(ISSN 0161-8202) Journal of ARACHNOLOGY PUBLISHED BY THE AMERICAN ARACHNOLOGICAL SOCIETY VOLUME 37 2009 NUMBER 2 THE JOURNAL OF ARACHNOLOGY EDITOR-IN-CHIEF: James E. Carrel, University of Missouri-Columbia MANAGING EDITOR: Douglass H. Morse, Brown University SUBJECT EDITORS: Ecology — Stano Pekar, Masaryk University; Systematics — Mark Harvey, Western Aus- tralian Museum and Ingi Agnarsson, University of Puerto Rico; Behavior — Linden Higgins, University of Vermont; Morphology and Physiology — ^Jeffrey Shultz, University of Maryland EDITORIAL BOARD: Alan Cady, Miami University (Ohio); Jonathan Coddington, Smithsonian Institution; William Eberhard, Universidad de Costa Rica; Rosemary Gillespie, University of California, Berkeley; Charles Griswold, California Academy of Sciences; Marshal Hedin, San Diego State University; Herbert Levi, Harvard University; Brent Opell, Virginia Polytechnic Institute & State University; Norman Platnick, American Museum of Natural History; Ann Rypstra, Miami University (Ohio); Paul Selden, University of Kansas; Matthias Schae- fer, Universitaet Goettingen (Germany); William Shear, Hampden- Sydney College; Petra Sierwald, Field Mu- seum; I-Min Tso, Tunghai University (Taiwan). The Journal of Arachnology (ISSN 0161-8202), a publication devoted to the study of Arachnida, is published three times each year by The American Arachnological Society. Memberships (yearly): Membership is open to all those interested in Arachnida. Subscriptions to The Journal of Arachnology and American Arachnology (the newsletter), and annual meeting notices, are included with membership in the Society. Regular, $55; Students, $30; Institutional, $125 . Inquiries should be directed to the Membership Secretary (see below). Back Issues: James Carrel, 209 Tucker Hall, Missouri University, Columbia, Missouri 65211-7400 USA. Telephone: (573) 882-3037. Undelivered Issues: Allen Press, Inc., 810 E. 10th Street, P.O. Box 368, Lawrence, Kansas 66044 USA. THE AMERICAN ARACHNOLOGICAL SOCIETY PRESIDENT: Paula Cushing (2007-2009), Zoology Department, Denver Museum of Nature & Science, Denver, CO 80205-5798 USA. PRESIDENT-ELECT: Rosemary Gillespie (2007-2009), Environmental Science, Policy & Management, Division of Organisms and Environment, University of California, Berkeley, CA 94720-3 1 14 USA. MEMBERSHIP SECRETARY: Jeffrey W. Shultz (appointed). Department of Entomology, University of Maryland, College Park, MD 20742 USA. TREASURER: Karen Cangialosi, Department of Biology, Keene State College, Keene, NH 03435-2001 USA. SECRETARY: Alan Cady, Dept, of Zoology, Miami University, Middletown, Ohio 45042 USA. ARCHIVIST: Lenny Vincent, Fullerton College, Fullerton, California 92634 USA. DIRECTORS: Elizabeth Jakob (2007-2009), Greta Binford (2007-2009), Matt Persons (2008-2010) PAST DIRECTOR AND PARLIAMENTARIAN: H. Don Cameron (appointed), Ann Arbor, Michigan 48105 USA. HONORARY MEMBERS: C.D. Dondale, H.W. Levi, A.F. Millidge. Cover photo: Marked female harvestman Chavesincola inexpectabilis with egg from southeastern Brazil. Photo by G. Machado. Publication date: 16 July 2009 ©This paper meets the requirements of ANSI/NISO Z39.48-1992 (Permanence of Paper). 2009. The Journal of Arachnology 37:127-134 Reproductive behavior of Chavesimcola mexpectabilis (Opiliones, Gonyleptidae) with description of a new and independently evolved case of paternal care in harvestmen Tais M. Nazareth: Programa de Pos-graduagao em Ecologia e Conservagao de Recursos Naturals, Universidade Federal de Uberlandia, CP 593, 38400-902 Uberlandia, MG, Brazil Glauco Machado': Departamento de Ecologia, Institute de Biociencias, Rua do Matao, trav. 14, n° 321, 05508-900, Sao Paulo, SP, Brazil Abstract. In this paper, we investigate the reproductive behavior of the gonyleptid Chavesincola inexpectahilis Soares & Soares 1946 (Heteropachylinae) and provide basic descriptive information about courtship, copulation, oviposition, and paternal care. Like most gonyleptids, males of C. inexpectahilis have a strong armature on the fourth pair of legs and use their spines and apophyses to fight other males and to repel them from their nesting sites. The mating pair interacts briefly before copulation, but the male touches the female both during and after penetration while she oviposits. The oviposition behavior differs markedly from that of other Laniatores: females hold the eggs on the chelicerae before depositing them on the substrate. After oviposition, the eggs are left under the guard of the male to defend against attack from cannibalistic conspecifics. Mapping the available data on reproductive biology of the Gonyleptidae on the phylogeny of the family, it is possible to infer that paternal care has evolved at least three times independently: once in the clade Progonyleptoidellinae + Caelopyginae, once in the Gonyleptinae, and once in the Heteropachylinae, which occupies a basal position within the group. Keywords: Copulation, courtship, evolution, Heteropachylinae, oviposition, sexual dimorphism The great majority of the harvestmen species reproduce sexually, although some species reproduce asexually by parthenogenesis (e.g., Phillipson 1959; Tsurusaki 1986). Fertilization is internal and the transfer of sperm may occur indirectly through spermatophores in representatives of the suborder Cyphophthalmi, or directly by means of a long and fully intromittent male genitalia in the suborders Eupnoi, Dyspnoi, and Laniatores (Machado & Madas-Ordonez 2007). Courtship before intromission is generally quick and tactile, but there are some cases in which males offer a glandular secretion produced in their chelicerae before copulation as a nuptial gift for their mates. Courtship during intromission, on the other hand, may be longer and involve leg tapping and rubbing. Copulation is often followed by a period of mate guarding in which the female is held or constantly touched by the male (see table 12.1 in Machado & Macias-Ordofiez 2007). Females may lay their eggs immediately or in the months after copulation, and the oviposition strategies seem to be related to the length of the ovipositor. Most species of the suborders Cyphophthalmi and Eupnoi have a long ovipositor and hide their eggs inside small holes in the soil, trunk crevices, or under stones. Representatives of the suborders Dyspnoi and Laniatores, constrained by their short ovipositor, lay their eggs on exposed substrates such as leaves, wood, and rocks (Machado & Madas-Ordonez 2007). The forms of parental care range from microhabitat selection for oviposition to active egg guarding by a parental individual. In most species, eggs are laid singly in shallow natural cavities or are covered by debris by the female. In some species, however, females lay eggs in a single large clutch and brood eggs throughout the embryonic development, remaining with the newly hatched nymphs for some days until they disperse (Machado & Raimundo 2001). Maternal care has been reported for many families of the suborder Laniatores, especially among the ‘ Corresponding author. E-mail: gIaucom@ib.usp.br Neotropical representatives of the superfamily Gonyleptoidea (see Machado & Warfel 2006). While maternal egg guarding is widespread among arach- nids, exclusive paternal care is present only in the order Opiliones (Machado et al. 2004). Male assistance has evolved in at least five families belonging to three non-closely related superfamilies of the suborder Laniatores: Travunioidea, Epedanoidea, and Gonyleptoidea (Machado 2007). Within Gonyleptidae, which comprises nearly 1,000 species and corresponds to the largest family of Laniatores, there are eight cases of paternal care recorded so far (Machado & Macias-Ordonez 2007). In this paper, we investigate the reproductive behavior of the gonyleptid Chavesincola inex- pectabilis Soares & Soares 1946 (Heteropachylinae) and provide basic descriptive information about courtship, copu- lation, oviposition, and paternal care of this species. This study is the first description of the reproductive biology of a representative of the subfamily Heteropachylinae and the results obtained here represent a new and independently evolved case of paternal care in gonyleptid harvestmen. METHODS In all, 9 females and 14 males of C. mexpectabilis were collected along the borders of a small (ca 8 ha) urban forest fragment in Santa Teresa city (19°58'S; 40°32'W; elev. 675 m), Espirito Santo state, southeastern Brazil. The individuals were found under rotting logs and piles of tree fern trunks discarded from a green house nearby. They were brought to our laboratory in the Natural History Museum at Universidade Estadual de Campinas (Sao Paulo state, Brazil) and were maintained in a communal terrarium (40 X 90 cm base, 20 cm height) containing soil, small pieces of tree fern trunks collected in the study site, and 10 artificial nests built in clay blocks (with 6 X 2 cm base, 3 cm height). Each mud nest had a central hole (1 cm in diameter and 2 cm depth) crossing the clay block from side to side. These blocks were placed against 127 128 THE JOURNAL OF ARACHNOLOGY Figures 1, 2. — 1. Marked female of the harvestman Cliavesiiicola mexpectahilis everting the ovipositor and manipulating the egg with the chelicerae while serapping the substrate of the nest with her first pair of legs. 2. Another marked female covering a recently laid egg with debris. Behind the female, it is possible to see the nest entrance (A) and the guarding male walking around while she is ovipositing (B). Both photos were taken through the glass wall of the terrarium. Scale bars = 5 mm. the glass wall of the terrarium so that it was possible to observe the harvestman behavior inside the nests through the glass (Figs. 1, 2). These mud nests simulated natural cavities in roadside banks, which are occupied by males of another Heteropachylinae species from Espirito Santo {Pseiidopucrolia sp.). Males of Pseiidopucrolia take care of the eggs laid by females inside these natural cavities, and the possession of nests is crucial for their reproductive success (Nazareth & Machado unpubl. data). During the study period, the abiotic conditions in the laboratory were (mean ± SD): temperature of 25.5 ± 1 .2° C, humidity of 82.0 ± 5.4%, and photoperiod of 13L:11D. Individuals were measured (dorsal scute width) and individually marked on their dorsal scute with colored dots NAZARETH & MACHADO— REPRODUCTIVE BEHAVIOR OF CHA VESINCOLA 129 of enamel paint. They were fed pieces of dead cockroaches and an artificial diet for ants (Bhatkar & Whitcomb 1970) three times a week. The mud nests were individually numbered and, at each observation, the identity of the individuals inside each nest was recorded. Behavioral data are based on nearly 50 h of ad libitum observations (sensu Altman 1974), of which 43 h were conducted at night (from 18:00 to 00:00 h) when individuals were more active. Nocturnal observations were made with a red lamp to avoid disturbing the animals (cf. Elpino-Campos et al. 2001; Pereira et al. 2004). Continuous recording (sensu Martin & Bateson 1993) was made of all relevant behavioral events such as fights between males, copulations, and ovipositions. Voucher specimens of males and females were deposited in the arachnological collection of the Museu de Zoologia da Universidade de Sao Paulo (MZSP), Sao Paulo state, Brazil. RESULTS Nesting. — Ten males were observed occupying and occa- sionally fighting for the ownership of the mud nests. No fight or any kind of aggressive interaction was observed between males outside the nests. Only two males were observed mating: the first one (Ml) achieved copulation after staying in the same mud nest for four consecutive days, and the second (M2), after five consecutive days. These males had a dorsal scute width of 4.99 mm (Ml) and 4.71 mm (M2), and were, respectively, the first and the third largest males in the terrarium (mean male size ± SD = 4.48 ± 0.27 mm; n = 14). On two occasions, as soon as an intruder male entered a mud nest occupied by one of these two males, a brief period of intense mutual tapping with the second pair of legs occurred. After that, the individuals turned their backs to each other and intertwined the fourth pair of legs, which bears many spines and tubercles. In this position, the males seemingly attempted to capsize each other by means of sudden upward movements in which each male brought its femur IV close to the body, pinching his opponent’s fourth pair of legs, a behavior known as “nipping 2” (sensu Willemart et al. 2009). This phase of nipping 2 lasted nearly 30 s in the two fights observed and, in both cases, resident males managed to pull the intruders out of the mud nests. Copulation. — All copulations occurred inside nests, and no sexual interaction between males and females were observed in other places of the terrarium. Most of the females (6 out of 9) were observed copulating at least once. One of them was observed copulating and laying eggs with Ml and M2 and another one was observed laying eggs twice with Ml. Ml copulated at least five times with four different females, resulting in a total of 228 eggs in his mud nest, and M2 copulated at least three times with three different females, resulting in a total of 83 eggs. After the hatching of all nymphs inside his nest, Ml left the mud nest and established a new nesting site under a piece of tree fem trunk (Fig. 3). After 1 1 days, 54 eggs covered by debris (Fig. 4) and in two different stages of embryonic development (according to Machado et al. 2004) were found attached to the undersurface of the tree fem trunk. Since there was no egg under the tree fem trunk before M 1 arrival, the presence of the clutch suggests that Ml copulated with two females or twice with the same female. Ml remained close to the eggs in the trunk nest until they hatched 16 days later. Just before copulation, the male approached the female frontally and intensely tapped her genital opening with his second pair of legs. Meanwhile, the male also gently touched the dorsum of the female with his first pair of legs {n = 2). In one case, touching behavior lasted 30 s and, in the sequence, the male (Ml) grasped the female pedipalps with his own pedipalps. The female raised the front of her body, exposing her ventral region to the genital opening of the male. In this position, the male everted his penis and penetrated the female’s genital opening. The other courtship lasted almost 1 h and, during all this time, the male (M2) touched the female as described above. During most of the courtship, the female bent the front of her body so that it was impossible for the male to penetrate her. Occasionally, she also put her venter in contact with the substrate, also preventing the male from touching her genital opening. Eventually, the male managed to grasp the female pedipalps with his pedipalps and then she spontaneously raised the front of her body allowing penetration. Both copulations lasted nearly 2 min, and during penetra- tion, the male performed intense leg tapping on the dorsum of the female using his first pair of legs, and simultaneously on the female’s hind legs and venter using his second pair of legs. Penetration was apparently terminated by the female when she was able to propel herself backwards with enough force to release herself from the grasp of the male’s pedipalps. Immediately after separation, males continued to tap the dorsum and venter of their partners with the second pair of legs for nearly 2 min. Oviposition. — After copulation, the female generally walked inside the mud nest for nearly 3 min {n = 7), always followed by the male, probably searching for a proper place for egg laying. In the first step of the oviposition, the female everted her ovipositor and placed its tip in contact with her chelicerae for up to 7 min. At the same time, the male, stood behind the female, repeatedly tapped her dorsum using his second pair of legs. Once every 3 min, the male also gently tapped the venter of the female (n = 2 ovipositions); it was not possible to see if the male touched the ovipositor. Next, the female released an egg, which was held on the chelicerae while she scraped the nest’s wall with her first pair of legs (Fig. 1). Every two or three scrapes of the nest’s wall, the female brought the leg to the mouth, probably to clean or humidify the tip of the leg; this process lasted from 7 to 13 min. In the sequence, the female put the egg on the scraped area using her chelicerae and rolled it on the substrate using the first pair of legs until the egg was completely covered by debris, a process that lasted up to 1 min (Fig. 2). After oviposition of each egg, the male walked around inside the nest until the female started to lay the next egg (Fig. 2). At this moment, the male resumed tapping the female using his second pair of legs, as described above. The whole process of oviposition lasted 2 to 4 days (mean ± SD = 2.6 ± 0.7; n = 8), and was interrupted by periods of rest (sensu Elpino-Campos et al. 2001), when both male and female did not interact with each other. After this period, the female abandoned the nest and the eggs were left under the male protection until they hatched 23-24 days later. The mean number of eggs laid in each oviposition was 38.9 (SD = 12.2; n — 8), and the intervals between the two oviposition events of each female ranged from 9 to 12 days (n = 8). 130 THE JOURNAL OF ARACHNOLOGY Figures 3, 4. — 3. Marked male of the harvestman Chavesincola inexpectahilis taking care of eggs laid on a piece of tree fern trunk. The dotted circles indicate the position of the eggs. 4. Detail of the clutch after the addition of more eggs. Note that the eggs are covered by debris (photos by B.A. Buzatto). Scale bars = 5 mm. Paternal care. — Non-guarding males and females were frequently seen walking around in the terrarium at night, and they were observed eating at least 10 times. Guarding males, on the other hand, rarely left their nests to forage at night; when they did (n = 2), they remained within 10 cm of the nest entrance. Additionally, unlike females that ate the cockroach pieces on the spot, guarding males and males that were defending mud nests without eggs took the food to their nests before consumption {n = 6). In one case, a non-guarding male was observed entering a mud nest and seemingly trying to remove some eggs with his pedipalps, probably as an attempt of cannibalism. The guarding male (Ml), which was 2 cm away from the nest entrance, attacked the intruder male using the first pair of legs and pedipalps. The non-guarding male left the nest NAZARETH & MACHADO— REPRODUCTIVE BEHAVIOR OF CHA VESINCOLA 131 without cannibalizing any egg, and was chased by the guarding male for nearly 30 s. After that, the guarding male returned to his nest and remained with the fourth pair of legs blocking the nest entrance for nearly one hour. DISCUSSION When males are in charge of egg brooding, they become a reproductive resource for females and some degree of sex-role reversal may be expected (Owens & Thompson 1 994; Parker & Simmons 1996). In such cases, male-male competition may be less intense and no sexual dimorphism is expected. Although most gonyleptids show strong sexual dimorphism, males being larger and more armed than females, this dimorphism in paternal species of the subfamilies Caelopyginae and Progo- nyleptoidellinae is very subtle. Females of many species have spiny legs and apophyses as long as those of males (e.g., Pinto- da-Rocha 2002), or in other cases, neither sex has any leg armature at all (e.g., Kury & Pinto-da-Rocha 1997). However, strong sexual dimorphism may be found among paternal species of the subfamily Gonyleptinae. In this subfamily, males of some species defend very specific sites (holes in roadside banks and trunks) as nesting sites, and leg armature seems to be involved in the defense of this scarce resource against other males (Machado et al. 2004). Males of C inexpectabilis also defend nesting sites and, as could be expected, males have strong armature on the fourth pair of legs. They use the spines and apophyses of these legs to fight other males and to repel them from the nesting sites. Similarly to males of Neosadocus sp., which also occupy holes in roadside banks as nesting sites (see figs. 2B, C in Machado et al. 2004), males of C. inexpectabilis use the heavily armed fourth pair of legs to block the entrance of their nests and to pinch intruder males. Most descriptions of courtship in harvestmen of the suborder Laniatores lack detailed information, such as which parts of the female body are touched by the male. Even though the courtship behavior of C. inexpectabilis follows the general pattern previously recorded for some gonyleptid harvestmen (see Machado & Macias-Ordonez 2007), here we provide additional information showing, for instance, that males intensively touch the genital opening of the female. It is possible that these touches stimulate the female to open her genital opening, a prerequisite for male intromission among Laniatores. Unreceptive females clearly avoid male touches on the genital opening by lowering the venter to the substrate. On the other hand, receptive females allow the males to grasp them with their pedipalps and raise the front of their bodies so that penetration can occur. The end of the copulation is also apparently determined by the females, when they are able to release themselves from the intromission and from the pedipalpal grasping. In species of Eupnoi, the female may reject intromission, but grasping seems harder to avoid because the male tightly hooks his long, sexually dimorphic pedipalps to the base of female’s legs 11 near the trochanter. Apparently, Eupnoi males rely more on the powerful grasping to initiate copulation with females, whereas Laniatores rely more on precopulatory courtship (discussion in Machado & Macias-Ordonez 2007). Post-copulatory courtship in C. inexpectabilis occurs as males tap on the dorsum and venter of females using their legs. Intense female stimulation both during and after copulation may be viewed as a male strategy to increase the number of eggs fertilized and also increase paternity (Eberhard 1996). Addi- tionally, the total time spent by ovipositing females inside a male’s nest may reach four days, quite a long period when compared to other harvestman species (e.g., Juberthie & Mufioz-Cuevas 1971; Mora 1990; Machado & Oliveira 1998; Willemart 2001). In Pseudopucrolia sp., another Heteropachy- linae species we are studying in our laboratory, males block the entrance of the nest with their bodies and also actively prevent females from leaving (Nazareth & Machado unpub. data). This coercive behavior, associated with repeated copulations, is possibly another male strategy to increase paternity and the number of eggs that one female will lay inside the nest. The oviposition behavior of C. inexpectabilis is markedly different from that of other Laniatores, including representa- tives of the family Gonyleptidae (e.g., Juberthie & Munoz- Cuevas 1971; Machado & Oliveira 1998; Willemart 2001). A unique behavioral feature is that females hold the eggs on the chelicerae before depositing them on the substrate. It is possible that females use secretions from the mouthparts to cover the eggs before their deposition on the substrate to promote the attachment of debris on them or to moisten them with anti-pathogenic compounds, as some centipedes do (Brunhuber 1970; Lewis 1981). The behavior of covering eggs with debris has been previously described for several harvestman species of the families Cosmetidae and Gonylep- tidae that present no care or exclusive maternal care (references in Willemart 2001). The only cases of egg covering reported so far for a paternal species occur in the tryaeno- nychids Karamea spp. (Machado 2007), which are not closely related to the Gonyleptidae (Giribet & Kury 2007). This behavioral trait, therefore, clearly evolved independently in these two families, but in both cases might be related to egg protection by providing camouflage and/or preventing dehy- dration (Willemart 2001; Elpino-Campos et al. 2001). Maternal egg-guarding is a costly behavioral strategy for iteroparous arthropods because it reduces lifetime fecundity by increasing the risk of death from predation and reducing foraging opportunities for guarding females during the long periods of care (Tallamy & Brown 1999; see also Buzatto et al. 2007). Reduction of foraging is one of the main costs paid by guarding females and, according to the “enhanced fecundity hypothesis,” exclusive post-zygotic paternal care may be viewed as a fitness-enhancing gift from males to females because it offers females two direct benefits: the cost-free care of their offspring and the freedom to forage for additional food (Tallamy 2001). After oviposition, eggs of C inexpect- abilis are left under the guard of the male, and females are released to forage and to produce more eggs. The intervals between two consecutive ovipositions ranged from 9 to 12 days, which is almost three times shorter than the interval between two ovipositions in the maternal gonyleptid Dis- cocyrtus oliverioi Soares 1945, which was also studied in captivity where food was always available (Elpino-Campos et al. 2001; G. Machado, unpub. data). This interval is also 10- 15 times shorter than the median interval between two ovipositions in three other maternal gonyleptids studied in the field (where food is supposed to be a limiting factor for female fecundity): Bourguyia hamata (Machado & Oliveira 132 THE JOURNAL OF ARACHNOLOGY Cosmetidae Metasarcinae Heteropachylinae Cobaniinae Bourguyiinae Pachylinae Pachylospeleinae Tricommatidae Mitobatinae Goniosomatinae Gonyleptinae Hernandariinae Sodreaninae Progonyleptoidellinae Caelopyginae Cosmetidae Metasarcinae Heteropachylinae Cobaniinae Bourguyiinae Pachylinae Pachylospeleinae Tricommatidae Mitobatinae Goniosomatinae Gonyleptinae Hernandariinae Sodreaninae Progonyleptoidellinae Caelopyginae Figure 5, 6. — Internal phylogeny of the family Gonyleptidae (modified from Kury 1994 and Pinto-da-Rocha 2002) showing the forms of parental care presented by each subfamily. Behavioral data were mapped using the program Winclada (Nixon 1999), using ACCTRAN (5) and DELTRAN (6) optimization. Since there are no data on the internal phylogeny of some groups, the following assumptions were made: (1) most species of Cosmetidae do not care for the eggs (see table 12.2 in Machado & Macias-Ordonez 2007) and the only case of maternal care reported so far in the family was considered as an autapomorphy (see Goodnight & Goodnight 1976 and Machado & Raimundo 2001 ); (2) although there is a great diversity in the forms of parental care within the subfamily Gonyleptinae (see table 12.2 in Machado & Macias-Ordonez 2007), paternal care was tentatively considered as the plesiomorphic state in order to investigate how this polarity assumption could affect the optimization of this behavioral trait on the tree; (3) the information for the Pachylinae was considered as polymorphic because cases of no care and maternal care are evenly distributed in the species of this subfamily (see table 12.2 in Machado & Macias-Ordonez 2007). NAZARETH & MACHADO— REPRODUCTIVE BEHAVIOR OF CHA VESINCOLA 133 2002), Goniosoma albiscriptum (Willemart & Gnaspini 2004), and Acutisoma proximum (Buzatto et al. 2007). Apparently, the reproductive rate of C. inexpectabiiis females is higher than females of species with maternal care, a likely consequence of their increased foraging rate, but experimental studies are necessary to address this question more carefully. By mapping the available data about reproductive biology on the internal phytogeny of the Gonyleptidae, it is possible to infer that paternal care has evolved two or three times independently in the family, according to the type of optimization (Figs. 5, 6). Since the clutch and the nesting site of paternal species from the subfamilies Gonyleptinae and Progonyleptoidellinae + Caelopyginae are remarkably differ- ent (see discussion in Machado et al. 2004), we believe that DELTRAN optimization, which favors convergence, is the most appropriate scenario for the evolution of male care in the gonyleptids (Fig. 6). According to both Figs. 5 and 6, all cases of paternal care in gonyleptids are derived from no care. For the Heteropachylinae, however, this evolutionary transition should be interpreted cautiously because there is no published information on the reproductive biology of the Andean subfamily Metasarcinae and of the basal monotypic subfamily Cobaniinae. Data on the reproductive behavior of these two subfamilies are crucial to provide both a robust hypothesis about the plesiomorphic form of egg assistance in gonyleptids and a more complete scenario of the transitions between different forms of parental care in the family. ACKNOWLEDGMENTS We are grateful to Thiago Gon?alves-Souza (Toyoyo) for his help in the fieldwork and for hosting us in Santa Teresa, to Bruno A. Buzatto for taking some photos used in this paper, to Ricardo Pinto da Rocha and Adriano B. Kury for sharing unpublished data on the internal phylogeny of the Gonylepti- dae, to Ariovaldo A. Giaretta for helping to map the behavioral characters, and to Rogelio Macias-Ordonez, Alfredo V. Peretti, Katia G. Facure, Roberto Munguia Steyer, Bruno A. Buzatto, Gail Stratton, and two anonymous reviewers for comments on the manuscript. TMN was supported by a fellowship from CAPES and GM has a research grant from Fundai?ao de Amparo a Pesquisa do Estado de Sao Paulo (02/00381-0). LITERATURE CITED Altmann, J. 1974. Observational study of behavior: sampling methods. Behaviour 49:227-265. Bhatkar, A. & W.H. Whitcomb. 