2007; revised and accepted 25 March 2008. 84 Journal of the Lepidopterists’ Society Journal of the Lepidopterists’ Society 62(2), 2008, 84-88 DIFFERENTIAL ANTENNAL SENSITIVITIES OF THE GENERALIST RUTTERFLIES PAPILIO GLAUCUS AND P. CANADENSIS TO HOST PLANT AND NON-HOST PLANT EXTRACTS R. J. Mercader Department of Entomology, Michigan State University, East Lansing, MI 48824, USA; email: mercade2@msu.edu. L. L. Stelinski Entomology and Nematology Department, University of Florida, Citrus Research and Education Center, 700 Experiment Station Road, Lake Alfred, FL 33850, USA, AND J. M. SCRIRER Department of Entomology, Michigan State University, East Lansing, MI 48824, USA. ABSTRACT. It is likely that olfaction is used by some generalist insect species as a pre-alighting cue to ameliorate the costs of foraging for suitable hosts. In which case, significantly higher antennal sensitivity would be expected to the volatiles of preferred over less or non-preferred host plants. To test this hypothesis, antennal sensitivity was measured by recording electroantennogram (EAG) responses from intact antennae of the generalists Papilio glaums L. and P. canadensis R & J (Papilionidae) to methanolic leaf extracts of primary, secondary, and non-host plants. EAGs recorded from antennae of P. glaucus were approximately four fold higher than those of P. canadensis in response to extracts of its most suitable host plant, Liriodendron tulipifera (Magnoliaceae). Likewise, EAG responses of P. canandensis to its preferred host, Populus tremu- loides (Salicaceae), were significantly higher than those of P glaucus, In addition, P. glaucus exhibited significantly higher (approximately three fold) EAG responses to its preferred host, L. tulipifera, than to its less-preferred hosts, Ptelea trifoliate. Sassafras alhidum, and Lindera ben- zoin. The results from this study indicate a significant divergence in the olfactory system of two closely related generalist butterfly species, in- cluding a strong specialization in the olfactory system of P. glaucus. Additional key words: Electroantennogram, olfaction, opposition. For insects with larvae that develop on a single host plant, female ovipositional choice determines larval habitat and therefore the likelihood of larval survival. However, a clear correlation between adult ovipositional preference and host suitability for larval growth has not been found in many systems (reviewed in Mayhew 1997), and ‘mistakes’ in which eggs are laid on plants toxic to the larvae are fairly common (Straatman 1962; Wiklund 1975; Chew 1977; Berenbaum 1981; Larsson & Ekbom 1995; Renwick 2002; Graves & Shapiro 2003). Such 'mistakes’ are believed to be rare for phytoehemically specialized species, but generalists, such as Papilio glaucus L. and P. canadensis R & J (Papilionidae), are known to regularly place a small fraction of their eggs on hosts toxic to their larvae in natural habitats (Brower 1958, 1959) and in controlled environments despite the presence of a suitable alternative (Scriber et al. 1991; Scriber 1993). In contrast, specialist herbivores may fail to oviposit on readily available suitable hosts; for example, Papilio palamedes’ geographic range is determined by female ovipositional preference and not the availability of hosts suitable for larval development (Lederhouse et al. 1992). One hypothesis that has been proposed to explain this observation and the higher abundance of specialist insects is that an increase in error rate should decision-making, host selection. be associated with an increase in polyphagy (Levins & MacArthur 1969). In recent years this idea has been updated in terms of neural limitations to include a prolonged decision-making time along with an increased error rate as costs of polyphagy (Bernays 2001; Janz 2003). Despite the common assertion that olfaction is an important sensory modality for orientation to host plants (Renwick & Chew 1994; Dicke 2000; Finch & Collier 2000), the importance of olfactory cues for oviposition- site location in day-flying butterflies has received relatively little attention compared with moths (reviewed in Hansson 1995; Honda 1995, but see Feeny et al. 1989; van Loon et al. 1992; Baur & Feeny 1995; Kroutov et al. 1999). In addition, the role of olfactory cues in butterfly host plant searching and acceptance behavior has received little attention relative to visual and/or contact cues (e.g. Rausher 1978; Stanton 1982; Scherer & Kolb 1987; Grossmueller & Lederhouse 1985; Thompson & Pellmyr 1991; Honda 1995; Weiss 1997; Sehoonhoven et al. 1998). Olfactory cues may play an important role in long and short-range searching behavior of pre-alighting generalist butterflies increasing their efficiency. Baur & Feeny (1995) found electroantennogram (EAG) evidence for evolutionary lability in the peripheral Volume 62, Number 2 85 olfactory system of three specialist butterflies, Papilio polyxenes, P. machaon Hippocrates , and P. troilus. If the peripheral olfactory system is labile, it may allow for adaptations in generalist species that allow for a functionally specialized behavior in areas where a primary host is abundant, while maintaining the flexibility to accept alternate hosts in areas where the primary host(s) are rare or not present. If olfactory cues are used to reduce decision-making time in generalist species, significantly higher sensitivity would be expected in the peripheral nervous system to primary hosts over less preferred hosts. We tested this hypothesis for the polyphagous P. glaucus by comparing its antennal sensitivity by EAG recordings with that ol its sibling species, P. canadensis , to extracts of preferred, secondary, and non-host plants. These two sister species can readily produce fertile hybrid offspring (e.g. Scriber 1998); P. canadensis males prefer P. glaucus females (Deering & Scriber 2002), and until recently, they were considered the same species (Hagen et al. 1991). However, despite their similarities they exhibit significant differences in host plant use. In particular, tulip tree, Liriodendron tulipifera (Magnoliaceae), the preferred host of P. glaucus , is toxic to P. canadensis larvae, while quaking aspen, Populus trenudoides (Salicaeeae), the preferred host of P. canadensis , is toxic to P. glaucus. For each species, antennal sensitivity was measured by recording EAG responses to plant extracts of four hosts and one non- host of P. glaucus , which included tulip tree and quaking aspen. Materials and Methods Insect source. Butterflies used in EAG studies were reared from eggs laid by wild-caught females on their natural host plants. P. canadensis females were collected from the first flight in the Battenkill River Valley area at the New York/Vermont border, U.S.A. and the larvae were reared to pupae in the field on sleeved tree branches of black cherry, Primus serotina (Rosaceae). P. glaucus females were collected in Lancaster Co. in southeastern Pennsylvania, U.S.A. and were also field- reared on black cherry. After eclosion, butterflies were fed a honey-water solution and stored at 4° C for a maximum of 6 days until they were tested. By using adults that had not encountered any hosts prior to our assays and were reared on the same common host we prevented any influence due to adult or larval induction of preference (reviewed in Mereader & Scriber 2005). Plant extracts. Leaves of tulip tree, L. tulipifera (Magnoliaceae), quaking aspen, P. tremuloides (Salicaeeae), hop tree, Ptelea trifoliata (Rutaeeae), sassafras. Sassafras albidum (Lauraceae), and spieebush, Lindera benzoin (Lauraceae) were collected from trees growing in Ingham Co. Michigan, U.S.A. in areas known to be pesticide free. For simplicity, hereafter hosts will be referred to by their common names. The detailed protocol for preparing plant extracts was described by Gok 98 % pure) dissolved in hexane (Aldrich) as a positive control. Hexanal was used as a standard positive control given that synthetic green leaf volatiles are known to elicit EAG responses in Papilio species (Bauer & Feeny 1995). Two milligrams of each plant extract, hexanal solution, and methanol or hexane solvents alone (20 pL total solution) were pipetted onto 1.4 x 0.5 cm strips of Whatman No. 1 filter paper. These were aged for 5 min in a fume hood to allow for solvent evaporation. Subsequently, strips treated with extract or volatile treatments were inserted into glass Pasteur pipettes. EAG measurements were recorded as the maximum amplitude of depolarization elicited by 1 mL puffs of air through EAG-cartridges directed over antennae ol live butterfly preparations. The time interval to expel 1 mL of stimulus odor or clean air was ca. 120 ms (Stelinski et al. 2003). Plant-extract or chemical stimuli were delivered through one arm of a glass Y-tube (each arm 2 cm in length, base 1 cm long, and 0.5 cm diameter) positioned approximately 5 mm from the antenna as carbon-filtered and humidified air was delivered at 50 mL/min into the second arm and onto the preparation via Tygon tubing. Male and female butterflies of each species and sex were 2-6 d post-eclosion when used for EAG assays. Butterflies were mounted on 5.0 cm diameter plastic Petri dishes with a clay strip (30 x 5 mm) placed over their wings and thorax. EAG recordings were conducted by removing the terminal tip of the club (< 0.5 mm) of the antenna used for recording with fine scissors, and the recording electrode was positioned directly over the severed end. The reference electrode was inserted into the head near the base of the antenna. EAGs were performed ca. 30 s following mounting of butterflies and terminated at most 2 min later. For each plant extract 86 Journal of the Leridopterists’ Society tested, EAGs were recorded from 8-10 insects of each sex and species. Plant-extract stimulations were presented to individual butterflies in random order, and control stimulations (filter paper impregnated with 20 pL of hexane or methanol) were delivered prior to each plant-extract stimulus presentation. Statistical analyses. Between-speeies, pairwise comparisons of EAG responses were performed separately for tulip tree and quaking aspen on female responses using Mann-Whitney U tests with a Bonferroni corrected significance level of a < 0.05. Within species, EAG responses for P. canadensis were log transformed and P. glaucus responses were square- root transformed to normalize the distributions and homogenize variance. Data were analyzed as repeated measures analysis of variance with individual butterfly as the subject, using Proc Mixed in the SAS System (SAS Institute 2000). The model included odor stimulus and sex as explanatory variables. Pair mean separations were performed for P. canadensis and P. glaucus using Tukey’s multiple comparisons test. Results EAG between species comparisons. There were significant differences between EAG responses of P. canadensis (y2 = 15.6, df = 2, P < 0.001) and P. glaucus 1 n >0.8 £ CD if) §0.6 if> CD l— ID 20-4 c 03 'td 4; 0.2 -| • P. glaucus BP. canadensis 1 ) * * - 1 1 B “1 1 1 < » 1 TT QA QA Leaf Fig. 1. Median EAG responses of P. glaucus and P. canadensis females to the extracts of tulip tree TT ( Liriodendron tulipifera) and quaking aspen QA ( Populus tremuloides) . Bars around the medians represent the inter-quartile ranges ( P. canadensis inter- quartile range for tulip tree is smaller than size of square). Pair- wise differences were analyzed for each extract using the Mann- Whitney U tests. Values within extract with an “ had a significant difference at Bonferroni corrected a < 0.05. (y2 = 1 1.3, df = 2, P = 0.003) to the extracts of tulip tree and quaking aspen (Fig. 1). The magnitude of EAG responses of P. glaucus was significantly greater to extracts of tulip tree than those of P. canadensis. In contrast, the magnitude of EAGs elicited by quaking aspen extracts was significantly higher for P. canadensis than P. glaucus. EAG within species comparisons. Within-species odor stimuli had a significant effect for P. glaucus (F = 50.1, df=6,102, P< 0.0001), and P canadensis (F = 24.67, df = 6,108, P< 0.0001). There was no significant sex-by-odor stimulus interaction for P. glaucus or P. canadensis ; therefore, male and female responses were combined for analysis of pair-wise differences (Tables I Table 1. Mean EAG responses ± SE of male and female P. glaucus. Data for males and females were combined for analy- sis given that there was no significant sex by odor stimulus in- teraction. Mean ± SE EAG antennal response (mV) to plant extracts Odor sources Males N Females N P < 0.05 Methanol 0.07 ± 0.01 8 0.09 ±0.02 10 C° Hexanal 0.70 ± 0.06 8 0.60 ±0.09 10 a Tulip Tree 0.85 ± 0.07 8 0.87 ± 0.07 10 a Quaking Aspen 0.26 ± 0.07 8 0.19 ±0.04 10 b Sassafras 0.23 ± 0.05 8 0.22 ± 0.04 10 b Spicebush 0.23 ± 0.04 8 0.30 ± 0.04 10 b I lop Tree 0.25 ± 0.04 8 0.28 ± 0.07 10 b “Significant differences in antennal responses to odorant stimuli are indicated by different lowercase letters (P < 0.05, Tukey’s 1ISD). Table 2. Mean EAG responses ± SE of male and female P. canadensis . Data for males and females were combined for analysis given that there was no significant sex by odor stimulus interaction. Mean ± SE EAG antennal response (mV) to plant extracts Odor sources Males N Females N P < 0.05 Methanol 0.14 ± 0.03 10 0.09 ± 0.02 9 d“ Hexanal 0.57 ± 0.07 10 0.60 ±0.09 9 a Tulip Tree 0.21 ± 0.03 10 0.24 ± 0.03 9 c Quaking Aspen 0.43 ± 0.07 10 0.37 ± 0.05 9 a Sassafras 0.30 ± 0.06 10 0.46 ± 0.09 9 abc Spicebush 0.35 ± 0.05 10 0.41 ± 0.04 9 ab Hop Tree 0.28 ± 0.05 10 0.29 ± 0.06 9 be “Significant differences in antennal responses to odorant stimuli are indicated by different lowercase letters (P < 0.05, Tukey’s HSD). Volume 62, Number 2 87 and 2). P. glaucus exhibited higher EAG responses to its preferred host, tulip tree, than to less-preferred hosts, hop tree, sassafras, and spicebush, and the non-host quaking aspen (Table 1). Responses to tulip tree were similar to those elicited by the hexanal positive control (Table 1). Likewise, P. canadensis exhibited a significantly higher EAG response to its preferred host, quaking aspen, than to hop tree or tulip tree (Table 2). Once again, responses to the preferred host were not different from those elicited by the synthetic standard (Table 2). However, EAGs elicited by two of the non- hosts, sassafras and spicebush, were not significantly different from those elicited by quaking aspen for P. canadensis. There was no difference between responses to methanol versus hexane solvents alone; hence, data are not shown for the latter negative control. Discussion Antennal sensitivity of P. glaucus was approximately three-fold higher to extracts of the preferred host, tulip tree, than to any other extract tested (Table 1). Conversely, tulip tree extract elicited a weaker antennal response from P. canadensis than the others tested (Table 2). Furthermore, EAGs recorded from P. canadensis to extracts of this species’ preferred host plant, quaking aspen, were greater than those from P. glaucus (Fig. 1). These results agree with the prediction that peripheral sensitivity to primary hosts should be greater than to less preferred hosts in generalist butterfly species if olfactory cues play a role in host finding behavior. It is notable that species-specific responses were recorded to preferred host plants despite the use of extracts of dried leaves in the current study, which may have limited our assay to higher molecular weight volatiles. This suggests that these butterfly species may use host-plant volatiles, at least as short-range cues, while foraging for suitable host plants, which agrees with field observations of P. glaucus females hovering, but not landing, on non-hosts while searching for oviposition sites (R. }. M. personal observations). Although P. glaucus is a highly polyphagous swallowtail species, females exhibit a distinct ovipositional preference for tulip tree throughout their range (Scriber et al. 1991; Mercader & Scriber 2005), even in populations where this host plant does not occur (Bossart & Scriber 1995). Congruently, antennal responses to tulip tree were approximately three times greater than to another major host (hop tree), two secondary hosts (sassafras and spicebush), and a non- host (quaking aspen). Although the EAG technique cannot distinguish between attractive versus deterrent olfactory stimuli, the heightened antennal sensitivities recorded in this study corresponded well with known host plant preferences of both species. It is important to note that although greater EAG responses were observed for females of P. canadensis for its most common host quaking aspen than all other hosts tested, these were not significantly different than those for the marginal host sassafras and non-host spicebush (Table 2). This lower specificity in P. canadensis relative to P. glaucus is not unique to the olfactory system. In ovipositional arenas that primarily test contact chemoreception, P. canadensis females have a significantly lower specificity than P. glaucus , including a high acceptance rate for the non-host tulip tree (Scriber et al. 1991; Mercader & Scriber 2007). This lower specificity in P. canadensis is likely to be due to the absence of plants in the Lauraceae (e.g. sassafras and spicebush), Magnoliaceae (e.g. tulip tree), and Rutaeeae (e.g. hop tree) where P. canadensis occurs, greatly reducing the selection pressure for higher specificity. Interestingly the divergence in antennal sensitivity between P. glaucus and P. canadensis was observed in both males and females (Tables 1 and 2). As males do not oviposit and these species do not mate on host plants, divergence in sensitivity to host plant odors does not have any clear advantage for the males of these two species. This similarity in the antennal sensitivities of males and females in both species may reflect a developmental similarity between males and females (with no adaptive function in males) or serve an unknown function. Heightened antennal sensitivity of P. glaucus to tulip tree relative to the other host extracts tested lends support to the hypothesis that olfactory cues may be used to reduce decision-making time during host-plant selection in this species. Pre-alighting cues are more likely involved in maximizing rates of oviposition than in host acceptance behavior (Thompson & Pellmyr 1991); therefore, it is likely that olfactory cues may be used to maximize P. glaucus' rate of landing on tulip tree wherever this preferred host is present. Furthermore, the higher sensitivity of P. canadensis to odors of quaking aspen relative to the other less-preferred plant species evaluated here adds further support to the hypothesis that host-plant location may be, in part, mediated by chemical signals in these two generalist, sister butterfly species. Further laboratory and field behavioral assays will need to be conducted to confirm this hypothesis. Acknowledgements We would like to thank William Iloutz and Howard Romack for providing adult butterflies and/or pupae. Earlier drafts of the 88 Journal of the Lepidopterists’ Society manuscript were significantly improved by comments from Ce- sar Rodriguez-Saona and Rufus Isaacs. 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Pellmyr. 1991. Evolution of oviposition be- havior and host preference in Lepidoptera. Ann. Rev. Entomol. 36: 65-89. vanLoon, J. J. A., W. H. Frentz & F. A. Vaneeuwijk. 1992. Elec- troantennogram responses to plant volatiles in two species of Pieris butterflies. Entomol. Exp. Appl. 62: 253-260. Weiss, M. R. 1997. Innate colour preferences and flexible colour learning in the pipevine swallowtail. Anim. Behav. 53: 1043-1052. Wiklund, C. 1975. The evolutionary relationship between adult oviposition preferences and larval host plant range in Papilio machaon L. Oecologia 18: 185-197. Received for publication 3 January 2007; revised and accepted 5 December 2007. Volume 62, Number 2 89 Journal of the Lepidopterists’ Society 62(2), 2008, 89-98 DESCRIPTION OF THE IMMATURE STAGES OF METHONA CONFUSA CONFUSA BUTLER, 1873 AND METHONA CURVIFASCIA WEYMER, 1883 (NYMPHALIDAE, ITHOMIINAE) FROM EASTERN ECUADOR Ryan I. Hill Department of Integrative Biology, 3060 Valley Life Sciences Building, University of California, Berkeley, CA 94720; email: riliill@berkeley.edu AND Luis A. Tipan La Selva Jungle Lodge, Mariana de Jesus E7-211 y Pradera, Quito, Ecuador ABSTRACT. Hei •e we describe the complete life history for Methona confusa confusa Butler, 1873 and Methona curvifascia Weymer, 1883 from eastern Ecuador. Each stage from egg to pupa is described and illustrated. Descriptions of first instar chaetotaxy and instar durations are also reported. Both species were found feeding on Brunfelsia grandiflora schultesii Plowman. Mature M. confusa larvae have 12 transverse bands that are all yellow in color, including one on segment A9 as observed for M. megisto and M. themisto. In contrast, M. curvifascia lacks a transverse band on segment A9, having 11 transverse bands in total that are white in middle segments and orange in anterior and posterior seg- ments. The pupa of M. confusa and M. curvifascia differs in the arrangement of spots on tire thorax dorsum. Additional key words: Brunfelsia grandiflora, chaetotaxy, egg clustering, Solanaceae. Butterflies in the genus Methona Doubleday, 1847 (Ithomiinae) are large, warningly colored butterflies illustrated in the original descriptions of both Batesian (M. confusa) and Mullerian (M. megisto) mimicry (Bates 1862; Muller 1879). Despite being involved in the conception of a theory that has generated a massive publication record, Methona biology is relatively poorly understood. The genus Methona is distributed across much of South America east of the Andes reaching its southern limit in southern Brazil, extreme northern Argentina and Uruguay (Forbes 1943; Mielke & Brown 1979; G. Lamas pers. comm.). In addition. Lamas (2004) indicates that two new subspecies of M. confusa are present in Panama. Host records have been published for four of the seven recognized species and Methona are apparently monophagous on the Solanaceae genus Brunfelsia (Brown 1987; Drummond 1976, 1986; Drummond & Brown 1987). However, only three species have any published information on immature stage morphology (Brown 1987; Brown & Freitas 1994; Drummond 1976; Motta 2003; Willmott & Freitas 2006), and a complete description of the immature stages has not been published lor any species in the genus. Here we report on the immature stages of two species of Methona from the upper Amazon basin in eastern Ecuador, Methona confusa confusa Butler, 1873 and M. curvifascia Weymer, 1883. Both of these sj^ecies are residents of the Amazon basin, however M. confusa is distributed more broadly, occurring throughout the whole basin (and including the populations in Panama mentioned above), and M. curvifascia is restricted to western Amazonia (G. Lamas jrers. comm.). We describe all early stages, report instar durations and provide detailed description of first instar chaetotaxy and briefly discuss differences in larval color pattern in the genus. Materials and Methods Observations were made from January to February 2007 in Provincia Sueumbios, Ecuador, in the forests surrounding Garzaeoeha (00°29.87’S, 76°22.45’W) and Challuacocha (00°26.29’S, 76°16.81’W). Early stages were reared in plastic cups and plastic bags under ambient conditions (22-30° C, 70-100% relative humidity) in a wood building with screen windows. Larvae were moved daily to a shaded environment under a nearby building to maintain ambient conditions. Observations were recorded daily and head capsules and pupal exuviae were collected. Larval specimens were boiled and subsequently stored and studied in 70% ethanol. Vouchers are deposited in the Essig Museum of Entomology at UC Berkeley. Descriptions other than first instar chaetotaxy are based on several individuals from a single clutch of eggs for M. confusa , and more than 10 individuals for M. curvifascia . First instar chaetotaxy follows nomenclature of Motta (2003), and Hinton (1946), Kitching (1984) and Peterson (1962) were also consulted. The number of specimens for which first instar chaetotaxy was examined is listed in Appendix 1. Host plant vouchers are deposited in the University and Jepson Herbaria at UC Berkeley (voucher number RIH-1424, UC accession # 90 Journal of the Lepidopterists’ Society UC1933451) and Herbario Nacional de Ecuador (voucher number RH01-1 17). Results Methona confusa confusa Butler, 1873 Hostplant. Brunfelsia grandiflora schultesii Plowman (Solanaceae), known locally as chiricaspi. The group of M. confusa eggs was found on an individual plant that also hosted eggs of M. curvifascia. Opposition. Not observed. Eggs occur in large clusters on the underside of fresh but mature sized leaves. One cluster of 46 un- hatched eggs (at 1.5 m) and a cluster of 18 hatched eggs were found. Plants with eggs were ~2m tall and located in shaded areas at gap edges. Egg. Figure 1A. Duration: Unknown. Eggs hatched four days after found in the field. Egg is white, adorned with 9-11 horizontal and 19-22 vertical ridges making many small rounded cells. Mean egg height = 1.23 mm (s.d. = 0.04, n = 3). Mean egg width = 0.98 mm (s.d. 0.03, n = 3). Mean axes ratio (height/width) = 1.26 (s.d. = 0.01, n = 3). 1st instar. Figure IB & C. Duration: 3 to 7 days. Mean head capsule width = 0.77 mm (s.d. = 0.02, n = 10). Head capsule and thoracic legs are black. Proleg shields are large and black. Anal plate is present and shiny black. Body is covered with short pale setae. Body is widest near the head and tapering posteriorly. Body is dark olive green with paler olive transverse bands. Body has pale transverse bands with slightly raised ridges within, ridge on A1 & A2 most pronounced. Larvae eat channels into the leaf from the margin consuming all layers of the leaf. See Appendix 1 for description of first instar chaetotaxy. An additional lateral body seta (Figure 2) was observed on the meso- and metathorax of the two larvae studied compared with the ithomiines studied by Motta (2003), including Methona themisto. This seta is assigned to the lateral group in descriptions (Appendix 1) because this keeps other setae consistent with adjacent segments and the lateral group has a third seta in some moth families (Hinton 1946). Thus, the top seta is inferred to be LI with the middle L2 and most ventral L3 (Fig. 2). Descriptions of characters involving setae LI and L2 on these segments should be treated with caution, as homology of LI and L2 may not have been correctly inferred. 2nd instar. Figure ID. Duration: 4 to 6 days. Mean head capsule width = 1.18 mm (s.d. = 0.04, n = 10). Like the previous instar with the following observations: body is brown and transverse bands are dirty white with tints of yellow. Segments T1-A9 have a transverse band making 12 bands total. The transverse band on segments A3-A6 leans slightly to the posterior. The transverse pale band is located in the posterior of each segment except Tl, which is pale anteriorly and almost entirely pale. Transverse ridges are more pronounced this instar. 3rd instar. Figure IE. Duration: 3 to 4 days. Mean head capsule width = 1.69 mm (s.d. = 0.04, n = 5). Like previous instar with the following observations: body a rich dark brown and transverse bands dirty white first day turning yellow subsequently. Rest on underside of leaf, sometimes with body straight, sometimes curled in a “J” (Fig. 1H). 4th instar. Figure IF. Duration: 4 to 7 days. Mean head capsule width = 2.50 mm (s.d. = 0.07, n = 5). Like previous instar with the following observations: transverse bands are yellow and slanting slightly toward the posterior. Transverse bands on A3-6 extend farthest ventrally and are not as pointed at their terminus. Transverse band on A9 is smaller than others, extending the shortest distance ventrally. Laterally, rounded protuberances form a fleshy shelf. Transverse ridges run across this shelf ending below it. The transverse ridges are generally located in the anterior of the transverse yellow band on each segment. Spiracle on Tl is located at posterior margin of yellow band, all other spiracles are anterior of yellow band. Body is covered in short pale pubescence. 5th instar. Figure 1G & IT. Duration: 8 to 12 days. Mean head capsule width = 3.39 mm (s.d. = 0.20, n = 4). Like previous instar with the following changes and observations: body dark brown, appearing black in some individuals, with yellow transverse bands. Area of yellow transverse band posterior to ridge fades to whitish on segments A3-6. Yellow bands fade slightly laterally. The day before pupating the yellow fades in all bands. Pupa. Figure II, J & K. Duration: 12 days. Pupa is pendant and bent near abdomen tip but not at abdomen-thorax junction. Pupa colored yellow with distinct black marks. Dorsally with two rows of thin black marks that are thinnest near head. Last segment before cremaster has these dorsal marks merged into wide line. Cremaster is black. Spiracles are outlined in thick black marks. Wing pad has costal margin marked with black. Wing pad posterior margin along thorax marked with black that breaks up into dots near spiracles. Center of wing pad has broken black lines. Ventrally is an inverted black mushroom-shaped spot anterior of cremaster that surrounds a pair of black tubercles. Ventrally at edge of wing pad two black marks merge together. Ocular caps marked with black that starts near eye and extends ventrally as a thick line. Ventrally central black marks over legs and black marks near base of antennae. Thorax slightly keeled with three black marks: anterior spot elongate and thickest toward head, middle one elongate and forked, and posterior one an elongate spot that is widest in the middle (Fig. 1J). The extent of dark markings is variable with some individuals with heavier dark markings (Fig. 1 J ) . Eves darken one to two days before eclosing. The day before eclosing black and gold appear in wing pad, then wing pad turns black, followed by abdomen. Pupa has unpleasant odor, as does freshly eclosed adult. Methona curvifascia Weymer, 1883 Hostplant. Bninfelsip grandiflora schultesii Plowman (Solanaceae). Oviposition. A female was observed ovipositing on a relatively small host from 11:30-12:30. The plant was 1.25-1.5 m tall and located in tall secondary growth with bright light but shaded by a thin canopy of leaves. The female flew to host leaves, tapped the upper surface of the leaf, and then would land on these leaves, occasionally opening her wings and antennating the leaf. She did this repeatedly for 40 minutes. She then landed on a leaf at 0.5 m and hung at its edge, curled her abdomen under and laid a single egg on the underside of the leaf. She was obscured from view after laying this egg but remained close to the site where she laid for - 3 min. She then flew to a nearby bird dropping and fed from it. Three eggs were found where she laid the one observed, so she may have laid all three in a few minutes although only the one was observed. Two other eggs were found on this plant along with a freshly hatched first instar. Egg placement with respect to the leaf border appeared somewhat variable and not confined to the leaf border, with eggs sometimes being closer to the leaf midvein than leaf border. Leaves chosen for oviposition varied from small younger growth (3-5 cm in length) to fresh nearly full-sized leaves (8-10 cm). Other plants hosting eggs/larvae were ~2 m tall and found in shaded areas at the edge of primary forest gaps. Egg. Figure 3A. Duration: 6 days (n = 1). Mean egg height = 2.05 mm (s.d = 0.10, n = 3). Mean egg width = 1.30 mm (s.d. = 0.05, n = 3). Mean axes ratio (height/width) = 1.58 (s.d. = 0.07, n = 3). Egg is white, widest two-thirds the distance from base but only slightly wider there. Egg adorned with 14-17 horizontal and 26-30 vertical ridges. The horizontal and vertical ridges make rounded cells that merge near the apex. Head capsule is visible at egg apex one day before hatching. 1st instar. Figure 3B. Duration: 3 days (n = 2) to 4 days (n = 2). Mean head capsule width = 0.90 mm (s.d. = 0.03, n = 2). When first hatched body is dark grey with paler grey transverse bands in anterior of Tl and posterior of segments T2-A8 making 11 pale bands in total. Within each pale band is a raised transverse ridge that crosses the dorsum. Body covered in short pale setae. Head capsule black and thoracic legs are black. Proleg shields large and black. Black sclerotized anal plate present. Second day and beyond, body dark brown with white to dirty white transverse bands (Fig. 3B). Transverse bands widest dorsally, more narrow laterally. Transverse band on Tl Volume 62, Number 2 91 Fig. 1. Methona confusa immature stages. A. Egg clutch. B. First instar, first day with very pale bands, feeding from leaf mar- gin. C. First instar > 1 d old with pale bands. D. Second instars, midmolt first instars and feeding damage. E. Third instar. F. Fourth instar. G. Fifth instar. H. Fifth instar in resting position. I. Ventro-lateral view of pupa. J. Dorsal view of two pupae showing range of variation in black markings. K. Dorso-lateral view of pupa illustrating detail near cremaster. 92 Journal of the Lepidopterists’ Society wider than other segments dorsally. Larvae eat little, to more than three quarters egg when first hatched. Larvae feed at leaf margin making channels into side of leaf (Fig. 3C & D). See Appendix 1 for description of first instar chaetotaxy. 2nd instar. Figure 3C. Duration: 3 days (n = 2) to 4 days (n = 1). Mean head capsule width = 1.33 mm (s.d. = 0.03, n = 4). Like previous instar with the following observations: First day, white transverse band of T1 and T2 now with yellow tints. Second day and beyond, transverse band on T1 is yellow and white transverse band on T2 and A8 with yellow tints. Raised ridges more pronounced this instar, widr T1 less pronounced than other segments. On segments A3-6 the transverse white band bends fonvard laterally and ends just above tire proleg. Spiracles dark. Spiracle on T1 surrounded by yellow in posterior of transverse band. Spiracles in other segments located at anterior margin of bands. 3rd instar. Figure 3D. Duration: 3 days (n = 6) to 4 days (n = 1). Mean head capsule width = 2.00 mm (s.d. = 0.15, n = 8). Like previous instar with the following observations: Body is very dark brown, some individuals appearing matte black. Non-white bands are more orange-yellow this instar. First day, transverse band on T2 and A8 more strongly colored than previous instar, and orange-yellow like Tl, band on T3-A7 white. Second day and beyond, transverse band on segments T3, A1 (only some individuals), and A7 develops yellow tints. Transverse bands extend farthest toward venter on segments A3-6. 4th instar. Figure 3H. Duration: 5 days (n = 3), 6 days (n = 2), 7 days (n = 2). Mean head capsule width = 2.70 mm (s.d. = 0.06, n = 12). Like previous instar with the following observations: Transverse band on Tl, T2 and A8 is orange and band on segments T3, Al, A2 and A7 is tinted orange this instar. Transverse band on A7 is wider than other bands except for that on Tl. 5th instar. Figure 31. Duration: 9 days (n = 1), 11 days (n = 7), 12 days (n = 4), 13 days (n = 1). Mean head capsule width = 3.70 mm (s.d, = 0.19, n = 6). Like previous instar with the following observations: Head capsule narrows dorsally with two subtle humps and has short dark setae. Clypeus area is pale grey and frontal sutures pale colored. Body is very dark browm appearing matte black in some individuals. Thorax has additional wrinkles between ridges dorsally. Pale body pubescence more pronounced on ridges. Transverse band on Tl & 2 is orange turning white just above leg where it ends without tapering. Transverse band on segment T3 is white, tinted with orange dorsally and ends above leg without tapering. Transverse band on Al & A2 tinted orange dorsally, and is white where terminates ventro- laterally in narrow point (Al narrower point than A2). Segments A3-6 with white transverse band that bends slightly to posterior just before terminating on fleshy bulge above proleg. A7 with white band tinted orange and terminating in rounded point ventro-laterally. A8 band is orange, but not as bright as Tl & T2, and turns white before tapering to a point ventro-laterally. Orange coloration becomes more extensive and white bands on A3-6 darken two to three days before pupating. Larvae in all instars rest on underside of leaf with head down near where feeding. Larvae tend to feed first at distal end of leaf and subsequently toward leaf base in later instars. Larvae raise thorax off substrate or curl into tight “J” when disturbed. Pupa. Figure 3E, F, G & J. Duration: 11 days (n = 4) to 12 days (n = 1). Pupa is pendant and bent near abdomen tip but not at abdomen- thorax junction. Pupa is yellow and marked with distinct black spots. Black marks develop within a couple hours of pupating. Head and thorax are slightly darker yellow than abdomen and wing pad. Abdomen dorsum with two rows of black marks that become larger and more rounded toward abdomen apex, and merge into thick line on A10. Laterally, abdomen has seven black spots over spiracles that increase in size toward abdomen apex. Lateral abdomen marks not in a straight line, with marks on A3 & A4 at wing pad margin out of line with the others. Wing pad has three black lines near its center, black spots along its dorsal margin that become lines basally, and black lines along its ventral margin. Ocular caps colored black, widi black extending into a line ventrally. Black cremaster. Thorax dorsum has a pair of anterior black spots, a single medial spot near the anterior pair and another spot posteriorly. Venter has rough upside down “T” near the cremaster that surrounds two black tubercles. Some variation observed in extent of dark spots