1970. Artificial diet for rearing various species of ants. Florida Entomologist 53:229-232. Brunhuber, B.S. 1970. 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The Journal of Arachnology 37:135-138 Reversed cannibalism, foraging, and surface activities of Allocosa alticeps and AUocosa hvasiliemis: two wolf spiders from coastal sand dunes Anita Aisenberg’, Macarena Gonzalez', Alvaro Laborda-, Rodrigo Postiglioni'^ and Miguel Simo^: 'Laboralorio de Etologia, Ecologia y Evolucion, Institute de Investigaciones Biologicas Clemente Estable, Avenida Italia 3318 CP 11600, Montevideo, Uruguay; ^Seccion Entomologia, Facultad de Ciencias, Igua 4225 CP 11400, Montevideo, Uruguay. E-mail: aisenber@iibce.edu. uy Abstract. Environments where prey availability is scarce or highly variable have been reported as potential settings for the occurrence of paternal investment and sex-role reversal (choosy males and competitive, courting females). AUocosa hrasiliensis (Petrunkevitch 1910) and AUocosa alticeps (Mello-Leitao 1944) are two sand-dwelling wolf spiders that construct burrows along the Uruguayan coastline. Both species present a reversal in typical sex roles and size dimorphism. In the present study, we investigated foraging behavior and population density of both species by performing monthly samplings at the field during one year. Both AUocosa are general and highly opportunistic predators, varying their diet according to prey availability. The three most represented common prey belonged to Araneae, Diptera, and Hymenoptera (Formicidae). There were high levels of cannibalism in A. hrasiliensis and, furthermore, males were observed frequently preying on conspecific adult females. Our discussion of the results based on hypotheses about food limitation and sex-role reversal contributes to our understanding of AUocosa species and establishes them as models for future evolutionary, behavioral, and ecological studies. Keywords: Lycosidae, Uruguay, prey, sex-role reversal, food limitation Environments with fluctuations in prey abundance and access to refuges or other resources have been reported as potential causes for the evolution of paternal care and sex-role reversed systems (Gwynne 1991; Karlsson et al. 1997; Lorch 2002). AUocosa hrasiliensis (Petrunkevitch 1910) and AUocosa alticeps (Mello-Leitao 1944) are two sympatric and synchronic wolf spider species that live in sandy coasts of Uruguay (Capocasale 1990; Costa 1995; Costa et al. 2006). Individuals reported in studies of Costa (1995), Sim6 et al. (2005), and Costa et al. (2006) as AUocosa sp. belong to AUocosa alticeps. The environment these AUocosa species inhabit can be considered harsh where prey abundance and weather condi- tions are highly variable. The changeable environment could be imposing unusual constraints on these species, affecting the sexual behavior of each gender and causing adaptations. Recent studies (Aisenberg et al. 2007; Aisenberg & Costa 2008) report a reversal in typical sex-roles and size dimor- phism for both spider species. Aisenberg & Costa (2008) reported that females are smaller than males, both in A. hrasiliensis (carapace width, females: 4.63 ± 0.49 mm; males: 5.76 ± 0.59 mm) and A. alticeps (carapace width, females: 2.94 ± 0.30 mm; males: 3.28 ± 0.54 mm). The reproductive sexual peak of A. hrasiliensis and A. alticeps takes place in January (Costa 1995; Costa et al. 2006). Females are the roving sexual aggressors that locate and court the males. Copulation takes place inside male burrows and after the final dismount, males abandon their burrows, leaving them to the females (Aisen- berg et al. 2007; Aisenberg & Costa 2008). AUocosa hrasiliensis and A. alticeps are the sole wolf spiders adapted to living in the Uruguayan coastline (Costa et al. 2006). Food limitation could be an important factor affecting female and male feeding strategies in both lycosid species. Furthermore, occasional field observations suggested the occurrence of cannibalism of females by males of A. hrasiliensis during the reproductive period (F.G. Costa and A. Aisenberg, pers. obs.), a phenomenon considered unknown for spiders (Elgar 1992; Wise 2006). Both AUocosa species inhabit areas that have been drastically reduced in the last sixty years (Costa et al. 2006), possibly affecting population densities and competition for prey or other resources. In the present work, we studied feeding and surface activities of A. hrasiliensis and A. alticeps with the hypotheses that both species are generalists with high levels of intraguild predation as adaptations for prey unpredictability and intraguild competition in these coastal habitats. Foraging samplings took place over the course of 1 1 mo (June 2007-April 2008) in the coastal sand dunes of Marindia (34°46'52.3"S, 55°49'29.6''W), Salinas (34°46'59.8"S, 55°49'51.5"W), Canelones, and Paso del Molino (34°16' 40.10"S, 55°14'00.80"W), Lavalleja, Uruguay. For 1 h after dark, four to six researchers using headlamps collected AUocosa spiders and any prey they had captured. Feeding spiders and prey were captured and taken to the laboratory for identification. Prey were identified to family, and in the case of Araneae and Hymenoptera, to genus. We considered any prey that was primarily consumed so that identification was unfeasible as “unidentified prey.” Surface activity was studied monthly in Salinas from June 2004 to April 2005, for 2 h after dark, using headlamps. We considered the presence of AUocosa individuals walking as an event of activity. We labeled as “sea-side” the side of the dune which faced the sea-front; the opposite side was designated as “land-side.” Spiders were sampled in four plots of 5 m x 5 m (two plots on the sea-side and two on the land-side) that were drawn parallel to the line of the coast, on the first line of dunes. Additionally, in the period between December 2004 and April 2005, we recorded surface activity as reported previously and related it to the presence/absence of vegetation on the plot. We identified and sexed the individuals in the field. Voucher specimens of both species were deposited in the 135 136 THE JOURNAL OF ARACHNOLOGY Lepidoptcra 5.7 Coleoptera 17.0 Hynienoptera 18.8 (Foiniicidae) Orthoptcra 1.9 I Dipfera 20.7 spp 83.3 spideis 16.7 Figure 1. — Prey captured by individuals of both AUocosa species, with the corresponding percentages (n = 50). Jun Jul Aug Set Oct Nov Dec Jan Feb Mar Apr Month arachnological collection of Seccion Entomologia, Facultad de Ciencias, Montevideo, Uruguay. We recorded 45 individuals of A. hrasiliensis and 9 individuals of A. ahiceps feeding during the sampling periods. Most of the diet consisted of spiders, represented mainly by other AUocosa individuals. AUocosa spiders also preyed on Diptera, Hymenoptera, Coleoptera, Lepidoptera, Homoptera, and Orthoptera individuals but in lower frequencies (Fig. 1). The Hymenoptera captured were worker Acromyrmex and Dorymirmex ants, caught on their trails. We found a high rate of intraguild predation in A. hrasiliensis (Fig. 2). Surprisingly, though females and large juveniles of A. hrasiliensis preyed on small conspecific juveniles and on adults or juveniles of A. alticeps, adult males of A. hrasiliensis preyed frequently on females of their own species (Fig. 2). We observed 37 AUocosa individuals on the land-side of the dunes and 9 on the sea-side. Surface activity and prey capture showed higher values between December and January, while a lower intensity of feeding was registered in the period between June and November (Fig. 3). We found 13 individuals of A. hrasiliensis in areas without vegetation and 5 in areas with vegetation. AUocosa alticeps did not show preference for areas with (n = 9) or without vegetation (n =6). The present results indicate that the diets of both AUocosa species are non-specific and highly opportunistic, according to prey availability. The occurrence of the three most represented prey (Araneae, Diptera, and Hymenoptera) was highly variable through the year. Both Diptera and Hymenoptera individuals were frequently caught by AUocosa spiders during their nuptial swarms or, in the case of ants, while working on Figure 2. — Number of males, females, and juveniles of A. hrasiliensis found feeding on individuals of A. alticeps, or individuals from their own species. Figure 3. — Phenology of prey capture (dotted line) and surface activity (black line) recorded for both AUocosa species (individuals foraging n = 50; individuals walking n = 213). their characteristic trails. The consumption of ants in natural conditions has been cited for web-building spiders, consisting mainly of winged ants but also walking individuals (Carico 1978; Nentwig 1987). Ground-hunting salticids, thomisids, gnaphosids, and oxyopids often show a high percentage of ants in their diets (Nentwig 1987; Foelix 1996) and members of the Zodariidae family are myrmecophages (Foelix 1996; Pekar 2004; Pekar et al. 2008). Moya-Larano & Wise (2007) reported Schizocosa spiders (Lycosidae) feeding on ants under labora- tory conditions. However, studies on lycosid spiders in the field report that Collembola, Diptera, Cicadina, Aphidina, and Araneae would be the main prey groups for this family (Nentwig 1987). Moya-Larano et al. (2002) provide a different list of prey (absence of ants) for Lycosa tarentula, another burrowing wolf spider. Ants are considered very abundant in coastal areas, especially during the summer (Costa et al. 2006) and though they can be considered small prey for a spider the size of an AUocosa, they are the most predictable prey. The consumption of ants by AUocosa spiders could suggest food limitation and the requirement of special adaptations to manage potentially dangerous prey. The lack of cannibalism and feeding on small prey in A. alticeps could be associated with a smaller size and, consequently, lower energetic requirements (Andersson 1994; Blanckenhorn 2005; Foellmer & Fairbairn 2005). On the other hand, cannibalism rates are high in A. hrasiliensis. Intraguild predation is considered widespread among wolf spiders (Fernandez-Montraveta & Ortega 1990; Wagner & Wise 1996; Moya-Larano et al. 2002). In general, studies report juveniles feeding on other juveniles, adults feeding on juveniles, or females feeding on males; overall large individuals feed on small ones (Polls 1981; Polls et al. 1989, 1997; Wise 2006). However, we found males of A. hrasiliensis cannibal- izing females of the same species, a phenomenon unexpected for spiders (Llgar 1998; Llgar & Schneider 2004; Wise 2006). Although we observed males cannibalizing females in just three instances (see Fig. 2), this fact is remarkable because observations of this kind in the field are very scarce even in studies with substantial field effort (Moya-Larano et al. 2002). The consumption of females by male spiders can be considered non-adaptive, in general, in terms of losing a potential mate. Males of A. hrasiliensis are sedentary, probably remaining inside their burrows without feeding for long periods (Costa et al. 2006; Aisenberg et al. 2007). So, after copulation and AISENBERG ET M^.—ALLOCOSA FORAGING & ACTIVITY before constructing a new burrow, they need to forage intensively. Considering the high concentration of Allocosa individuals in some areas and the fact that copulations take place exclusively inside male burrows (Aisenberg et al. 2007), females could turn into a good meal for a hungry and large recently-copulated male without a burrow. This would mean no risks to male paternity, as copulated females stay inside the burrows after copulation and until the emergence of spider- lings (Aisenberg et al. 2007). The cannibalism level increases with increasing size-differences (Polis 1981; Polls et al. 1989, 1997; Elgar 1998; Buddie et al. 2003; Wise 2006), so the larger size in Allocosa males compared to females could be favoring this atypical male strategy. Furthermore, males of sex-role reversed species are expected to be choosy (Gwynne 1991; Andersson 1994). Male selection with regard to female size has been cited in Lycosa tarantula (L. 1758), another role reversed wolf spider species (Moya-Larano et a!. 2003; Huber 2005). Males of A. brasiliensis could exhibit extreme mate choice based on female reproductive or nutritional status: copulate with the female or eat her (Elgar 1992). Adaptive foraging (Newman & Elgar 1991), mistaken identity (Gould 1984), and aggressive-spillover (Arnqvist & Henriksson 1997) hypotheses, already tested in other spider species, require further testing in A. brasiliensis. A. brasiliensis was more highly represented in our samplings compared with A. alticeps. This could mean that the first species is more abundant, in contrast to the findings of Costa et al. (2006) based on results of pitfall trapping in the same areas, or the fact that it is less sedentary. Furthermore, A. brasiliensis individuals could be more easily detected due to their larger size or their greater presence in more open areas compared with A. alticeps. However, present surface activity data needs more exhaustive field work, recording not only surface activity but also burrow density, presence of individ- uals inside open / closed burrows and marking - tracking of individuals. In the last century, the coastal landscape of the Rio de la Plata and Atlantic Ocean in Southern Uruguay has decreased considerably, especially due to urbanization (Costa et al. 2006). Simo et al. (2005) reported the occurrence of Allocosa spiders strictly associated with the presence of sand dunes. The current results suggest that the highest surface and foraging activities of both Allocosa spiders coincide with the summer of the Southern hemisphere. During this season, coastal areas are most critically affected by tourism, which could also be impacting negatively on critical phases of the spiders’ life cycle. This fact may be considered for adequate management plans for these areas. Simo et al. (2005) also postulated Allocosa species as potential biological indicators of human effects on coastal ecosystems, as Marshall et al. (2000) did for Geolycosa, another burrowing wolf spider species of coastal areas. Both Allocosa species seem to be more abundant on the land-side of the dunes, probably because these areas are more protected against the strong winds typical of the Uruguayan coastline. On the other hand, the burrows of the sea-side could be closed off by the spiders due to the strong wind beating on this side of the dune, thus escaping observers’ detection. This behavior has been reported for another wolf spider inhabitant of coastal sand dunes (Gwynne & Watkiss 1975). Allocosa brasiliensis seems to be more closely associated with areas 137 without vegetation compared to A. alticeps, though we need further studies to confirm the trend. Marshall (1997) reported that Geolycosa xera arboldi McCrone 1963, another burrowing wolf spider inhabitant from sand dunes, was more directly associated with areas without vegetation. In the first decades of the twentieth century, dunes of the Uruguayan coast were fixed by human plantation of exotic vegetation as Acacia longifoHa, Pinus spp. and EiicaUptus spp., especially on the land-side of the dunes (Costa 1995). Areas with exotic vegetation are associated with invader spider species, so the exclusion of A. brasiliensis from these areas could be a mechanism to avoid competition for resources, interference competition, and intraguild predation. 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E-mail: maria.chatzaki@gmail.com Petros Lymberakis: Natural History Museum of Crete, University of Crete, P.O. Box 2208, 71409 Irakleio, Greece Plamen Mltov: Department of Zoology and Anthropology, Faculty of Biology, University of Sofia, 8, Dragan Tsankov Boulevard, 1164 Sofia, Bulgaria Moysis Mylonas: Natural History Museum of Crete, Department of Biology, University of Crete, P.O. Box 2208, 71409 Irakleio, Greece Abstract. The harvestman fauna was studied along an altitudinal gradient on the southern slope of Lefka Ori Mountains, Crete, Greece for one year. Four sampling areas were defined at 800, 1200, 1600, and 2000 m elevation and they were sampled with pitfall traps that were emptied at monthly intervals. In total, six species were collected: Histricostoma creticum (Roewer 1927), Lacinius imularis Roewer 1923, Graecophalangium cretaeum Martens 1966, Opilio insulae Roewer 1956, Rafalskia cretica (Roewer 1923) and Leiobunum ghigii Di Caporiacco 1929. Species richness was the same (5 spp.) at the three lower zones and then declined to three species at 2000 m. Catches were more than double at this elevation. Differences of phenological patterns were observed among species and among altitudinal zones within the same species. High activity during spring and autumn and a summer recession were characteristic of most taxa. Opiliones did not seem strongly affected by the severe harshness of climatic conditions at higher elevations, as observed in other taxa, indicating a strong physiological tolerance and/or behavioral adaptation in order to withstand environmental stress. Keywords: Elevation, harvestmen, Mediterranean, pitfall traps, seasonal variation Mountain ecology has long been the focus of interest for many plant and animal ecologists. Mani (1968) compiled the earlier entomological literature of the alpine zone, which consists mainly of anecdotal, casual, or incidental observa- tions. The lack of knowledge on quantitative and functional aspects of the soil fauna of Mediterranean-type ecosystems has been noted by Di Castri & Vitali-Di Castri (1981). However an effort has been made to try to clarify patterns of ecological variation in this kind of ecosystems in Greece. Stamou et al. (1984) studied the soil macrofauna succession in Mt. Olympus and Legakis (1986) analyzed the arthropod soil fauna of Mt. Hymettos. Despite their contribution to the knowledge of ecological variation of the fauna in Mediterranean-type ecosystems, these studies covered few taxonomic groups and did not account for seasonal variation of the fauna. Sfenthourakis (1992) gave a clearer picture of the variation of isopod assemblages along altitudinal gradients in three mountain systems of Greece. Lymberakis et al. (2003) discussed the altitudinal variation of Oniscidean communities and Chatzaki et al. (2005a) presented the variation of spider communities along altitudinal gradients on the island of Crete, Greece, as far as both composition and activity on a year-long basis are concerned. The phenology of the same group is presented in a separate paper (Chatzaki et al. 2005b). Studies on the Opiliones of Crete have been predominantly faunistic and taxonomic. Only the work of Martens (1966) contains some phenological and chorological notes concerning the opilionid fauna of the island. As a whole, publications about the biology, ecology, and zoogeography of the group on Crete are lacking. In view of this, the present paper is the first that concentrates on the comparison of the phenology of Opiliones at different elevations, as well as on the seasonal and altitudinal variation in species composition in Lefka Ori (White Mountains) on Crete. Lefka Ori is one of the most important insular mountain systems of the Mediterranean region, because of its high elevations, the large area it covers and its unique topography and climate (see climate paragraph in Methods). The results presented here may serve as a basis for further comparison of the phenological peculiarities of the Opiliones in the region, as well as for the evaluation and analysis of the adaptive strategies of the endemics and their related species in other parts of the Balkan Peninsula. METHODS Description of the study area. — The Lefka Ori mountain system, situated at the western part of the island, is the most massive mountain range of Crete, including 56 peaks over 2000 m, the highest of which reaches an elevation of 2453 m. The study was carried out on the southern slope of Lefka Ori, just above the plateau of Anopolis found at ca 600 m (Fig. 1). Four sampling sites were selected, according to the main vegetation types, along the altitudinal gradient from 800 m to 2000 m: 1 . An old mature forest of Fim/s brutia Ten. at 800 m above sea level (a.s.l.) (35°14'N, 24°5'E), with very little under- story, consisting mainly of Quercus cocdfera L. shrubs and phrygana species, such as Euphorbia acanthothamnos Heldr. & Sart. ex Boiss., Sarcopoterium spinosum (L.), Verbascum spinosum L. and Drimia maritima (L.). 2. A Cupressus sempervirens L. mature forest at 1200 m a.s.l. (35°15'N, 24°6'E), composed mainly of Cupressus sempervirens, Quercus cocdfera and a few Acer semper- virens L. trees. The understory is practically absent in this vegetation type. 3. A plateau immediately over the timberline at 1600 m a.s.l. (35°16'N, 24°5'E), partially covered with crawling 139 140 THE JOURNAL OF ARACHNOLOGY scrubs of Juniperus oxycedrus L. subsp. oxycedrus and Berheris cretica L., accompanied by Primus prostrata Labill., R/ummus saxatilis Jacq. subsp. primifoUa (Sm), Acantholinwn idicimmi (Willd. ex Schult.), Satureja spinosa L. and Anchusa caespitosa Lam. 4. A rocky site at 2000 m a.s.l. (35°17'N, 24°3'E), where the dominant scrub plants are: Berheris cretica. Primus prostrata. Astragalus august ifolious Lam., and Satureja spinosa. Plants are largely restricted to areas offering wind shelter. The area, as the whole of the island, is a karstic landscape. The ground is largely covered by stones and rocks that provide shelters for ground living animals. Human presence in the area dates from antiquity. For the last two centuries, but especially at present, intense grazing by sheep and goats takes place over the entire area. A main effect of overgrazing and the consequent trampling of the soil is that densities of most arthropod taxa have been reduced (Legakis 1986). Climate. — The climate of Crete is typical Mediterranean with 5-7 mo of arid, hot and dry summers, alternating with shorter periods of rainy, mild winters. There are not sufficient meteorological data, especially for the high mountains. Mean annual temperatures fall 6° C per 1000 m rise in elevation (Grove et al. 1991). The range of temperatures is much narrower near the coast than in the mountains, due to the more maritime character of the former (Strid et al. 1995). Although there are no records of the precipitation above 900 m, Rackham & Moody (1996) estimated that at the top of Lefka Ori the annual precipitation must be as high as 2000 mm. In southern Lefka Ori, snow above 1400 m can be several meters high, but the water disappears into the porous crystalline limestone immediately after the snow melts in May. Bonnefont (1972) estimated that snow cover lasts in Crete for 2-3 mo at 1500 m and for 5-6 mo at 2000 m. Over 2000 m, the snow cover lasts generally from November to May and then melts progressively. Small, localized spots can persist until August. During the summer months, the lower atmo- spheric pressure and the lack of trees combined with other physical characteristics (strong winds, thin soils, large proportion of precipitation as snow), favor high insolation and create dry and hot conditions (Shanks 1956; Mani 1990). This harsh landscape (because of snow cover in winter and high aridity in summer) forms the “High Desert” of the Lefka Ori highlands, a term coined by Rackham & Moody (1996) to describe a unique environment that does not exist to such extreme in any other Mediterranean mountain. Sampling. — Harvestmen were collected by pitfall trapping. Forty plastic cups (10 cm depth X 7 cm diam.) were used per site. Though these dimensions are rather small, traps were capable of catching large arthropods (scorpions, large spiders, centipedes), and even lizards. Traps were placed in a straight- line ca 3 m apart. Lfndiluted ethylene glycol was used as a preservative. To diminish passive filling of the traps with organic matter and water, as well as predation from larger animals and vandalism, they were partially covered with stones, without reducing access to them by the animals. Traps were emptied at approximately 30-day intervals from August 1990 to March 1992. However, the results presented here include data from one full collecting year (March 1991 to February 1992), since data from the other years did not deviate from the results obtained from the main collecting CHATZAKI ET AL.