on the thorax (Fig. 3E & F). Eyes become dark one to two days before eclosion, followed by black and yellow visible in wing pad. Pupa becomes nearly black just before eclosing. Discussion Our observations provide several early stage characters useful for distinguishing Methona confusa and M. curvifascia at this site. Larval coloration differs between these two species with M. confusa exhibiting 12 transverse bands similarly yellow in color along the body, whereas M. curvifascia has 1 1 bands with those in the middle of the body white, and those at either end orange. The pupa of these two species can be distinguished by the black spots on the thorax dorsum. M. confusa s anterior spot consists of a single spot and its middle spot is “Y” shaped, whereas M. curvifascia’ s anterior spot is split into two small spots and its middle spot is round. Observed variation in the extent of black markings on the pupa is illustrated in Fig. 1} and Fig. 3E&F and does not appear to pose problems for identification using the aforementioned pupal characters. M. confusa eggs are laid in clusters and are shorter (p = 0.002, t = 13.1, n =3, see above descriptions for means) and narrower (p = 0.001, t = 10.7, n =3, see above descriptions for means) than M. curvifascia . M. confusa eggs are also relatively more rounded with a lower axes ratio than M. curvifascia (p = 0.013, t = 8.2, n = 3, see above descriptions for means). Fig. 2. Schematic of Methona confusa first instar chaetotaxy for meso- and nretathoracic segments illustrating additional lat- eral seta (L3). Arrangement of body setae on other segments for M. confusa otherwise resembles M. themisto (Figure 19.3 in Motta 2003) except for characters 92 and 93 which are de- scribed in Appendix 1. Volume 62, Number 2 93 Fig. 3. Methona curvifascia immature stages. A. Egg. B. First instar. C. Second instar on leaf showing feeding damage of young instars. D. Third instar. E. Dorsal view of freshly formed (< 1 d) pupa. F. Dorsal view of pupa exhibiting mature coloration. Note variation in black mark on thorax. G. Lateral view of pupa. H. Fourth instar showing feeding position and damage. I Fifth instar showing resting behavior. J. Ventral view of pupa. 94 Journal of the Lepidopterists’ Society Observations made here also allow comparison of larval morphology within and among Methona species. The M. confusa confusa larvae observed here are similar to the M. confusa psamathe larva figured in Brown (1987). Although it is difficult to see, the larva in Brown (1987) Figure 8X appears to have a transverse band on segment A9 making 12 transverse bands in total. The presence of a transverse band on A9 in M. confusa is a trait shared with M. megisto and M. themisto illustrated in Brown & Freitas (1994) and was identified as a synapomorphy of Methona in Willmott & Freitas (2006)(Table 2, character 49:1). However, M. curvifascia lacks the transverse band on A9 indicating that not all Methona have this character. M. curvifascia is placed as the basal Methona species in a molecular phylogenetic study (Hill unpublished) suggesting that absence of a transverse band on A9 is the plesiomorphic condition, and evolution of the extra band on A9 occurred after M. curvifascia diverged from the rest of the group. Methona curvifascia also maybe divergent in egg shape with a mean axes ratio observed here just outside of the range indicated for M. themisto (Brown & Freitas 1994) and significantly different than M. confusa as mentioned above. Aside from the characters just discussed, observations made here are congruent with most of the synapomorphies for Methona larvae listed in Table 2 of Willmott & Freitas (2006). The pupa of both M. confusa and M. curvifascia exhibit the sharp curve along the dorsum in the posterior half of the abdomen (character 55:1). The following characters, with their states indicated in parentheses, are also the same for M. confusa and M. curvifascia as listed for M. megisto and M. themisto in Willmott & Freitas (2006): 22(1), 54(0), 56(0), 59(1). Willmott & Freitas (2006) report that M. megisto and M. themisto lay eggs at the border of leaves and this is indicated as a synapomorphy for the genus (table 2, character 9:1), however obseivations on egg placement for both Methona species reported here seem to conflict with this character state. M. confusa lay eggs in clusters that covered a large portion of the leaf, including the middle of the leaf (Fig. 1), although scoring this species for this character seems inappropriate because of its elusterdaying habit. It is likely that M. confusa lays eggs while resting on the topside of the leaf, but given its cluster laying behavior it would be interesting to confirm this. M. curvifascia opposition location does not seem confined to the leaf border, although this may be a result of relatively small host leaf size observed here, because on hosts with larger leaves, laying from the leaf top and curling the abdomen underneath would result in eggs placed near the border. Thus, it may be Fig. 4. Adult Methona reared in this study. A. Male M. con- fusa dorsum. B. Male M. confusa venter. C. Female M. curvi- fascia dorsum. D. Female M. curvifascia venter. M. confusa were identified by vein Sc coalescing with Rl, presence of dorsal hindwing costal “hair pencils” in females, and male last abdomi- nal tergite not produced and block-like or spine-like. M. curvi- fascia were identified by vein Sc not coalescing with Rl, absence of dorsal hindwing costal “hair pencils” in females and male last abdominal tergite produced and narrowing into a spine-like process. useful to re-evaluate this character by focusing more on female opposition behaPor and less on the resulting egg position. Methona confusa is the first species in the genus to be observed laying eggs in clusters. In addition to the observation here, A. Freitas observed a female Methona in Acre, Brazil laying a cluster of 12 eggs. The female escaped after oPpositing but was likely M. confusa (A. Freitas, pers. comm.). Cluster-laying has been found to be relatively rare in ithomiines, but it is widely distributed across their phylogeny, being present in 12 genera (including Methona) (Brown & Freitas 1994; Drummond 1976; Haber 1978; Hill 2006; Willmott & Freitas 2006). Indeed, using the tribal classification of Willmott & Freitas (2006), only the tribes Tithoreini and Oleriini lack any cluster-laying species. In addition to Methona , the genera Hi/pothyris, Episcada, Ithomia, and Pteronymia contain cluster-laying species as well as species known to lay eggs singly (Brown & Freitas 1994; Willmott & Freitas 2006; Hill pers. obs.). This suggests life history studies on additional ithomiine species could reveal cluster-laying species in other genera presently known to only lay solitary eggs. Volume 62, Number 2 95 Some ithomiine species that are documented laying eggs in clusters also exist in solitary-laying populations, and this may lie the case with M. confusa as well. In contrast to the M. confusa immatures studied here. Brown (1987) illustrated a single M. confusa larva from Venezuela suggesting it was solitaiy. Of course. Brown's (1987) larva could have been part of a cluster of eggs that had dispersed at some larval stage only appearing to be more or less solitary. Larvae studied here were confined to bags and so no observations on dispersal of a larval group were made. It would be interesting to confirm whether M. confusa populations vary in cluster- laying because this would be an additional example of intraspecific variation similar to what has been observed in two other ithomiine species. Gilbert (1969) observed Mechanitis menapis saturate laying eggs in clusters in Costa Biea, but Drummond (1976) found M. menapis mantineus laying single eggs in western Ecuador. Similarly, Gilbert (1969) reported Hypothyris euclea valora (called H. e. leucania) laying eggs in clusters in Costa Rica, and Drummond (1976) observed H. euclea intermedia (called H. e. peruviana) laying single eggs at Limoneocha. In contrast to Drummond’s (1976) observation we have observed H. euclea intermedia laying eggs in clusters at Garzacocha. Such intraspecific variation could be a fruitful area for investigating hypotheses for cluster-laying in ithomiines (Clark & Faeth 1998; Courtney 1984; Haber 1978; Stamp 1980; Vasconcellos-Neto 1986; Young & Moffett 1979), and indicates the continuing importance of immature stages to understanding ithomiine biology. Acknowledgements We are grateful to the communities of Sani Isla and Pilche, and to the management and staff of Sani Lodge and La Selva Jungle Lodge for facilitating this work. RIH would like to thank T. Carlson for discussion about Brunfelsia. We thank the Museo Eeuatoriano de Ciencias Naturales in Quito for granting per- mits, and G. Bernes, M Medeiros and two anonymous review- ers for their comments on the manuscript. This work was sup- ported in part by fellowships from the UC Berkeley Department of Integrative Biology and the Margaret C. Walker Fund for teaching and research in systematic entomology. Literature Cited Bates, II. W. 1862. Contributions to an insect fauna of the Amazon valley. Lepidoptera: Heliconidae. Transactions of the Linnean So- ciety of London. XXI 1 1 : 495-566. Brown, K. S. JR. 1987. Chemistry at the Solanaceae Ithomiinae in- terface. Annals of the Missouri Botanical Garden. 74: 359-397. , & A. V. L. Freitas. 1994. |uvenile stages of Ithomiinae: overview and systematics (Lepidoptera: Nymphalidae). Tropical Lepidoptera. 5: 9-20. Clark, B. R., & S. H. Faeth. 1998. The evolution of egg clustering in butterflies: A test of the egg desiccation hypothesis. Evolutionary Ecology. 12: 543-552. Courtney, S. P. 1984. The evolution of egg clustering by butterflies and other insects. American Naturalist. 123: 276-281. Drummond, B. A., III. 1976. Comparative ecology and mimetic rela- tionships of Ithomiine butterflies in eastern Ecuador. Ph.D. the- sis. University of Florida, Gainesville. . 1986. Coevolution of ithomiine butterflies and solanaceous plants. Pp. 307-327. In W. G. D'Arcy (ed.), Solanaceae biology and systematics. Columbia University Press, New York. , & K. S. Brown, JR. 1987. Ithomiinae (Lepidoptera: Nymphal- idae): Summary of known larval food plants. Annals of the Mis- souri Botanical Garden. 74: 341-358. Forbes, W. T. M. 1943. The genus Thyridia (Lepidoptera, Ithomi- inae). Annals of the Entomological Society of America. 36: 707-716. Gilbert, L. E. 1969. Some aspects of the ecology and community structure of ithomid butterflies in Costa Rica. Pp. 61-92. In, Or- ganization for Tropical Studies Report, Organization for Tropical Studies. Haber, W. A. 1978. Evolutionary ecology of tropical mimetic butter- flies (Lepidoptera: Ithomiinae). Ph.D. thesis. University of Min- nesota. Hill, R. I. 2006. Life history and biology of Forbestra olivencia (Bates, 1862) (Nymphalidae, Ithomiinae). Journal of the Lepi- dopterists1 Society. 60: 203-210. Hinton, H. E. 1946. On the homology and nomenclature of the setae of lepidopterous larvae, wi th some notes on the phylogeny of the Lepidoptera. Transactions oi the Royal Entomological Society' of London. 97: 1-37. Kitching, I. ) 1984. The use of larval chaetotaxy in butterfly system- atics, with special reference to the Danaini (Lepidoptera: Nymphalidae). Systematic Entomology. 9: 49-61. Lamas, G. 2004. Atlas of Neotropical Lepidoptera - Checklist: Part 4A Hesperioidea - Papilionoidea. Gainesville, FL, Scientific Publish- ers. Mielke, O. H. H., & K. S. Brown, JR. 1979. Suplemento ao catalogo dos Ithonriidae Americanos de R. Ferreira d'Almeida (Lepi- doptera). (Nymphalidae: Ithomiinae). Centro de Recursos Au- diovisual da UFPr, Curitiba. Motta, P. C. 2003. Phylogenetic relationships of Ithomiinae based on first-instar larvae. Pp. 409-429. In C. I,. Boggs, W. B. Watt, & P. R. Ehrlich (eds. ), Butterflies: ecology and evolution taking flight. University of Chicago Press, Chicago. Muller, F. 1879. Ituna and Thyridia • a remarkable case of mimicry in butterflies. Proceedings of the Royal Entomological Society of London: xx-xxix. Peterson, A. 1962. Larvae of insects. I. Lepidoptera and plant infest- ing Hymenoptera. Edwards Brothers, Inc., Columbus, Ohio. Stamp, N. E. 1980. Egg deposition patterns in butterflies - why do some species cluster their eggs rather than deposit them singly. American Naturalist. 115: 367-380. Vasconcellos-Neto, J. 1986. Interactions between Ithomiinae (Lep- idoptera: Nymphalidae) and Solanaceae. Pp. 364-377. In W. G. D'Arcy (ed.), Solanaceae: Biology and systematics. Columbia University Press, New York. Willmott, K. R., & A. V. L. Freitas. 2006. Higher-level phylogeny of the Ithomiinae (Lepidoptera: Nymphalidae): classification, pat- terns of larval hostplant colonization and diversification. Cladis- tics. 22: 297-368. Young, A. M., & M. W. Moffett. 1979. Studies on the population bi- ology of the tropical butterfly Mechanitis isthmia in Costa Rica. American Midland Naturalist. 101: 309-319. Recieved 10 October 2007; revised and accepted for publica- tion 15 April 2008. Please see Appendix on next page 96 Journal of the Lepidopterists’ Society Appendix 1. First instar chaetotaxy of Methona confusa and Methona curvifascia. For the reasons given in Hill (2006), characters are listed as text here rather than as states of Motta’s (2003) characters. No larval specimens of M. curvifascia were preserved for study of body chaetotaxy. Descriptions of body setae are based on two larvae for M. confusa. Descriptions of head, lab rum and mandible chaetotaxy are based on two head capsules for M. curvifascia and five head capsules for M. confusa. Character # of Motta M. confusa M. curvifascia Head capsule 1 Seta Cl equidistant to frontal and anteclypeal sutures 2 Seta C2 nearer to Cl than to a medial imaginary line 3 Seta C2 same length at Cl 4 Seta FI undoubtedly more dorsal and medial to C2 5 Seta FI nearer to C2 than it is to coronal bifurcation 6 Seta FI subtly nearer to frontal suture than to imaginary medial line Seta Cl equidistant to frontal and anteclypeal sutures Seta C2 nearer to Cl than to a medial imaginary line Seta C2 same length at Cl Seta FI undoubtedly more dorsal and medial to C2 Seta FI nearer to C2 than it is to coronal bifurcation Seta F 1 subtly nearer to frontal suture than to imaginary medial line 7 Puncture Fa aligned with seta FI 8 Distance between Fa punctures subtly longer than distance between Fa and seta FI 9 Puncture AFa, and setae AF1 and AF2 all present 10 Puncture AFa slightly medial of line connecting setae AF1 and AF2 11 Puncture AFa equidistant to setae AF1 and AF2 (or subtly nearer to AF1) 12 Setae AF1 and AF2 similar in length 13 Seta AF2 subtly above level of coronal suture bifurcation 14 Distance of seta AF2 to coronal suture same as distance of AF1 to frontal suture 15 Puncture Aa below imaginary line connecting AF1 and A2 16 Puncture Aa nearer to A2 than to AF1 17 Seta A3 posterior to imaginary line between stemma iv and PI; distance of A3 to the imaginary line less than distance of A3 to stemma iv 18 Seta A1 slightly closer to stemma i than ii and aligned to slightly above stemma i 19 Seta A2 aligned with imaginary line between stemma ii and AF1 20 Seta A3 not much longer in length than A2 and LI 21 Puncture Pa ventral to slightly ventral to imaginary line connecting setae A2 and A3. 22 Puncture Pa nearer to seta A2 than to A3 23 Puncture Pb aligned with, to slightly medial of, imaginary line between setae PI and P2. 24 Puncture Pb closer to seta P2 than PI 25 Setae PI and P2 equidistant to coronal suture 26 Setae PI and P2 same length to PI slightly longer 27 Puncture La much closer to seta LI than A3, and less than 1/3 distance between LI and A3 28 Alignment of puncture La and setae LI and A3 somewhat aligned to forming a very obtuse triangle 29 Seta Ol nearly in line with stemmata i and iv, equidistant to ii and iii; Ol slightly closer to iv than i. Puncture Fa subtly above seta FI Distance between Fa punctures similar to that between Fa and FI Puncture AFa, and setae AF1 and AF2 all present Puncture AFa in line with to slightly medial of line connecting setae AF1 and AF2 Puncture AFa equidistant to setae AF1 and AF2 (or subtly nearer to AF2) Setae AF1 and AF2 similar in length Seta AF2 subtly above level of coronal suture bifurcation Distance of seta AF2 to coronal suture same as distance of AF1 to frontal suture Puncture Aa above imaginary line connecting AF1 and A2 Puncture Aa nearer to A2 than to AF1 Seta A3 posterior to imaginary line between stemma iv and PL distance of A3 to the imaginary line less than distance of A3 to stemma iv Seta A1 closer to stemma i than ii and aligned to slightlv above stemma i Seta A2 aligned with imaginary line between stemma ii and AF1 Seta A3 not much longer in length than A2 and LI Puncture Pa ventral to slightly ventral to imaginary line connecting setae A2 and A3. Puncture Pa nearer to seta A2 than to A3 Puncture Pb aligned with, to medial of. imaginary line between setae PI and P2. Puncture Pb closer to seta P2 than PI Seta P2 slightly farther from coronal suture than is seta PI Setae PI and P2 same length to PI slightly longer Puncture La much closer to seta LI than A3, and less than 1/3 distance between LI and A3 Alignment of puncture La and setae LI and A3 somewhat aligned to forming a verv obtuse triangle Seta Ol nearly in line with stemmata i and iv, equidistant to ii and iii; Ol slightly closer to iv than i Volume 62, Number 2 97 Appendix 1. Continued Character # of Motta M. confusa M. curvifascia Head capsule (cont.) 30 31 32,33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 Labrum 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 Angle formed between 02 and stemmata iv and v less than 90° Seta 02 equidistant to stemmata iv and v Seta 02 longer than Ol and 03, with Ol and 03 similar lengths Seta 03 aligned with stemma v and "groove" Puncture Oa ventral (toward antennal socket) to imaginary line between stemma i and seta A1 Puncture Ob aligned to stemma v and 03, and forming a triangle with stemma v and 02. Puncture Ob equidistant or nearer to stemma v relative to 02, and farthest from 03 501 in ventral end of antennal socket so that distance of SOI to end of antennal socket is less than 1/2 distance between SOI and S03 502 ventral to imaginary line connecting stemmata v and vi 502 equidistant to slightly closer to stemma vi relative to stemma v 503 posterior to line between stemma vi and SOI SOa between suture and imaginary line joining S03 and Gl, SOa same distance from suture as Gl SOa falls on line between S02 and nearest point of maxillary (ventral )suture, SOa is subtly closer to the suture than to S03 and much closer to the suture than to S02 SOb near antennal socket; distance of SOb to antennal socket about 1/2 that of S03 to antennal socket SOb nearer, to slightly nearer, to S03 than to stemma vi Gl subtly closer to maxillary (ventral suture) relative to groove Ga aligned to line joining