— OPILIONES ON CRETE 141 Table 1 . — Species overall presence and total activity (transformed into number of individuals per 100 trap-days) at different elevations of Lefka Ori Mountain, Crete. Elevation Species 800 m 1200 m 1600 m 2000 m Lacinius insularis 2.82 21.06 31.94 Opilio insulae 6.48 20.97 6.56 32.10 Graecophalangium cretaeum 4.32 5.73 6.01 9.70 Rafakkia cretica 13.00 6.53 0.84 Histricostoma creticum 0.36 0.16 Leiobunum ghigii 1.27 0.26 Total 26.98 34.66 34.73 73.74 year. le the phenology graphs the last two months are presented jointly (J-F) because there was no intermediate emptying of the traps. Data analysis. — Opiliones were identified by P. Mitov. Vouchers are deposited at the Natural History Museum of the University of Crete and some are kept in the private collection of P. Mitov. As some traps were destroyed or the number of collecting days per sampling period was not equal, sampling effort was unequal too. In order to render the data from four sites and eleven sampling periods comparable, catches from each sample were transformed into catches per 100 trap-days. RESULTS Descriptive analysis and correlation. — Six species, belonging to the families Phalangiidae (4 spp.), Nemastomatidae (1 sp.) and Sclerosomatidae (1 sp.) and represented by 2226 specimens were collected. Species are: Histricostoma creticum (Roewer 1927), Lacinius insularis Roewer 1923, Graecopha- langium cretaeum Martens 1966, Opilio insulae Roewer 1956, Rafakkia crelica (Roewer 1923) { = Metaplatybunus rhodiensis Roewer 1924), and Leiobunum ghigii Di Caporiacco 1929. The latter is recorded from Crete for the first time. Two out of the three species that reach the elevation of 2000 m -L. insularis and G. cretaeum- are endemic to the island. All other species may be characterized as Ponto-Mediterranean faunistic elements (sensu De Lattin 1949, 1967). Species overall presence and total catches at the study sites are presented in Table 1. The number of species is five at the three lower elevations and decreases to three at 2000 m. Species overall catches increase gradually from 800 to 1600 m and become highest (more than double) at 2000 m. The qualitative and quantitative composition among the first three elevations is different. Lacinius insularis presents statistically significant positive correlation with elevation (Pearson’s coefficient R = 0.378, P = 0.03), while Rafakkia cretica presents significant negative correlation with increasing elevation (Pearson’s coefficient R = -0.396, P = 0.023). The remaining species do not present any significant correlation with elevation. Phenology. — The phenology of the dominant species at each site is presented in Figs. 2-5. Lacinius insularis was absent at the 1200 m site. Since its total catches seem to significantly increase at the two higher elevations (Table 1, Fig. 2) and sampling was equally intense at all sites, its absence from 1200 m cannot be accidental. At the other elevation, L. 800 m ■o Q. § ■o •E 1 o £ 0,5 -i ' 1 m Df 0j ■ — ,, m ^ M — g I 1 1 » M M N D J-F 1600 m m fB ■H O. 2 > ■u c a> E 14 ■ 12 ■ 10 8 6 - 4 M A - W aa M J J A S 0 N D J-F 2000 m 2.14 m MAMJ J ASONDJ-F Figure 2. — Phenology of males (m), females (0 and juveniles (j) of L. insularis at different elevations. Grey horizontal bars indicate the months during which the sites were covered by snow. insularis shows one peak of activity in autumn, represented mainly by female individuals. This peak is shifted from November at 800 m to September at 2000 m, thus the peak of activity occurs one month earlier at each higher altitudinal zone. Juveniles become mainly active in late spring (May at 800 m and 1600 m) and then again in autumn (September- October at 800 m and November at 1600 m). At the 2000 m site juveniles become very active in June and then again in October. This probably means that individuals of this species hibernate as immatures and remain inactive for a longer period, until the following summer when they mature (August) 142 THE JOURNAL OF ARACHNOLOGY 800 m 800 m 1600 m E 6 I 5 m 4 > C O , H H A M J J N D J-F 2000 m Figure 3. — Phenology of males (m), females (0 and juveniles (j) of O. insulae at different elevations. Grey horizontal bars indicate the months during which the sites were covered by snow. 1200 m Figure 4. — Phenology of males (m), females (f) and juveniles (j) of R. cretica at different elevations. Grey horizontal bars indicate the months during which the sites were covered by snow. and are ready to reproduce in September-October. Activity under the snow cannot be excluded, but is not demonstrated by the present data. Opilio insulae presents a clearer phenological plasticity along the altitudinal gradient (Fig. 3). At the two lower elevations it presents a peak of activity in May-June, while at the two higher elevations it presents autumn peaks, in October at 1600 m and in August-September at 2000 m. At the first three elevations, juveniles present higher activity during September-October and at 2000 m during August. Rafalskia cretica presents an early or mid spring peak of activity (Fig. 4), thus almost avoiding O. insulae and L. insukiris. Very few immature individuals were found at the 1600 m site and none at 2000 m. At the two lower elevations the peaks of adult catches are shifted from April-May at 800 m to May-June at 1200 m, while immatures are active for a longer period at 800 m (December-March) than at 1200 m (March-April). In a similar pattern, G. cretaeum presents a peak in March- April at 800/1200 m which is shifted towards April-May at 1600 m (Fig. 5). Interestingly at 2000 m there are two almost equal peaks of activity of all stages simultaneously present, one in June and another one in October. Although this might indicate a double generation per year or overlapping CHATZAKI ET AL.— OPILIONES ON CRETE 143 m n 7 a E 6 5 O 4 M J 800 m B m Df H j 1200 m MAMJ JASONDJ-F 1600 m MAMJ JASONDJ-F 2000 m MAMJ JASONDJ-F Figure 5. — Phenology of males (m), females (f) and juveniles (j) of G. cretaeum at different elevations. Grey horizontal bars indicate the months during which the sites were covered by snow. generations, the number of individuals caught is too small to allow such an interpretation. Alternatively this double peak of activity might also correspond to the same single generation of harvestmen that become active only under the most favorable conditions with optimum temperatures and humidity (i.e., June and October), while in the meantime (summer) they remain totally inactive due to their inability to tolerate the high aridity and high temperatures of this season. Hisiricostoma creticum is restricted to the two lower elevations (800 and 1200 m), whereas L. ghigii to the two intermediate ones (1200 and 1600 m). Low numbers of both of them do not allow us any phenological interpretation. However, individuals of the former species were caught throughout the year (June, October, December and January at 800 m and April, July, November at 1200 m), while the latter species showed a tendency to be more active during autumn months (October-November). At the two lower elevations, catches of all species cease for one (1200 m) or two (800 m) summer months (August and July-August, respectively). At the three higher elevations snow covered sampling traps for four (1200 m and 1600 m) and seven (2000 m) winter-spring months. DISCUSSION Phenology. — At Lefka Ori at all elevations harvestmen seem to have an annual life-cycle, as inferred from the phenological patterns (Figs. 2-5). The favorable period in which Opiliones at high elevations of Crete reach maturity is either in early autumn or in spring. Maturity lasts one to two months and is followed or preceded by a juvenile peak. In general, immature activity is expanded for a longer period within a year than mature stages, even at the highest elevations, where immature activity overlaps with that of adults (see for example G. cretaeum and O. insulae at 2000 m as extreme cases). This is common in species inhabiting high elevation habitats and signifies the shortening of biological processes within the year (Pinto-da-Rocha et al. 2007). The phenological patterns of L. insularis, R. cretica, and G. cretaeum do not change along the altitudinal gradient, but their peaks of activity are shifted to one or two months later (R. cretica and G. cretaeum) or earlier (L. insularis) at the two higher elevations. It is important here to note that seasons are not the same at each elevation. Although lack of meteorolog- ical data allows us only to speculate, we suggest that with the rise of elevation, winter lasts longer and “squeezes out” spring and autumn. In this respect, at 2000 m October represents late autumn and June represents spring, whereas at 800 m, October and June are still very warm and dry months corresponding to “summer,” taking into account the high mean temperatures encountered in both periods. Hence Opiliones find similar climatic conditions in March-April at 800 and 1200 m and in May-June at 1600 and 2000 m (i.e., G. cretaeum) or in October-November at 800 m and in Septem- ber at 2000 m (i.e., L. insularis). The case of phenological differentiation of O. insulae (Fig. 3) is more pronounced, changing its peaks of activity from spring (800-1200 m) to late summer/autumn (1600- 2000 m). The latter pattern is one of the very few cases of late summer activity peak among all taxa studied, at all elevations. The only species that presents the same pattern of activity at Lefka Ori is Zelotes creticus Kulczynski 1903, an endemic spider species that belongs to the family Gnaphosidae (Chatzaki et al. 2005b). This phenomenon may indicate that at the two higher elevations both mating and egg laying occur 144 THE JOURNAL OF ARACHNOLOGY in the same period; i.e., in late summer - beginning of autumn, while at the lower ones these processes are interrupted by the hot dry summer, during which animals remain inactive. Martens (1966) noted that the reproductive period of this harvestman in the seacoast region of Crete is from mid-April- June. This difference of phenological patterns between lowland and highland populations may possibly be justified by the climatic heterogeneity of the Cretan landscape. An alternative explanation however might be a taxonomic divergence of the species masked by morphological similarity, a phenomenon that has been observed in various taxa (e.g., Parmakelis et al. 2003 and references therein). This hypothesis remains to be further tested with molecular data. A partitioning of the favorable periods, with peaks of activity following one another may be observed. As a result, activity periods of R. cretica and G. cretaeum never coincide with those of O. insulae and L. insukiris. This seems to be the rule concerning arthropod phenology, since it has also been observed in spiders of the family Gnaphosidae (Chatzaki et al. 2005b) on Crete and in other Mediterranean regions (Urones et al. 1995) as well as in temperate ecosystems (Enders 1976; Toft 1976; Uetz 1977). Cloudsley-Thompson (1962) places water conservation as the prime physiological problem for the survival of terrestrial invertebrates. Harvestmen are especially susceptible to water loss and this is a prime factor limiting species distributions (Hillyard & Sankey 1989; Pinto-da-Rocha et al. 2007). The fact that no harvestmen were caught for two months at 800 m and for one month at 1200 m, indicate a severe stress due to high aridity, which these animals try to overcome by ceasing their activity. Things are even more pronounced at the two higher elevations, the “High Desert” according to Rackham & Moody (1996). On one hand, snowfall and snow cover during four (1600 m) and seven months of the year, respectively, and on the other hand the extreme aridity of summer months (especially August at 2000 m), considerably restrict the suitable period for exploitation by harvestmen. A compromise between water conservation, limited time for activity, and avoidance of other species might result in the, still mysterious, mid-summer peak of activity of O. insulae. Kinetic activity and species richness. — Janzen (1973), Janzen et al. (1976), Wolda (1987), and McCoy (1990) reported a decrease of the number of species and individuals of certain arthropod groups, mainly insects, along an altitudinal gradient. Almeida-Neto et al. (2006) showed a decline of both species richness and abundance of Opiliones in elevational gradients of three mountains in Brazil. Our data partially agree with the decrease in the number of species, but as far as kinetic activity of Opiliones is concerned a total increase in the number of individuals is observed, which is due to the impressive increase of three species, namely L. insukiris, O. insulae and to a lesser extent G. cretaeum. Similarly, species of other taxa (i.e., members of the spider family Gnaphosidae (Chatzaki et al. 2005a), as well as some other families (Lycosidae, Dictynidae, Thomisidae, and Philodromidae (Chatzaki, unpublished data)) tend to reach extremely high numbers of individuals at the higher elevations of the same mountain system. According to Hagvar et al. (1978) harvestmen is the largest group (as far as numbers of individuals are concerned) found among other predatory arthropod communities at high elevations in Norway. The difference between our results and those found by Almeida-Neto et al. (2006) as far as the abundance along elevation is concerned, may lie in the fact that the range of climatic conditions that the tropical arthropods can tolerate cannot be as wide as that tolerated by animals of the temperate zone. Therefore, Opiliones of Crete should be able to reach higher elevations (hence harsher conditions) and create denser populations there. Another point which may partly explain this difference is the methodological bias caused by the fact that the authors measured the abundance of species as revealed by hand collecting and we measured the activity/abundance in the sense of pitfall catches. In view of the identification of the origin of the mountain harvestmen of Crete, one has to follow the paleohistory of the island formation. The mountains of Crete have a very short history dating back to the beginning of the Pliocene (Meulenkamp et al. 1994) with an estimated elevation of 2000 m since then, which was accelerated mostly during the Pleistocene (Meulenkamp 1971). This newly and rapidly formed mountain landscape gave rise to new niches on the island. Taking into consideration that Crete was isolated from the mainland since the early Pliocene (Meulenkamp et al. 1988), the only available fauna to occupy the newly formed high elevations would be the already existing lowland species. This is true for several taxa studied until now. Studies on ground beetles (Trichas 1996), terrestrial snails (Vardinoyan- nis 1994) and plants (Greater 1972) agree that life of the Cretan mountains is mainly composed of derivatives of lowland endemics and a small number of old relicts. Chatzaki et al. (2005a) found one endemic but mostly non endemic high elevation specialists and lowland species which altogether compose the main part of the arachnofauna above 1600 m. Among the 20 species of Opiliones recorded on the entire island (Giltay 1932; Roewer 1927, 1940; Martens 1965, 1966; Gruber 1998), only six are reported above 800 m at Lefka Ori mountains. Three species increase in numbers of individuals along the altitudinal gradient and dominate at higher elevations (Table 1), two of which are Cretan endemics (L. insularis and G. cretaeum). At 2000 m, the latter are the only opilionid residents together with O. insulae. There are no high- elevation specialists, like in ground spiders, since all three of them are also found in the lowlands of Crete (for detailed references see Roewer 1927; Giltay 1932; Martens 1966). In conclusion, harvestmen of the high elevations of Crete show a high tolerance to the extreme climatic conditions found in these environments and do not seem to be limited by them in order to create viable populations. The six species recorded above 800 m at Lefka Ori mountains represent the most physiologically tolerant species among the lowland residents of the island and none of them is a high-elevation specialist. Most species tend to exploit the favorable period of April-June and September-October. At the higher elevations, biological cycles are compressed to a narrow period in which climatic conditions allow the animals to be active. ACKNOWLEDGMENTS This work was conducted in the framework of the PhD thesis of the second author. We are deeply grateful to the CHATZAKI ET AL.— OPILIONES ON CRETE 145 editor of the journal Soren Toft, Christian Komposch, and an anonymous reviewer for important comments and suggestions that improved the final version of this paper. 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The Journal of Arachnology 37:147-150 The chemical defenses of a stylocellid (Arachnida, Opiliones, Stylocellidae) from Sulawesi with comparisons to other Cyphophthalmi Tappey H. Jones', William A. Shear--"* and Gonzalo Girlbet-'*: 'Department of Chemistry, Virginia Military Institute, Lexington, VA 24450, USA; ^Department of Biology, Hampden-Sydney College, Hampden-Sydney, VA 23901, USA; ^Museum of Comparative Zoology, Harvard University, Cambridge, MA 02138, USA Abstract. Two female specimens of an uiidescribed species of stylocellid harvestman (Opiliones, Stylocellidae) from Sulawesi were extracted in methanol, and compounds in the product were identified by means of gas chromatography-mass spectrometry. Nineteen significant peaks were found, 12 of which were identified, indicating the presence of naphthoquinone, 2-tridecanone, 2-tetradecanone, 2-pentadecanone, 6-methy 1-1, 4-naphthoquinone, 6-methyl- 1,4-naphtha- lenediol, and at least four unsaturated ketones. The spectrum differed both qualitatively and quantitatively from previously published data on Siro exilis Hoffman 1963 and Cyphophthalmus duricorius Joseph 1868 (Sironidae). Keywords: Siro, Cyphophthalmus, exocrine products, ketones, naphthoquinones Harvestmen (Opiliones) have a distinctive pair of prosomal exocrine glands opening to the surface via ozopores usually placed in the vicinity of the second leg pair. These glands produce a variety of substances (Table 1) effective in defense (e.g., Juberthie 1961a, 1961b, 1976; Eisner et al. 1971; Jones et al. 1977), and possibly with other functions as well. The secretions of approximately 48 species have been studied in detail up to this time. The results have been effectively reviewed by Gnaspini & Hara (2007; also see Hara et al. 2005), who presented a table listing the species and the compounds recorded from them. Forty-six compounds have been identi- fied to date (numerous others are present but have not been identified), falling into the broad classes of long-chain alcohols and ketones, hydroxyquinones, phenols, and, more rarely, an alkaloid (nicotine), an amine (N,N-dimethyl-B-phenylethyl- amine), terpenoids (camphene, limonene) and bornyl esters (bornyl acetate and propionate) (Gnaspini & Hara 2007). A number of these compounds are unique or rare in nature, particularly in animals. Typically a single species produces a mixture of compounds, and the mixture of both identified and unidentified molecules may be characteristic of the particular taxon (family, genus, species) involved (Hara et al. 2005). Although the defensive chemistry of Cyphophthalmi, Eupnoi and Laniatores has been studied for some species, that of many important higher taxa (i.e., Dyspnoi) remains completely uninvestigated. Our preliminary results (unpub- lished data) suggest that travunioids, only a single species of which has been examined so far, are extraordinarily diverse in their chemistry. Studies of Grassatores have concentrated on just a few families (Cosmetidae, Gonyleptidae, Manaosbiidae, Stygnommatidae) in this very diverse assemblage. Defensive chemistry of the basal and divergent harvestman suborder Cyphophthalmi (e.g., see Giribet et al. 2002 for the phylogenetic placement of Cyphophthalmi) is interesting because if the composition of defensive secretions is of any phylogenetic use, its ancestral state in this group could be used to polarize the characters higher in the tree and to optimize the ancestral state of the defensive substances in Opiliones. But to date, only two cyphophthalmid species, Siro exilis Hoffman Corresponding author. E-mail: wshear@hsc.edu 1963 of North America, and Cyphophthalmus duricorius Joseph 1868 of Europe, have been studied for their defensive secretions (Raspotnig et al. 2005). Both of these species belong to the same family, Sironidae, while members of the other extant five families remain unexamined. The secretions of the two sironids consist of complex arrays of ketones and naphthoquinones. In this paper, we present data on the secretion of an undescribed species probably belonging to the genus Stylo- cellus, family Stylocellidae. This species will be named and described in a forthcoming revision of the family by Ronald Clouse. In the current phylogenies of cyphophthalniids, the families Stylocellidae or Pettalidae appear basal within the suborder (Giribet & Boyer 2002; Boyer et al. 2007), although their stable placement is still debatable. The family Stylocelli- dae is found exclusively in Southeast Asia, from Southern China and Northeast India to the western side of New Guinea, in the Indonesian province of Irian Jaya, and the Philippine island of Palawan (Shear 1993; Clouse & Giribet 2007; Giribet et al. 2007; R. Clouse, in progress). The species we studied was collected on the Indonesian island of Sulawesi and is tentatively assigned to the genus Stylocellus, although the whole generic systematics of the family is currently under re- examination (R. Clouse & G. Giribet, in progress). METHODS Two female Stylocellus specimens of a yet undescribed species (Fig. 1) were collected alive in Bantimurung, Sulawesi (5°02'33"S, 119°44'08"E; 348 m elev.; Giribet locality number 512; collected 28 June 2008, R. Clouse, G. Giribet, C. Rahmadi, leg.; MCZ DNA101938), shipped alive to WAS, and extracted in about 0.5 ml of USP methanol. The extracts of the two specimens were pooled. The code number 06-172 was assigned to the specimens, both of which will be placed as vouchers in the Museum of Comparative Zoology, Cam- bridge, Massachusetts. The analysis of the extract was performed by THJ. Gas chromatography-mass spectrometry was carried out in the El mode using a Shimadzu QP-5000 GC/MS equipped with an RTX-5, 30 m X 0.25-mm i.d. column. The instrument was programmed from 60° C to 250° C at 10°/min. Identification of components was accomplished 147 148 THE JOURNAL OF ARACHNOLOGY Table 1. — Distribution of defensive secretions in Opiliones. For more detail, see Gnaspini & Hara (2007). Taxon Suborder Cyphophthalmi Suborder Eupnoi Family Phalangiidae Family Sclerosomatidae Suborder Dyspnoi Suborder Laniatores Infraorder Insidiatores Infraorder Grassatores Compounds Ketones, naphthoquinones naphthoquinones ketones, alcohols unknown Amines, bornyl esters, terpenes, alkaloids Phenols, methylquinones Benzoquinones, phenols Figure 1. — One of the extracted specimens of stylocellid from Bantimurung, Sulawesi, photographed alive by G. Giribet. Specimen is about 6.5 mm in body length. using NIST/EPA/NIH mass spectral library on CD-rom, version 1.7 (1999) and the NIST/EPA/NIH mass spectral library version 2.0d (2005). RESULTS AND DISCUSSION After gas chromatography-mass spectrometry examination, 19 significant peaks were identified in the extract. The major peaks in our analysis indicate the presence of naphthoquinone, 2-tridecanone, 2-tetradecanone, 2-pentadecanone, 6-methyl- 1 ,4-naphthoquinone, 6-methyl- 1,4-naphthalenediol, and at least four unsaturated ketones, two of which correspond to peaks S and U as noted by Raspotnig et al. (2005), so we have adopted their identification of these compounds (Table 2). Table 3 compares the analyses of the three species of Cyphophthalmi. Chloronaphthoquinones are unique in the two sironid species (and as exocrine products of arthropods [Raspotnig et al. 2005]), but were not found in the stylocellid species; likewise undecan-2-one, 6-tridecen-2-one and 7- tridecen-2-one were absent from the secretion of Stylocellus. It is possible that these unusual compounds came from the males included in the sironid samples. Furthermore, the stylocellid secretion contained 6-methyl- 1,4-naphthalenediol, a reduction product of 6-methyl- 1,4-naphthoquinoine, which did not occur in the two sironids. The percent composition of compounds that were common to the three species shows strong differences between the stylocellid and the two sironids, and to a lesser degree, between Siro exiiis and Cyphophthaimus dmicorius. While 2-tridecanone made up about 20% of the secretion of both sironids, it comprised 50% in the stylocellid; 1,4-naphthoquinone and 6-methyl- 1,4-naphthoquinone were found in very small amounts in the stylocellid, but at values from about 12% to nearly 18% in the sironids; pentadecan-2- one was at 13.8% for the stylocellid, but less than 1.7% for the sironids, and so on. In summary, we may observe that while the composition of the secretions of the three species is similar in constituting mixtures of ketones and naphthoquinones, there are signifi- cant differences both qualitative and quantitative. Some of the differences could be due to the fact that we extracted our animals in methanol, while Raspotnig et al. (2005) used hexane. Raspotnig et al. (2005) found that the composition of the secretions in the two species studied by them was highly consistent from individual to individual within species, as has been reported in numerous previous studies of harvestman defensive secretions. This lessens our concern about the small size of our sample versus the much larger samples taken by Raspotnig et al. (2005); the remote location and relative rarity of the animals we studied makes it unlikely that large samples will be available in the near future. The secretion of the stylocellid seems to contain fewer compounds than found by Raspotnig et al. (2005), who identified at least tentatively all of the 24 peaks they found (as opposed to our 19). However, very small peaks were not considered by us. Nevertheless, the observation about the Table 2. — Compounds and percent composition identified in methanol extract of whole females of the stylocellid from Sulawesi. See also Fig. 2. Peak Compound Relative % Peak code in Raspotnig, et al. 2005 1 1 ,4-Naphthoquinone 2 E 2 2-Tridecanone 42 J 3 6-methylnaphthoquinone 1 L 4 Methyl branched 2-tridecanone 8 M or N 5 2-Tetradecanone 9 M or N 6 6-Methyl- 1 ,4-naphthalenediol 1 Not detected 7 2-Pentadecadienone 8 S 8 2-Pentadecenone 7 U 9 Unsaturated ketone a 3 ? 10 Unsaturated ketone b 3 ? 11 2-Pentadecanone 14 w 12 2-Methoxy- 1 ,4-naphthoquinone 3 Not detected JONES ET AL.