Gl and 03 Ga nearer to 03 VI nearer to “V” group than P2 Stemmata all similar diameter Similar distance between stemma i, ii, iii and iv Stemma v closer to vi than to iv Seta M2 aligned or slightly basal to LI M2 aligned, to slightly dorsal, of line between Ml and L2 Ml shifted slightly dorsal relative to M2 Distance between Ml's greater than distance between Ml to M2 M2 longer than M 1 Puncture S located basal to Ml and M2 Puncture S equidistant to Ml and M2 Angle between the lines that connect Ml and M2, and Ml and the puncture S is 40° - 70° Puncture equidistant to subtly nearer to Ml and M2 relative to posterior border Puncture S basal to widest point of labrum M3 on the distal border of the labrum L2 nearer to LI than L3 LI level to widest point of labrum Less sclerotized region near the labium notch and to Ml and M2 Less sclerotized basal patches absent Internal border of the Iabral lobe smoothly curved Basal angle of labrum notch obtuse Angle formed between 02 and stemmata iv and v less than 90° Seta 02 equidistant to stemmata iv and v Seta 02 longer than Ol and 03, with Ol and 03 similar lengths Seta 03 aligned with stemma v and "groove" Puncture Oa ventral (toward antennal socket) to imaginary line between stemma i and seta A1 Puncture Ob aligned to stemma v and 03, and forming a triangle with stemma v and 02 Puncture Ob nearer to stemma v than 02 and farthest from 03 501 in ventral end of antennal socket so that distance of SOI to end of antennal socket is less than 1/2 distance between SOI and S03 502 ventral to imaginary line connecting stemmata v and vi 502 equidistant to slightly closer to stemma vi relative to stemma v 503 posterior to line between stemma vi and SOI SOa between suture and i magi nan line joining S03 and Gl, SOa same distance from suture as Gl SOa falls on line between S02 and nearest point of maxillary ventral) suture, SOa is subtly closer to the suture than to S03 and much closer to the suture than to S02 SOb near antennal socket; distance of SOb to antennal socket about 1/2 that of S03 to antennal socket SOb nearer, to slightly nearer, to S03 than to stemma vi Gl equidistant to groove and maxillary (ventral) suture Ga aligned to line joining Gl and 03 Ga nearer to 03 VI nearer to “V” group than P2 Stemmata all similar diameter Similar distance between stemma i, ii, iii and iv Stemma v closer to \i than to iv Seta M2 aligned to LI M2 basal to line between Ml and L2 Ml aligned to slightly dorsal of M2 Distance between Ml's greater than distance between Ml to M2 M2 longer than Ml Puncture S located basal to Ml and M2 Puncture S equidistant to Ml and M2 or a little closer to M2 Angle between the lines that connect M 1 and M2, and Ml and the puncture S is 40° - 70° Puncture equidistant to Ml and M2 relative to posterior border Puncture S basal to widest point of labrum M3 on the distal border of the labrum L2 nearer to LI than L3 LI level to widest point of labrum Less sclerotized region near the labrum notch and to Ml and M2 Less sclerotized basal patches absent Internal border of the labial lobe smoothly curved Basal angle of labrum notch obtuse 98 Journal of the Lepidopterists’ Society’ Appendix 1. Continued Character # of Motta M. confusa M. curvifascia Labruni (cont.) 70 Ratio of notch length (= depth) to overall labral length (labral lobe to base) ~ 0.3; ratio of labral notch width, as measured between apices of lobes, to labral length ~ 1.1 Ratio of notch length (= depth) to overall labral length (labral lobe to base) ~ 0.4; ratio of labral notch width, as measured between apices of lobes, to labral length ~ 1.1 71 Ratio of labrum width (between Lis) to length (labral lobe to base) ~ 2 Ratio of labrum width (between Li’s) to length (labral lobe to base) ~ 2 Mandible 72 Fewer than three small molar teeth Fewer than three small molar teeth 73 Incisors 2 and 3 similar lengths Incisors 2 and 3 similar lengths 74 Body Lateral grooves radiating from each side of 4th incisor, one on outside more subtle than others, 4 grooves in total Lateral grooves radiating from each side of 4th incisor, one on outside more subtle than others, 4 grooves in total 75 No tubercles present on the thorax 77 Average seta length less than segment width 78,79 Crochets arranged in a circle on segments A3-6, but A10 arranged in a semicircle; all crochet lengths similar 80 Prolegs with more that 14 crochets on average 81 Cervical sclerite absent on XD1 and XD2 and D1 82 Seta D1 shorter than XD1 mid XD2, XD1 mid XD2 are equivalent in length 83,87 Setae SD2 and SD1 aligned on Tl, SD2 shifted posterior of SD1 on T2- A8, and SD2 shifted slightly posterior of SD1 on A9 84 On segment Tl setae LI and L2 slightly dorsal of spiracle with L2 between LI and spiracle; on T2 and T3, L2 is at level of abdominal spiracles; on Al- A8 LI and L2 below spiracle 85,91 Setae D1 and D2 are equivalent lengths 86 Seta SD2 closer to D2 than to SD1 88 Seta SD2 ventral and posterior to D1 and D2 89,94 Seta SD1 longer than LI and equivalent to L2 on segment Tl; on T2 & T3 setae SD1 and L2 equivalent and shorter than LI and L3 (which are same length); SD1 equivalent to LI and L2 on abdomen 90 Seta L2 present on segments T1-A8 92 Seta SD2 and D1 equivalent lengths and longer than D2 93 SD2 shorter than SD1 on Tl; SD2 longer than SD1 on T2 & T3; SD2 shorter than SD1 on abdomen 95 LI shorter than L2 on TL LI longer than L2 on T2 and T3 with L3 equivalent to LI; LI and L2 equivalent on abdomen 96 Additional SV seta on A2 only 97 A9 with one less seta (LI or L2) than A7 and AS 98 Epiproct setae Dl, D2, SD1 and LI similar lengths 99 PI and SP1 setae present on A10 Volume 62, Number 2 99 Journal of the Lepidopterists’ Society 62(2), 2008, 99-105 EXPERIMENTAL DESIGN AND THE OUTCOME OF PREFERENCE-PERFORMANCE ASSAYS, WITH EXAMPLES FROM MITOURA BUTTERFLIES (LYCAENIDAE) Matthew L Forister Dept, of Natural Resources & Environmental Science / MS 186, 1000 Valley Road, University of Nevada, Reno, U.S.A; email: mtorister@cabnr.unr.edu ABSTRACT. Investigations into adult host preference and the performance of larvae on different host plants have played a central role in ecological and evolutionary plant-insect research. Here I present two sets of experiments that address aspects of the experimental design of pref- erence-performance assays, using a well-studied system ol lycaenid butterflies. First, I compare results from sequential, no-choice opposition assays to previous results reported from simultaneous choice tests with Mitoura nelsoni. Second, I describe an experiment in which the larvae of two closely related species (M. nelsoni and Mitoura muiri) were reared in parallel on plants in the laboratory and in the field to assess the po- tential influence of environmental conditions on performance. Results from the no-choice preference assays are consistent with previous results, suggesting that, at least in this system, the two types of experimental design lead to similar conclusions. The experiment rearing larvae in the field and in the laboratory revealed a significant effect of environment on pupal weights, but did not detect a species by environment interac- tion. Thus for pupal weights, a laboratory-based study is sufficient to compare performance between M. nelsoni and M. muiri. However, a species by environment interaction was observed for development time, which has implications for host-associated speeiation in this group that would not have been detected in a solely laboratory-based study. Additional key words: Callophrys, specialization, choice test, no-choice test. Preference-performance assays are used to address a range of questions in the ecology and evolutionary biology of herbivorous insects (Dethier 1954; Thompson 1988; Jaenike 1990; Waekers 2007; Craig & Itami 2008). Preference refers to the choices made by ovipositing females or feeding individuals for different host plant species (Singer 2000), and performance refers to the development of juvenile stages on specific hosts. The questions addressed by preference- performance experiments may be as simple as; will a species of insect accept a particular species of plant as a host, and is the same plant a suitable host for larval development? Questions may also involve genetic variation and correlations among genetic elements: in particular, is preference for a particular plant species genetically correlated with the ability of larvae to utilize the same host species (Via 1986; Thompson 1988; Mayhew 1997). Experiments involving preference and performance are also central to the practice of biocontrol, in which behavioral and physiological host range must be determined before the release of a control agent (Marohasy 1998). These and related topics have been reviewed by many authors, including Jaenike (1990), Thompson & Pellmyr (1991), Craig & Itami (2008), and Berenbaum & Feeny (2008). The goal of this paper is to address two methodological and experimental issues involved in preference- performance assays: choice versus no-choice preference tests, and the influence of laboratory versus field conditions on performance experiments. Two of the more common ways in which preference assays can be constructed include choice and no-choice tests (for a review of other experimental designs and related issues not discussed here, see Courtney et al. (1989), Singer & Lee (2000), Barton Browne & Withers (2002), Singer et al. (2002) Van Driesche & Murray (2004), and Mereader & Scriber (2007)). In choice tests, host plants are presented to an adult female or group of females in an array and the response is typically the number of eggs laid on the different plants in a set amount of time. In a no-choice assay, the behavioral response (oviposition) is scored with plants in isolation, often sequentially, with plants being presented one after the other to adults. Simultaneous choice tests have been criticized as being unrealistic, as different host plant species may not be in immediate physical proximity in the wild (Singer et al. 1992). On the other hand, an argument can be made that simultaneous choice tests are conservative: the juxtaposition of plants in an experimental arena could make it more difficult for an ovipositing female to make a choice (since information gathered from volatile plant cues may be overlapping or mixed). In any event, the two types of test, choice and no- choice, potentially provide different and complementary information (Withers & Mansfield 2005). Consider a simple, hypothetical scenario: two host plants (A and B) are used by a particular insect herbivore. In choice tests, plant A is overwhelmingly preferred to the exclusion of plant B, but in no-choice tests both plants receive a comparable number of eggs from ovipositing females. It might be the case that the volatile and tactile cues that characterize plant A are sufficiently more stimulating to ovipositing females such that B is ignored in the presence of A. While in the absence of A, B is recognized as a suitable host and will 100 Journal of the Lepidopterists’ Society be utilized. The choice test tells us not only what could happen in the wild when plants are interdigitated or in very close proximity, but it tells us something about the inherent ranking of host cues by the herbivore (e.g. Thompson 1993). The no-choice test on the other hand might give a clearer picture of what could happen in the wild as a female moves from one isolated patch of plants to another. Choice tests are more common in the literature, perhaps because they are logistically more efficient. What is not often tested (and which I address here with one butterfly species) is how often the results from choice and no-choice tests provide different lines of information (as in the hypothetical example above), or how often results are congruent or redundant. Preference experiments are often rather contrived in that females are typically presented plants under artificial conditions (cages or preference arenas), and in arrays or sequences that they might never encounter in the field (though more realistic preference tests have been conducted, e.g. Singer & Thomas 1988). In contrast to this, performance experiments need not be quite so highly abstracted from natural conditions: it is possible to rear larvae in the field by confining them to small cages or bags. Despite this, the majority of performance experiments have addressed the performance of larvae in laboratory conditions, often with larvae reared singly in petri dishes (Zalueki et al. 2002). Whatever measure of performance is taken (pupal weight, development time, etc.), it seems intuitively obvious that results may be biased by laboratory conditions. For example, the architecture oi a given species of plant might provide a microclimate that allows larvae to feed throughout the heat of the day, resulting in faster development than on a host that does not have the same architecture (Alonso 1997). This effect would only be apparent if larvae were reared in the field. Other environment-dependent effects could include interactions with predators and parasitoids. I used two species of lyeaenid butterflies, Mitoura nelsoni Boisduval and Mitoura muiri H. Edwards, to address these issues in the design of preference and performance experiments. The oviposition behavior of M. nelsoni females in choice tests has been previously described: they have consistent preferences for their host incense cedar ( Calocedms decurrens Torrey), laying the most eggs on that host in both four-way and two-way choice tests involving other hosts of Mitoura in Northern California (Forister 2004, 2005a). Here I ask if the preferences of M. nelsoni females for incense cedar are expressed in no-choice tests as a willingness to lay eggs on incense cedar and a reticence to lay eggs on an alternate host when encountered in isolation. The larval performance of M. nelsoni and M. muiri on multiple hosts, as expressed in pupal weight and survival, has been previously described (Forister 2004, 2005a). Here I focus on one host, a host of M. muiri, and ask if differences between the two butterfly species in performance on that host are consistent between laboratory and field environments. Materials and Methods Butterflies and plants. M. nelsoni and M. muiri are part of a complex of host-specific lyeaenid butterflies in North America associated with plants in the family Cupressaceae which have been the focus of recent investigations into the ecology of speciation (Nice & Shapiro 2001; Forister 2004, 2005a, 2005b). M. nelsoni is found in association with incense cedar at low to middle elevations in mesic forests from southern British Columbia to Baja California. M. muiri is an edaphic- endemic associated with cypress hosts (primarily MacNab cypress, Cupressus macnabiana A. Murray, and Sargent cypress, Cupressus sargentii Jepson) on low elevation, ultramafic soils such as serpentine in California ( Gervais & Shapiro 1999). The experiments described here used M. nelsoni adults in preference experiments, and cateipillars of both species in performance experiments. The M. nelsoni adults consisted of wild-caught and laboratory- reared individuals. Wild-caught individuals were taken from the following locations in 2004 on the west slope of the Sierra Nevada Mountains near interstate 80: Drum Powerhouse Road and Fang Crossing (see Forister 2004 for more details on these locations). Faboratory-reared adults were part of a colony that was being maintained for other experiments at the University of California, Davis. These individuals were the offspring of females collected from a number of populations in the Sierra Nevada and North Coast Ranges in the previous season. Farvae used in performance experiments were generated from individuals reared and mated in the laboratory. For both M. nelsoni and M. muiri , larvae were pooled from multiple lines without regard to genetic background within species. In other words, M. nelson larvae were the product of matings between M. nelsoni adults from a number of locations throughout California (and the same for M. muiri). These matings are described in detail in Forister (2005a). Three host plant species were involved in these experiments: incense cedar (the host of M. nelsoni), Sargent cypress and MacNab cypress (hosts of M. muiri). For preference experiments, incense cedar and Sargent cypress were collected from Goat Mountain in the North Coast Range of California, where the two hosts grow sympatrically. For the performance experiments, MacNab cypress was used both in the field Volume 62, Number 2 101 and through collection from one location, Knoxville Public Lands, also in the North Coast Range. Preference assays. In order to assess the oviposition behavior of M. nelsoni in no-choice assays, females were confined individually with sprigs of host plants in oviposition arenas (cylinders of wire mesh, 3600 cm3). They were exposed to one host for 24 hours, and then switched to the other host for 24 hours (the two hosts, as mentioned above, were incense cedar and Sargent cypress). The switch from one host to the other was done in the early morning of the second day, before butterflies were active. Experiments were only conducted for 48-hour periods because previous experience with Mitoura butterflies had shown that females become considerably less vigorous and egg- laying begins to drop off after 48 hours when they are kept in a greenhouse in full sun (Forister, pers. obs.). At the start of the experiment, each female was haphazardly assigned to one of two groups, with one group being confined first with incense cedar, and the second group being confined first to Sargent cypress. Sugar water was applied to the cages as an artificial nectar source that was readily consumed by butterflies throughout the experiment. The number of eggs on plants was counted at the end of each interval as a measure of host preference ( Mitoura butterflies very rarely ovisposit on any surface in preference arenas other than the host plants; and if eggs were found on the side of the cage they were not counted). Results from preference assays were analyzed in two ways. First, the number of eggs laid by each female on the two hosts was treated as a pair in a nonparametric Wileoxon matched-pairs test. This analysis addressed the question: which host received more eggs without reference to the order of the hosts? Second, a Wileoxon rank-sum test was used to ask: does the first host encountered affect the number of eggs laid on incense cedar? In this case, each female is represented by one data point (the number of eggs laid on cedar), and females are identified as belonging to either the treatment that received incense cedar first or Sargent cypress first. Performance assays. The goal of performance assays was to ask if differences in performance between the two butterfly species observed in the laboratory (Forister 2004, 2005a) are also observed in the field. To address this question, ten trees of MacNab cypress, the