— CHEMICAL DEFENSES OF A STYLOCELLID 149 Figure 2. — Gas chromatographic profiles of whole-body methanol extract of stylocellid from Sulawesi. Identified peaks are numbered; numbers correspond to those in Table 2. number of compounds would hold on the basis of the number of peaks alone. Aside from 5-methyInaphthoquinone, not found in the sironids, the stylocellid secretion is strongly dominated by ketones and lacks the diversity of naphthoqui- nones found in sironids. If stylocellids are in fact a sister group to other Cyphophthalmi, this observation suggests that the other naphthoquinones, especially the chlorinated ones, have been added in the course of evolution (or lost in the stylocellid lineage), and perhaps that ketones dominated the secretion of the common ancestor of extant Opiiiones. Quinones may have been co-opted later from substances used by all arthropods to sclerotize cuticular proteins. However, proper polarization of this character requires first, a well-resolved cyphophthalmid phytogeny and second, a proper outgroup comparison. Since Opiiiones’ putative outgroups (Solifugae, Scorpiones, and Pseudoscorpiones; e.g., Wheeler & Hayashi 1998; Giribet et al. 2002; Shultz 2007) do not have this type of secretion, polarization may be difficult. Raspotnig et al. (2005) pointed out that while both ketones and naphthoquinones are found in Cyphophthalmi, scleroso- matids (suborder Eupnoi) secrete only ketones (and alcohols) and the single phalangiid studied (also an Eupnoi), naphtho- quinones. In Laniatores, Grassatores utilize phenols and methylquinones, while the single member of Insidiatores analyzed uses an eclectic mix of N.N-dimethyl-B-phenyleth- ylamine, nicotine, bornyl esters, and terpenes. But again we must point out the strong bias in the data. Most of the species studied have been either Grassatores or sclerosomatids (see Table 1), while Dyspnoi and the Eupnoi family Caddidae, two groups of significant phylogenetic importance, have not been studied chemically at all. In addition, no study has yet been made of the effect of collection method on the results of the analysis of harvestman secretions. In our studies, we are using methanol to extract whole bodies of live specimens, but other solvents, such as hexane, have been used in other laboratories. In some studies, live animals are induced to secrete their defensive compounds, which are collected either by micropipettes, fine glass tubing, or by absorption on filter paper. Thus we do not know if differences between species where different collection methods were used are real, or artifacts of the different methods. Certainly the fact that in many species the secretion of the repugnatorial glands is mixed with regurgitate from the gut could play a role if the already-mixed secretion is collected. The question of sample size also arises; here we used pooled extract from only two animals of the same sex, while Table 3. — Comparison of percent composition of secretions of the stylocellid from Sulawesi with those of Siro exilis and Cyphophtlialmus duricorius (data from Raspotnig et al. 2005). Confidence limits are given for the data on S. exilis and C. duricorius because Raspotnig et al. (2005) were reporting on individual extractions from 26 and 95 adult specimens respectively (for a complete list of compounds identified from the sironid species, see Raspotnig et al. [2005]). Our measurements are single points representing a pooled extraction of two adults due to the smaller collections of stylocellids when compared to the two sironid species studied previously. Bold figures represent the largest amount if significant differences are present; if two figures in a row are in bold type, there was no statistically significant difference between those two. Compound Stylocellid S. exilis C. duricorius 2-Tridecanone 50.0 20.28±3.79 20.21±3.56 7-Tridecen-2-one not identified 15.47±1.35 18.98±2.38 1 ,4-Naphthoquinone 1.7 14.01±1.9 17.61 ±2.82 6-Methyl- 1 ,4-naphthoquinone !.0 13J8±L43 12.15±2.03 Undecan-2-one not identified 0.57±0.19 9.71 ±1.59 4-Chloro- 1 ,2-naphthoquinone not identified 1L83±1.68 7.09±2.44 6-Tridecen-2-one not identified 4.13±0.92 4.02 ±1.03 Pentadecan-2-one 13.8 1.68±0.5 0.01 ±0.02 6-Methyl-4-chIoro- 1 ,2-naphthoquinone not identified 4.30±1.24 0.36±0.51 2-Tetradecanone 8.0 1.10±0.33 0.65±0.25 Pentadecadieone 8.0 3.0±0.86 0.04±0.06 Pentadecenone 6.7 4.5±1.14 0.03±0.05 6-Methyl- 1 ,4-naphthalenediol 0.7 not identified not identified 150 THE JOURNAL OF ARACHNOLOGY Raspotnig et al (2005) used large samples including both sexes and juveniles, and analyzed each extract individually. Previous studies of harvestman defensive secretions vary greatly as to the numbers of individuals sampled, and in some cases numerous animals were used, but the samples were pooled for analysis. In the near future we hope to carry out a study using different collection methods and solvents on numerous individuals of the same species, in order to compare the effects of these methods. Hara et al. (2005) were able to map the composition of the secretions of 22 gonyleptids (Laniatores, Grassatores) on a phylogenetic tree constructed from other characters. They found rampant homoplasy, but noted that closely related compounds (pairs of phenols and quinones) could easily be transformed into one another by oxidation or reduction, suggesting that “families” of such compounds could represent synapomorphies as transformation series of compound families. However, their final conclusions were ambiguous. On the one hand, it appeared that the great diversity of the secretions did not allow the recognition of phylogenetic lineages, with some exceptions, but on the other, they hoped that the analysis of more species and the addition of metabolic sequences and interchangeable compounds might provide phylogenetic information in the future. We agree with this position and plan to continue to analyze opilionid defensive secretions not only in a search for molecules new to science or new to arthropods, but also with the expectation that more data will help to clarify phylogenetic relationships within Opiliones. ACKNOWLEDGMENTS Ron Clouse and Cahyo Rahmadi participated in the collecting trip to Sulawesi that yielded the specimens for this study. Support for fieldwork was made possible by a Putnam Grant from the Museum of Comparative Zoology. Permits were awarded by LIPI. The work of WAS was supported by a grant from the Professional Development Committee of Hampden-Sydney College. LITERATURE CITED Boyer, S.L., R.M. Clouse, L.R. Benavides, P.J. Schwendinger, I. Kuranarathna & G. Giribet. 2007. Biogeography of the world: a case study from cyphophthalmid Opiliones, a globally distributed group of arachnids. Journal of Biogeography 34:2070-2085. Clouse, R.M. & G. Giribet. 2007. Across Lydekker’s Line — first report of mite harvestmen (Opiliones : Cyphophthalmi : Stylo- cellidae) from New Guinea. Invertebrate Systematics 21:207-227. Eisner, T., A.F. Kluge, J.E. Carrel &. J. Meinwald. 1971. Defense of phalangid: liquid repellent administered by leg dabbing. Science 173:650-652. Giribet, G. & S.L. Boyer. 2002. A cladistic analysis of the cyphophthalmid genera (Opiliones, Cyphophthalmi). Journal of Arachnology 30:110-128. Giribet, G., G.D. Edgecombe, W.C. Wheeler & C. Babbitt. 2002. Phylogeny and systematic position of Opiliones: a combined analysis of chelicerate relationships using morphological and molecular data. Cladistics 18:5-70. Giribet, G., P.P. Sharma & D.B. Bastawade. 2007. A new genus and species of Cyphophthalmi (Arachnida: Opiliones) from the north- eastern states of India. Zoological Journal of the Linnean Society 151:663-670. Gnaspini, P. & M.R. Hara. 2007. Defense mechanisms. Pp. 374-399. hi Harvestmen: The Biology of Opiliones. (R. Pinto-Da-Rocha, G. Machado & G. Giribet, eds.). Harvard University Press, Cam- bridge, MA. Hara, M.R., A.J. Cavalheiro, P. Gnaspini & D.Y.A.C. Santos. 2005. A comparative analysis of the chemical nature of defensive secretions of Gonyleptidae (Arachnida: Opiliones: Laniatores). Biochemical Systematics and Ecology 33:1210-1225. Jones, T.H., J. Meinwald, K. Hicks & T. Eisner. 1977. Characteriza- tion and synthesis of volatile compounds from the defensive secretions of some “daddy longlegs” (Arachnida: Opiliones: Leiobummi spp.). Proceedings of the National Academy of Sciences USA 74:419^22. Juberthie, C. 1961a. 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Zoological Journal of the Linnean Society 150:221-265. Wheeler, W.C. & C.Y. Hayashi. 1998. The phylogeny of extant chelicerate orders. Cladistics 14:173-192. Manuscript received 11 June 2008, revised 12 January 2009. 2009. The Journal of Arachnology 37:151-159 Natural history of coastal Peruvian solifuges with a redescription of Chinchippus peruvianiis and an additional new species (Arachnida, Solifugae, Ammotrechidae) Alessandro Catenazzi': Department of Biological Sciences, Florida International University, Miami, Florida 33199, USA Jack O. Brookhart and Paula E. Cushing: Department of Zoology, Denver Museum of Nature and Science, 2001 Colorado Blvd., Denver, Colorado 80205-5798, USA Abstract. Two species of Chinchippus (Ammotrechidae) were studied in central Peru. Both species are endemic to the hyper-arid coastal desert and appear to derive most of their energy and nutrients from maritime prey, such as intertidal amphipods feeding on beach-cast algae or as arthropod scavengers feeding upon seabird and pinniped carcasses. Data on the spatial distribution of the two species were obtained from analyzing stomach contents of one common predator, the gecko Phyllodactylus angustidigitus, and suggest that both species are more abundant in insular than in mainland habitats. We redescribe Chinchippus peruvianiis Chamberlin 1920, known only from a female specimen and describe the male for the first time while C. viejaensis is recognized as new. The new species is distinguished from C. peruvianiis by its darker coloration, smaller size, and differences in cheliceral dentition. Keywords: Camel spiders, coastal desert, ecology, Gekkonidae, Peru, Phyllodactylus angustidigitus, taxonomy In the process of investigating the ecology of terrestrial organisms in the coastal desert and guano islands of central Peru, we have come across a series of Chinchippus peruvianiis Chamberlin 1920 and a closely related species. These solifuges, along with other terrestrial predators, thrive in places that could be defined as a barren land of gravel, sand, and granitic outcrops - a moonscape where arachnids and lizards somehow manage to survive, reproduce, and colonize new habitats. The western coast of South America is among the driest places on Earth (Dietrich & Perron 2006), where arid conditions have persisted for the last 14 million years (Alpers & Brimhall 1988). Facing this hyper-arid ecosystem is one of the world’s most productive marine ecosystems, the Peru-Chile cold current (Tarazona & Amtz 2001). The stark contrast in productivity promotes the exchange of energy and nutrients between these two adjacent ecosystems, and marine-derived resources subsidize terrestrial predators along the Peruvian coast (Catenazzi & Donnelly 2007a) and in other coastal deserts (Polls & Hurd 1996). In this study we describe the taxonomy and natural history of the two Chinchippus species and explore their distribution in relation to the availability of marine-derived resources. The genus Chinchippus was established by Chamberlin (1920) based on a single female from the Peruvian island of Chincha. He considered it to belong to the African family Daesiidae. Roewer (1934) included Chinchippus in the ammotrechid subfamily Saronominae based on the segmenta- tion of legs I, II, and IV and the palpal spination. Munia (1976) tentatively included it with the saronomines although its placement was still based on Chamberlin’s sole female. Based on Chamberlin’s single female, the genus Chinchippus can be recognized by: all the legs having a single tarsal segment, no claws on leg I, stridulating ridges on the mesal ' Current address: Department of Integrative Biology, University of California, Berkeley, 3060 Valley Life Sciences Bldg ^3140, Berkeley, California 94720-3140, USA. E-mail: acatenazzi(^gmail.com surface of the chelicera, lateral plates of the “rostrum” shorter than the median plates, and a recurved cephalothorax. METHODS One of us (AC) conducted fieldwork at the Paracas National Reserve (PNR; 13°51'S, 76°16'W), ~19 km S of the Chinchas islands, in the Peruvian Region of Ica (Fig. 1 ). This reserve protects 335,000 ha of coastal waters and subtropical Peruvian coastal desert, including a variety of arid and hyper-arid terrestrial habitats. The PNR includes the Paracas Peninsula, which forms the southern edge of Paracas Bay, and the islands of Sangayan and La Vieja. The coastal topography is extremely heterogeneous and includes sandy, gravel, pebble and boulder beaches; cliffs; wind-shaped landforms; and uplifted ancient beaches. The climate is characteristic of the arid coastal desert of Peru and northern Chile and receives less than 2 mm of rain per year (Craig & Psuty 1968). Temperatures are mild and range between an average high of 22.9° C in February to an average low of 16.3°C in August (Environmental Resources Management 2002). Using the methods of Muma (1951), Brookhart & Muma (1981, 1987), Muma & Brookhart (1988), and Brookhart & Cushing (2004), we measured total length; length of palpus, leg 1, leg IV; length and width of chelicera and propeltidium; width of base of fixed finger; and length and width of female genital operculum using Spot Basic^"^ with an Olympus SZX12 microscope at 25 X magnification. All measurements are in millimeters. Ratios used previously by Brookhart & Cushing (2002, 2004) were computed. These ratios are as follows: A/CP: the sum of the lengths of palpus, leg I, and leg IV divided by the sum of length of chelicera and propeltidium indicating length of appendages in relation to body size. Long- legged species have larger A/CP ratios. Because there is no fondal notch, the cheliceral width/fixed finger width ratio is used to indicate whether the fixed cheliceral finger of the male is thin or robust in relation to the size of the chelicera. Genital operculum length/genital operculum width represents the 151 152 THE JOURNAL OF ARACHNOLOGY Figure 1. — Map of study localities and spatial variation in the frequency of occurrence of Cliincliippus peruvianus and C viejaensis in stomach contents of the gecko Pliyllodactyhis angustidigitus at Isla Sangayan (3 sites), the Paracas Peninsula (10 sites) and Isla La Vieja (4 sites), central Peru. The diameter of circles represents frequency of occurrence ranging from 0% (Yumaque, Paracas Peninsula) to 100% (gull colony, Isla Sangayan). relative size of the female genital operculum in terms of length and width. Species determinations were based on a combina- tion of color comparisons, the shape and dentition patterns of the male chelicerae, palpal setation, and color patterns of the propeltidium, palpus, and legs. The shape of the female chelicerae and the female genital operculum margin were observed using the method of Brookhart & Cushing (2004). Cheliceral dentition patterns were based on the method of Maury (1982) in which, for example, PT-1-2-AT indicates one primary tooth, two intermediate teeth, and one anterior tooth. Collections from which material was borrowed or deposited include the Museum of Comparative Zoology at Harvard University, Cambridge, Massachusetts, USA (MCZ); the Museo de Historia Natural, Universidad Nacional Mayor de San Marcos, Lima, Peru (MUSM); and the Denver Museum of Nature and Science, Denver, Colorado, USA (DMNS). We collected solifuges by using pitfall traps or by opportunistically collecting during nocturnal walks. We used pitfall traps consisting of plastic cups 9 cm in diameter and 10 cm deep filled with a mix of water and detergent along the southern end of Paracas Bay between 6-9 January and 6-11 April 2003. During the January trapping, a series of three pitfall traps were placed near sandy/muddy beaches in coastal dunes and the adjacent desert at 5, 10, and 15 m distance from shore. During the April trapping, we placed pitfall traps near shelly beaches along transects at 0, 0.1, 1, 10 and 100 m from shore. We installed three transects, each one composed of three lines of pitfall traps. In addition to pitfall trapping, we also counted and measured solifuges in 90 1-m^ plots in the intertidal zone in March 2003. Count data of the March and April trapping period were reported by Catenazzi & Donnelly (2007a). Here we report count data from the January trapping, as well as solifuge size-distribution data from the March trapping along the shelly beach. Means are reported ± SE and statistical tests are considered significant at E < 0.05. We include anecdotal observations on predators, prey, and behavior of solifuges observed in the field. Some of these observations were captured in photographs and video and are available online at http://acatenazzi.googlepages.com/chinch- ippus. We relied upon stomach content examinations of the gecko Phyllodactylus angustidigitus Dixon & Huey 1970, a common and ubiquitous reptile in the coastal desert, to better understand the distribution of solifuges in the coastal desert of the Paracas Peninsula and the islands of Sangayan and La Vieja. These geckos feed opportunistically on any live terrestrial arthropod of appropriate size, including beach hoppers, centipedes, arachnids, and insects (Catenazzi & Donnelly 2007a), and do not masticate their prey, facilitating the task of identifying prey remains in the stomachs. Stomach contents were obtained by inserting a small catheter through the esophagus and by flushing the geckos’ stomachs with water (Catenazzi & Donnelly 2007a). Stomach contents (n = 814) were collected from Isla La Vieja (4 sites), Isla Sangayan (3 sites), and the Paracas Peninsula (10 sites). We considered whole prey items only to calculate frequency of occurrence of Chinchippus prey with respect to number of geckos sampled and with respect to total number of prey items in all pooled stomach contents. CATENAZZI ET AL.— COASTAL PERUVIAN SOLIFUGES 153 Figures 2-8. — Chinchippus peruvianus. 2-5. Female holotype, Peru: Ica: Islas Chinchas, 26 October 1919, R. C. Murphy (MCZ 519): 2. Propeltidium, dorsal; 3. Palp, dorsal ; 4. Right chelicera, ectal; 5. Genital operculum, ventral. 6-8. Male, Peru, Cangrejal, 25 March 2003, A. Catenazzi: 6. Propeltidium, dorsal; 7. Right chelicera, ectal; 8. Right chelicera, mesal. Scale bars = 1 mm. TAXONOMY Family Ammotrechidae Roewer 1934 Subfamily Saronominae Roewer 1934 Genus Chinchippus Chamberlin 1920 Chinchippus peruvianus Chamberlin 1920 (Figs. 2-8) Chinchippus peruvianus Chamberlin 1920:36-37. Material examined. — Type: PERU: Ica Region: female holotype, Islas Chinchas (13°37'37"S, 76°23'21"W), 26 Octo- ber 1919, R.C. Murphy (MCZ 519). Other material: PERU: Ica: Paracas Bay: 2 ? (1 from the stomach content of Phyllodactylus angustidigitus), Cangrejal (13°51'03"S, 76°17'08"W, 1 m elev.), 3 March 2003, A. Catenazzi (DMNS); 2 c? same data except 25 March 2003, A. Catenazzi (DMNS); 2 $, La Aguada (13°51'44"S, 76°16'15"W, 2 m elev.), 7 January 2003, A. Catenazzi (DMNS); 1 cJ, Museo de Paracas Julio C. Tello (13°52'00"S, 76°16'26"W, 13 m elev.), 25 March 2003, A. Catenazzi (DMNS). Diagnosis. — Chinchippus peruvianus is larger and lighter than C. viejaensis and differs by cheliceral dentition (compare Figs. 7 and 1 1). 154 THE JOURNAL OF ARACHNOLOGY Figure 9. — Habitus of Chinchippus viejaensis. Male (left), female (right). Description. — Female holotype: Color: Overall color as described by Chamberlin (1920): chelicera mottled violet- brown; propeltidium violet-brown with a pale creamy light ovate area highlighted by a lighter median band extending from eye tubercle to posterior end of propeltidium; eye tubercle dark (Fig. 2); mesapeltidium, metapeltidium white; abdomen creamy yellow with a median mottled violet-brown stripe dorsally and creamy grey laterally and dorsally; palp darker on tarsus and femur, metatarsus and tibia pale to white, coxa creamy yellow (Fig. 3). Legs II, III, IV light, violet-brown on tarsus, metatarsus, and apical parts of the tibia, darker on distal end of tibia as in femur, coxa creamy white. Leg I creamy white except for femur, which is a light violet; malleoli white. Chelicera: fondal teeth graded HI, L IV, II; Fixed finger teeth arranged 1-PT-l-MT-l-AT; movable finger teeth ar- ranged PT-1-2-AT (Fig. 4; however, Chamberlin illustrates PT-l-AT). Six to seven stidulatory ridges on the nieso dorsal aspect of the chelicerae. Palp: tarsus/metatarsus ratio 3:1. Legs: leg IV tarsus with ventral paired setae arranged 2-2-2- 2-2-1. Abdomen: genital operculum: clavate with anterior arms thick, median edge recurved forming a deep central cavity accessing the genital opening, posterior edge straight (Fig. 5). Male: Color: color pattern, including appendages, the same as in the female except the median pale stripe extends only from the posterior third of the propeltidium towards the ocular tubercle (Fig. 6); chelicera mottled dorsally and ectally coalescing anteriorly (Fig. 7); pale ventrally; malleoli white. Chelicera: fondal teeth graded I-III-II-IV ectally and I, III, II mesally; fixed finger teeth arranged PT-1-2-MT-1-AT; movable finger teeth arranged PT-1-2-AT (Figs. 7, 8). Flagellum a broadly elliptical structure attached to the fixed finger above the primary tooth, slightly to the dorsal edge. The attachment appears to be a concave structure. No setae or fringes are seen on the edges of the flagella. It bears some resemblance to the flagella of Saronomus capensis (Kraepelin 1899) (Maury 1982:127-130, figs. 1-8). Six to seven stridula- tory ridges on the mesal dorsal aspect of the chelicerae (Fig. 8). Palpus: tarsal/metatarsal ratio of 3.4:1 (Fig. 3). Legs: all legs with a single tarsal segment; leg I with no claw and slightly enlarged (bulbous) tarsus; leg IV with ventral spination of 2-2-2-2-2-1. Dimensions. — Female holotype: total length 10.0, cheliceral length 5.0, cheliceral width 1.7, propeltidium length 1.75, propeltidium width 3.15, palpus length 10.0, first leg length 8.0, fourth leg length unknown (damaged). Ratios: A/CP (cannot be computed), genital operculum length/width 0.5. Females (n = 4): total length 12.0-16.0, cheliceral length 4.4-4.85, cheliceral width 1.37-1.65, propeltidium length 1.8- 2.2, palpus length 12.0-16.0, first leg length 10.0-11.0, fourth leg length 16.0-20.0. Ratios: A/CP 7.26-7.48. Males (n = 3): total length 12.5-13.5, cheliceral length 2.5- 2.7, cheliceral width 0.88-0.99, propeltidium length 1.7-1.75, propeltidium width 1.8-2. 2, palpus length 12.5-13.5, first leg length 9.0, fourth leg length 17.0-19.0. Ratios: A/CP 10.3- 11.7. Chinchippus viejaensis new species (Figs. 9-14) Material examined. — Types: PERU: Ica Region: male holotype, Isla La Vieja, Reserva Nacional de Paracas, CATENAZZI ET AL.— COASTAL PERUVIAN SOLIFUGES 155 Figures 10-14. — Chinchippus viejaemis. Male holotype and female allotype, Peru: Isla La Vieja, 15 September 2003, A. Catenazzi. 10-13. Male holotype: 10. Propeltidium, dorsal; 11. Right chelicera, ectal; 12. Right chelicera, mesal; 13. Palpal tarsus, metatarsus, dorsal. 14. Female allotype, right chelicera, ectal. Scale bars = 1 mm. 14°26'28.4"S, 76°12'28.4"W, 230 m, 15 September 2003, A. Catenazzi (DMNS). Allotype female collected with holotype (MUSM). Etymology. — Named for the type locality, Isla La Vieja, Peru. Diagnosis. — This species can be differentiated from C. peruvianus by its darker coloration, smaller size, and differences in cheliceral dentition (compare Figs. 7 and 11). Description. — Male holotype'. Color: pale mottled violet to dark violet-brown overall; palpal tarsus, metatarsus, tibia, and 156 THE JOURNAL OF ARACHNOLOGY Table 1 . — Average number of Chinchippus peruvianus captured per pitfall trap near sandy and muddy beaches at Paracas Bay between 6- 9 January 2003. Position = distance from the mean high tide level. Traps = number of pitfall traps. No males were captured in pitfall traps. Position Traps Females Immatures 5 m 27 0.11 ± 0.06 0.04 ± 0.04 10 m 27 0.07 ± 0.05 — 15 m 27 0.07 ± 0.05 0.04 ± 0.04 Total 81 0.09 ± 0.03 0.02 ± 0.02 apical two thirds of the femur dark violet-brown dorsally, creamy white ventrally; legs I and II dusky brown and violet- brown at the tibia-femur; legs III and IV violet-brown dorsally on tibia, fibula, and apical portion of tarsus; propeltidium darker violet-brown with a very pale ovate area and a median thin, pale stripe extending from eye tubercle to the posterior of the propeltidium (Figs. 9, 10); chelicerae mottled violet-brown (Figs. 9, 11); abdomen with dorsal violet-brown patches on each sternite dorsally (Fig. 9); malleoli white. Chelicera: fondal teeth three of equal size, fixed finger dentition 1-P-M-l-A, movable finger dentition P-1 -A. Six to seven stridulatory ridges found on posterior mesal surface of chelicerae (Figs. 11, 12). Flagellum of C. viejaensis similar to C. peruviamis with a slightly narrow anterior opening and perhaps a more medial attachment above the primary tooth. No setae or fringe is visible. The cheliceral dentition pattern shows some similarity to Ammotrechida gervaisii (Pocock 1895) (Roewer 1934:600) but has no mesal tooth and no fringed flagellum. Palpus: metatarsus/tarsus ratio 3:1; no spine-like setae (Fig. 13). Legs: leg I with no claw; leg IV with 2-2-1 spine-like setae on the ventral aspect of the tarsus and 2-2-1 -1 on the metatarsus. Female allotype: Color: very similar to the male with the abdominal tergites a lighter color (Fig. 9). The median pale ovate area of the propeltidium is lighter in the female. Leg III femur creamy white. Chelicera: fixed finger dentition P-l-M-l-A; movable finger P-1 -A; fondal teeth III, I, II, IV ectally and mesally; 5-6 stridulatory ridges on the dorsal mesal aspect (Fig. 12). Palps: metatarsi/tarsi ratio of 2.5:1. Legs: leg IV with 2-2-1 spine-like setal pattern on ventral aspect of the tarsus and 2-2- 1-1 on the metatarsus. Abdomen: genital operculum similar to C. peruvianus with a deep central cavity forming the entrance to the genital orifice. Dimensions. — Male holotype: length 10.0, cheliceral length 2.35, cheliceral width 0.97, propeltidium length 1.54, propelti- dium width 2.02, palpus length 10.0, first leg length 7.0, fourth leg length 8.0. Ratios: A/CP 6.84. Female allotype: total length 1 1.0, cheliceral length 2.75, cheliceral width 1.04, propeltidium length 1.51, propeltidium width 2.27, palpal length 7.0, first leg length 6.0, fourth leg length 6.5. Ratios: A/CP 5.64. TAXONOMIC REMARKS Previous to this study, Chinchippus was a monotypic genus based on a single female specimen. The identification of the associated male and a second species supports Chamberlin’s (1920) erection of this genus. These two species are in the subfamily Saronominae based on tarsal segmentation of the 12 1 10 - 0 2 4 6 8 10 12 14 16 18 20 22 Body length (mm) Figure 15. — Size distribution of Chinchippus peruvianus (n = 101 individuals) at Cangrejal, Paracas Bay, Peru between 26-28 March 2003. 4'*’ leg and the flagellar structure. These characters differen- tiate the species in the Saronominae from all species currently placed in the Ammotrechinae. The genita! opercula of the two species of Chinchippus are very similar and differ from other members of the subfamily Saronominae or Ammotrechinae. ECOLOGICAL RESULTS AND DISCUSSION Chinchippus peruvianus. — Murphy (1925) collected the holotype in a building of the Compania Administradora del Guano on the Islas Chinchas and reported that individuals hunt for invertebrates attracted to a light source at night. We made most of our collections and observations on the natural history of C. peruvianus at the Paracas Peninsula and on Isla Sangayan. The species appeared to be extremely abundant along a 2 km stretch of shelly beach near the southern end of Paracas Bay (Catenazzi & Donnelly 2007a, b), where individuals could easily be found by lifting rocks, empty shells, and dried marine wrack near shore. At Isla Sangayan, we observed C. peruvianus under rocks and carcasses of South American sea lions (Otaria Jlavescens Peron 1816). Results of the January pitfall trapping near sandy/muddy beaches of Paracas Bay (Table 1) suggest that C. peruvianus was more frequent along shelly beaches (see results in Catenazzi & Donnelly 2007a) than it was along sandy or muddy beaches (see below for data from gecko stomach content analyses in support of this hypothesis). Body length distribution for individuals captured on shelly beaches during March 2003 (Fig. 15) averaged 9.4 ± 0.4 mm for the population including immatures (n = 101); the maximum body length was 20.2 mm for a female. Chinchippus peruvianus was found in stomach contents of P. angustigitus from most sites in the Paracas Peninsula and from all sites on Isla Sangayan (Tables 2, 3; Fig. 1). Note that the site with 100% frequency of occurrence of C. peruvianus (gull colony on Sangayan) was based on the stomach contents of only three geckos. At the Paracas Peninsula, near-shore sites along Paracas Bay and Lagunillas Bay had the highest frequencies of occurrence, possibly because of the large amount of beach-cast macro-algae supporting abundant CATENAZZI ET AL.— COASTAL PERUVIAN SOLIFUGES 157 Table 2. — Spatial variation in the frequency of occurrence of Chinchippus peruvianus in stomach contents of the gecko Phyllodactylus anguslidigitus in 10 sites of the Paracas Peninsula, Peru between March and December 2003. See Fig. 1 for site locations. Geckos = number of stomach contents examined; Occurrence = frequency of C. peruvianus in the geckos’ stomach contents; Prey items = number of all prey items in the geckos’ stomach contents; Frequency = frequency of C. peruvianus relative to the number of prey items. Site Geckos Occurrence Prey items Frequency Paracas Peninsula Cangrejal 127 22.8% (29) 607 5.8% (35) La Aguada 43 4.7% (2) 383 0.5% (2) Sequion 33 12.1% (4) 218 1.8% (4) Yumaque 17 -(0) 81 — (0) Talpo 41 -(0) 171 -(0) Lagunillas 41 9.8% (4) 235 2.1% (5) Playa roja 24 8.3% (2) 166 1.2% (2) Los Viejos (beach) 54 -(0) 421 -(0) Los Viejos (desert) 38 5.3% (2) 96 2.1% (2) Arquillo 18 5.6% (1) 123 0.8% (1) Isla Sangaydn Lobera 178 1.1% (2) 1408 0.2% (3) Gull colony 3 100.0% (3) 65 20.0% (13) Lomas 27 3.7% (1) 164 0.6% (1) Total 644 7.8% (50) 4138 1.6% (68) populations of intertidal arthropods and/or because these beaches were easily accessible to both solifuges and geckos. Frequencies of occurrence were low at coastal sites near cliffs (Arquillo, Yumaque, Playa Roja) or sites that are exposed to the ocean (Los Viejos, Talpo), similarly to distribution patterns found in P. angustidigitus geckos (Catenazzi & Donnelly, unpublished data). Chinchippus peruvianus readily excavates burrows in fine sand when disturbed. The burrowing behavior included biting, raking, and plowing sand at irregular intervals. However, C. peruvianus was also found in pebble beaches and in coarse soil where other microhabitats replace burrows (e.g., dried macroalgae, sea lion and seabird carcasses; A. Catenazzi pers. obs.). Other species of South American solifuges seem to be associated with vegetation cover and soil characteristics: for example Xavier & Rocha (2001) detected a preference of Mummucia mauryi Rocha (in Xavier & Rocha 2001) for areas covered by Opuntia inamoena (Cactaceae) during the dry season, whereas Rocha & Carvalho (2006) and Martins et al. (2004) noted that white sandy soils where solifuges can easily excavate their burrows may facilitate colonization by Mum- mucia taiete Rocha & Carvalho 2006 and M. coaraciandu Pinto-da-Rocha & Rocha 2004 respectively. In the case of C. peruvianus (as well as C. viejaensis, see below), vegetation cover is unlikely to explain distribution patterns because it is extremely scarce and absent at most sites. Island and coastal habitats colonized by the two Chinchippus also differ widely in soil types (A. Catenazzi, pers. obs.; see habitat descriptions). The higher frequency of occurrence of these solifuges in places that receive marine-derived energy and nutrients, such as beaches with stranded marine macroalgae colonized by arthropods or insular seabird colonies with arthropod scavengers and ectoparasites suggests that food availability in the hyper-arid Peruvian coastal desert may explain distribution patterns. Seasonal activity can be inferred from results of the geckos’ stomach contents, by assuming that the feeding preference of geckos did not vary seasonally. Solifuges at Paracas Bay were most frequent in the geckos’ stomachs during the austral summer (Table 3), and their frequency of occurrence with respect to the total number of prey items from March to December 2003 (including data from December 2004) followed a polynomial curve (y = 0.42x^ — 6.8x + 27.7, R = 0.81) with a minimum predicted value for August (0.3% Table 3. — Seasonal variation in the frequency of occurrence of Chinchippus peruvianus in stomach contents of the gecko Phyllodactylus angustidigitus at Paracas Bay, Peru during March-December 2003. See Fig. 1 for site location and Table 2 for table headings; * includes 15 stomach contents collected in December 2004. Month Geckos Occurrence Prey items Frequency March 12 25.0% (3) 40 10.0% (4) April 35 20.0% (7) 130 7.7% (10) May 43 18.6% (8) 176 5.7% (10) June 27 18.5% (5) 188 2.7% (5) July 9 -(0) 35 -(0) August 16 6.3% (1) 143 0.7% (1) September 3 -(0) 89 -(0) October 8 25.0% (2) 65 3.1% (2) November 13 — (0) 112 -(0) December* 19 26.3% (5) 55 9.1% (5) Total 170 18.2% (31) 990 3.7% (37) 158 THE JOURNAL OF ARACHNOLOGY Table 4. — Spatial variation in the frequency of occurrence of Chinchippus viejaensis in stomach contents of the gecko Phyllodacty- his cmgustidigitus in four sites of Isla La Vieja, Peru. See Fig. 1 for site locations and Table 2 for table headings. Site Geckos Occurrence Prey items Frequency AB 22 54.5% (12) 449 3.6% (16) AC Pan de Azucar 19 15.8% (3) 96 3.1% (3) Beach 36 2.8% (1) 224 0.4% (1) Zona guanera 14 7.1% (1) 182 0.5% (1) Total 91 18.7% (17) 951 2.2% (21) frequency of occurrence). This seasonal activity contrasts with the phenology described by Martins et al. (2004) for M. coarackmdu in the Brazilian Cerrado, where surface activity based on pitfall traps was negatively correlated with monthly temperature. However, higher temperatures in the Cerrado were associated with higher rainfall, which could also intluence solifuge activity. Rainfall in Paracas is negligible throughout the year, but average night temperatures can be low in the austral winter and could limit solifuge activity. The diet of C. peruvianus is dependent upon marine sources of energy and nutrients. In the case of supratidal populations, such as those in Paracas Bay (Tables 3 and 4), solifuges rely on intertidal algivores for their diet (Catenazzi & Donnelly 2007a). The beach hopper Trcmsorchestia chiliensis (Amphi- poda, Talitridae) was the most common prey item based on field observations (5 out of 10 feeding events). Analyses of stable carbon isotopes also suggested that these beach hoppers were an important prey item for C. peruvianus (Catenazzi & Donnelly 2007a, b). Insular solifuge populations likely feed on ectoparasites and other arthropods found in detritus or on the carcasses of seabirds and pinnipeds because the Islas Chinchas (type locality of C. peruvianus) are entirely devoid of vegetation, and Isla Sangayan has scant vegetation that occupies a tiny fraction of the island (the site Lomas in Fig. 1); both islands are mostly cliff-bound. For solifuge populations near the Otaria flavescens colony on Isla Sangayan (site Lobera in Fig. 1 ), arthropod scavengers of pinniped carcasses are likely to be important dietary items, as suggested by the high carbon and nitrogen stable isotope values (albeit only two individuals were analyzed, with 6'^C = — 14.64%o and 14.89%o and 5'^N = 24.33%o and 25.58%o; A. Catenazzi, unpubl. data). Nitrogen isotopic values increase on average by 3.4%o for each trophic interaction and therefore can be used to estimate the trophic position of an organism (Post 2002). Nitrogen isotopic values of O. flavescens on Sangayan average 17.44 ± 0.35%o (Catenazzi & Donnelly 2008); therefore, isotopic values of C. peruvianus are consistent with the idea that solifuges feed on scavengers of O. flavescens', (i.e., that they are two trophic positions above O. flavescens). Natural predators of C. peruvianus at the Paracas Peninsula and Sangayan include, in addition to P. cmgustidigitus, the scorpion Brachistosternus ehrenhergii (Gervais 1841), the spider Odo sp. (Zoridae), as well as conspecific individuals. Cannibalism is likely to be common, because these solifuges occur at high density in the first meters from shore. We observed cannibalism in the field on two occasions, and all captive encounters of pairs of C peruvianus resulted in one individual devouring the other one. The tsunami that followed a 7.8 magnitude earthquake on 15 August 2007 modified the coastal landscape in Paracas Bay. High waves removed many of the supralitoral dunes where C. peruvianus specimens had been collected for this study. It is possible that the flooding of the supratidal areas caused a decline in local populations because Catenazzi & Donnelly (2007a) noted that most C. peruvianus are found in the supratidal zone within 1 m from the high mean tide level. However, C peruvianus could recolonize supratidal areas from sections of beach that were protected from the tsunami by a steeper slope of the beach and/or by the presence of rocks and other topographic features. Chinchippus viejaensis. — This species has only been collected from Isla La Vieja (also called Isla Independencia) in central Peru. This island (area 60.86 ha) is located in Independencia Bay, approximately 6 km west of the mainland and 1.6 km north of a smaller island, Santa Rosa (Fig. 1). Both La Vieja and Santa Rosa are guano islands where hundreds of thousands of seabirds, mainly guanay cormorants {Phalacro- corax bougainvillii Lesson 1837) and Peruvian boobies {Sula variegata Tschudi 1843), used to congregate. At the time of our visits between July and November 2003, La Vieja did not have any breeding colony of these two guano bird species. However, the upper parts of the island were interspersed with nests of kelp gulls {Larus dominicanus Lichtenstein 1823) and Peruvian diving-petrels {Pelecanoides garnotii Lesson 1828). Most specimens of Chinchippus viejaensis were collected in pitfall traps and stomach contents of the gecko P. angustidi- gitus from a small ridge on the southern slope of the island (type locality, site AB on Fig. 1). The slope measures ~12° and is exposed towards the south. The ground is covered with coarse pebbles (16-32 mm grain size) interspersed with a few granitic outcrops. No plants grow along the ridge or neighboring areas; however, a thin lichen crust covered some rocks along the top of the ridge. Predominant winds carry ocean aerosols and moisture towards the ridge, which may explain the presence of lichens in an otherwise unproductive environment. During our November visit, we observed many nests of kelp gulls; most nests had been built between rocks along the ridge. Based on our observations, we can document predation of this solifuge by P. angustidigitus only. Chinchippus viejaensis was found in 18.7% of the gecko stomach contents from the entire island, and in 54.5% (12 in 22) of the stomach contents collected at the AB site (Fig. 1, Table 4). Additional sampling locations (Fig. 1) included the western slope of the island (site AC), the beach near Pan de Azucar, and the guano area north of Pan de Azucar. Site AC is similar to site AB in being a barren slope with granitic outcrops and seabird nests. However, this slope is steeper (19°) and exposed to the west. The ground is composed of fine to medium pebbles with some large rocks and several granitic outcrops. Subterranean nests of Peruvian diving-petrels occupy areas of very fine pebbles and coarse sand, whereas nests of kelp gulls (much less frequent than at the AB site) are located among rocks in the granitic outcrops, along with very few plants of Solanum murphyi I.M. Johnst. (four plants within a 2.25-ha quadrant plot). The beach near Pan de Azucar is made of coarse pebbles CATENAZZI ET AL.— COASTAL PERUVIAN SOLIFUGES Table 5. — Seasonal variation in the frequency of occurrence of Chinchippus viejaensis in stomach contents of the gecko Phyllodacty- lus angustidigitus at Isla La Vieja, Peru. See Fig. 1 for location and Table 2 for table headings. Month Geckos Occurrence Prey items Frequency July 35 14.3% (5) 403 1.2% (5) September 38 18.4% (7) 376 2.4% (9) November 18 27.8% (7) 172 4.1% (7) Total 91 18.7% (17) 951 2.2% (21) frequently littered with marine wrack including kelp and crustacean carcasses. The ground adjacent to the beach is a gentle slope that abuts on a shallow depression to the northwest of Pan de Azucar beach. Much of this slope had been used by guanay cormorants and Peruvian boobies for nesting ground because at the time of our visit, the ground was covered with ~30 cm of guano. Similarly, guano birds once occupied the guano area to the north of the mentioned shallow depression. It is likely that invertebrates feeding upon seabirds or consuming detritus associated with seabird activity (e.g., regurgitates, feathers, guano, etc.) are important prey items for this solifuge species because the extreme aridity and scant primary productivity of the island supports very few herbivores. In support of this hypothesis, the occurrence and frequency of C. viejaensis in the gecko stomach contents almost doubled at the onset of the breeding season of kelp gulls in November (Table 5). ACKNOWLEDGMENTS We thank the Reserva Nacional de Paracas for logistic support, the National Institute for Natural Resources (INRENA) for issuing research and collecting permits, PROABONOS and its staff for authorizing our visit to Isla La Vieja, and J. Carrillo for field assistance. We thank anonymous reviewers who provided helpful comments that improved the manuscript. AC was funded by a Florida International University (FIU) Dissertation Year Fellowship and by grants from the Organization for Tropical Studies, the PADI Foundation, the American Museum of Natural History, the FIU Graduate Student Association and the Tinker Field Research Grant. PEC and JOB were supported by National Science Foundation grant DBI-0640245 awarded to PEC. 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Bronn, ed.), Vol. 5 (IV) (4) (4— 5):481-723, Akademische Verlagsgesellschaft M.B.H., Leipzig. Tarazona, J. & W. Arntz. 2001. The Peruvian coastal upwelling system. Pp. 229-243. In Coastal Marine Ecosystems of Latin America. (U. Seeliger & B. Kjerfve, eds.). Springer-Verlag, Berlin. Xavier, E. & L.S. Rocha. 2001. Autoecology and description of Mummucia mauryi (Solifugae, Mummuciidae), a new solifuge from Brazilian semi-arid Caatinga. Journal of Arachnology 29:127-134. Manuscript received 25 March 2008, revised II December 2008. 2009. The Journal of Arachnology 37:160-169 Estimating biomass of Neotropical spiders and other arachnids (Araneae, Opiliones, Pseudoscorpiones, Ricinulei) by mass-length regressions Hubert Hofer; Department of Zoology, Staatliches Museum fiir Naturkunde Karlsruhe, Erbprinzenstrasse 13, D-76133 Karlsruhe, Germany. E-mail: hubert.hoefer@smnk.de Ricardo Ott: Museu de Ciencias Naturals, Fundagao Zoobotanica do Rio Grande do Sul, Porto Alegre, Brazil Abstract. We sampled 505 specimens of 7 arachnid orders (313 Araneae, 65 Opiliones, 111 Pseudoscorpiones, 10 Ricinulei, 3 Schizomida, 1 Thelyphonida, 2 Scorpiones) in natural forest and agroforestry sites in central Amazonia to analyze fresh and dry mass to body length relations. The low number of schizomids, scorpions, and thelyphonids did not allow statistical analyses, but the raw data are given, because these represent the first data published for these groups from Amazonia. For all other orders general mass-length relationships for ecological studies were determined. Non-linear regressions with a power model proved to describe the relations very well and are highly significant for all taxa and groups analyzed. The resulting equations can thus be used to estimate biomass of large samples of arachnids from Amazonia based on individual body length measurements. Linear regressions of mass to length with log-transformed data also described the relation adequately, but using the resulting equations to estimate biomass of the whole spider sample caused a higher bias. This is because small biases of mass-length relation of the largest spider individuals are exponentiated. However, linear regressions behaved better for spiders smaller than 8 mm. The ratio of dry to fresh mass was around 0.3 for spiders; 0.4 for pseudoscorpions, schizomids, and thelyphonids; 0.44 for opilionids; and 0.53 for Ricinulei. A second sample of 99 spiders from a South Brazilian Atlantic Forest revealed similar mass-length relations, but a different dry to fresh mass ratio. For spiders, the usefulness of general equations to determine the biomass of bulk samples from ecological studies with certain precision requirements was further explored by using the equations from the two datasets crosswise, regarding the resulting bias and by applying equations to a further dataset from an ecological investigation. In conclusion and accordance to former studies, general equations derived from mass-length regressions of bulk samples including many specimens of different families and guilds are appropriate for an estimation of the biomass of bulk samples from ecological studies. Equations from mass-length regressions from the literature, resulting from spider samples in temperate regions, should not be used to estimate biomass of samples from neotropical spider assemblages, especially when absolute biomass is of interest and when precision is required. They underestimate biomass of tropical assemblages due to a strong bias in mass-length relation of tropical spiders larger than 10 mm. Depending on the distribution of large spiders in samples, considerable biases in single samples could affect ecological analyses. Resumo. Analisamos as relagoes entre comprimento corporal e massa fresca e seca de 505 especimes de sete ordens de aracnideos (313 Araneae, 65 Opiliones, 111 Pseudoscorpiones, 10 Ricinulei, 3 Schizomida, 1 Thelyphonida, 2 Scorpiones) coletados em llorestas e agroflorestas na Amazonia Central. Devido ao numero baixo de Schizomida, Scorpiones e Thelyphonida nenhuma analise estatistica foi possivel e os dados brutos sao apresentados a serem os primeiros dados publicados destes grupos para a Amazonia. Para as outras ordens analises de regressao foram feitas. Regressoes nao- lineares de modelo potencial demonstraram excelente descriqao para as relagoes, sendo altamente significativas para os taxons e grupos analisados. Os coeficientes obtidos nestas regressoes poderao servir de base para o calculo de biomassa em amostras da Regiao Amazonica que contenham grande mimero de aracnideos, utilizando-se como medida somente o comprimento total de cada individuo. Utilizando-se dados logaritmicamente transformados, regressoes lineares de massa- comprimento tambem descreveram adequadamente a relaqao. Todavia a utilizaqao destes coeficientes, para estimar exclusivamente a biomassa da amostra total de aranhas, apresentou resultados tendenciosos em funqao do efeito forte da relaqao exponencial a desvios pequenos em aranhas de grande porte. Regressoes lineares apresentaram um comportamento estatistico mais favoravel apenas para aranhas com menos de 8 mm de comprimento corporal. A relagao obtida para massa seca em relaqao a massa fresca foi de cerca de 0.3 para aranhas, cerca de 0.4 para Pseudoscorpiones, Schizomida e Thelyphonida, 0.44 para Opiliones e 0.53 para Ricinulei. Uma segunda amostragem de 99 aranhas na regiao meridional da Mata Atlantica brasileira revelou rela?6es de massa-comprimento similares, porem, com uma relaqao diferenciada de massa seca a massa fresca. Para a ordem de aranhas a utilidade de equagoes gerais para a determinagao da biomassa de amostras ecologicas com devida precisao foi analisada aplicando coeficientes resultando de amostragens de outras regioes. Concluimos que coeficientes de regressoes de massa-comprimento sao apropriados para uso em relagao a assembleia inteira de aracnideos, desde que as amostras contenham especimes de varias familias e guildas diferentes. Os coeficientes obtidos na regressao da grande amostragem da Regiao Amazonica podem ser usadas para a assembleias de aranhas da Mata Atlantica, porem nao e aconselhavel uso reciproco, mais especificamente para estimativas de massa seca. A utilizagao de coeficientes de regressoes de massa-comprimento disponiveis atualmente na literatura, resultante de amostragens em > regioes temperadas, deveria ser evitada para a estimativa de biomassa em amostras de assembleias de aranhas neotropicais. Estes coeficientes subestimam a biomassa de assembleias tropicais devido a uma grande distorgao na relagao entre massa e comprimento corporal em aranhas maiores do que 10 mm. Desta maneira analises ecologicas podem ser altamente influenciadas pcla distribuigao de grandes aranhas entre as amostras individuais com distorgao dos resultados. Keywords: Arachnida, mass-length relationship, Brazil 160 HOFER & OTT— ESTIMATING BIOMASS OF SPIDERS 161 Biomass data (in the sense of the weight of living animals per unit area, Bornebusch 1930; Edwards 1966) for arthropods are needed in many ecological studies, especially when these aim to analyze the role and functions of these abundant animals in ecosystems and food webs. Biomass of soil fauna is of special interest in studies of nutrient cycling involving the role of the fauna in decomposition and organic matter transformation. The importance of soil fauna has long been recognized and their function is also being studied more frequently in Neotropical ecosystems (Lavelle et al. 1997, 2001; Barros et al. 2003, 2006; Mathieu et al. 2004). The context in which we needed to estimate biomass of arachnids and other arthropods was given by two projects in the Brazilian-German research programme SHIFT (Studies on Human Impact on Forests and Floodplains in the Tropics) studying the quantitative contribution of soil fauna to decomposition in central Amazonian natural forests and different agroforestry systems (Hofer et al. 2001; Hanagarth et al. 2004; Martins et al. 2004; Brown et al. 2006). Biomass can be obtained by direct weighing of individual living arthropods with analytical balances, but this is a very time consuming task and for very active animals it is difficult or impossible to obtain precise data. Certainly direct weighing is not a practical method in the field and for larger samples in laboratories. Most specimens in ecological studies are trapped and killed in fluids such as ethanol and it is difficult to measure preserved animals on a balance. Also, weighing fresh weight of preserved animals may provide incorrect estimations as body weight may be altered during preservation. For most studies dry mass is easier to obtain, but drying specimens or bulk samples to a constant weight, usually at 65° C or more, makes it impossible to later identify them due to their fragility. An alternative method is to use statistically verified relation- ships of mass with easily measurable body dimensions, such as body length or width, to estimate the biomass of each specimen. Body length might even be measured in the field or estimated with live animals so animals may not even need to be collected. Regressions using a power model (mass = a (size)’’) usually adequately describe mass-length-relations for most arthropods (Rogers et al. 1976, 1977; Schoener 1980; Sample et al. 1993; Edwards 1996). They have also been shown to provide useful data for spiders from temperate regions (Breymeyer 1967; Norberg 1978; Clausen 1983; Edwards 1996; Henschel et al. 1996a; Lang et al. 1997; Edwards & Gabriel 1998). Spiders and to a lesser extent other arachnids (opilionids, pseudoscorpions) are abundant in all terrestrial environments and are often included in functional ecological studies due to their well defined position in the food web as (arthropod) predators and their usefulness to indicate habitat quality (Jocque 1981; Chen & Wise 1999; Wise et al. 1999; Lawrence & Wise 2000, 2004; Wise 2004). As Henschel et al. (1996a) state, it is useful and possible to use general equations for arachnid orders (e.g., spiders and opilionids) to estimate the biomass of single specimens for the whole assemblage, notwithstanding the different species-specific mass-length relationships. They suggest their equations are valid for other regions and habitats in Europe, at least for community studies involving numerous families, genera and species. Our main interest was to derive an equation for a general relationship to estimate biomass of bulk samples to compare soil fauna biomass at different sites in tropical South America. Thus we sampled 505 specimens of spiders and other arachnids from one location in central Amazonia and analyzed mass- length relations of this large collection (first data set) in order to obtain valid equations for the biomass estimates we needed for our studies of Amazonian forest and agroforestry systems. We tested whether these equations reliably estimated biomass of bulk samples of spiders or if different equations were necessary for different functional groups (e.g., wandering versus web building spiders), size classes (tiny spiderlings versus large mygalomorphs), or spiders with an extraordinary body shape (like Micrathena or Deinopis). A second sample of spiders (second data set) was obtained from another region and large scale forest ecosystem of Brazil, e.g., in the southern part of the Brazilian Atlantic Forest (Mata Atlantica) and analyzed in the same way. Having two large data sets on spiders at hand and given the numerous data for this arachnid order in the literature, we explored the usefulness and limitations of general equations to determine the biomass of bulk samples from ecological studies with the required precision. This was done in three steps: 1 . Determin- ing which biases would be introduced when using equations from outside the Neotropical region for the Amazonian sample; 2. Determining the bias introduced by using the equations from the first data set (Amazonia) for the second data set (Atlantic Forest) and vice versa; 3. Determining the bias introduced by applying different equations for data from one ecological study in Amazonia and one ecological study in the Atlantic forest (application data sets) and looking for an effect of the bias on the conclusions of these studies. METHODS Mass-length relations were analyzed using specimens sampled in primary and secondary forests and