host of M. muiri, were selected at a field site that has been studied previously (Knoxville, see Forister 2004). Trees were selected haphazardly within a small area (approximately 100 square meters), and caterpillars of both M. muiri and M. nelsoni were reared to pupation simultaneously on these trees in the field and on cuttings from these trees brought back to the laboratory. Caterpillars in the laboratory were reared in groups of five in large drinking cups nested within smaller cups so that the cut ends of branches could be pushed through holes in the larger cup and into water held in the smaller cup. Upon pupation, pupae were weighed on a Mettler Toledo microbalance to the nearest hundredth of a milligram. Caterpillars that became part of the field component were reared initially in the laboratory through the first instar. They were then transferred to the field, where they were reared to pupation in groups of five in spun mesh bags enclosing tree branches. Each of the ten trees in the field had two bags (one M. muiri bag and one M. nelsoni bag). Caterpillars in bags were checked weekly and moved to new branches on the same trees when foliage had been depleted. Upon pupation, pupae were removed from bags, brought back to the laboratory and weighed. In addition to pupal weight, survival and days to pupation were recorded for both the laboratory and field-reared individuals. Analyses of variance (ANOVA) using restricted maximum likelihood (REML) mixed models were used to analyze results from performance experiments (Littell et al. 1996). Fixed factors in models included species, location (field or laboratory), and an interaction between species and location. Random factors were tree, and interactions between tree and species, and between tree and location. Rearing group is not included in models because values within groups (for pupal weight, development time and survival) were simply averaged prior to analysis (individuals within groups are not statistically independent). No transformations were found to be necessary to meet the assumptions of ANOVA for pupal weight or development time. Residual error from analysis of survival (the fraction of individuals surviving to pupation within each rearing group) was highly non-normal (even following arcsine transformation) due to the large number of groups in which survival was 100%. Therefore, two separate nonparametric Wileoxon rank- sum tests were performed to compare survival between the two species in the laboratory and in the field. IMP- IN software, version 7.0 (SAS Institute, Cary, NC, U.S.A.), and Kaleidagraph, version 3.6 (Synergy Software, Reading, PA, U.S.A.), were used for nonparametric analyses (both for survival data and preference results, described above), and PROC MIXED in SAS, version 9.1 (SAS Institute, Cary, NC, U.S.A.), was used for REML analyses of variance. Results Preference assays. A total of 45 M. nelsoni females were tested in no-choice assays using incense cedar, the 102 Journal of the Lepidopterists’ Society host of M. nelsoni, and Sargent cypress, the host of M. muiri. As can be seen in Fig. la, females laid a majority of their eggs on incense cedar in these no-choice assays (T = 4.63, P < 0.0001). The behavior of females was not influenced by the order in which plants were presented to them: a comparable number of eggs was laid on incense cedar regardless of whether that host was presented first or second in sequence (Fig lb; Tj = - 1.11, P = 0.26). Performance assays. A total of 185 larvae were reared to pupation in 39 rearing groups (20 in the laboratory and 19 in the field; larvae from one M. muiri group in the field escaped). As has been observed in previous work (Forister 2004), M. nelsoni individuals develop to pupal weights that are greater than M. muiri (on average 10% greater), even though the host in question is the natal host of M. muiri. The results reported here demonstrate that this difference (a) cedar cypress (b) cedar cypress Fig. 1. Results from preference assays illustrated as box plots. The same data is shown in two different ways in (a) and (b): data shown in (a) is the fraction of eggs laid on the two hosts, while (b) shows the influence of experimental sequence in sequential no-choice assays on oviposition behavior. In other words, in (b), the data shown is the fraction of eggs laid on incense cedar for females which were exposed to that plant first (the left box), and for females which were exposed to cypress first (the right box). (between the two species) is not affected by rearing environment (Fig 2a, and note the insignificant species by location interaction in Table 1). In contrast, rearing environment did have a differential effect on the development time of the two species: in the field, M. muiri individuals reach pupation 4.62 days earlier than M. nelsoni individuals (Fig. 2b, Table 2). In general, larvae of both species developed more slowly in the field, and this might be because they did not feed at night: when checking the bags in the early morning, I found larvae to be inactive, while larvae in the laboratory are capable of feeding throughout the night. There were no significant differences between the (a) (c) Fig. 2. Means and standard errors from assays of performance in the laboratory and in the field. Statistical results for pupal weight (a) and days to pupation (b) are shown in Tables 1 and 2 respectively. See text for more details related to survival (c). Volume 62, Number 2 103 survival of M. nelsoni and M. muiri caterpillars in the laboratory (T; = 0, P = 1.0) or in the field (T; = 0.51, and P = 0.61) (Fig. 2c). Differences among individual trees had a significant effect on pupal weight (Table 1), but not on development time (Table 2). Although tree had an effect on pupal weight, this was not influenced by rearing environment, nor was there a significant species by tree interaction. In other words, larvae of both species did better on certain trees, and this was true whether larvae were reared in the field or on cuttings from the same trees in the laboratory. In order to better visualize the influence of individual trees on pupal weight. Fig. 3 shows the correlation between weights of larvae reared in the field and in the laboratory. One outlier has been excluded from the relationship shown in Fig. 3: one M. nelsoni rearing group had high mean pupal weight in the field (80.03 mg), but unusually low weight in the laboratory (66.9 mg). With the outlier excluded, the correlation is significant: Pearson product- moment correlation of 0.73, P = 0.0006; with the outlier included the correlation is 0.37, P = 0.12. Discussion M. nelsoni females express a clear preference for their natal host, incense cedar, in both choice tests (Forister 2004, 2005a), and no-choice tests, as reported here (Fig. 1). Choice tests are more efficient from the point of view ol experimenter effort: there is less manipulation in choice tests, as plants do not need to be changed part way through the test (as compared to a no- choice design with sequential replacement of hosts). The results reported here suggest that, at least in the Mitoura system, choice tests provide equivalent T) .£ o> E O) '55 3 75 a 3 a 85 80 75 70 65 60 55 70 75 80 85 90 95 100 105 110 Pupal weight (mg), lab Fig. 3. Comparison of pupal weights in the field versus pupal weights in the laboratory. Each point corresponds to a group of larvae reared on foliage from a single plant in the field and in the laboratory. Each host plant is shown twice, once as the host of M. muiri larvae and once as the host of M. nelsoni larvae (circles are M. muiri , squares are M. nelsoni). A single outlier was excluded, see text for details. information to no-choice tests. There are two important caveats to this conclusion. First, these results should not be used to infer that the two types of choice test are equivalent in other systems. Rather, the results reported here highlight the utility ol exploring both types of assay, and the possibility that in some systems choice tests may be sufficient. Second, while it is true that choice and no- choice assays with M. nelsoni lead to similar conclusions about the relative ranking of the two hosts by ovipositing females, there may be situations in which no-choice tests would still be uniquely useful. For example, no- choice tests could be used to survey for variation among Table 1. Results from analysis of pupal weights. Degrees of freedom and F ratios are reported for fixed effects, covariance estimates and standard errors for random values are shown in bold text. i effects. Significant P Source NDF DDF F P Species 1 26.3 16.8 0.00040 Location 1 26.3 97.3 < 0.0001 Species x location 1 26.3 0.21 0.65 Covariance SE P Tree 19.0 13.1 0.0095 Tree x species 0 - - Tree x location 0 Table 2. Results from analysis of development time (days to pupation). Degrees of freedom and F ratios are reported for fixed effects, covariance estimates and standard errors for ran- dom effects. Significant P values are shown in bold text. Source NDF DDF F P Species 1 13.3 13.2 0.00300 Location 1 26.3 97.3 < 0.0001 Species x location 1 26.3 0.21 0.24 Covariance SE P Tree 0.354 0.93 0,33 Tree x species 0.945 1,36 0.24 Tree x location 0.836 1.34 0.26 104 Journal of the Lepidopterists’ Society females in preference for a less preferred host, while such variation could potentially be harder to detect in choice tests where females always spend a majority of their time ovipositing on the preferred host. No-choice tests could also be used to study factors (such as egg load) which may influence “motivation” and lead to the acceptance of an otherwise less-preferred host (Singer etol. 1992). With the performance results reported here, it is apparent that a comparison between the two species for at least one element of lanal performance (pupal weight) is not greatly influenced by rearing environment. M. nelsoni pupae are bigger than M. muiri pupae, and individuals reared in the laboratory are bigger than individuals reared in the field (Fig. 2a), but being reared in the laboratory or the field does not change the relative sizes of M. nelsoni and M. muiri pupae. The foliage quality of individual trees was also consistent across rearing environments (Fig. 3). The vast majority of performance experiments are done in the laboratory (Zalucki et al. 2002), thus the results reported here are heartening: not only may laboratory performance (as measured by pupal weight) be an accurate reflection of performance in the field (at least in the absence of natural enemies), but intraspecific variation in plant quality may in some cases also be reasonably studied under laboratory conditions. Osier et al. (2000) reported a similar consistency between performance in the laboratory and in the field on particular plant genotypes using gypsy moth larvae and quaking aspen clones. The performance results reported here are also interesting in the light of a scenario of host-associated speciation that has been described in Mitoura. Differences in host preference are believed to be a key mechanism in the diversification of this group (Nice & Shapiro 2001; Forister 2004, 2005a), as has been suggested for a number of other phytophagous insect systems in which adults mate and oviposit on their host plants (Berlocher & Feder 2002; Dies & Mallet 2002). Divergent host preferences are expected to evolve in association with host-specific larval adaptations, particularly when divergence is in sympatry or parapatry (Fry 2003) (which appears to be the case for Mitoura). M. nelsoni fits this model nicely: females have strong preferences and larvae attain considerably larger pupal weights on incense cedar (larger than M. nelsoni larvae reared on other hosts of Mitoura in northern California, and larger than other Mitoura larvae reared on incense cedar). In contrast, M. muiri females have strong host preferences but M. muiri larvae do not attain greater pupal weights or have higher survival on their natal cypresses relative to M. nelsoni larvae on the same hosts. The present study suggests a previously undetected component of local adaptation in M. muiri-. faster development than M. nelsoni on MacNab cypress in the field. Why this difference would only be manifest in the field is not known, though one possibility is that M. muiri larvae may be able to feed over a slightly wader range of temperatures than M. nelsoni larvae. Faster growth may reduce exposure to natural enemies (Williams 1999), or extreme climatic events (Fordyce & Shapiro 2003). In particular, faster development at low elevations in the diy, inner North Coast Range of California might allow larvae to pupate before temperatures become unfavorably high (three days before the end of the experiment, a maximum daily temperature of 40 degrees Celsius was recorded at the field site). Although the adaptive significance of faster development in the field is unknown, this is a difference between M. nelsoni and M. muiri that would not have been observed in a solely laboratory-based study. Acknowledgements I thank S. L. Thrasher for assistance in rearing larvae and tending to adult butterflies, and A. M. Shapiro, E. A. Leger and G. W. Forister for help in collecting plants and butterflies. Literature Cited Alonso, C. 1997. Choosing a place to grow. Importance of within- plant abiotic microenvironment for Yponorneuta mahalebella . En- tomol. Exp. Appl. 83: 171-180. Barton Browne, L., & T. M. Withers. 2002. 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Evolution of feeding preferences in phy- tophagous insects. Evolution 8: 33-54. Dres. M., & J. Mallet. 2002. Host races in plant-feeding insects and their importance in sympatric speciation. Phil. Trans. R. Soc. Lond. B Biol. Sci. 357: 471-492. Fordyce. J. A., & A. M. Shapiro. 2003. Another perspective on the slow-growth/high-mortality hypothesis: chilling effects on swal- lowtail larvae. Ecology 84: 263-268. Forister, VI. L. 2004. Opposition preference and larval performance within a diverging lineage of lycaenid butterflies. Ecol. Entomol. 29: 264-272. . 2005a. Influence of host plant phenology on Mitoura nelsoni (Lepidoptera : Lycaenidae). Ann. Entomol. Soc. Am. 98: 295-301. . 2005b. Independent inheritance of preference and perfor- mance in hybrids between host races of Mitoura butterflies (Lep- idoptera : Lycaenidae). Evolution 59: 1149-1155. Volume 62, Number 2 105 Fry, J. D. 2003. Multilocus models of sympatric speciation: Bush ver- sus Rice versus Felsenstein. Evolution 57: 1735-1746. Gervais, B. R., & A. M. Shapiro. 1999. Distribution of edaphic-en- demic butterflies in the Sierra Nevada of California. Global Ecol. Biogeography 8: 151-162. Jaenike, J. 1990. Host specialization in phytophagous insects. Ann. Rev. Ecol. Syst. 21: 243-273. Littell, R. C., W. W. Milliken, & R. D. Wolfinger. 1996. SAS sys- tem for mixed models computer program, version By Littell, R. C„ W. W. Milliken, and R. D. Wolfinger, Cary, NY. Marohasy, [. 1998. The design and interpretation of host-specificity tests for weed biological control with particular reference to in- sect behaviour. Biocontrol News and Information 19: 13-20. Mayhew, R J. 1997. Adaptive patterns of host-plant selection by phy- tophagous insects. Oikos 79: 417P28. Mercader, R. J., & J. M. Scriber. 2007. Diversification of host use in two polyphagous butterflies: differences in opposition specificity or host rank hierarchy? Entomol. Exp. Appl. 125: 89-101. Nice, C. C., & A. M. Shapiro. 2001. Population genetic evidence of restricted gene flow between host races in the butterfly genus Mi- toura (Lepidoptera : Lycaenidae). Ann. Entomol. Soc. Am. 94: 257-267. Osier, T. L., S. Y. Hwang, & R. L. Lindroth. 2000. Effects of phyto- chemical variation in quaking aspen Populus tremuloides clones on gypsy moth Lyinantria clispar performance in the field and laboratory. Ecol. Entomol. 25: 197-207. Singer, M. C., D. Vasco, C. Parmesan, C. D. Thomas, & D. Ng. 1992. Distinguishing between preference and motivation in food choice: An example from insect opposition. Anim. Behav. 44: 463-471. . 2000. Reducing ambiguity in describing plant-insect interac- tions: "preference", "acceptability" and "electiPty". Ecology Lett. 3: 159-162. & ]. R. Lee. 2000. Discrimination within and between host species by a butterfly: implications for design of preference ex- periments. Ecology Lett. 3: 101-105. , C. Stefanescu, & I. Pen. 2002. When random sampling does not work: standard design falsely indicates maladaptive host pref- erences in a butterfly. Ecology Lett. 5: 1—6. Thompson, J. N. 1988. Evolutionary ecology of the relationship be- tween opposition preference and performance of offspring in phytophagous insects. Entomol. Exp. Appl. 47: 3-14. . 1993. Preference hierarchies and the origin of geographic spe- cialization in host use in swallowtail butterflies. Evolution 47: 1585-1594. , & O. Pellmyr. 1991. Evolution of opposition behaPor and host preference in Lepidoptera. Ann. Rev. Entomol. 36: 65-89. Van Driesche, R. G., & T. J. Murray. 2004. OverPew of testing schemes and designs used to estimate host ranges. In R. G. Van Driesche, and R. Reardon (eds . ) , Assessing host ranges for para- sitoids and predators used for classical biological control: a guide to best practice. Forster Health Technology Enterprise Team. Via, S. 1986. Genetic covariance between opposition preference and larval performance in an insect herbivore. Evolution 40: 778-785. Wackers, F. L„ J. Romeis, and P. van Rijn. 2007. Nectar and pollen feeding by insect herbivores and implications for multitrophic in- teractions. Ann. Rev. Entomol. 52: 301-323. Williams, I. S. 1999. Slow-growth, high-mortality - a general hypoth- esis, or is it? Ecol. Entomol. 24: 490-495. Withers, T. M., & S. Mansfield. 2005. Choice or no-choice tests? Effects of experimental design on the expression of host range. In M. Hoddle (ed. ), Proceedings, 2nd international symposium of biological control of arthropods. USDA Forest SerPce, West Vir- ginia. Zalucki, M. P, A. R. Clarke, & S. B. Malcolm. 2002. Ecology and behaPor of first instar larval Lepidoptera. Ann. Rev. Entomol. 47: 361-393. Received for publication 24 November 2007; revised and accepted 15 April '2008 . 