tree plantations within the area of the Brazilian Agricultural Research Corporation EMBRAPA in central Amazonia near Manaus (02°53'47"S, 59°59'45"W) (first data set). Sampling took place in May 1999 with the aim to obtain as many differently sized and shaped specimens from as many taxa as possible. Specimens were captured alive by hand and stored individu- ally in vials during transport to the laboratory. They were killed by freezing for about one hour and then weighed to obtain fresh mass to the nearest 0.001 mg with a Sartorius MP2 microbalance. Body length, in dorsal view from the anterior edge of the prosoma (excluding chelicerae) to the posterior edge of the opisthosoma (excluding spinnerets), was measured with a graduated eyepiece to the nearest 0.01 mm. Numbers of specimens measured for each order and lower taxonomic levels are given in Table 1 (first data set). Lastly specimens were oven-dried for 24 h at 105° C, cooled to room temperature, and weighed to obtain dry mass. Only three of the ten Ricinulei specimens were dried because of their rarity in museum collections. The resulting ratio dry/fresh mass for these specimens was used to calculate the dry mass for the seven other specimens. From three other arachnid orders too few specimens were caught to calculate regressions (Schizo- mida: 3, Thelyphonida: 1, Scorpiones: 2). Results are presented in Tables 1, 2 and in Figure 3. A second data set including 99 spiders from a South Brazilian Atlantic Forest (Mata Atlantica) (Reserva do 162 THE JOURNAL OF ARACHNOLOGY Table 1. — Number of specimens measured and weighed for length-mass regression, mean and range of body length (minimum and maximum in brackets) from seven arachnid orders. Order/Infraorder/Family First data set (Amazonia) Second data set (Mata Atlantica) Specimens Length (mm) Specimens Length (mm) Araneae 313 4.83 (0.56-36.0) 99 7.08 (1.35-28.0) Infraorder Mygalomorphae 43 3.17 (0.78-19.1) Infraorder Araneomorphae; Amaurobiidae 1 1 8.27 Anapidae 1 1.07 Anyphaenidae 2 6.87 (6.83-6.92) Araneidae 8 1.89 (0.81-3.40) 18 5.77 (2.69-10.67) Corinnidae 11 5.85 (1.85-13.9) 2 4.57 (4.52^.62) Ctenidae 74 12.43 (1.30-36.0) 18 16.52 (4.23-28.0) Deinopidae 1 16.50 Linyphiidae 9 1.60 (1.20-1.90) 1 2.30 Lycosidae 2 16.85 (7.69-26.0) Mysmenidae 3 1.73 (1.35-2.40) Ochyroceratidae 24 1.40 (0.56-2.40) 1 1.83 Oecobiidae 2 1.75 (1.70-1.80) Oonopidae 68 1.46 (0.67-2.50) 1 2.31 Palpimanidae 4 3.04 (1.52^.00) Pholcidae 8 2.00 (1.07^.30) 6 2.79 (1.92-3.94) Pisauridae 2 4.67 (3.96-5.40) 1 4.61 Salticidae 39 3.40 (1.12-6.60) 4 4.86 (3.65-5.77) Scytodidae 5 2.57 (1.60-3.10) Selenopidae 1 5.00 Sparassidae 3 6.00 (5.90-6.10) 2 5.58 (3.56-7.60) Tetragnathidae 2 5.86 (4.33-7.40) Theridiidae 5 1.33 (1.00-2.00) 20 3.02 (1.63-10.0) Theridiosomatidae 3 0.75 (0.62-0.83) 3 1.91 (1.49-2.69) Thomisidae 1 7.60 Trechaleidae 3 12.53 (5.29-25.0) Uloboridae 1 5.38 Zodariidae 4 3.60 (2.00-4.50) Zoridae 4 4.23 (3.85^.81) Opiliones 65 2.12 (0.57-6.90) Pseudoscorpiones 111 1.38 (0.86-2.10) Ricinulei 10 4.46 (2.10-5.60) Schizomida 3 1.62 (1.45-1.88) Scorpiones 2 16.30 (3.60-29.0) Thelyphonida 1 7.00 Cachoeira, Antonina, Parana: 25°25'S, 48°40'W) was ob- tained in 2007. Spiders (Table 1) were sampled manually at night and during the day along trails in secondary forests. Weighing and measuring procedures were the same as described above. Tests for the effects of the bias from different equations were done with two application data sets: one from Amazonia, where spiders were sampled from 16 replicate sites of each of 7 different plantation systems (EMBRAPA central Amazonia) by means of large soil cores; and one from the Atlantic Forest, where 10 litter samples (1 m^) were taken in each of three different regeneration stages of a sub-mountain forest (Schmidt et al. 2008). From both collections all spider specimens {n = 441 and 276) were individually measured (body length), so that coefficients from different regression equations could be applied to estimate the total biomass per site. Data were analyzed with Statistica 7.1 (StatSoft 2005) and graphs prepared with SigmaPlot® 8.0.2 (SPSS 2002). RESULTS Analyses of mass-length relations. — Mass-length relation- ships (for both fresh and dry mass) for the arachnid orders with enough specimens sampled in the Amazonian habitats (first data set) are very well correlated with a regression model of the non-linear (power) form: mass = a (length)*’. Determination coefficients are usually > 0.9 (Tables 3, 4) and type I error probabilities are very low (< 0.001) for both parameters, with the exception of the rare Ricinulei (« = 10, P = 0.15 for coefficient a). The mass-length relationship is almost equally well de- scribed with a linear model using logarithmic data for length and weight {In (mass) = a + b /h (length)). Note that power regression results are often presented in double-logarithmic plots, but the model parameters are not the same for a power model calculated on raw data and a linear model calculated on log-transformed data. In our dataset the linear model represents the most abundant small spiders better because HOFER & OTT— ESTIMATING BIOMASS OF SPIDERS 163 Table 2. — Ratios dry/fresh mass for arachnid orders. Order Ratio dry/fresh mass Family Guild First data set (Amazonia) Second data set (Mata Atlantica) Araneae 0.29 0.21 f4ygalomorphae hunting 0.29 Araneomorphae Anyphaenidae hunting 0.25 Amaurobiidae hunting 0.12 Corinnidae hunting 0.29 0.27 Ctenidae hunting 0.26 0.19 Lycosidae hunting 0.19 Oonopidae hunting 0.34 0.19 Oxyopidae hunting 0.24 Palpimanidae hunting 0.32 Pisauridae hunting 0.28 0.22 Salticidae hunting 0.28 0.21 Scytodidae hunters 0.29 Selenopidae hunting 0.16 Sparassidae hunting 0.26 0.19 Thomisidae hunting 0.18 Trechaleidae hunting 0.20 Zodariidae hunting 0.34 Zoridae hunting 0.20 Anapidae web-building 0.24 Araneidae web-building 0.25 0.22 Deinopidae web-building 0.16 Linyphiidae web-building 0.33 0.19 Mysmenidae web-building 0.20 Ochyroceratidae web-building 0.31 0.19 Oecobiidae web-building 0.29 Pholcidae web-building 0.27 0.20 Tetragnathidae web-building 0.29 Theridiidae web-building 0.28 0.21 Theridiosomatidae web-building 0.29 0.18 Uloboridae web-building 0.18 Opiliones 0.41 Pseudoscorpiones 0.38 Ricinulei 0.53 Schizomida 0.37 Scorpiones 0.30 Thelyphonida 0.39 Table 3. — Regression coefficients (a, b) and coefficient of determination in regressions of fresh mass to body length (left: power model: mass [mg] = a body length [mm]*’, right: linear model: In mass [mg] = a + /« body length [mm] b) for arachnids from Amazonia (first data set) and Mata Atlantica (second data set) {n = sample size, se = standard error, = coefficient of determination). All regressions are highly significant (P < 0.001). Power model Linear model n a ± se b ± se a ± se b ± se R- Mata Atlantica: all Araneae 99 0.066 ± 0.025 3.160 ± 0.118 0.98 - 2.166 ± 0.175 2.872 ± 0.097 0.90 Amazonia: all Araneae 313 0.169 ± 0.009 2.899 ± 0.016 0.99 - 2.058 ± 0.029 2.980 ± 0.020 0.99 Araneae < 2.5 mm 225 0.085 ± 0.010 3.288 ± 0.081 0.94 - 1.958 ± 0.037 2.746 ± 0.053 0.92 Ctenidae 74 0.177 ± 0.020 2.886 ± 0.034 0.99 - 1.758 ± 0.096 2.894 ± 0.039 0.99 Oonopidae 68 0.131 ± 0.007 2.682 ± 0.076 0.94 - 2.039 ± 0.042 2.666 ± 0.099 0.96 Hunting spiders 253 0.169 ± O.OIO 2.899 ± 0.018 0.99 - 2.108 ± 0.023 3.017 ± 0.015 0.99 Web-building 60 0.072 ± 0.011 3.710 ± 0.114 0.97 - 1.784 ± 0.092 2.255 ± 0.169 0.75 Opiliones 65 0.147 ± 0.028 3.622 ± 0.105 0.98 - 0.899 ± 0.048 2.984 ± 0.060 0.97 Pseudoscorpiones 111 0.156 ± 0.006 2.453 ± 0.071 0.92 - 1.892 ± 0.027 2.515 ± 0.073 0.91 Ricinulei 10 0.225 ± 0.146 2.760 ± 0.387 0.93 - 1.907 ± 0.192 3.014 ± 0.130 0.98 164 THE JOURNAL OF ARACHNOLOGY Table 4. — Regression coefficients (a, b) and coefficient of determination in regressions of dry mass to body length (left: power model: mass [mg] = a body length [mm]^, right: linear model: In mass [mg] = a + /« body length [mm] b) for arachnids from Amazonia (first data set) and Mata Atlantica (second data set) {n = sample size, se = standard error, R" = coefficient of determination). All regressions are highly significant (P < 0.001). Power model Linear model n a ± se b ± se a ± se b ± se Mata Atlantica: all Araneae 99 0.0067 ± 0.005 3.413 ± 0.245 0.96 - 3.860 ± 0.224 2.950 ± 0.092 0.93 Amazonia: all Araneae 313 0.0165 ± 0.001 3.242 ± 0.014 0.99 - 3.213 ± 0.029 2.902 ± 0.021 0.98 Araneae < 2.5 mm 225 0.028 ± 0.003 3.180 ± 0.079 0.94 - 3.121 ± 0.038 2.680 ± 0.054 0.92 Ctenidae 74 0.017 ± 0.002 3.232 ± 0.029 0.99 - 3.197 ± 0.096 2.921 ± 0.039 0.99 Oonopidae 68 0.050 ± 0.003 2.459 ± 0.094 0.90 - 3.162 ± 0.046 2.767 ± 0.108 0.95 Hunting spiders 253 0.0165 ± 0.001 3.242 ± 0.016 0.99 - 3.237 ± 0.025 2.926 ± 0.016 0.99 Web-building 60 0.017 ± 0003 3.881 ± 0.123 0.97 - 2.997 ± 0.093 2.199 ± 0.172 0.74 Opiliones 65 0.042 ± 0.009 3.879 ± 0.119 0.98 - 1.862 ± 0.049 3.069 ± 0.062 0.97 Pseudoscorpiones 111 0.057 ± 0.003 2.589 ± 0.103 0.86 - 2.967 ± 0.037 2.771 ± 0.100 0.87 the few large spiders have a very high influence in the power model (Fig. 1). Flowever the fresh biomass of the whole sample (313 spiders) with a mean length of 4.83 mm when estimated with the power model was closer to the observed biomass (99.8%) as when estimated with the linear model (95.7%). The same is true for dry mass estimation (power: 97.6%, linear: 86.9% of observed mass). Because different bulk samples might predominantly consist of either small or large spiders, often influenced by the sampling method, it might be useful to use either the linear model or the power model. In some cases it might even be useful to split a sample by size and use the linear model for spiders < 8 mm and the power model for spiders > 8 mm. Therefore, we present the coefficients of both models (Tables 3, 4). The 313 Amazonian spiders that were measured and weighed represent a large spectrum in terms of size, shape, and taxonomic and functional groups. This dataset includes tiny orb-weavers like Theridiosomatidae and Anapidae; tiny, but long-legged Ochyroceratidae; tiny, but short-legged wandering spiders like Oonopidae; median-sized jumping spiders; very small to large mygalomorphs; large ctenid hunters; as well as large, long-legged pholcids (Table 1). Very few spider specimens (the smallest spider an ochyroceratid, one ctenid, and most of the long-legged ochyroceratids) lay Figure 1 . — Ln-In plot of dry mass (mg) vs. body length (mm) relationship of spiders in the first data set from Amazonia, showing the bias of a power model regression for small spiders as compared to a linear model regression with a bias for large spiders. HOFER & OTT— ESTIMATING BIOMASS OF SPIDERS outside the 95% confidence limits of our regressions and their exclusion did not lead to considerable changes in the model parameters. Nevertheless we calculated separate regressions for small spiders, the families Ctenidae and Oonopidae, the main hunting (or wandering) guilds; and web-building spiders because these groups might be of special interest in ecological studies (see also below); and because they always received high determination coefficients and significances (Tables 3, 4). The strong correlations in some cases caused very high PRESS values (> 30,000 for fresh mass and > 500,000 for dry mass vs. length of spiders). The PRESS value (Predicted Residual Error Sum of Squares) is a gauge of how well a regression model predicts new data and often a hint to overfitting of a dataset, resulting in decreased usefulness for other datasets. To test this, we split the whole Amazonian data set by a random procedure in one learn- and one test dataset (cross-validation). For both fresh mass and dry mass the regression line of the test dataset was well inside the 95% confidence limits of the learn dataset. This shows that the strong correlation is not a result of overfitting and conse- quently the resulting formulae should be useful for an estimation of fresh or dry mass of bulk spider samples from the same region (central Amazonia). The other three orders (Opiliones, Pseudoscorpiones, Ricinulei) for which regression analyses were possible were much more uniform in size and shape (Table 1). Power and linear models performed equally well and the coefficients are presented in Tables 3, 4. Mass-length relationships of these orders and also the single specimens of Schizomida, Scor- piones, and Thelyphonida are presented in Figure 3. The mass-length regressions for spiders collected in the Mata Atlantica (second data set) were also strongly correlated and highly significant, but coefficients were slightly different (Tables 3, 4). Only one subadult deinopid and a twig-like Argyrodes specimen lay outside the 95% confidence limits, but they did not influence the coefficients of the power model, which produced very good estimates of fresh and dry mass (99.5% of observed value) for the whole sample. The linear model in contrast produced a considerable underestimate of fresh and dry mass (70.2% resp. 73.4%). Ratio dry/fresh mass. — Fresh mass and dry mass of spiders were strongly correlated (R~ = 0.99, P < 0.001) in both data sets; the ratio dry/fresh mass was on average 0.293 ± 0.055 for Amazonian spiders and 0.208 ± 0.06 for spiders from the Atlantic forest. There was no significant difference in ratios for the two main hunting and web-building spider guilds (Mest P - 0.4). Anapids (tiny orb weavers) show the smallest ratio (0.24), oonopids and zodariids (small hunters, mostly strongly chitinized) the highest ratio (0.34) (Table 2). The highest variation of dry/fresh mass ratio occurred in the lowest range of body size, which is considered an effect of the decreasing precision of both length and weight measurements with decreasing size of the spiders. There was no correlation between length and the ratio dry/fresh mass. The ratio dry/fresh for opilionids was 0.44 ± 0.06 and for pseudoscorpions 0.38 ± 0.06. Both correlations are strong (i?" > 0.95) and highly significant (P < 0.01). Mean ratio dry/fresh for the three ricinuleid specimens was 0.53, and for the other arachnids between 0.30 and 0.39 (Table 2). 165 General usefulness of equations. — Regarding the statistics of mass-length relationships, one certainly gets good estimates of biomass by length measurements for the Amazonian fauna using the coefficients from our equations. But how large would be the bias when using coefficients from other samples for our data or our coefficients for other data? When using coefficients derived from spider samples from temperate regions (taken from the literature) the estimate of the total biomass of our sample of 313 spiders produced serious biases from the observed mass: 56% (fresh) and 58% (dry mass) with coefficients from the linear model of Edwards & Gabriel (1998; spiders from Massachusetts, USA); 43% (dry mass) with coefficients from the power model of Breymeyer (1967; spiders from Europe); 25% (dry mass) using the coefficients from the power model of Henschel et al. (1996a; spiders from Germany); 23% (fresh mass) from the power model of Norberg (1978; spiders from spruce in Sweden). These strong biases are caused by the relatively high number of spiders with a length over 12 mm (e.g., Ctenidae) and some very large individuals (24-36 mm) in our samples and the underestimation of these large spiders by formulae from temperate spider faunas (Fig. 2), which only represent spiders up to a length of 10 mm (Henschel et al. 1996a) or 8 mm (Norberg 1978). The equation of Rogers et al. (1977) from spiders (0.7-12 mm) collected from a shrub-steppe in south- central Washington suited our data set better (105% of observed dry mass). To answer the question whether our equations are generally applicable to samples from spider assemblages in the Neotropics we tested our Amazonian equation on a spider sample (second data set) from another forest Brazilian ecosystem (Mata Atlantica) situated further south, geograph- ically in the subtropics, and vice versa. When applying the Amazonian coefficients, the fresh biomass of the Atlantic Forest spiders was relatively well estimated (1 13% with power model, 110% with linear model), but the dry mass estimate was considerably overestimated (143% and 121%). This is most probably caused by the lower ratio dry/fresh mass (0.21) for the spiders sampled in the Atlantic Forest in comparison with the spiders from Amazonia (0.29) (Table 2). When using the coefficients from the Mata Atlantica data set for the Amazonian data set the following biases (underestimation) resulted for fresh respectively dry mass: 84.5% / 66.4% by power, 62.6% / 52.4% by linear model. To obtain an idea of the effect of such biases we used one application data set from Amazonia. Fig. 4 shows box plots with means, medians and variances (percentiles) of spider biomass samples from different plantation systems, calculated with different coefficients. For most (5) systems the biomass of spiders per plot estimated with the equation from Henschel et al. (1996a) was higher than the biomass calculated with our own coefficients and showed comparable relations between medians and means and similar variance. This is due to overestimation of the dominant small spiders (< 4 mm) by the Henschel equation (s.a.). In each of the systems 4 and 6, however, one larger spider (8 mm) was sampled, and these are underestimated by the Henschel equation. In consequence, for these two systems the relative position of the means change depending on the equation used. However, due to the generally high variance of spider abundance between the 166 THE JOURNAL OF ARACHNOLOGY Figure 2. — Ln-ln plot of dry mass (mg) vs. body length (mm) relationship of spiders in the first data set from Amazonia, showing the bias when using regression coefficients (a, b) from the literature (all linear models). Figure 3. — Ln-ln plot of dry mass (mg) vs. body length (mm) relationship for other arachnid orders from Amazonia (regression lines for opilionids and pseudoscorpions from linear models). HOFER & OTT— ESTIMATING BIOMASS OF SPIDERS 167 100 10 ro E o CO .i ® 0,01 0,001 different agroforestry systems Figure 4. — Estimated total biomass of spiders in samples from seven different agroforestry systems in Amazonia (application data set), showing biases caused by applying regression coefficients originating from a data set from a temperate region in comparison with the coefficients originating from the first (Amazonian) data set. Box plots show the median (thin line), the mean (thick broken line), 25th and 75th percentiles (box), 10th and 90th percentiles (whiskers) and outliers (circles). I I caic. by formula Hofer & Ott I J calc, by formula Henschel et al 1 1 1 — 1 1 1 1 1 r replicates there are no significant differences between the systems, no matter if tested on means (ANOVA) or ranks (Kruskall-Wallis) and by both equations. We also applied the coefficients derived from the Amazo- nian data set in comparison with the coefficients derived from the Atlantic Forest data set to a second application data set: 30 litter samples taken in three different regeneration stages of an Atlantic sub-mountain forest (Schmidt et al. in press). Mean dry mass values of spiders calculated by the Amazonian formula were 2.8, 5.3, 14.4 mg m~^ and calculated by the Atlantic forest formula 1.5, 2.9, 8.6 mg m“^. Biomass values were significantly different (overestimated by the Amazonian formula, paired /-test P < 0.01), but ANOVA for the effects of the regeneration stage on biomass gave no significant effect. DISCUSSION Mass-length regressions are a formidable solution for estimating biomass without having to destroy the specimens or handle them tediously on a microbalance, which is time- consuming and expensive. Literature and our investigation show clearly that this can be made with one measurement of body length, which can be precisely taken with a micrometer eyepiece or a vernier caliper, even for live arthropods. In view of the very high determination coefficients and very low error probabilities, power regressions of length to estimate fresh or dry mass absolutely satisfy the needs, and no further effort is necessary to estimate volume by measurements of several body dimensions. A model should also not be overfitted (see below) since it would lose its applicability to new datasets. As mass is expected to be proportional to length cubed, in regression formulae the power (b) in a uniformly proportioned series of animals is expected to be close to 3. The fresh mass of spiders generally followed this relation, whereas dry mass of spiders and fresh and dry mass of opilionids increased with a power greater than 3. For pseiidoscorpions, ricinuleids, and the oonopid spiders the power was less than 3. Schoener (1980) explained a power smaller than 3 for insects by a trend of longer species tending to be thinner. For our data set we suppose this to be due to different body densities (mass per volume), because all three groups represent more strongly chitinized rather than thinner animals in comparison to the other groups. When the aim is to estimate the biomass of bulk samples including many different spider species of different sizes and shapes, one formula can be used for all spiders, although a few very extraordinary shapes (e.g., very long and thin like some Argyrodes or Deinopis) may lie outside acceptable confidence limits. Especially for tropical soil fauna communities where most specimens are not readily identifiable, often not even to genus or family level, it is desirable, if not necessary, to have one regression equation covering the taxonomic level to which the organisms can be identified (sorted) easily, which most often is the order level for arthropods (Schoener 1980; Sample et al. 1993; Henschel et al. 1996b). Although not appearing very different, the coefficients given by other authors for estimation of spider biomass from length measurements when applied to our data produced slightly different values for single specimens, which result in consid- 168 THE JOURNAL OF ARACHNOLOGY erable biases for bulk samples. The adequate precision of a single mass-length regression depends upon the scientific question, and especially the variance included in the data set (e.g., how many different taxa with different body shapes were included and how strong the abundances vary in reality and in samples). As more mass-length relations of different speci- mens/species are included, the coefficient of determination gets smaller, but unless it remains large enough to explain a considerable portion of the variation (> 0.8) and as long as the probability of being wrong in concluding that the coefficient is not zero remains small (P < 0.05), the regression model gains in predictability. In community ecology data sets, the variances in inverte- brate abundance between different samples and study sites are usually high (standard deviation >100% of the mean) and thus precision of regression factors to calculate the biomass of groups of the community must not be very high, thus allowing relatively fast and rough measures. However, a systematic bias towards certain samples should be avoided. The comparison of the coefficients extracted from the two different models fitting our own data has already shown a possible cause for such a bias: a different proportion of very small or very large spiders in different samples treated with the same equation. In our tests, bias due to the “wrong” equation used for an estimation of biomass did not produce different ecological results. If no equation for the spider assemblage of interest is available, coefficients from an equation based on samples from other regions can be used if the size distributions do not differ strongly, which is obviously the case comparing spider assemblages from temperate and tropical regions. Attention must be given to individual, very large spiders in a sample, which in addition to its already problematic outlier position can produce a king-size bias due to the power effect of the regression. But this should be resolved by statistical proce- dures in the ecological study. We have shown that it is difficult if not impossible to estimate biomass from different studies (regions) using the same equation and compare the absolute values. Even within the Neotropical rainforest realm, considerable bias can result from the estimation with non-autochthonous coefficients. We conclude from our results that our equations from the Amazonian sample are useful for biomass estimation of bulk arachnid samples from ecological studies in Amazonian rain- forests and, with some restrictions, also for other neotropical forest spider assemblages. As these are often rich in species, which are represented by several developmental stages, it is valuable to have an idea of the distribution of size classes in the samples. If a wide range of sizes is represented, including spiders larger than 15 mm, the coefficients of the power functions should be used. If only smaller spiders were collected, which is often the case in soil or litter samples, the coefficients of the linear models would be more adequate or the equation resulting from the subsample of spiders < 2.5 mm should be used. We also present the coefficients for specific (abundant) taxa (ctenids, oonopids) and the guilds of hunting and web-building spiders, which can be used in studies of these specific groups. ACKNOWLEDGMENTS The sampling in Amazonia was conducted within the framework of the program SHIFT, the sampling in Parana within the framework of the program MATA ATLANTICA. Both Brazilian-German research programs were funded by the German Federal Ministry for Education and Research (BMBF) and the Brazilian Council on Research and Technology (CNPq). We thank the Brazilian institutions EMBRAPA Amazonia Occidental (Manaus) and Federal University of Parana (UFPR) and the NGO Society for Wildlife Research and Environmental Education (SPVS) for the permission to use their sites and laboratories. We are very grateful to our friend Wemer Hanagarth for assistance in sampling the arachnids in Amazonia and we thank Florian Raub and Ludger Scheuermann for their help in sampling the spiders in the Mata Atlantica. LITERATURE CITED Barros, E., J. Mathieu, S.C. Tapia-Coral, A.R.L. Nascimento & P. Lavelle. 2006. Soil macrofauna communities in Brazilian Amazo- nia. Pp. 43-55. In Soil Biodiversity in Amazonian and Other Brazilian Ecosystems. (F.M.S. Moreira, J.O. Siqueira & L. Brussaard, eds.). CAB International Publishing, Wallingford, UK. Barros, E., A. Neves, E. Blanchart, E.C.M. Fernandes, E. Wandelli & P. Lavelle. 2003. Development of the soil macrofauna community under silvopastoral and agrosilvicultural systems in Amazonia. 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The Journal of Arachnology 37:170-177 Two new species of the spider genus Ochyrocem (Araneae, Ochyroceratidae) from Mexico Alejandro Valdez-Mondragon: Coleccion Nacional de Aracnidos, Departamento de Zoologia, Institute de Biologi'a, Universidad Nacional Autonoma de Mexico, Apto. Postal 70-153, C. P. 04510, Ciudad Universitaria, Mexico D. F. E-mail: lat_mactans@yahoo.com. mx Abstract. Two new species of the spider genus Ochyrocera Simon 1891 are described from Mexico. Ochyrocera Juquila new species was collected under moist rotten logs and hollow trunks on a thick bed of pine needles in oak-pine forests located in a mountain range south of the city of Oaxaca at 1400-2700 m elev. Ochyrocera juquila resembles O. qidiiquevittata Simon 1891 from the Island of St. Vincent, in the angular shape of the embolus, which in the new species is V-shaped and in O. quinquevittata is L-shaped. Ochyrocera chiapas new species, was collected under rotten trunks and hollow trunks in abundant leaf litter in the tropical, humid Lacandona rainforest region located in eastern Chiapas, near the border with Guatemala. The species occurs at 160-260 m elev. Ochyrocera chiapas resembles O. arietina Simon, 1891 from the island of St. Vincent, in the similar shape of the embolus and distal apophysis of the cymbium, but in O. chiapas the embolus is more strongly curved and directed toward the distal part of the tibiae forming a “D”; in O. arietina the embolus is not as strongly curved as in O. chiapas. In both species, males and females were collected near each other; the females carried their egg sacs with their chelicerae. A key to the four known Mexican species is provided. Resumen. Dos nuevas especies del genero de aranas Ochyrocera Simon 1891 son descritas para Mexico. Ochyrocera juquila nueva especie, fue colectada bajo troncos podridos y troncos huecos en una capa gruesa de agujas de pino, en bosque de pino-encino, en un sistema montanoso al sur de la ciudad de Oaxaca entre 1400-2700 m elev. Ochyrocera juquila esta relacionada con Ochyrocera quinquevittata Simon 1891 de la Isla de San Vicente, en la forma angular del embolo, el cual en la nueva especie es en forma de “V”, y en O. quinquevittata en forma de “L”. Ochyrocera chiapas nueva especie, fue colectada en bosque tropical, bajo troncos podridos y en troncos huecos con mucha hum.edad, y abundante hojarasca, en la region de la selva Lacandona localizada al este de Chiapas, cerca de la frontera con Guatemala, localizada entre 160-260 m elev. Ochyrocera chiapas esta relacionada con Ochyrocera arietina Simon, 1891 de la Isla de San Vicente, en la forma similar del embolo y apofisis distal del cimbio, pero en O. chiapas el embolo es mas fuertemente recurvado y dirigido hacia parte distal de la tibia formando una “D”, en O. arietina el embolo no esta fuertemente recurvado como en O. chiapas. En ambas especies, machos y hembras fueron colectados cercanamente entre ellos; las hembras cargaban sus sacos de huevos con los queliceros. Se presenta una clave de identificacion para las cuatro especies mexicanas. Keywords: Haplogynae, taxonomy, Oaxaca, Chiapas The spider family Ochyroceratidae Page 1912 has 14 genera and 155 species (Platnick 2008). Edwards et al. (2003) reported four genera from the western hemisphere: Fageicera Dumi- trescu & Georgescu 1992 and Speocera Berland 1914, recorded only from Cuba, Ochyrocera Simon 1891 in the Caribbean region and Brazil, and Theotimci Simon 1893 restricted to the Caribbean region. Ochyroceratids are lucifugous spiders which live in leaf litter and detritus in mesic habitats, and many occur in caves as troglophiles (Gertsch 1973, 1977; Brignoli 1973; Lopez & Lopez 1997; Hormiga et al. 2007). These spiders spin tiny tangled webs in wall crevices and under litter (Gertsch 1973). Ochyrocera species build small, rather flimsy sheet webs with silk lines extending above the sheet that appear to serve as structural lines overlaid with finer silk lines running parallel to each other. The sheet is probably made of silk from the linearly arranged brush of posterior lateral spinneret aciniform gland spigots (Hormiga et al. 2007). There is limited published information available on the life history of these spiders (Edwards et al. 2003; Hormiga et al. 2007); some species of the family are parthenogenetic (Edwards et al. 2003). Simon (1893) included Theotima and Ochyrocera in the Leptonetidae, where they remained until Page (1912) erected the family Ochyroceratidae (Paquin & Ubick 2005). The genus Ochyrocera has 22 species, mostly from the Neotropical region (Brignoli 1974, 1978; Hormiga et al. 2007; Platnick 2008). In the New World, the genus is found in Florida, Central America, and parts of South America. Some species are distributed in the West Indies, like Puerto Rico, where two undescribed, sympatric species of Ochyrocera occur in forest leaf litter (Simon 1891; Edwards et al. 2003; Paquin & Ubick 2005; Platnick 2008). Recently Hormiga et al. (2007) described Ochyrocera cachote from Hispaniola. Two species have been recorded from Mexico: Ochyrocera fagei Brignoli 1974 from Teopisca, Chiapas and O. simoni O. Pickard-Cambridge 1894 from Teapa, Tabasco. The objective of this contribution is to describe two new species recently collected in Mexico. Ochyrocera juquila new species and O. chiapas new species are the third and fourth species of the genus Ochyrocera from Mexico. METHODS The specimens, preserved in 80% ethanol, were examined with a Nikon SMZ645 stereoscope. A Nikon Coolpix SIO VR camera was used to photograph the dorsal view of the prosoma and opisthosoma of male and female specimens, and the internal genital area of females. The photographs were edited in Adobe Photoshop 7.0 to make the illustrations. The specimens were then processed in order that photomicro- graphs could be taken with an HITACHI S-2460N scanning electron microscope (SEM). All measurements of the descrip- tions are recorded in millimeters and SEM photomicrographs 170 VALDEZ-MONDRAGON— TWO NEW SPECIES OF GENUS OCHYROCERA 171 are noted in microns. The map was done using Microsoft Encarta Encyclopedia and was edited in Adobe Photoshop 7.0. The specimens are deposited in the Coleccion Nacional de Aracnidos (CNAN) of the Institute de Biologia, Universidad Nacional Autonoma de Mexico, Mexico D. F. (IBUNAM) and the American Museum of Natural History (AMNH), New York, USA. Abbreviations used in the description are: ALE, anterior lateral eyes; ALS, anterior lateral spinnerets; AME, anterior median eyes; B, bulb of the palp; C, cymbium; DAC, distal apophysis of cymbium; E, embolus; PLE, posterior lateral eyes; PLS, posterior lateral spinnerets; PMS, posterior median spinnerets; S, spermathecae. TAXONOMY Family Ochyroceratidae Page 1912 Genus Ochyrocera Simon 1891 Type species. — Ochyrocera arietina Simon 1891 Ochyrocera iuqmlm new species Figs. 1-10 Type material. — MEXICO: Oaxaca: 1 5 mm); chelicerae uniformly brown A. maculosus Small species (< 5 mm); chelicerae pale or slightly darkened distally ................................. A. aurioculatus Australomimetus audax (Hickman 1929) new combination (Figs, la-e, 10) Mimetus audax Hickman 1929:107-110, figs. 4A-D, plate XVII; Hickman 1967:50-52, figs. 87-89, plate IX fig. 1; Roewer 1942:1021; Bonnet 1957:2917; Platnick 1997:228. Material examined. — Type; AUSTRALIA: Tasmania: ho- lotype ?, Launceston (41°27'S, 147°10'E), 25 April 1928, V.V. Hickman (QVM 13:7338; Old type No. 36), examined. Other material examined: AUSTRALIA: Victoria: 1 ¥, Warby Range State Park, 10 km W. of Wangaratta (36°18'S, 146°11'E), 28 July 2000, M. Scholes (QM S54173). Etymology. — Derived from the Latin audax (=bold, dar- ing). The type-specimens were collected in webs of Latrodectus hasselti - the red back spider - where they were found to prey upon the offspring of the host (Hickman 1929). Diagnosis. — Medium-sized species (carapace length 2. 8-3.0 mm) distinguished from other congeners by a combination of the following characters: leg formula I IV II III, presence of a conspicuous creamy, serrated and triangular folium situated distomedially on the opisthosoma (Fig. Id), epigynum oval; strongly sclerotized with 2 large, circular genital openings and a broad medial septum (Figs, la, b). Description. — The male is unknown and the female was described by Hickman (1929, 1967). New drawings of the epigynum are provided since the originals are poor (Figs. la-c). Affinities. — This species was described by Hickman (1929) within Mimetus based on 3 female specimens. A holotype was not designated and generic placement was not justified or discussed. Only one female could be found in the collection of QVM; the other females are probably lost. Mimetus audax was not included or even mentioned in the revision of the Queensland and New South Wales fauna in which the genus Australomi- metus was described (Heimer 1986). However, the female genital and somatic characters easily allow referal to Australomimetus. The species is probably a member of a group of rather robust taxa with a conspicuous whitish folium in a distomedial position on the opisthosoma (Harms & Harvey, in press). This folium is frequently present in many species from the Australian eastcoast such as A. burnetti Heimer 1986, A. harteyensis Heimer 1986, A. raveni Heimer 1986 or A. robustus Heimer 1986 and is also very conspicuous in A. audax. Females of this group also share strongly sclerotized genitalia and large genital openings. Adult specimens possess a strongly maculated integument of the legs. The position of A. audax within this group seems equivocal, due in part to the unusual leg formula and the reduction of the epigynal javelined scape with a broadened ectal tip, which is another prominent feature of this species group and is highly conspicuous in A. robustus, A. hartleyensis and A. raveni (e.g., see Heimer 1986: fig. 14). This scape shows reductive tendencies in several species (e.g. A. mendax) and is almost 194 THE JOURNAL OF ARACHNOLOGY Figure 4. — Tasmanian records for Australomimetus awioculatus. completely absent in A. aiidax. Australomimetus audax is also very similar to A. mendux new species in somatic appearance and morphology and both species were probably mixed up by Hickman (1967). On the one hand, A. audax was included and figured in his guide to the common spiders of Tasmania although this species is rare and only the type specimens were known for a long time. On the other hand, A. mendax is much more common in Tasmania and it seems possible that this species was initially left undescribed because Hickman didn’t recognize that there are two similar species. Distribution. — Australomimetus audax is apparently rare and was only known from the three specimens described by Hickman (1929) until another female was found in Victoria. This extends the distribution range for this species to include the Australian mainland (Fig. 10). Australomimetus awioculatus (Hickman 1929) new combination (Figs. 2a-f, 3, 4) Mimetus aurioculatus Hickman 1929:110-114, figs. 6 A-C, 7 A-C; Roewer 1942:1021; Bonnet 1957:2917. Material examined. — Type: AUSTRALIA: Tasmania: ho- lotype lateral carinae continuous, costate-granular, fully developed PRENDINI ET AL.— REDESCRIPTION OF RHOPALURUS ABUDl 219 Figure 10. — Rhopalurus princeps (Karsch 1879), c? (AMNH), metasomal segments I-V and telson: A. Dorsal aspect. B. Lateral aspect. C. Ventral aspect. Scale bar = 1 mm. on segment I, obsolete, granular, restricted to the posterior two-thirds of segment 11, absent on segments III-V. Segment V, dorsosubmedian carinae absent; dorsolateral and ventro- lateral carinae continuous, costate-granular, granules subeq- ual; ventrosubmedian carinae obsolete, granular, reduced to anterior half of segment; ventromedian carina continuous, costate-granular, granules subequal, without posterior bifur- cation. Telson: Vesicle globose, height/length 52-57%, with flat dorsal surface and rounded ventral surface, slightly com- pressed anteroventrally (Table 1); slightly narrower than metasomal segment V, width 59-60% (d) or 63-77% (?) of segment V. Subaculear tubercle absent (Fig. 8B). Ventrolat- eral and ventrosubmedian carinae absent; ventromedian Carina continuous, granular. Vesicle surfaces with scattered granules, sparse microsetae, and fewer than 16 macrosetae. Aculeus long, 73-80% (d) to 81-85% (?) of vesicle length (Table 1), strongly curved. Male hemispermatophore: Flagelliform, flagellum gradually tapering along its length, folded against shaft (Fig. 11); basal process lobate longitudinally; distal process terminating adjacent to base of flagellum (in dorsal aspect), rib-like and extending longitudinally; distal lobe represented by shelf at base of flagellum; median lobe not developed; internobasal inflection absent; external lobe present, separated from distal process; with small, longitudinally-oriented costate process. Female reproductive system: Ovariuterine network compris- ing three longitudinal and ten transverse tubules, forming eight “cells.” Geographic variation: The single male specimen from Cabo Flaso is similar to those from the track between Boca de Yuma and Punta Faustino. Ontogenetic variation: As in other species of Rhopalurus, male closely resembles female until the final instar; however, juveniles and subadults may be sexed by examination of the pectines and genital aperture. Sexual dimorphism: In addition to aforementioned charac- ters, adult males are proportionally longer than adult females. The increased length of the male is attributed mainly to the longer metasomal segments, which sum to 72-78% of the total length of males, but to 69-72% of the total length of females. Adult males are slightly more slender than adult females: 220 THE JOURNAL OF ARACHNOLOGY Figure 1 1 . — Rhopcilwus ahudi Armas & Marcano Fondeur 1987, •£ (AMNH), left hemispermatophore: A. External aspect. B. Anterior aspect (left view rotated right 90° around longitudinal axis). Abbreviations: F, flagellum; DL, distal lobe; DP, distal process; BP, basal process; IL, internal lobe; ILP, internal lobe process. Scale bars = 0.5 mm. sternite VII length is 37-51% greater than its width in males and 49-53% greater in females (Table 1). The coloration of adult females is similar to but darker than that of adult males. Distribution. — Rhopalwns ahudi was described from Ca- tuano, Isla Saona, off the southeast coast of the DR (Armas & Marcano Fondeur 1987). No new records of this species have been reported in the literature since the original description (Armas & Marcano Fondeur 1987; Armas et al. 1999; Teruel 2005, 2006). The records reported here, therefore, represent the first for this species on mainland Hispaniola. Based on published records and those obtained during our expedition, R. ahudi appears to be restricted to humid coastal forest in the southeast of mainland DR and Isla Saona (Fig. 1). Rhopalurus princeps inhabits dry scrub in the central part of Hispaniola, including the valley of the Yaque del Norte River, the Neiba Valley, the Sierra de Baoruco, Sierra de Martin Garcia, and Sierra de Ocoa (Teruel 2006). Rhopalurus honettii is restricted to dry spiny forests south of the Sierra de Baoruco in the western part of mainland DR and Isla Beata, the type locality. The plotted locality data agree with Teruel’s (2006: 51, fig. 12) map illustrating the approximate distributions of the three species. Ecology. — Rhopalurus ahudi is probably restricted to humid forests, a habitat not previously reported for any Rhopalurus species. Although collections were made on the western and eastern sides of Barque Nacional del Este during the course of our expedition, no specimens were found on the western side, which is drier and dominated by dense, spiny forest. Whereas the South American species of Rhopalurus appear to be restricted to savannas (Lourenfo 1996, 2008), those of the Caribbean are also found in other vegetation zones, including forest (Armas 2001). During our expedition, R. ahudi was collected in lowland coastal humid forest on limestone, R. honettii in dry spiny forests on limestone, and R. princeps in dry scrub on mixed substrata. All specimens of R. ahudi were collected at night using UV light detection. None were found during the day, unlike R. honettii, which was commonly found sheltering between slabs of rock (though never under bark or wood), and R. princeps, which was found under bark, wood and stones, as well as in dead and dry agave plants. The holotype of R. ahudi was collected from under a stone (Armas & Marcano Fondeur 1987). ACKNOWLEDGMENTS We are grateful to the Department de Investigaciones de la Subsecretaria de Areas Protegidas y Biodiversidad, Govern- ment of the Dominican Republic, for Permit Number 01496 to collect and export scorpions from the country. Kelvin Guerrero kindly assisted with the permit application and provided valuable advice on collecting in the DR (he was the first to observe R. ahudi in the Barque Nacional del Este). We thank the following for assistance with the study of materia! at their institutions: Peter Jager and Julia Altmann (SMF), Jason Dunlop and Shahin Nawai (ZMB), Hieronymus Dastych (ZMH); the following for donating specimens to L. Prendini that were examined during the course of this study: Santos Bazo Abreu, Dietmar Huber, Siegfried Huber, Adriano Kury, Charles Siederman, Rolando Teruel Ochoa, Alex Tietz, Rick C. West; and the following for the participating in fieldwork during which specimens, examined during the course of this study, were collected: Camilo I. Mattoni, Ricardo Pinto-da- Rocha, Humberto Yamaguti. The 2004 field expedition to the DR, during which the series of R. ahudi and comparative material of R. honettii and R. princeps was collected, was funded by a Genomics Postdoctoral Research Fellowship PRENDINI ET AL.— REDESCRIPTION OF RHOPALURUS ABUDI 221 from the AMNH to E.S. Volschenk and National Science Foundation grant EAR 0228699 to L. Prendini. Fieldwork by C.I. Mattoni in Brazil and by L.A. Esposito in the DR, during which other material examined for this study was collected, was funded by grants from the National Science Foundation (EAR 0228699) and the Richard Lounsbery Foundation to L. Prendini. We thank Steve Thurston (AMNH) for assistance with preparing the plates for this contribution, and Mark Harvey and an anonymous reviewer for comments on a previous draft of the manuscript. While at the AMNH, E.S. Volschenk was supported by a Genomics Postdoctoral Research Fellowship, supplemented by a grant from the Richard Lounsbery Foundation to L. Prendini; L.A. Esposito was supported by a National Science Foundation GK-12 Fellowship, a City University of New York MAGNET Fellowship, and a City University of New York/NSF AGEP Fellowship. LITERATURE CITED Armas, L.F. de. 1999. Quince nuevos alacranes de La Espanola y Navassa, Antillas Mayores (Arachnida: Scorpiones). Avicennia 10/11:109-144. Armas, L.F. de. 2001. Scorpions of the Greater Antilles, with the description of a new troglobitic species (Scorpiones: Diplocentri- dae). Pp. 245-253. In Scorpions 2001. In Memoriam Gary A. Polis. (V. Fet & P.A. Selden, eds.). British Arachnological Society, Burnham Beeches, Buckinghamshire, UK. Armas, L.F. de. & E. de J. Marcano Fondeur. 1987. Nuevos escorpiones (Arachnida: Scorpiones) de Republica Dominicana. Poeyana 356:1-24. Armas, L.F. de., J.A. Ottenwalder & K.A. Guerrero. 1999. Escorpiones de las Islas Saona, Beata y Catalina, Republica Dominicana (Arachnida: Scorpiones). Cocuyo 8:30-32. Borelli, A. 1910. Scorpion! nuovi o poco noti del Brasile. Bollettino dei Musei di Zoologia ed Anatomia Comparata della Reale Universita di Torino 25(629): 1-8. Fet, V., B. Gantenbein, A.V. Gromov, G. Lowe & W.R. Lourenyo. 2003. The first molecular phylogeny of Buthidae (Scorpiones). Euscorpius 4:1-10. Fet, V. & G. Lowe. 2000. Family Buthidae C.L. Koch, 1837. Pp. 54-286. In Catalog of the Scorpions of the World (1758-1998). (V. Fet, W.D. Sissom, G. Lowe & M.E. Braunwalder, eds.). New York Entomological Society, New York. F16rez, E. 2001. Escorpiones de la familia Buthidae (Chelicerata: Scorpiones) de Colombia. Biota Colombiana 2(l):25-30. Francke, O.F. 1977. Two emendations to Stahnke’s (1974) Vaejovi- dae revision (Scorpionida, Vaejovidae). Journal of Arachnology 4:125-135. Gonzalez-Sponga, M.A. 1996. Guia para identificar escorpiones de Venezuela. Cuadernos Lagoven, Caracas. 204 pp. Hjelle, J.T. 1990. Anatomy and Morphology. Pp. 9-63. In The Biology of Scorpions. (G.A. Polis, ed.). Stanford University Press, Stanford, California. Kovafik, F. 1998. Stifi [Scorpions]. Madagaskar, Jihlava. 175 pp. [in Czech] Lamoral, B.H. 1979. The scorpions of Namibia (Arachnida: Scorpionida). Annals of the Natal Museum 23:497-784. Lenarducci, A.R.I.P., S.M. Lucas & R. Pinto-da-Rocha. 2005. Descrigao de uma nova especie de Rhopalunis Thorell, 1876 (Scorpiones: Buthidae) do nordeste brasileiro. Biota Neotropica 5(la):l-8. Louren9o, W.R. 1979. A propos de la veritable identite des genres Rhopalunis Thorell, 1876 et Centruroides Marx, 1889 (Scorpiones, Buthidae). Revue Arachnologique 2:213-219. Louren^o, W.R. 1982. Revision dii genre Rhopalunis Thorell, 1876 (Scorpiones, Buthidae). Revue Arachnologique 4:107-141. Louren^o, W.R. 1984. Complementary notes on the systematics of the genus Rhopalunis for the Caribbean area (Scorpiones, Buthidae). Revista Brasileira da Biologia 44:169-170. Lourenfo, W.R. 1986. Biogeographie et phylogenie des scorpions du genre Rhopalunis TXwxqW, 1876 (Scorpiones, Buthidae). Memoires de la Societe Royale Beige d’Entomologie 33:129-137. Lourengo, W.R. 1996. The biogeography of scorpions. Revue Suisse de Zoologie 103:437^48. Louren^o, W.R. 2002. Nouvelles additions a la faune de scorpions neotropicaux (Arachnida). Revue Suisse de Zoologie 109:127-141. Lourenijo, W.R. 2007. New considerations on the taxonomic status of the genus Physoctomis Mello-Leitao, 1934 (Scorpiones, Buthidae). Boletin de la Sociedad Entomoldgica Aragonesa 40:359-365. Lourengo, W.R. 2008. The geographic pattern of distribution of the genus Rhopalunis Thorell, 1876 in the Guayana-Amazon region (Scorpiones: Buthidae). Euscorpius 73:1-14. Lourengo, W.R., R.L.C. Baptista & A.P. de L. Giupponi. 2004. Troglobitic scorpions: a new genus and species from Brazil. Comptes Rendus Biologie 327:1151-1156. Lourengo, W.R. & J.L. Cloudsley-Thompson. 1995. Stridulatory apparatus and the evolutionary significance of sound production in Rhopalunis species (Scorpiones: Buthidae). Journal of Arid Environments 31:423-429. Lourengo, W.R. & R. Pinto-da-Rocha. 1997. A reappraisal of the geographic distribution of the genus Rhopalunis Thorell (Scor- piones, Buthidae) and description of two new species. Biogeo- graphica 73:181-191. Pocock, R.I. 1904. On a new stridulating-organ in scorpions discovered by W.J. Burchell in Brazil in 1828. Annals and Magazine of Natural History (Series 7) 13:56-62. Prendini, L. 2001a. 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On the taxonomy and distribution of Rhopalwus kiticcmcici Thorell, 1876 (Scorpiones: Buthidae), with description of a new species of the genus. Euscorpius 68:1-12. Teruel, R. & A.K. Tietz. 2008. The true identity of Rhopalwus pintoi Mello-Leitao, 1932, with notes on the status and distribution of Rhopalwus crasskauda Caporiacco, 1947 (Scorpiones: Buthidae). Euscorpius 70:1-14. Vachon, M. 1973 [1974]. Etude des caracteres utilises pour classer les families et les genres de scorpions (Arachnides). 1. La trichobo- thriotaxie en arachnologie. Sigles trichobothriaux et types de trichobothriotaxie chez les scorpions. Bulletin du Museum National d’Histoire Naturelle, Paris (3) 140:857-958. Volschenk, E.S. 2005. A new technique for examining surface morphosculpture of scorpions. Journal of Arachnology 33: 820-825. Volschenk, E.S., C.I. Mattoni & L. Prendini. 2008. Comparative anatomy of the mesosomal organs of scorpions (Chelicerata, Scorpiones), with implications for the phylogeny of the order. Zoological Journal of the Linnean Society 154:651-675. Manuscript received 8 July 2008, revised 5 November 2008. Appendix 1. Currently recognized species and subspecies of Rhopalwus Thorell 1876 and related genera, with countries, departments (Colombia, Haiti), provinces (Cuba, Dominican Repub- lic), regions (French Guiana) and states (Brazil, Venezuela) of known distribution (data from Gonzalez-Sponga 1996; Fet & Lowe 2000; Florez 2001; Teruel 2006; Teruel & Roncallo 2008; Teruel & Tietz 2008; Louren^o 2008; this study). New records reported in this study are marked with an asterisk. Physoctonus dehilis (C.L. Koch 1840): Brazil (Bahia, Ceara, Pernambuco*, Piaui). This species was originally placed in the non- buthid genus Vaejovis C.L. Koch 1836. It was transferred to Rhopalwus by Borelli (1910) and remained there until Lourengo (2002) resurrected the genus Physoctonus Mello-Leitao 1934, earlier synonymized with Rhopalwus by Francke (1977). Rhopalurus ahudi Armas & Marcano Fondeur 1987: Dominican Republic (La Altagracia, La Romana). Rhopalurus acromelas Lutz & Mello 1922: Brazil (Bahia, Ceara, Tocantins, Maranhao*, Pernambuco, Piaui). Rhopalurus agamemnon (C.L. Koch 1839): Brazil (Bahia, Ceara, Tocantins, Mato Grosso, Pernambuco, Piaui). Rhopalurus amazonicus Lourenfo 1986: Brazil (Para). Rhopalurus honettii Armas 1999: Dominican Republic (Pedernales). Rhopalurus carihensis Teruel «& Roncallo 2008: Colombia (Atlan- tico. La Guajira, Magdalena), Venezuela (Zulia). Lourengo (2008) suggested that this species might be more appropriately recognized as a subspecies of Rhopalurus laticauda Thorell 1876. Rhopalurus crasskauda Caporiacco 1947: Brazil (Amazonas*, Roraima), Guyana. This species was synonymized with Rhopalwus pintoi Mello-Leitao 1933 by Lourengo (1982) and reinstated by LourenQo (2002). Teruel & Tietz (2008) demonstrated that R. pintoi is a distinct species but questioned whether R. crassicauda can be regarded as distinct from R. laticauda. In our opinion, R. crassicauda is probably a junior synonym of R. laticauda. Loureng:o (2008) rejected the suggestion that R. cras.sicauda may be synonymous with R. laticauda, suggesting instead that it might be a subspecies of the latter. Lourengo (2008) also created two new subspecies of R. crassicauda. The distinction between R. laticauda, R. crassicauda and its two subspecies warrants further investigation. Rhopalurus crassicauda kourouensis Lourenfo 2008: French Guiana (Kourou). Rhopalurus crassicauda paruensis Lourenfo 2008: Brazil (Para). Rhopalurus garridoi Armas 1974: Cuba (Guantanamo). Rhopalurus giharae Teruel 2006: Cuba (Holguin). Rhopalurus granulimanus Teruel 2006: Cuba (Holguin). i Rhopalurus guanambiemis Lenarducci, et al. 2005: Brazil (Bahia). Rhopalurus junceus (Herbst 1800): Cuba (Camaguey, Cienfuegos, Ciego de Avila, Granma, Guantanamo, Havana, Holguin, Isla de la Juventud, Las Tunas, Matanzas, Pinar del Rio, Santiago de Cuba, Sancti Spiritus, Villa Clara). Records of this species from Haiti and Venezuela (see, e.g. Fet & Lowe 2000: 220) are probably erroneous (Armas 2001:248). Rhopalurus lacrau Louren^o & Pinto-da-Rocha 1997: Brazil (Bahia). Rhopalurus laticauda Thorell 1876: Colombia (Arauca, Boyaca, Casanare, Cesar, Meta, La Guajira, Magdalena, Norte de Santander, Vichada), Venezuela (Amazonas, Anzoategui, Apure, Aragua, ( Barinas, Bolivar, Carabobo, Cojedes, D.F., Falcon, Guarico, Lara, Merida, Miranda, Monagas, Nueva Esparta, Portuguesa, Sucre, Tachira, Vargas, Yaracuy, Zulia). Rhopalurus melloleitaoi Teruel & Armas 2006: Cuba (Granma). Rhopalurus pintoi Mello-Leitao 1933: Brazil (Roraima), Guyana, I ?Venezuela (Bolivar). This species was relegated to a subspecies of R. laticauda by Louren^o (1982) until reinstated by Lourengo (2002). Teruel (2006) suggested that it might be a senior synonym of Rhopalurus piceus Lourengo & Pinto-da-Rocha 1997 and this was confirmed by Teruel & Tietz (2008). Lourengo (2008) agreed with the recognition of R. pintoi as a distinct species, but suggested that R. piceus may yet prove to be valid. We agree with the decision of Teruel & Tietz (2008). Rhopalurus princeps (Karsch 1879): Dominican Republic (Azua, S| Barahona, Baoruco, Independencia, Montecristi, Pedernales, Pera- jl via), Haiti (Departement du FOuest). Records of this species from Cuba (listed by Fet & Lowe 2000:221) are erroneous. Rhopalurus rochae Borelli 1910: Brazil (Bahia, Ceara, Paraiba, Pernambuco, Piaui, Rio Grande de Norte, Sergipe*). Borelli (1910) ! named the species after Francisco Diaz da Rocha, but his original i spelling was rochae. Fet & Lowe (2000) noted that the correct spelling j is rochai and changed it accordingly. Although the corrected spelling has been adopted by others (e.g., Teruel 2006:52), we use Borelli’s (1910) original spelling. Troglorhopalurus translucidus Lourengo, et ai. 2004: Brazil (Bahia). j In our opinion, this monotypic genus is a junior synonym of Rhopalwus. As twice noted by Lourengo et al. (2004:1153, 1156), when comparing Troglorhopalurus with Rhopalurus: “It may be that al) modifications presented by the new troglobitic scorpion are the i result of adaptation to a cave dwelling life.” Appendix 2. Material examined for comparison with Rhopalwus ahudi Armas & Marcano Fondeur, 1987. Specimens are deposited in the following collections: American Museum of Natural History (AMNH), New York, USA, incorporating the Alexis Harington (AH) Collection; Natur-Museum Senckenberg, Frankfurt (SMF), Germany; Zoologisches Museum der Humboldt-Universitat, Berlin (ZMB), Germany; Zoologisches Museum der Universitat Hamburg (ZMH), Germany. Reference numbers (ESV and LP), provided on labels with the specimens, correspond to entries in the specimen databases of the author with the corresponding initials. i Physoctonus dehilis (C.L. Koch, 1840): BRAZIL: Pernambuco: Exu, 5 km N, 4 October 1977, L.J. Vitt, 1 ? (AMNH), 18 January 1978, L.J. Vitt & K.E. Streilein, 1 $ (AMNH); Exu, 18 km N, 5 March ' 1977, L.J. Vitt, under leaf of granite on boulder, caatinga habitat, 1 9 (AMNH); Fazenda Batente, 13 km E Exu, 10 November 1977, L.J. Vitt & K.E. Streilein, 1 ? (AMNH); Fazenda Caterino, 10 km NE Exu, 9 July 1977, L.J. Vitt, 1 ? (AMNH), 25 September 1977, L.J. J Vitt, 1 9 (AMNH). Rhopalurus acromelas Lutz & Mello, 1922: BRAZIL: Maranhao: Municipio de Loreto: Santa Barbara, on shore of Rio Parnoiba, June 1962, G. Eiten, 1 8 (AMNH). Pernambuco: Exu, 10 km N, 13 March PRENDINI ET AL.