106 General Notes Journal of the Lepidopterists’ Society Journal of the Lepidopterists' Society 62(2), 2008, 106-107 NOCTUA COMES IN ONTARIO: AN INTRODUCED CUTWORM (NOCTUIDAE: NOCTUINAE) NEW TO EASTERN NORTH AMERICA Additional key words: grape, tobacco, Palaearctic. The Lesser Yellow Underwing, Noctua comes Hiibner, [1813], is an Old World cutworm moth that was introduced in North America in the Vancouver, British Columbia, area around 1982 (Neil 1984; Copley & Cannings 2005). It has since spread eastward in British Columbia as far as the Okanagan Valley, and south into Washington and central Oregon, and continues to expand, but has not yet crossed the Continental Divide (Lafontaine 1998, J. Donald Lafontaine pers. com.). The slow expansion of Noctua comes in the Pacific Northwest in the last twenty- five years is in stark contrast to the spread of its highly invasive congener, the Large Yellow Underwing, Noctua pronuba (L.), which was introduced in Halifax, Nova Scotia, around 1979 and in the same time period has traversed the continent (Neil 1981, Powell 2002), quickly becoming abundant in most areas. On 15 August 2006 I collected a fresh male Noctua comes (Fig. 1) at a mercury vapor light in my garden in urban Toronto, Ontario, Canada (43.674°N, 79.337°W). On 25 September 2006 a second worn male was collected at the same location. The specimens are deposited in the Canadian National Collection (CNC), Ottawa, and the identification was confirmed by J. Donald Lafontaine. This is the first report of Noctua comes in eastern North America. In 2007, two additional specimens (both female) were collected at the same location on 24 September and 26 September (specimens in collection of the author). Despite regular moth collecting in downtown Toronto for a number of Fig. 1. Noctua comes , male, Toronto, Ontario, Canada, 15 August 2006. years, yielding over 150 species of noctuid moths, Noctua comes has not previously been detected. Its sudden appearance suggests that it has only recently become established here and, over 3000 km east of its known North American range, undoubtedly represents a separate introduction from the Palaearctic or a secondary introduction from the Pacific Northwest. With the increasing number of recent introductions of Old World noctuids in the Northeast (Mikkola & Lafontaine 1994, J. Donald Lafontaine pers. com.), and the proximity to the Great Lakes-St. Lawrence Seaway (and the port of Toronto a few kilometers away), a European origin seems more likely. Dual introductions on the Atlantic and Pacific coasts, often almost simultaneously, have been noted with some frequency in the Lepidoptera (Ferguson 1996, Mikkola & Lafontaine 1994, Miller 1999, Powell & Passoa 1991). Larvae of Noctua comes teed at night on a wide variety of herbaceous plants in open areas including weedy species, cultivated plants, and grasses (Poaeeae) and in spring also climb to feed on low woody plants (Lafontaine 1998, Waring et al. 2003). It is a minor pest of grape (Vitis L.) (Vitaceae) and tobacco ( Nicotiana L.) (Solanaceae) in the western Palaearctic, and larvae were recently found feeding on developing grape buds in vineyards in Washington (Sannino & Espinosa 1999, James 2007). Additional records from Toronto are expected and the species should be watched for in southern Ontario, southwestern Quebec and the Great Lakes states. Noctua comes can be distinguished from Noctua pronuba by its smaller size (forewing length = 16 to 21 mm) and by the presence of a conspicuous black discal spot on the hindwing. The living moth with wings closed may suggest a species of Abagrotis Smith more than a small pronuba. Diagnostic characters of the genitalia and larvae of Noctua comes and N. pronuba are provided by Lafontaine (1998). Additional Palaearctic species of Noctua are illustrated in Fibiger (1993, 1997). The early stages of Noctua comes are described and illustrated by Sannino & Espinosa (1999). It has a single brood annually and overwinters as a larva; the flight season extends from July to September, with extreme dates in June and October in the Pacific Northwest and the British Isles. Specimens from British Columbia in Volume 62, Number 2 107 the CNC from February and March are labeled “from nursery” and were likely reared from larvae found in greenhouses. Thanks to Don Lafontaine and George Balogh for helpful comments on a draft of this note, and to Jocelyn Gill (CNC) for photographing the specimen for the figure. Literature Cited Copley, C.R. & R.A. Cannings. 2005. Notes on the status of the Eurasian moths Noctua pronuba and Noctua comes (Lepidoptera: Noctuidae) on Vancouver Island, British Columbia. | Entomol. Soc. Brit. Columbia. 102:83-84. Ferguson, D.C. 1996. The introduction and spread of Chloroclystis rectangulata (L.) (Geometridae), and its first reported occur- rences in the United States. J. Lepid. Soc. 50:145-148. Fibiger, M. 1993. Noctuidae Europaeae. Vol.2, Noctuinae II. Ento- mological Press, Sor0. . 1997. Noctuidae Europaeae. Vol. 3, Noctuinae III. Entomo- logical Press, Sor0. James, D. 2007. Hop, grape, and red currant entomology: grape in- sects and mites. http://www.goodbugs.prosser.wsu.edu/ grapes.htm [accessed 30 May 2007], Lafontaine, |.D. 1998. Noctuoidea, Noctuidae (part), Noctuinae (part-Noctuini). In R.B. Dominick et al. (eds.). The moths of North America, Fasc. 27.3. The Wedge Entomological Research Journal of the Lepidopterists’ Society 62(2), 2008, 107-108 Foundation, Washington, D.C. Mikkola, K. & J.D. Lafontaine. 1994. Recent introductions of ri- parian noctuid moths from the Palaearctic region to North America, with the first report of Apamea unanimis (Hiibner) (Noctuidae: Amphipyrinae). J. Lepid. Soc. 48:121-127. Miller, W.E. 1999. A new synonomy in Dichrorampha that reveals an overlooked immigrant record lor North America (Tortricidae). J. Lepid. Soc. 53:74—75. Neil, K. 1981. The occurrence of Noctua pronuba (L.) (Noctuidae) in Nova Scotia: a new North American record. J. Lepid. Soc. 35:248. . 1984. Noctua conies, a noctuid new to North America (Lepi- doptera: Noctuidae: Noctuinae). Can. Entomol. 116:479-480. Powell, J.A. 2002. Noctua pronuba reaches the Pacific coast. News of the Lepid. Soc. 44:111.120. & S. Passoa. 1991. Rapid colonization of the western United States by the Palearctic moth, Agonopterix alstroemeriana (Oe- cophoridae). J. Lepid. Soc. 45:234-236. Sannino, L. & B. Espinosa. 1999. On the morphology ol Noctua comes (Lepidoptera: Noctuidae). 11 Tabacco. 7:35-43. Waring, R, M. Townsend, & R. Lewington. 2003. Field guide to the moths of Great Britain and Ireland. British Wildlife Publish- ing, Hampshire. 432pp. Jeffrey P. Crolla, 413 Jones Ave., Toronto, Ontario, Canada M4J 3G5; email: crollaj@rogers.com Received for publication 3 July 2007, revised and accepted 4 December 2007. ERYNNIS FUNERALIS OVIPOSITS ON EXOTIC ROBINIA PSEUDOACACIA IN WESTERN ARGENTINA Additional key words: Fabaceae Butterflies are adapting to exotic host plants world- wide, including high elevations in the Andes (Shapiro 2006) and in the South American subantarctic (Shapiro 1997). This note reports the apparently widespread use of the naturalized North American tree Black Locust ( Robinia pseudoacacia L. (Fabaceae)) as an oviposition substrate and presumptive host plant of a presumably native skipper in western Argentina. In late afternoon on 24 January 2008 at Chos Malal, Neuquen Province, I watched a female Erynnis funeralis (Scudder & Burgess) lay three eggs in succession on coppice growth of R. pseudoacacia in town. Alerted to this behavior, I then observed another female in a different part ol town lay one egg on this plant three hours later. I subsequently saw repeated instances of oviposition, always on growth less than 4m tall and often in shade, at Las Lajas, Neuquen; in the city of Mendoza, Mendoza Province; and around Calingasta and Barreal, San Juan Province, all over the next three weeks, for a total of >30 ovipositions by at least 8 different females. Though the species was common, I never observed oviposition on other substrates. Pastrana (2004) includes this plant as a host based on Aravena (1983), adding that that record might be based on reared material provided by J. Williamson from the Province of LaPampa. Scott (1986) lists this as a host of E. zarucco (Lucas), at that time considered conspecific, in the United States. He also lists Robinia neomexicana A. Gray as a host of E. funeralis. Although alfalfa ( Medicago sativa L., Fabaceae) is the most widely-cited host of E. funeralis in both the United States and Argentina, and is regularly visited as a nectar source, I have never seen any trace of oviposition or pre- oviposition behavior directed toward it in 30 years’ experience in Argentina. Black Locust is widely naturalized, having escaped from urban cultivation in Argentina, and is routinely found as a participant in synthetic woody riparian communities recruited from the horticultural flora in irrigated zones in the arid and semiarid west. Erynnis funeralis is a consistent inhabitant of these communities as well as appearing in urban gardens and parks; its distribution in western Argentina is broadly concordant with that of Robinia pseudoacacia . A significant element of the western regional fauna is similarly restricted to 108 Journal of the Lepidopterists’ Society irrigated zones in association with naturalized weedy hosts (Shapiro, unpublished data). It is not known if this butterfly is native to the region or is itself naturalized; it is the only member of its genus in the Southern Cone of South America. Literature Cited Aravena, R.O. 1983. Insectos que perjudician a la flora lenosa de la Provincia de La Pampa. V Congreso Forestal Argentino, La Pampa, 4: 315-321. Pastrana, J.A. 2004. Los Lepidopteros Argentinos: Sus Plantas Hospedadoras y otros Substratos Alimenticios. South American Bio Control Lab, USDA and Sociedad Entomologica Argentina, Buenos Aires. 334 pp. Journal of the Lepidopterists' Society 62(2), 2008, 108-110 Scott, J.A. 1986. The Butterflies of North America. Stanford Univer- sity Press, Stanford, CA. 583 pp. Shapiro, A.M. 1997. Impactos antropogenicos sobre la fauna de mari- posas (Lepidoptera, Rhopalocera) de Patagonia austral y Tierra del Fuego. Anales Instituto de la Patagonia (Punta Arenas, Chile), sec. Cs. Nat. ,25: 117-126. . 2006. Use of an exotic weed as an oviposition substrate of the high-Andean Pierid Phulia nymphula. J. Lepid. Soc. 60: 100-101. Arthur M. Shapiro, Center for Population Biology, University of California, Davis, CA 95616; e-mail : amshapiro@ucdavis.edu Received for publicationl9 February 2008, revised and accepted 3 July 2008. COMMENTS ON LARVAL SHELTER CONSTRUCTION AND NATURAL HISTORY OF URBANUS PROTEUS LINN., 1758 (HESPERIIDAE: PYRGINAE) IN SOUTHERN FLORIDA. Additional key words. Egg stacking, hostplant, oviposition, clutch size. The Bean Leaf Roller ( Urbanus proteus Linn.) is a common and widespread skipper (Hesperiidae) found from southern United States south to Argentina (Smith et al. 1994). Early observations on its natural history (Seudder 1889) have been supplemented with details from various parts of its range (Greene 1970, 1971a; Kendall 1965; Moss 1949; Riley 1975; Skinner 1911; Smith et al. 1994; Young 1985), particularly in Florida (Quaintance 1898), where it is a pest on leguminaceous crops (Green 1971b; Quaintance 1898; Watson &Tissot 1942) and where there exist documented seasonal movements (Urquhart & Urquhart 1976). Like most skippers (Greeney & Jones 2003), the larvae of U. proteus construct and live in shelters made from the leaves of the food plant, but only two authors have described or pictured these shelters in any detail (Quaintance 1898; Young 1985). In fact, detailed knowledge of larval shelter construction for most skippers is weak or nonexistent for all but one widely distributed North American species, Epargyreus clams Cramer, 1775 (Jones et al. 2002; Lind et al. 2001;Weiss et al. 2003). As shelters may prove useful in resolving phylogenies (Greeney & Jones 2003), here we present our observations of shelters from a population of U. proteus in southern Florida. We made observations at Burns Lake Campground (25°53’N, 81°13’W) in Big Cypress National Preserve, Collier County, Florida. On 30 December 2005, at 14:15, we observed a female U. proteus ovipositing on the under surface of a leaflet of Vigna luteola (Jacq.) Bentham (Leguminaceae). She laid three dull yellow eggs in an evenly spaced row, and then flew out of sight. Tl lis observation prompted us to search foliage of other V. luteola plants, and resulted in the discovery of 26 additional clutches of hatched and unhatched eggs. At hatching, larvae consume only the top portion of the eggs (pers. obs.), and we were able to use the remaining egg fragments to determine clutch size from all 27 clutches (mean = 2, SD = 1 .1, range = 1-5). Most clutches were located on the under surface of mature leaves (n = 24), but occasionally on leaf petioles (n = 3). Within a clutch, eggs were placed adjacent to (touching) or up to 1 mm from other eggs. One exception was a clutch of three eggs found stacked end to end such that only the bottom egg was attached to the leaf surface (Fig. 1). Similarly, Quaintance (1898) reported a clutch size of 1-6 and noted that eggs were frequently laid in a stacked fashion, 3-4 eggs high. Young (1985), however, recorded only single egg clutches in Costa Rica. In addition to the eggs, we found a total of 50 larvae representing the following instars: 36 first, 8 second, 3 third, 2 fourth, and 1 fifth. We removed larvae from their shelters and carefully determined their ages using the prior experience of HFG with the larvae of related species. We also watched as 3 first-instars constructed new shelters after removal from their original shelters. By examining shelter construction and comparing our observations to previously constructed shelters, we determined that larvae build 3-5 shelters as they develop, and that these belong to three shelter types. First through third instars were found inside shelters built by excising a small triangular portion of the leal margin and creasing it into a tent-shaped lid (Greeney & Jones 2003; group III, type 10, two-cut stemmed Volume 62, Number 2 109 Fig. 1. Three Urbanus proteus egg shells found on the under side of a leaf. Burns Lake Campground, Collier County, Florida, Decem- ber 2005. shelters). These U. proteus shelters were, in faet, very similar to shelters described for E. clarus (Weiss et al. 2003), and would be considered the same type under the classification of Greeney & Jones (2003). Like E. darns, the shelter cuts of U. proteus were always oriented in a distinct fashion in relation to the leaf base; the longest cut always being distal. The most obvious and consistent difference we found was the lack of a “notch” in the cut closest to the leaf petiole in shelters built by U. proteus larvae (Fig. 2). Early instar shelters were still “tented” into a distinct peak, however, by pinching together (using multiple silk ties) a small section along the margin of the shelter lid. The result was a shelter similar in appearance to that built by E. clarus, but arrived at by slightly different means (ie. without the notch). Fourth instars were found, one each, inside a shelter created by silking two leaves together (group I, type 4, two-leaf shelter) and one formed by silking several leaves together (group I, type 3, multi-leaf shelter). We found the single fifth instar feeding adjacent to several leaves silked together (group I, type 3 shelter) at around 17:45. Our observations bring to light several important aspects of egg laying and larval shelter building. Firstly, species building superficially similar shelters may use slightly different cut patterns or construction techniques to arrive at the finished product. Therefore, shelters will prove useful in testing phylogenetic hypotheses (Greeney & Jones 2003) only if we examine shelters and their construction in much more detail than previously reported (but see Greeney & Warren 2003, 2004; Lind et al. 2001; Weiss et al. 2003). Secondly, our observations, and observations of late instars of other pyrgines (HFG unpublished) and coeliadines (Common & Waterhouse 1972), suggest that there may be little difference between the “type 3" and “type 4” shelters distinguished by Greeney & Jones (2003); these shelter types being defined by the number of leaves included in the shelter. In later larval stadia, U. proteus silks together two or more leaves or leaflets into a silk-lined pocket (this study, Quaintance 1898). Young (1985), however, observed only two leaves used in late instar shelter construction. We conclude, therefore, that the number and arrangement of the various leaf parts used is likely related to the relative size and shape of the host plant leaves rather than to any innate shelter building behavior. In other words, larvae simply spin silk, pulling leaves (or parts thereof) around themselves until they are sufficiently covered. Similarly, E. clarus shows variation in the number of leaves used in late instar shelters, varying with size of the host plant leaf (M. Weiss pers. comm.). Based on these observations , we suggest that “type 3” and “type 4” shelters, as defined by Greeney & Jones (2003), should be merged into one “multi-leaf’ shelter type, regardless of whether the shelter includes two or more leaves or leaflets. Finally, the three egg shells we found stacked end to end showed a different emergence pattern than described in previous observations of lepidopteran opposition . Several species of the nymphalid genus Hamadryas Hiibner are also known to deposit eggs one on top of another, sometimes in chains of more than 10 eggs (Muyshondt & Muyshondt 1975b, 1975c). To Fig. 2. Comparison of cut patterns for first instar shelters of (a) Epargyreus clarus (redrawn from Weiss et al. 2003) and (b) Urbanus proteus. Large arrow points towards the base of the foodplant leaf to show orientation of shelters. Small arrows point to (a) position of notch made by E. clarus to aid in tenting the shelter and (b) position of silk laid down to pinch shelter into a tented peak by U. proteus. Journal of the Lepidopterists’ Society 110 emerge from the eggs, Hamadryas larvae create an opening in the side of the egg. Previous discussions on patterns of egg laying and larval emergence in nymphalids suggest an evolutionary signifigance to the correlation between side emergence and egg stacking: side emergence being necessary to avoid damaging eggs laid above (Muyshondt & Muyshondt 1975a). Our observation of egg stacking in U. proteus showed emergence from the top, suggesting that emergence from the side of the egg is not a necessary adaptive response to eggs laid in stacks. While we were unable to clearly illustrate top-emergence in Figure 2, our direct observations show that this was indeed the case. Figure 2 also shows that the eggs of U. proteus were not laid directly centered above the egg below, as illustrated in Muyshondt & Muyshondt (1975b, 1975c) for Hamadryas. It is possible that this means of attaching stacked eggs represents an alternative adaptation allowing eggs to be laid in stacks. Acknowledgements We thank Diane and Terry Sheldon for their hospitality dur- ing our visit to Florida. The work of HFG is supported in part by the Hertzberg Family Foundation, the Population Biology Foundation, Nature 6c Culture International, and a Rufford Small Grant. The PBNHS continues to provide inspiration and support for our natural history research. The clarity and content of this manuscript were greatly improved by the suggestions of Annette Aiello and Martha Weiss. This is publication number 96 of the Yanayacu Natural History Research Group and is ded- icated to Daniel Sheldon for his encouragement of K. Sheldon’s academic and scientific pursuits. Literature Cited Common, I. F. B. & D. F. Waterhouse. 1972. The butterflies of Aus- tralia. Angus & Robertson Publ., London, U. K. Greene, G. L. 1970. Head measurements and weights of the bean leaf roller, Urbanus proteus (Hesperiidae). J. Lepid. Soc. 24: 47-51. . 1971a. Instar distributions, natural populations, and biology of tire bean leaf roller. Florida Entomol. 54: 213-219. . 1971b. Economic damage levels of bean leaf roller populations on snap beans. ]. Econ. Entomol. 64: Greeney, H. F. & M. T. Jones. 2003. Shelter building in the Hesperi- idae: a classification scheme for larval shelters. J. Res. Lepid. 37: 27-36. . 6c A. D. Warren. 2003. Notes on the natural history of Eantis thraso (Hesperiidae: Pyrginae) in Ecuador. J. Lepid. Soc. 57: 43-46. . & A. D. Warren. 2004. Natural history and shelter building be- havior of Noctuana haematospila (Hesperiidae) in Ecuador. J. Lepid. Soc. 59: 6-9. Jones, M. T., I. Castellanos, 6c M. R. Weiss. 2002. Do leaf shelters always protect cateqrillars from invertebrate predators? Ecol. En- tomol. 27: 753-757. Kendall, R. O. 1965. Larval foodplants and distribution notes for twenty-four Texas Hesperiidae. J. Lepid. Soc. 19: 1-33. Lind, E. M., M. T. Jones, J. D. Long, & M. R. Weiss. 2001. Ontoge- netic changes in leaf shelter construction by larvae of Epargyreus clams (Hesperiidae), the silver-spotted skipper. J. Lepid. Soc. 54: 77-82. Moss, A. M. 1949. Biological notes on some “Hesperiidae” of Para and the Amazon (Lep. Rhop.). Acta Zool. Lilloana 7: 27-79. Muyshondt, A. & A. Muyshondt Jr. 1975a. Notes on the life cycle and natural history of El Salvador. IB. - Hamadryas februa (Nymphalidae-Hamadryadinae). J. New York Entomol. Soc. 83: 157-169. . 6c A. Muyshondt Jr. 1975b. Notes on the life cycle and natural history of El Salvador. II B. - Hamadryas guatemalena Bates (Nymphalidae-Hamadryadinae). J. New York Entomol. Soc. 83: 170-180. . 6c A. Muyshondt Jr. 1975c. Notes on the life cycle and natural history of El Salvador. Ill B. - Hamadryas amphinome L. (Nymphalidae-Hamadryadinae). J. New York Entomol. Soc. 83: 181-191. Quaintance, A. L. 1898. Three injurious insects. Bean leaf-roller, Corn delphax, Canna leaf-roller. Florida Ag. Exp. Sta. Bull. 45: 53-71. Riley, N. D. 1975. A field guide to the butterflies of the West Indies. Demeter Press, Boston, Massachusetts. Scudder, S. H. 1889. The butterflies of the eastern United States and Canada with special reference to New England. Published by the author, Cambridge, Massachusetts. Skinner, H. 1911. The larger boreal American Hesperiidae, including Eudamus, Erycides, Pyrrhopyge and Megathymus. Trans. Am. Entomol. Soc. 37: 169-209. Smith, D. S., L. D. Miller, & | Y. Miller. 1994. The butterflies of the West Indies and south Florida. Oxford Univ. Press, New York. Urquhart, F. A. & N. R. Urquhart. 1976. Migration of butterflies along the Gulf Coast of northern Florida. J. Lepid. Soc. 30:59-61. Watson, J. R. 6c A. N. Tissot. 1942. Insects and other pests of Florida vegetables. Florida Ag. Exp. Sta. Bull. 370: 33-35. Weiss, M. R., E. M. Lind, M. T. Jones, J. D. Long, 6c J. L. Maupin. 2003. Uniformity of leaf shelter construction by larvae of Epar- gyreus clams (Hesperiidae), the silver-spotted skipper. J. Insect Behav. 16: 465-480. Young, A. M. 1985. Natural history notes on Astraptes and Urbanus (Hesperiidae) in Costa Rica. J. Lepid. Soc. 39: 215-223. Harold F. Greeney & Kimberly S. Sheldon, Yanayacu Biological Station l? Center for Creative Studies, Cosanga. Ecuador do 721 Foch y Amazonas, Quito, Ecuador; email: revmmoss@yahoo.com. Received for publication 18 August 2007; revised and accepted 5 December 2007 Obituary Journal of the Lepidopterists' Society 62(2), 2008, 111-113 IAN FRANCIS BELL COMMON (23 JUNE 19] 7 TO 3 JUNE 2006) Ian Francis Bell Common (23 June 1917 to 3 June 2006) was an outstanding Australian entomologist who exerted a major influence on studies of Lepidoptera, not only in Australia but throughout the World. I first met Ian in 1968 when he was researching his book Butterflies of Australia and commenced work with him at Commonwealth Scientific and Industrial Research Organisation (CSIRO), Division of Entomology, Canberra, in November 1970. We worked on many moth groups in the Australian National Insect Collection (ANIC) and our close working association continued through Ians retirement until his death. He was brought up in Toowoomba, Queensland, and after a spell in his father’s business he entered Toowoomba Grammar School and matriculated in 1937. He attended the University of Queensland graduating with a B.A. (first class honors in Philosophy) in 1941 and B. Agr. Sc. in 1945 (with honors in 1947). In 1941 he volunteered for military service but was rejected because of his feet. In 1944 Ian was appointed Research Officer with the Queensland Department of Agriculture and Stock and was employed part time by CSIR Division of Economic Entomology to work under Ken Key to study clothes moths infestations in Brisbane wool stores. A letter to Ian survives from this time written by A.J. Turner, then doyen of Australian amateur lepidopterists, outlining the different clothes moths (Tineidae) known from Australia. Ian continued at the University of Queensland graduating with a M.A. in Philosophy in 1946 and M. Agr. Sei. in 1953. He was later, in 1969, awarded a D. Agr. Sei. by the University of Queensland. From December 1944 to June 1945 Ian worked for the Queensland Department of Agriculture and Stock at Biloela and then at Rockhampton from 1945 to 1948. He worked principally on cotton pests, tomato pests and the yellow-winged locust. His interest in Lepidoptera had developed early and he had tales of collecting with school friends around Toowoomba and there are many specimens dating from his early collecting now in the ANIC. For many years there was a small cabinet, made by Ian from silky oak wood, housing immaculate, minute, reared Graeillariidae collected in his youth but now all incorporated into the main collection. In his formative years and the early years of his career Ian worked without close contact with other lepidopterists. Few people could have developed the skills and knowledge that Ian possessed without close contact with experienced, practising lepidopterists. That he overcame this immense challenge so successfully is a measure of the man. Although this close contact with other lepidopterists was denied to him he greatly admired F.A. Perkins then lecturer in entomology at the University of Queensland. While at university Ian met Jill Dowzer who had come to Brisbane from Rockhampton to study Arts. They met again when Ian was posted to Rockhampton and married in 1946. They had two daughters Frances and Jennifer. Throughout his subsequent life Ian was to benefit greatly from the constant support of his wife and family. Ian was appointed to CSIR Division of Economic Entomology on 25 May 1948 after a State- 112 Journal of the Lepidopterists’ Society Commonwealth tussle eventually involving Prime Ministerial intervention. He became Technical Secretary to A.J. Nicholson, Chief of the Division, and able to work half time on the taxonomy of Lepidoptera. In 1947 the Turner collection had been transferred to the Division of Economic Entomology which was an added incentive for Ian to join the Division. In 1951 he transferred to hill-time work on cereal crop and pasture caterpillars. Ian’s early years with the Division were marked by a series of world-class revisions of pest moth groups in Australia including Heliothis, Agrotis, Persectania, Pectinophora, Scirpophaga and Epiphyas. Also at this time he published his classic work on the bogong moth migration. In the 1960s he started revisionary work on the Australian tortricine Tortricidae following his work on Epiphyas and Merophyas. He later worked extensively on the higher classification of the Lepidoptera culminating in the internationally widely acclaimed Lepidoptera chapter in Insects of Australia (1970 and revised with E.S. Nielsen in 1991). In this phase ol his work he described one new superfamily (Immoidea), two new families (Carthaeidae and Lophocoronidae), one new subfamily (Munychryiinae) and a new tribe (Epitymbiini). Following this he commenced work on the Oecophoridae, the favorite subject of his long and abiding interest in the vast Australian fauna. Throughout his career he also published on relevant or interesting items as they arose. Altogether he published about 100 papers and seven books on Lepidoptera. The first books were the Jacaranda guides Australian Moths (1963, revised 1966) and Australian Butterflies (1964). In 1972 Butterflies of Australia (with D.F. Waterhouse) was published with a revised edition in 1981. This book revolutionized butterfly studies in Australia empowering a growing band of butterfly enthusiasts to make many original discoveries and observations. In retirement he published the massive and internationally significant Moths of Australia (1990), which was awarded the Whitley Medal by the Royal Zoological Society of New South Wales and his three great volumes on the Genera of Australian Oecophorinae (1994, 1997, 2000). Besides his huge published contribution, Ian built the ANIC Lepidoptera collection from Turner’s small, but important and well-identified, collection into the immense resource it is today. Ian’s exquisitely preserved, set and labelled specimens ol microlepidoptera still dominate the ANIC collection. He recognized the need to maintain a good working collection and, whatever the backlog of accessions, always kept a well-sorted, named, core so that the ANIC could maintain an identification capacity across the whole Order. The good order, high curatorial standards and large holdings of the collection attracted many top overseas Lepidopterists to Australia and greatly facilitated their work on the Australian fauna. With Ken Key, Ian helped in establishing protocols for the high quality maintenance and operation of the collection well in advance of their time. To build a collection of a little-known and neglected fauna Ian recognized the need for field work and in the 1960s he and Murray Upton embarked on a series of renowned trips to various parts of Australia. They employed extensively the MV light for the first time and the expeditions were highly efficient camping trips dedicated to collecting with not a minute wasted. They were planned to the last can of beans and nothing was allowed to go wrong. These trips opened the eyes of many to how little was actually known of the fauna. Ian and Murray experimented with many light and sheet combinations and designed very efficient light traps which effectively separated out beetles and other rugged insects from the fragile moths permitting for the first time trapped moths to be obtained in good condition in a warm country where insect activity is intense. Ian introduced the extensive study of moth genitalia to Australia and helped develop new staining techniques and a protocol for the successful mounting of moth wings for detailed study of the venation. He participated in a 1953 expedition with C.B. Williams of Rothamstead, England, to the Pyrenees to observe insect migration followed by an academic year at the University of Cambridge and four weeks at the British Museum (Natural History) to study types. In 1966 he spent an additional six months at the BM(NH) photographing types of Australian microlepidoptera and dissecting the types of numerous Australian Oecophoridae. In 1979 he visited many overseas colleagues and collections and also gave the presidential address at the Lepidopterists’ Society meeting in Fairbanks, Alaska. Through his career Ian attracted dedicated colleagues who helped each other and obtained by synergy more than each could have attained individually. The Lepidoptera unit settled into a three person team, Ian as scientist, an experimental officer (Ted Edwards) and an assistant (most notably Vanna Rangsi), which achieved an efficiency now no longer possible when scientists have to grovel for funding and assistants are seen as short-term and dispensable. On Ian’s retirement this synergy was maintained by his successor Ebbe Nielsen, who encouraged and facilitated some of the most productive projects of Ian’s life. This was continued by Marianne Horak following Ebbe’s Volume 62, Number 2 113 untimely death. Ian had been one of Marianne’s mentors who, with John Dugdale, encouraged her on an entomological career specializing in Tortricidae. Ian always maintained close collaborations with people who used his identifications and advice, overseas colleagues, the State museums, agriculture departments and amateur lepidopterists. He was a wonderful lepidopterist; as ‘at home’ in telling stories of moths and collecting with amateur lepidopterists as he was with discussing the higher classification with distinguished overseas colleagues. He was a master in all branches of the subject. Ian achieved the rank of Chief Research Scientist in July 1974 and retired to Toowoomba in June 1982 when he became an Honorary Fellow of the Division of Entomology and in 2003 became an Emeritus Fellow. In retirement he continued to collect for the ANIC and his later books and papers were retirement projects strongly supported by CSIRO Entomology and the Australian Biological Resources Study. Throughout his career Ian remained a dedicated scientist and had no aspirations to enter administration. He was a member of the Entomological Society of Queensland from 1938 and Secretary 1939-40 and a foundation member of the Australian Entomological Society in 1965 becoming Vice-President in 1969-72, President in 1980-81 and an Honorary Life Member in 1987. He was a member ol the Lepidopterists’ Society from 1949 and was Vice-President 1957, 1st Vice- President 1965, President in 1978-79 and became an Honorary Life Member in 1987. He was a foundation member of the Ecological Society of Australia, a member of the Linnean Society of New South Wales from 1956 and a Fellow of the Royal Entomological Society from 1966. He was also an honorary member of the Sociedad Hispano-Luso-Amerieano de Lepidopterologia from 1982. Ian’s work was most widely recognized through the award of the Karl Jordan Medal by the Lepidopterists’ Society in 1996 for his contributions to the study of Lepidoptera and he became an Officer of the Order of Australia in 2001 for his outstanding contributions to entomology, science and education in the community. He received the Jacob Iliibner Award for Lepidoptera Systematic^ from the Association for Tropical Lepidoptera in 2003. Ian was a wonderful person to work with. He saw the implications and ramifications of taxonomic work (and much else) veiy clearly and often well beyond the view of many contemporaries. He was scholarly, dedicated, thorough, meticulous (a word he employed) as well as very hard-working. Few minutes were wasted. He took great pains to excel in all he did. Yet with this he was indulgent of neophytes provided they had application, interest and enthusiasm. He was courteous, quietly spoken and modest. He was approachable, open handed with his immense knowledge and respectful of other views. He greatly valued the critical faculty which could separate the sound from the unsound. He could also express himself concisely and cuttingly when he found foolishness. Jill Common has kindly made information on Ian’s early years available. A manuscript, an article in The Canberra Times on 29 January 2000 and an article in Qantas in January 2001 all by Brad Collis have been helpful. Biographies in The Lepidopterists' Society- Commemorative Volume (1945-1973), in Biologue No. 24 by Ted Edwards and in Murray Upton’s A Rich and Diverse Fauna have been of great assistance. A useful manuscript source was the nomination for the award of the Order of Australia prepared by the late Ebbe Nielsen. A manuscript biographical note by Ian himself was most valuable. Further biographical sources can be found in Murray Upton’s book. Other original sources are housed in the ANIC Archives. A complete list of Ian’s publications may be found in Greg Daniels’ Bibliography of Australian Entomology 1687-2000. E.D. (Ted) Edwards, CSIRO Entomology, GPO Box 1700, Canberra, ACT 2601, Australia ; email: Ted. Edwa rds @csi ro .an Received for publication 30 September 2006 , revised and accepted 29 Maj 2008. 114 Book Review Journal of the Lepidopterists’ Society Journal of the Lepidopterists’ Society 62(2), 2008, 114-115 LOS LEPIDOPTEROS ARGENTINOS: SUS PLANTAS HOSPEDADORAS Y OTROS SUSTRATOS ALIMENTICIOS. By Jose A. Pastrana. 334 pp. ISBN 987-21319-0-2. $40 US to purchasers in Mercosur countries; $70 to SEA members, otherwise $80 US elsewhere. South American Biological Control Laboratory USDA-ARS and Sociedad Entomologica Argentina, Buenos Aires (from whose Web site it may be purchased). Publication date: 2004. My favorite professor in graduate school was the late William L. (“Bill ”) Brown, Jr., who had a gift for telling it like it was. He frequently admonished his students to beware of what he called “validation by frequency of citation”, the process whereby errors become institutionalized by mere repetition. It is a process that is nowhere more common or more deleterious than in the listing of host plants for phytophagous insects. Back in 1983 I published a note in a Mexican journal, identifying several such errors which had crept into the Mexican literature from ours. The worst offenders in this regard are omnium-gatherum compilations, which rarely exercise discretion in evaluating the included material and all too frequently do not trace the source. Sometimes one compilation will incorporate all the dubious material from previous ones, compounding the problem. In 1972 Arthur Allyn arranged for the posthumous publication of Harrison M. Tietz’s card file of life-history information on North American Lepidoptera, which he had been accumulating during his decades as a faculty member at Pennsylvania State University. By the time it appeared it was 20 years out-of-date, but still valuable. Because it was well-referenced if uncritical, it at least allowed one to trace a variety of howlers that had crept into widespread usage — including some ol the ones I figured in my 1983 Mexican article. I still use my well- thumbed and -annotated copy, but often wish someone would publish a detailed critical addenda and corrigenda. I am not holding my breath, and the science has moved on. Now history has repeated itself in Argentina. Like Tietz, the late Jose Pastrana accumulated a bibliographic file over several decades, but died (at age 87) before it could be prepped for publication. The task of organizing it, standardizing the format and modernizing the taxonomy was shared by several colleagues: Karen Braun, Guillermo Logarzo, Hugo Cordo and Osvaldo Dilorio. They consulted a variety of specialists, including John Brown, Adriana Chalup, Don Davis, Fernando Navarro, Patricia Gentili, Gerardo Lamas (who reviewed the butterflies), Alma Solis and Maria Elvira Villagran. But the task clearly overwhelmed them, and the result is very much less than satisfactory. Plere is Dilorio speaking (my very rough translation): “When the Catalog of Phytophagous Insects of Argentina was being prepared, we found the unpublished manuscript of Pastrana. .. How to determine which records pertained to Argentina? The manuscript mentioned the host plants of each species of Lepidoptera without any indication of localities or bibliographic references to the sources. . ..For a certain number of plants the original source was never determined, though one could detect a certain pattern of repetition of data by Pastrana himself. . ..In future editions or addenda we can add the missing information and corroborate the corresponding plant-insect associations.” In other words, all the usual problems are present here, and more so than in Tietz. And as will become plain, even when sources are documented, they are often inaccessible, so that a real critical evaluation is not possible. Undoubtedly some taxonomic groups are in better shape than others. Since I work on the Argentine Pierini and have published more life-history and biological information on this fauna than anyone else (virtually none ot which is cited by Pastrana! — though my work overlapped his active years), and this group is better- known than most, I have chosen to illustrate the nature and magnitude of the problems by working through the couple of pages devoted to my own little group. My evaluations are based on my own 30 years of work in Argentina, and to save space I will not cite the various pertinent Shapiro publications. What is important is how reliable the data in Pastrana are. If what follows is at all representative. . . p.201: The only given host plants of Hypsochila wagenknechti wagenknechti (Ureta) are “Asteraceas: Aplopappus bailahuen; Senecio sp..” The bug is a Crucifer-feeder, and this hoary error is based on old records of nectar sources, ultimately going back to Ureta himself (?). p.203: Tatochila autodice autodice (Huebner): In addition to legitimate hosts (glucosinolate plants, i.e. Brassieaceae and Tropaeolaceae) many other, dubious records are cited: “Fabaceas ( Medicago sativa ) (Berg, 1895, Anonymous 1930, Lizer and Trelles 1941) (Hayward 1969, ex Joergensen, Biezanko 1959, Viana and Williner 1974); Solanaceas: Cestrum elegans (Biezanko 1959), C. noctumum (Biezanko 1959), C. parqui (Giaeomelli 1915, ex Burmeister 1878), (Lizer Volume 62, Number 2 115 and Trelles 1941, Biezanko 1959, H award 1969, Viana and Williner 1974); C. corymbosum (Berg 1875, Giacomelli 1915, Hayward 1969). Tatochila autodice blanchardii Butler: Lists only Tropaeolaceae, omitting the perfectly valid records on Brassicaceous hosts. Tatochila mercedis mercedis (Eschholtz): Oddly, the text lists the distributional records from the Province ol Neuquen as “doubtful,” when hardly anything in this book is similarly qualified. But the records are accurate! Pluilia nymphula (Blanchard): “Tropaeolaceae: Tropaeolum polyphyllum (Reed, Hayward 1969 ex Reed).” A glucosinolate specialist but apparently confined to plants in rosette growth form, making this exuberant herb highly unlikely. p.204: Tatochila orthodice (Weymer) is recorded from “Brassicaceas”: Brassica sp.; Cheiranthes (sic) annus (sic)(Hayward 1969), Lobulana maritime (Hayward 1969); Tropaeolaceas: Tropaeolum sp. (Hayward 1969). Despite the high degree of specificity, all of these records are wrong. The true host plant of this species remains undetermined but is almost certainly Fabaceous; it is not a feeder on glueosinolate- containing plants. And Cheiranthus are chemically odd, and normally avoided by Pierines. Tatochila stigmadice (Staudinger): again listed on “Brassicaceas (Hayward 1969)”, again incorrectly. Tatochila tlieodice tlieodice (Boisduval) is listed on Tropaeolaceae, attributed to Giacomelli (1915). ft is strictly a legume feeder. Theochila maenacte maenacte (Boisuval) is claimed to be a Brassicaceous feeder (Biezanko, Ruffinelli and Carbonell 1957; Hayward 1969, from them). It isn't. Again, its true host remains unknown but is suspected to be Fabaceous. Most of these errors show clear trains of repetition, eventually converging to Hayward (who in his later years committed many errors, some of which I have documented elsewhere) and thence to Pastrana. There is a clear tendency to assume that "if it’s a White, it eats Crucifers.” In South America this does not work. The attribution of Brassicaceous hosts to Legume feeders is actually repeated in the entry for Colias vautliieri (Guerin), which lists “Brassicaceas (Havrylenko 1949)!" (It also lists alfalfa, which this species does not eat, attributing the record to Crouzel and Salavin 1969.) The very persistent records of Tatochila autodice on the Solanaceous plant Oestrum are a special problem that needs to be dealt with definitively one way or the other. The chemistry is so outrageously different that the association must be viewed as highly unlikely at best. This volume is valuable for its huge bibliography of often very obscure references, most of which, alas!, are unobtainable via interlibrary loan services within the United States (I’ve tried). (The most obscure ones cited below are just as cited by Pastrana, if you feel inclined to push the envelope of your favorite retrieval system.) II you can't get them, you can just go ahead and cite them like everybody else, and keep the old errors in circulation to continue to confound those of us trying to study the interaction of coevolution and phylogeny! I consider it a sign of Divine intervention that Braby and Trueman (2006) did not consult this porqueria when they compared host relationships to molecularly- inferred Pierid phylogeny. May others with similar objectives do the same! Literature Cited Anonymous. 1930. Principales parasitos que danan el cultivo de la al- falfa en la Republica Argentina. Ministerio de Agricultura, Sec- cion Propaganda e Iniormes, Direction General de Agricultura y Defensa Agricola, Section Policia de los Vegetales. Mayo, 645: 1-19. Berg, C. 1875. Lepidopteros patagonicos observados en el \laje de 1874. IV. Acta Academia Nacional de Ciencias Exactas, Universi- dad Cordoba 1: 63-101. — . 1895. Revision et description des especes argentines et chili- ennes du genre Tatochila Butler. Anales Museo Nacional de His- toria Natural, Buenos Aires, 4: 217-255. Biezanko, C.M. 1959. Pieridae da Zona Missioneira de Rio Grande do Sul. IB. Arquiv. Ent.. Pelotas (B): 1-12. — . A. Ruffinelli & C.S. Carbonell. 1957. Lepidoptera del Uruguay. Revista Fac. Agron., Montevideo 46: 3-149. Braby,M.F. & J.W.H. Trueman. 2006. Evolution ol larval host plant association and adaptive radiation ol Pierid butterflies. Journal ol Evolutionary Biology 19: 1677-1990. Burmeister, IPG. 1878. Description physique de la Republique Ar- gentine, d’apres des observations personelles et etrangeres. 5. Lepidopteres, 1. partie contenant les diurnes, erepusculaires et Bombycoides. Buenos Aires, vi + 526pp. Crouzel, I.S. de, & R.J. Salavin. 1969. Llave dilematica para el re- conocimiento de los cineo estadios larvales de la "isoca de la al- falfa," Colias lesbian (F. ). DIA, Buenos aires, 256: 1-5. Giacomelli, E. 1915. El genero Tatochila: lo que sabemos y lo que ig- noramus tie el. Anales Museo Nacional de Historia Natural, Buenos aires, 26: 403—415. Havrylenko, D. 1949. Insectos del Parque Nacional Nahuel Huapi. Ministerio de Obras Publicas, Administration General tie Par- ques Nacionales y Turismo. Buenos Aires. 53pp. Hayward, K. 1969. Datos para el estudio de la ontogenia tie Lepi- dopteros argentinos. Miscel. Inst. Miguel Lillo, Tucuman, 31: 1-142. Lizer, Y. & C. Trelles. 1941. Insectos y otros enemigos de la quinta. Enciclopedia Agropecuaria Argentina 2: 214pp. Shapiro, A.M. 1983. Errores en plantas huesped de mariposas mexi- canas como una extrapolacion de la literature estadounidense. Revista Mexicana de Lepidopterologia 8: 47-48. Tietz, H.M. 1972. An Index to the Described Life Histories, Early Stages and Hosts of the Maerolepidoptera of the Continental United States and Canada. 2 vol. Allyn Museum of Entomology, Sarasota, FL. 1041pp. Viana, M.J. & G.J. Williner. 1974. Evaluation de la fauna entomo- logica y aracnologica de las Provincias Cuyanas — Primer Comu- nicacion. Acta Cientif., Ser. Ent. 5: 1—39. ARTHUR M. SHAPIRO, Center for Population Biology, University of California, Davis, CA 95616. e- mail: amshapiro@ucdavis.edu 116 Journal of the Lepidopterists’ Society Journal of the Lepidopterists’ Society 62(2), 2008, 116-117 PAPILLONAGES: UNE HISTOIRE CULTURELLE DU PAPILLON. By Nicolas Witkowski. 143 pages, copiously illustrated; 10.5” x 11.75”. ISBN 978-2-02-089821-8; EUR 40.00. Seuil, Paris. Publication date: 4 October 2007. This is, to my knowledge, only the second attempt at a coffee-table “cultural history” of butterflies. I reviewed the other (Manos-Jones,2000) in the News (Shapiro, 2001). Its author was a mere autodidact-amateur. Nicolas Witkowski, however, is a French Intellectual and as such his book demands a more formal review. Witkowski is a professor of physics, a cultural historian, a popularizer of science and a translator of Stephen Jay Gould. His forte is drawing connections between scientific themes and cultural trends, a very French Intellectual thing to do. In this visually stunning book he displays his virtuosity. To quote from the jacket blurb (all the translations that follow are my own): Symbol of beauty and lightness, but also emblem of the soul and of metamorpho- sis, the butterfly has always fluttered be- tween triviality and seriousness, fickleness and profundity, debauchery and meta- physics. Eveiy epoch has conferred its own particular “take” on this ambiva- lence. Our own has placed the butterfly somewhere between chaos theory and a snarky tattoo the butterfly has always adapted itself to the feel of the times and offers a faithful mirror of our most secret anxiety. . . The book is divided into nine chapters, each accompanied by sidebars and digressions and a wonderful array of color (often full-page) illustrations. Because of Witkowski’s cross-cutting style, only veiy rough descriptions of the chapters are possible: I: The invention of the butterfly. Very early representations of the butterfly in art, from cave paintings through the Middle Ages and across cultures and continents. Inaddition to an illuminated manuscript (by Jean Bourdiehon, the most distinguished ol the French artists who incorporated Lepidoptera in such work — butterflies are much commoner in books produced in Ghent and Bruges, but this is after all a French product!), this chapter reproduces a veiy rare Yuan Chinese scroll (circa early 14th Century) with an anatomically-correct swallowtail. Almost all butterflies in East Asian art are highly stylized; this is an extraordinary exception. II: The ephemeral and the immortal. Butterflies in European Renaissance art, from the “busy” bouquets ol the Dutch still-life masters to idealistic and romantic works of the Masters. (Includes a two-page spread of a bilateral gynandromorph ol Cymothoe sangaris and a discussion ol its sexual resonances.) III. The woman chrysalid. Focuses on the life and work of Maria Sibylla Merian, reproducing several of her exuberant plates and contrasting their dynamism with the usual static portrayal of insects in isolation. Also discusses Albert Seba and reproduces a painting by Jan van Kessel in which mounted butterflies and other insects are being displayed. IV. A butteifly. A study of the butterfly-and-moth- haunted dreanrseapes of the artist Henry Fuseli. V. Cunning hunts. An examination of butterfly collecting as it developed beginning in the 17th Century, particularly in the tropics, and how it fed into the Darwinian revolution. Includes a reproduction of a tropical swallowtail from Alexander Marshall’s famous manuscript (1660) in the Philadelphia Academy of Natural Sciences. VI. The moral of the butteifly. Female entertainers at the turn of the century dressed as butterflies; cartoons and illustrations ol fables and nursery rhymes; Jean- Ilenri Fabre as raconteur of true-life butterfly fables; Dante Gabriel Rossetti as romanticizer. The strangest thing in this chapter is a macabre painting by Felicien Rops, a fantasy melding butterfly, woman, and death. VII. Nabokov’s blues. Familiar territory thanks to recent books by others. Includes some of V.N. s fanciful butterfly sketches (juxtaposed with contemporary butterfly tattoos), and the Meadow Brown with the birds head from Hieronymus Bosch, featured in a Fife magazine article about Nabokov in 1947. VIII. The wings of chance. A riff on Edward Lorenzs butterfly metaphor in chaos theory, now almost universally known but seldom understood. IX. Under the sign of the butteifly. A summing-up. The tone is best conveyed by a fairly extended quote which, however, still falls within the boundaries of “fair use:” At the end of this personal voyage transformed into a cultural history, I rediscover in the Western approach to the butterfly the old drama of which Goethe was the harbinger: What can one learn of Nature by analysis? What more can one get besides a cadaver impaled on a blue steel pin? What remains of tire magic of flight under the frozen gaze of the researcher? The quarrel is as old as modern science — more or less four centuries — but today it takes on a new sense: the era of great butterfly massacres is at hand [referring to the crisis of biodiversity — A.M.S.]. . ..What science is worth what one sacrifices. . .the art of seeing, of loving that which one sees? The beauty of the butterfly, immediate, presenting itself to passive contemplation, is irremediably destroyed by Volume 62, Number 2 117 any effort at analysis. How can one be fully satisfied by such fleeting joy? How can one resist the temptation to capture ... and to crush between one’s fingers the object of one’s love? There are the important questions that underlie our everlasting interest in the wonderful butterfly, the precious “little soul” that always causes us such pain because we cannot catch it. If you are not used to the flowery, intricate idiom of French intellectual discourse this passage may turn you off. Even if you are used to it, it may do so. Whether or not you take all the pretentiousness seriously (and you are permitted, as a mere Anglo-Saxon, to dismiss it as airy twaddle), this is a magnificent book and highly recommended for your coffee table, whether or not you read French. I say this even though it omits any reference to my favorite cultural touchstone for the butterfly in French — the nursery rhyme that goes “Faites pipi sur le gazon/pour embeter les papillons.” (Make weewee on the grass, it drives the butterflies nuts.) The last image in the book is very disquieting and in the tradition of Medieval Memento mori. It is a two- page photograph, larger than life, of a box of Dermestidized 19th-Century Morpho specimens - glorious pieces of blue wings and meticulously handwritten labels adrift in a sea of beetle frass. Make of it what you will. Literature Cited Manos-Jones, M. 2000. The spirit of butterflies: Myth, magic and art. Harry N. Abrams, Inc., New York. 144 pp. Shapiro, A.M. 2001. [Review of Manos-Jones.] News of the Lepi- dopterists’ Society 43 (1): 16-17. Arthur M. Shapiro, Center for Population Biology, University of California , Davis , CA 95616. e-mail: amshapiro@ucclavis.edu EDITORIAL STAFF OF THE JOURNAL Brian Scholtens, Editor Biology Department College of Charleston, 58 Coming Street, Charleston, South Carolina 29424-001 1, USA scholtensb@cofc.edu Peg Toliver, Layout Editor Natural Imprints 706 Lake Road Eureka, Illinois 61530 naturimp @ mtco . com Phil DeVries, Book Review Editor Department of Biological Sciences University of New Orleans, 2000 Lakeshore Dr. New Orleans, Lousiana 70148-0001 USA pdevries@uno.edu Associate Editors: Gerardo Lamas (Peru), Kenelm W. Philip (USA), Robert K. Robbins (USA), Felix Sperling (Canada), David L. Wagner (USA), Christer Wiklund (Sweden), Carla Penz (USA), Andrew Warren (USA), NOTICE TO CONTRIBUTORS Contributions to the Journal may deal with any aspect of Lepidoptera study. 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A purchase order for reprints will accompany proofs. Page charges: For authors affiliated with institutions, page charges are $50 per Journal page. For authors without institutional support, page charges are $25 per Journal page. For authors who are not members of the Society, page charges are $75 per Journal page. Authors unable to pay page charges for any reason should apply to the editor at the time of submission for a reduced rate. Authors of Book Reviews and Obituaries are exempt from page charges. Correspondence: Address all matters relating to the Journal to the editor. Address book reviews directly to the book review editor. PRINTED BY THE ALLEN PRESS, INC., LAWRENCE. KANSAS 66044 U.S.A. SMITHSONIAN INSTITUTION LIBRARIES III III III CO 9C o CO CO 144 5, 373 CONTENTS Comparative Studies on the Immature Stages and Developmental Biology of Five Argynnis spp. (surgenus Speyeria ) (Nymphalidae) from Washington David G. James 61 Hawkmotii Fauna of a Northern Atlantic Rain Forest Remnant (Sphingidae) Jose Araujo Duarte Junior and Clemens Schlindwein 71 Notes on Papilio machaon alias ka (Papilionidae) Populations Near Fairbanks, AK Shannon Murphy 80 Differential Antennal Sensitivities of the Generalist Butterflies Papilio glaucus and P. canadensis to Host Plant and Non-host Plant Extracts R. J. Mercader, L. L. Stelinski and J. M. Scriber 84 Description of the Immature Stages of Methona confusa confusa Butler, 1873 and Methona curvifascia Weymer, 1883 (Nymphalidae, Ithomiinae) from Eastern Equador Ryan I. Hill and Luis A. Tipan 89 Experimental Design and the Outcome of Preference-Performance Assays, with Examples from Mitoura Butterflies (Lycaenidae) Matthew L. Forister 99 General Notes Noctua comes in Ontario: An Introduced Cutworm (Noctuidae: Noctuinae) New to Eastern North America Jeffrey P. Crolla 106 Erynnis funer,\lis Oviposits on Exotic Robinia pseudoacacia in Western Argentina Arthur M. Shapiro 107 Comments on Larval Shelter Construction and Natural History of Urbanus proteus Linn., 1758 (Hesperiidae: Pyrginae) in Southern Florida Harold F. Greeney and Kimberley S. Sheldon 108 Obituary Ian Francis Bell Common (1917—2006) by E. D. (Ted) Edwards 111 Book Reviews Los Lepidopteros Argentinos: Sus Plantas Hospedadoras y Otros Sustratos Alimenticios. By Jose A. Pastrana. Arthur M. Shapiro 114 Papillonages: Une Histoire Culturelle du Papillon. By Nicolas Witkowski. Arthur M. Shapiro 116 © This paper meets the requirements of ANSI/NISO Z39. 48-1 992 (Permanance of Paper).