— REDESCRIPTION OF RHOPALURUS ABUDI 223 1977, L.J. Vitt, rocky habitat within thorn scrub forest, 1 ?, 1 subad. $, 4 juv. (AMNH), 14 March 1977, L.J. Vitt, rocky habitat in thorn scrub, 1 4, 1 $ (AMNH [ESV7532]); Exu, 10 km NE, 28 April 1977, L.J. Vitt, 1 c3, 1 9, 2 subad. 9, 2 subad., 1 juv. (AMNH), 25 September 1977, L.J. Vitt, 1 (J, 1 9 (AMNH [ESV7244]); Exu, 15 km NE, 14 May 1977, L.J. Vitt, high caatinga, under bark of tree, 1 subad. 9 (AMNH); Exu, 20 km E, 30 March 1977, L.J. Vitt, 1 juv. J (AMNH); Fazenda Caterino, 10 km NE Exu, 9 July 1977, L.J. Vitt, 1 subad. S (AMNH), 1 August 1977, L.J. Vitt, 1 juv. (AMNH). Rhopalums agamemnom (Herbst, 1800): BRAZIL; Bahia: Salvador, February 1972, Weinkselbaum, 1 9 (AMNH [ESV7405]). Rhopalums honettli Armas, 1999: DOMINICAN REPUBLIC: Pedernales Province: Parque National Jaragua; Cabo Rojo, 17°53'45.2"N, 71°39'35.8"W, 9 July 2004, E.S. Volschenk & J. Huff, 15 m, dry cactus and spiny forest on limestone karst, hand collected at night with blacklights, 3 A 10 9, 4 subad., 2 juv. (AMNH [ESV6005]), 1 (J(AMNH [ESV7126]), 1 <3, 1 9 (AMNH [ESV7127]), 1 subad. 3 (AMNH), 1 juv. 3 (AMNH [LP 3267]); Road to Fondo Paradi, 1.8 km from Highway 44, 17°48.692'N, 7U26.600'W, 12 January 2004, J. Huff, 302 ft, found between rocks, 1 9 (AMNH [LP 2471]), 1 9 (AMNH [LP 3265]); Track into park, between Manuell Goa and Oviedo, 17°48'41.5"N, 71°26'35.9"W, 9 July 2004, E.S. Volschenk & J. Huff, 83.3 m, deciduous forest and thorny scrub, hand collected from between stones during the day and with blacklights at night, 13 3, 7 9, 1 subad., 1 juv. (AMNH [ESV6011]), I 3, 1 9 (AMNH [ESV71 12]), 1 3 (AMNH [ESV7129]), 1 juv. (AMNH [LP 3266]). Rhopalums carihensis Teruel & Roncallo, 2008: COLOMBIA: Magdalena Department: Bahia de Guairaca, Tayrona Park, 31 October 1985, H.-G. Muller, 1 9 (SMF 37027); Pozo Colorado, I I km W Santa Marta, 18-30 April 1968, B. Malkin, 1 9, 1 subad., 19 first instars (AMNH); Puente de Los Clavos, 15 km E Pueblo Bello, Sierra Nevada de Santa Marta, 13 June 1968, B. Malkin, 1500 m, 1 subad. 9 (AMNH); Santa Marta, 29 June-31 July 1966, 2 9 (SMF 39120). Rhopalurus crassitauda Caporiacco, 1947: BRAZIL: Amazonas: Rio Branco, Amazonasgebiet, 1912, E. Ule, 1 juv. 9 (ZMB 14867). Roraima: Mt. Roraima, 2 3, 1 9, 1 subad. (AMNH 29180). Rhopalurus junceus (Herbst, 1800): CUBA; July 2007, C. Hamilton, 1 juv. (AMNH [LP 7009]); ‘Antillen?,’ 1 3, 2 9 (ZMB 7370); ‘Portorico’, Stahl, 2 9, 1 juv. (ZMB 7280 [ESV7001]); Gundlach, 2 9 (ZMB 2637), 1 3, 1 9 (ZMB 7380 [ESV7224]), 1 juv. (ZMB, 7343); Arroyo Bermejo, near Fibacoa, 31 May 1967, Kleiderschrank, 1 3 (ZMB 31020), 15 June 1967, im zelt. wiese auf sandboden, 1 9 (ZMB 31021), June 1967, 1 juv. (ZMB 31022); 1 3 (ZMH), Santiago de las Caballeros, P. Thumb, 1936. Havana Province: Havana, 1 9 (AMNH), April 1941, Dr E. Weiss, 1 9, 1 subad. (AMNH). Holguin Province: August 2000, Heist, captive bred, 1 juv. (AMNH [LP 1928]); near Banos [Banes], May 1918, 2 3 (AMNH); Guardalavaca, 29 March 1993, W. Altmann, captive bred, 1 3 (AMNH [LP 1565]); Mountains near Guisa, October 1936, P. Thumb, 1 9, 28 juv. (ZMH); Moa, September 1937, P. Thumb, 1 3 (ZMH), 1938, P. Thumb, 4 9 (ZMH). Isla de la Juventud Province: Isle of Pines, 1 3 (AMNH). Pinar del Rio Province: Guanahacabiies, Akad.-stat. El Beral, December 1967, G. Peters, 1 subad. (ZMB 31023); Sierra de Anafe, 23 February 1947, M. Barro, 2 subad. (AMNH); Vinales Valley, 1940, Osorio, 1 9 (AMNH). Santiago de Cuba Province: La Socapa, 10 km SW of Santiago de Cuba, 9 April 1999, R. Teruel, 1 3 (AMNH), 1 9 (AMNH [LP 1509]), 1 juv. 9 (AMNH [LP 1517]), 1 9 (AMNH [LP 1518]); Santiago de Cuba, 1 3, 2 juv. (AMNH). Sancti Spiritus Province: Trinidad, August 1978, B. Acosta, 1 3 (AMNH AH 4514 [ESV7041]). Rhopalurus lacrau Louren^o & Pinto-da-Rocha, 1997: BRAZIL: Bahia: Municipio Itaete: Trail between Caves “Lapa do Bode” and “Lapa Escondida,” 12°56'9.1"S, 41°3'56.2"W, 21 January 2007, C.I. Mattoni, R. Pinto-da-Rocha & H. Yamaguti, under rocks, 2 9 (AMNH), 1 subad. 9, 4 juv. (AMNH [LP 7637]). Rhopalurus lalicauda Thorell, 1876: 2 9 (ZMB 14865); “Mexico,”: Drv. Hubl, 1 3 (ZMB 14866). VENEZUELA: F. Kummerow, 1 3, 1 9 (ZMB 8226). San Jose de Guaviare, December 1955, Meden, 1 9 (SMF 39252). Aragua: Maracay, 1 subad. 3 (SMF 29208), Fahrcn- holz, 1 3, 1 9, 1 subad. (SMF 8876/218). Bolivar: Ciudad Bolivar, 20 February 1903, 2 9 (ZMH); La Paragua, M.A. de Verde, 1 3 (AMNH); Upata, February 1973, A. Bordes, 1 9 (AMNH). Carahoho: Valencia, F. Kummerow, 29 December 1904, 1 9 (ZMB 31024), September 1958, H. Ardelt, 2 9 (ZMH). Distrito Federal: Caracas, March 1999, C. Siederman, 2 9, 20 first instars (AMNH [ESV7444]), 2001, C. Siederman, 1 3 (AMNH [LP 2462]). Guarico: Calabozo and San Fernando de Apure (about halfway between), 30 November 1967, M.A. de Verde, 1 9 (AMNH); ‘Hato Masaguarat,’ 45 km S Calabozo, 7 April 1978, Y. Lubin, 1 3 (AMNH [ESV7816]). MMda: Merida, 2 3, 3 9 (SMF 5712/27). Miranda: Guatire, 29 April 2004, R.C. West, under rocks, dry forest, 1 3 (AMNH [LP 2845]), 1 9 (AMNH); Hda. Santa Rosa, 3 km N Guatire, 10 January 1973, M.A. Gonzalez- Sponga, 450 m, 1 3, 1 9, 2 juv. (AMNH). Nueva Esparta: Isla Margarita, N of Peninsula de Macanao, 11°02.618'N, 64°21.542'E, 4 September 2005, S. Huber, 1 9 (AMNH [LP 4221]). Trujillo: Valera region, N, October 2005, S.E. Bazo Abreu, 1 9 (AMNH [LP 5504]), 1 9 (AMNH [LP 5505]). Rhopalurus pintoi Mello-Leitao, 1933: GUYANA: Roraima Prov- ince: Rununui region, SW Guyana, near Venezuelan border, ex A. Tietz, March 2008, 1 juv. 3 (AMNH [LP 8278]). Rhopalurus princeps (Karsch, 1879): DOMINICAN REPUBLIC: Independencia Province: Isla Cabritos, 18°30.019'N, 71°43.228'W, 7 January 2004, J. Huff, 110 ft, under rock, coral, 1 3, 1 9, 16 juv. (AMNH), 5 3, 3 9, 3 subad., 1 juv. (AMNH), 3 juv. (AMNH [LP 2470]), 1 subad., 2 juv. (AMNH [LP 3260]); Ranger station for Parque Nacional Isla Cabritos, 18°33'45"N, 71°41'50"W, 8 July 2004, E.S. Volschenk & J. Huff, -19 m, dry forest, hand collected from under stones and logs, and with blacklights, 3 3, 7 9, 5 subad., 2 juv. (AMNH [ESV6006]), 1 subad. 3 (AMNH), 1 subad. (AMNH [LP 3264]); Parque Nacional Isla Cabritos, behind Ranger Station, 18.56287°N, 71.69762°W, 8 August 2005, L. Esposito, -23 m, mixed dry forest with succulents, UV detection, 35°C, 2 3, 8 9, 1 subad. 9, 32 first instars (AMNH), 2 3, 1 subad. 9 (AMNH), 1 3 (AMNH [LP 5102]); Parque Nacional Sierra de Baoruco, road between Rabo de Gato and Duverge, 18“19'38''N, 17°33'55''W, 7 July 2004, E.S. Volschenk & J. Huff, 447 m, arid thorny scrub, hand collected from under stones and in dead and dry agaves, 3 3, 3 9, 3 juv. (AMNH [ESV6033]), 1 juv. (AMNH), 1 9 (AMNH [LP 3263]); Puerto Escondido, Sierra de Baoruco, 18°19.762'N, 71°33.502'W, 6 January 2004, J. Huff, 1592 ft, under dead agave, 1 3, 3 9, 1 juv. (AMNH), 1 juv. (AMNH [LP 3261]); Road to Puerto Escondido, 18°20.376'N, 71°33.345'W, 6 January 2004, J. Huff, 1388 ft, under rocks in gravel quarry, 1 9 (AMNH), 1 juv. (AMNH [LP 3262]). Pedernales Province: Manuel Goja, 3.9. km N, 9 May 1998, D. Huber, 1 3 (AMNH [LP 1566]); Oviedo to Pedernales, 1 1.5 km N, 8 May 1998, D. Huber, 1 3 (AMNH [LP 1516]). HAITI: Departement de 1’ Quest: Port-au-Prince, Ehrenberg, holotype 3 (ZMB 1 16). Rhopalurus rochae Borelli, 1910: BRAZIL: Bahia: Municipio Ceraima: Guanambi, 7 km S, 14°17'5.6"S, 42°47'2.2''W, 24 January 2007, C. Mattoni, R. Pinto-da-Rocha & H. Yamaguti, 533 m, UV sampling, modified savanna, cloudy and raining, 1 juv. (AMNH [LP 7638]); Fazenda du Fabiano, 8 km NE Guanambi, 14°10' 17.6"S, 42°43'56.4"W, 24 January 2007, C. Mattoni, R. Pinto-da-Rocha & H. Yamaguti, 539 m, under rocks, rocky hill and surrounds, open savanna modified, 1 3, 2 juv. (AMNH [LP 7639]), 1 9 (AMNH); Guanambi, 16 km SE, 14°17'19"S, 42°41'31.1''W, 25 January 2007, C. Mattoni, R. Pinto-da-Rocha, H. Yamaguti, 559 m, UV sampling and under leaf litter, banana plantation and surrounds, 1 juv. (AMNH [LP 7655]). Paraiha: Soledade, 07°02.118'S, 36°27.311'W, 16 March 1999, A. Kury & A. Giupponi, 575 m, 1 3 (AMNH [LP 1581]), 1 9 (AMNH [LP 1582]), 1 3 (AMNH [LP 1775]). Pernambuco: Escola 224 THE JOURNAL OF ARACHNOLOGY Aquicola, Exu, 30 March 1977, L.J. Vitt, caatinga, 1 I > II > III; male pedipalp without articulated tegular apophysis; lateral apophysis present; embolus a thin, curved spine and mostly covered by tegulum in ventral view; epigynum variable, often protruding scape-like posteriorly. The gently sloping margins of the cephalic area, small size and the lack of a distinct color pattern are here considered synapomorphies for Zoica. Lehtinen & Hippa (1979) reported a dorsal abdominal scutum in males, which we cannot confirm for the species described here or for any of the three species known from Australia (McKay 1979; VWF unpublished data). Zoica carolinensis new species (Figs. 1, 2, 5-8, 13) Types. — Holotype male. Federated States of Micronesia, Caroline Islands, Ponape (= Pohnpei), E of Kolonia, 6°57'50"S, 158°12'30"E, 7 June 1973 J.A. Beatty, J.W. Berry (BPBM); paratype female, data as holotype (BPBM). Other material examined. — FEDERATED STATES OF MICRONESIA: Caroline Islands: 1 female, Pohnpei, E of Kolonia, 6°57'N, 158°15'E, 7 June 1973; 2 males, 9 females, 2 females with eggsac, 5 juveniles, Pohnpei, Sokehs Island, 6°57'N, 158°1 1 'E, 8 June 1973, J.A. Beatty, J.W. Berry (BB); 1 male, 3 females, same data (WAM T80644). Diagnosis. — Zoica carolinensis is similar to Z. wauensis Lehtinen & Hippa 1979 from Papua New Guinea as illustrated in Lehtinen & Hippa (1979), in particular in regard to the structure of the male pedipalp. However, the basal part of the embolus is more exposed in Z. wauensis and the median tegular lobe is narrower and longer than that in Z. carolinensis. Both species differ considerably in the shape of the female epigynum, which is highly prominent in Z. wauensis (see Lethinen & Hippa 1979), but is flat in Z. carolinensis. Unfortunately, we were not able to compare specimens of both species as material of Z. wauensis, including the type material, could not be located in the Zoological Museum, University of Turku, Finland, where it is supposed to be housed (S. Kopponen, personal communication to VWF). Zoica carolinensis differs from Z. pacifica, the second species described here, by the presence of a median tegular lobe in the male pedipalp (absent in Z. pacifica) and the lack of a posterior lip of the epigynum (present in Z. pacifica). Description. — Male (based on holotype): Carapace: dorsal profile straight in lateral view; uniformly yellow-brown with gray pigmentation, centrally somewhat lighter, black around eyes (Fig. 1). Eyes: row of AE as wide as row of PME; row of AE very slightly procurved. Sternum: yellow, with some gray pigmentation marginally; brown macrosetae mainly margin- ally. Labium: yellow-brown. Chelicerae: yellow-brown with indistinct gray pigmentation, basally slightly darker; few whitish setae. Pedipalp (Figs. 5, 6): lateral apophysis tapering and bent dorsally at tip; embolus covered by terminal apophysis both of which are behind median tegular lobe (Fig. 5). Abdomen: yellow-brown with dense olive-gray pigmentation (Fig. 1); venter yellow. Legs: leg formula IV > I > II > III; uniformly yellow; spination of leg I; femur: 2 dorsal (only 1 on right leg), 1 apicoprolateral; tibia: 2 ventral pairs; metatarsus: 3 ventral pairs. Female (based on paratype): In all details like male (Fig. 2), except row of AE straight. Epigynum (Figs. 7, 8): ventral view: FRAMENAU ET Ah.—ZOICA OF THE PACIFIC REGION 227 Figures 1-4. — Zoica spp. 1. Z. carolinensis, holotype male from Ponape, E of Kolonia (Caroline Islands, Micronesia) (BPBM); 2. Z. carolinensis, paratype female from Ponape, E of Kolonia (Caroline Islands, Federated States of Micronesia) (BPBM); 3. Z. pacified, holotype male from Majuro Islet (Majuro Atoll, Marshall Islands) (BPBM); 4. Z. pacifica, female from Majuro Islet (Majuro Atoll, Marshall Islands) (BPBM). TL: (1) 1.63 mm; (2) 1.77 mm; (3) 1.92 mm; (4) 1.84 mm. weakly sclerotized with narrow posterior openings (Fig. 7); dorsal view: fertilization ducts form slightly bent tubes (Fig. 8). Measurements: Male holotype (female paratype): TL 1.63 (1.77), CL 0.90 (0.92), CW 0.65 (0.65). Eyes: AME 0.02 (0.03), ALE 0.03 (0.04), PME 0.06 (0.08), PLE 0.06 (0.06). Row of eyes: AE 0.15 (0.17), PME 0.14 (0.17), PLE 0.24 (0.26). Sternum (length/width) 0.54/0.42 (0.46/0.44). Labium (length/ width) 0.10/0.12 (0.15/0.10). AL 0.79 (0.81), AW 0.73 (0.63). Legs: lengths of segments (femur, patella/tibia, metatarsus, tarsus = total length): Pedipalp 0.38, 0.31, - , 0.36 = 1.06; leg I 0.63, 0.77, 0.48, 0.36 - 2.25; leg II 0.60, 0.67, 0.44, 0.35 = 2.05; leg III 0.56, 0.60, 0.48, 0.33 = 1.96; leg IV 0.77, 0.90, 0.73, 0.40 = 2.80 (Pedipalp 0.31, 0.38, - , 0.32 = 1.01; leg I 0.69, 0.83, 0.48, 0.36 = 2.36; leg II 0.65, 0.69, 0.44, 0.35 = 2.13; leg III 0.60, 0.63, 0.46, 0.33 = 2.02; leg IV 0.79, 1.02, 0.69, 0.38 = 2.88). Variation: S (?) (range, mean ± SD): TL 1.61-1.82, 1.73 ± 0.1 1; CL 0.88-0.96, 0.92 ± 0.04; CW 0.63-0.69, 0.67 ± 0.03; n = 3 (TL 1.73-2.28, 2.00 ± 0.17; CL 0.90-1.15, 1.00 ± 0.07; CW 0.65-0.84, 0.75 ± 0.05; n = 12). Etymology. — The specific epithet is an adjective derived from the Caroline Islands, where the species is found. Distribution. — Known only from Ponape (= Pohnpei) in the Caroline Islands, Eederated States of Micronesia (Fig. 13). Zoica pacifica new species (Figs. 3, 4, 9-13) Types. — Holotype male. Republic of the Marshall Islands, Majuro Atoll, Majuro Islet, 7°05'N, 171°08'E, 2 August 1969, J.W. Berry, breadfruit/coconut litter (BPBM); paratype female, same data as holotype (BPBM). Other material examined. — REPUBLIC OF THE MAR- SHALL ISLANDS: Majuro Atoll: 1 male, 3 females, 5 228 THE JOURNAL OF ARACHNOLOGY Figures 5-8. — Zoica caroliiwnsis new species. Male holotype and female paratype from Ponape, E of Kolonia (Caroline Islands, Federated States of Micronesia) (BPBM). 5. Left pedipalp, ventral; 6. Left pedipalp, retrolateral; 7. Epigynum, ventral view; 8. Epigynum, dorsal view. Scale bars: (5, 6) 0.26 mm; (7, 8) 0.17 mm. juveniles, 7°07'N, 171°2rE, 30 July 1969, J.W. Berry, grassy meadow (BB); 1 male, 2 females, 4 juveniles, Arniel Islet, 7°06'N, 171°22'E, 1 August 1969, J.W. Berry, grassy area in coconut forest, litter (BB); 1 male, 2 females, 5 juveniles, Dalap Islet, 7°06'N, 171°22'E, 1 August 1969, J.W. Berry, coconut/pandanus litter (BB); 1 male, 2 females, 5 juveniles, Long Island, 6 mi from Laura, 7°05'N, 171°08'E, 24 March 1980, J.A. Beatty, under coconut husks (BPBM); 1 female, 1 juvenile. Long Island, 6 mi from Laura, 7°05'N, 171°08'E, 24 March 1980, J.A. Beatty, from dead coconut leaves (BPBM); 2 females, 5 juveniles, Majuro Islet, 7°05'N, 171°08'E, 2 August 1969, J.W. Berry, breadfruit/coconut litter (BB); 3 females, Majuro Village, 7°06'N, 171°22'E, 24 July 1968, J.W. Berry, wet tropical forest, litter (BB); 1 female, 2 juveniles, Uotjaa Islet, 7°07'N, 171°21'E, 26 July 1968, J.W. Berry, Scaevolci litter (BPBM); 2 females, 3 juveniles, Uotjaa Islet, 7°07'N, 171°21'E, 26 July 1968, J.W. Berry, coconut litter (BPBM); 2 females, 1 juvenile, Uotjaa Islet, 7°07'N, 171°21'E, 26 July 1968, J.W. Berry, grass litter (BB); 6 females, 6 juveniles, Uotjaa Islet, 7°()7'N, 171°21'E, 25 July 1968, J.W. Berry, under coconut litter (BB). Diagnosis. — Zoica pacifica differs from all other species of Zoica by the absence of a median tegular lobe in the male pedipalp and the presence of a long posterior lip of the female epigynum. Description. — Male (based on holotype): Carapace: dorsal profile straight in lateral view; uniformly yellow-brown with gray pigmentation, black around eyes (Fig. 3); light-brown macrosetae around eyes, one large bristle that is bent dorsally centrally below AML, two macrosetae below ALE. Eyes: row of AE as wide as row of PME; row of AE straight. Sternum: long yellow-brown macrosetae mainly marginally. Labium: yellow-brown. Chelicerae: yellow-brown. Pedipalp (Figs. 9, 10): cymbium tip with two ventral macrosetae, lateral apophysis with mesal protrusion (Fig. 9); terminal apophysis with two round lobes and a pointed tip. Abdomen: yellow- brown with dense olive-gray pigmentation (Fig. 3); venter yellow. Legs: leg formula IV > I > II > III; uniformly yellow; spination of leg I: femur: 2 dorsal (only 1 on right leg), 1 apicoprolateral; tibia: 2 ventral pairs; metatarsus: 1 ventral. Female (based on paratype): In all details like male (Fig. 4), except leg spination: femur: 2 dorsal, 1 apicoprolateral; tibia: 2 FRAMENAU ET AL.—ZOICA OF THE PACIFIC REGION 229 03 0 ro o w Figures 9-12. — Zoica pacifica new species. Male holotype and female paratype from Majuro Islet (Majuro Atoll, Marshall Islands) (BPBM). 9, Left pedipalp, ventral; 10, Left pedipalp, retrolateral; 1 1, Epigynum, ventral view; 12, Epigynum, dorsal view. Scale bars: (9, 10) 0.20 mm; (1 1, 12) 0.19 mm. ventral pairs; metatarsus: 3 ventral pairs. Epigynum (Figs. 11, 12); ventral view: weakly sclerotized with long posterior lip (Fig. 11); dorsal view: spermathecal heads slightly wider than spermathecal stalks, fertilization ducts long and curved (Fig. 12). Measurements: Male holotype (female paratype): TL 1.92 (1.84), CL 0.98 (1.00), CW 0.71 (0.73). Eyes: AME 0.02 (0.03), ALE 0.03 (0.03), PME 0.07 (0.08), PEE 0.06 (0.07). Row of eyes: AE 0.16 (0.17), PME 0.16 (0.17), PEE 0.25 (0.27). Sternum (length/width) 0.46/0.44 (0.54/0.48). Labium (length/ width) 0.12/0.16 (0.12/0.16). AL 0.79 (1.02), AW 0.56 (0.65). Legs: lengths of segments (femur, patella/tibia, metatarsus, tarsus = total length): Pedipalp 0.38, 0.34, - , 0.36 = 1.09; leg I 0.71, 0.81, 0.54, 0.40 = 2.46; leg II 0.69, 0.75, 0.50, 0.35 = 2.28; leg III 0.63, 0.69, 0.52, 0.33 = 2.17; leg IV 0.83, 1.00, 0.75, 0.42 = 3.00 (Pedipalp 0.29, 0.34, - , 0.31 = 0.94; leg I 0.71, 0.81, 0.50, 0.38 = 2.40; leg II 0.65, 0.75, 0.46, 0.36 = 2.23; leg III 0.63, 0.69, 0.48, 0.35 = 2.15; leg IV 0.81, 1.06, 0.73, 0.44 = 3.03). Variation: S (9) (range, mean ± SD): TL 1.65-1.92, 1.76 ± 0.11; CL 0.90-0.98, 0.94 ± 0.04; CW 0.65-0.71, 0.68 ± 0.03; n = 4 (TL 1.77-2.30, 2.03 ± 0.18; CL 0.92-1.11, 1.02 ± 0.05; CW 0.69-0.81, 0.75 ± 0.03; n = 12). Etymology. — The specific epithet is an adjective derived from pacificus (Latin - peaceful) and refers to the Pacific region, where the species is found. Distribution. — Only known from Majuro Atoll in the Republic of the Marshall Islands (Fig. 13). ACKNOWLEDGMENTS We are especially grateful for the Academic Research Grants from Butler University awarded to JWB which helped support the field work. The U.S. Department of Energy (formerly the Atomic Energy Commission) provided travel 230 THE JOURNAL OF ARACHNOLOGY 120”E 130°E 140"E 150“E 160°E 170“E 120”E 130°E 140°E ISO'E 160°E 170“E Figure 13. — Records of Zoicci carolinensis new species (black circle) and Zoica pacifica new species (gray circle). funds for the work in the Marshall Islands. Two travel grants from the Indiana Academy of Science to JWB were of material assistance. Elizabeth Ramsey Berry’s contribution to all phases of the fieldwork in the Pacific and at home have been invaluable. The staff at the Bishop Museum, Honolulu, has been of assistance in many ways over a period of decades. This study was compiled while VWF received funds through the Australian Biological Resources Study (ABRS) to Mark Harvey (Western Australian Museum) and Andy Austin (The University of Adelaide) for a revision of the wolf spider fauna of Australia (2002-2005) and to VWF and Nikolaj Scharff (University of Copenhagen) for a revision of the orb- weaving spider fauna (Araneinae) of Australia. The senior author acknowledges, in particular, the support of his mentor Mark Harvey during studies at the Western Australian Museum. LITERATURE CITED Berland, L. 1924. Araignees de la Nouvelle-Caledonie et des lies Loyalty. Nova Caledonia, Zoologie 3:159-255. Berland, L. 1929. Araignees (Araneida). Pp. 35-78. In Insects of Samoa and Other Samoan Terrestrial Arthropoda. British Museum of Natural History, London. Berland, L. 1934. Araignees de Polynesie. Annales de la Societe Entomologique de France 103:321-336. Berland, L. 1938. Araignees des Nouvelles-Hebrides. Annales de la Societe Entomologique de France 107:121-190. Buchar, J. 1997. Lycosidae aus Bhutan. I. Venoniinae und Lycosinae (Arachnida: Araneae). Entomologica Basiliensia 20:5-32. Dondale, C.D. 1986. The subfamilies of wolf spiders (Araneae: Lycosidae). Actas X Congreso Internacional de Aracnologia, Jaca, Espaha 1:327-332. Framcnau, V.W. 2006. The wolf spider genus Venatrix Roewer: new species, synonymies and generic transfers (Araneae, Lycosidae). Records of the Western Australian Museum 23:145-166. Framenau, V.W. 2007. Revision of the new Australian genus Artoriopsis in a new subfamily of wolf spiders, Artoriinae (Araneae: Lycosidae). Zootaxa 1391:1-34. Gertsch, W.J. 1973. The cavernicolous fauna of Hawaiian lava tubes, 3. Araneae (spiders). Pacific Insects 15:163-180. Hippa, H. & P.T. Lehtinen. 1983. The zemtheres group of Zoicinae (Araneae, Lycosidae) and a relimitation of the subfamily. Annales Zoologici Fennici 20:151-156. Karsch, F. 1880. Mittheilung iiber die von Herrn Dr. O. Finsch wahrend seiner polynesischen Reise gesammelten Myriapoden und Arachniden. Sitzungsberichte der Gesellschaft naturforschender Freunde Berlin 5:77-83. Kronestedt, T. 1975. Studies on species of Holarctic Pardosa groups (Araneae, Lycosidae). 1. Zoologica Scripta 4:217-228. Lehtinen, P.T. & H. Hippa. 1979. Spiders of the Oriental-Australian region. 1. Lycosidae: Venoniinae and Zoicinae. Annales Zoologici Fennici 16:1-22. McKay, R.J. 1979. The wolf spiders of Australia (Araneae: Lycosidae): 10. A new species of the genus Flanona Simon. Memoirs of the Queensland Museum 19:231-235. Murphy, N.P., V.W. Framenau, S.C. Donnellan, M.S. Harvey, Y.C. Park & A.D. Austin. 2006. Phylogenetic reconstruction of the wolf spiders (Araneae: Lycosidae) using sequences from the 12S rRNA, 28S rRNA, and NADHl genes: implications for classification, biogeography, and the evolution of web build- ing behavior. Molecular Phylogenetics and Evolution 38:583- 602. Platnick, N.I. 2008. The World Spider Catalog, Version 9.0. American Museum of Natural History, New York. Online at http://research.amnh.org/entomology/spiders/catalog/INTR01. html (Accessed October 2008). Simon, E. 1898. Histoire Naturelle des Araignees, 2. Roret, Paris. Pp. 193-380. Simon, E. 1899. Ergebnisse einer Reise nach dem Pacific (Schauins- land 1896-1897). Arachnoideen. Zoologische Jahrbiicher, Abthei- lung fiir Systematik, Geographic und Biologic der Thiere 12:41 1-J37. Simon, E. 1900. Arachnida. Fauna Hawaiiensis, or the zoology of the Sandwich Isles: being results of the explorations instituted by the Royal Society of London promoting natural knowledge and the British Association for the Advancement of Science and carried on with the assistance of those bodies and of the Trustees of the Bernice Pauahi Bishop Museum at Honolulu. Cambridge FRAMENAU ET AL.—ZO/CA OF THE PACIFIC REGION 231 University Press, Cambridge, UK. Volume 2 Part 5 (ad part): 443-519. Thorell, T. 1895. Descriptive Catalogue of the Spiders of Burma, Based Upon the Collection Made by Eugene W. Oates and Preserved in the British Museum. British Museum of Natural History, London. 397 pp. Yoo, J.-S. & V.W. Framenau. 2006. Systematics and biogeography of the sheet-web building wolf spider genus Venonia (Araneae: Lycosidae). Invertebrate Systematics 20:675-712. Mcmusa ipt received 7 July 2008, revised 24 November 2008. 2009. The Journal of Arachnology 37:232-237 Plant nectar increases survival, molting, and foraging in two foliage wandering spiders Robin M. Taylor; Center for Life Sciences Education, Ohio State University, 260 Jennings Hall, 1735 Neil Avenue, Columbus, Ohio 43210, USA. E-mail; taylor.69@osu.edu Richard A. Bradley; Department of Evolution, Ecology, and Organismal Biology, Ohio State University, 384 B Morrill Hall, 1465 Mt. Vernon Avenue, Marion, Ohio 43302, USA Abstract. We predicted that because plant nectar is high in energy, it is likely to provide multiple benefits to spiders that spend a substantial amount of energy foraging. In three laboratory experiments, we tested the effects of dietary extrafloral nectar on the survival, molting, and activity of two foliage wanderers, Cheiracanthium mildei L. Koch 1864 (Miturgidae) and Hihana velox (Becker 1879) (Anyphaenidae), both highly active, quick-moving nocturnal foragers. Extrafloral nectar contributed significantly to survival and molting in prey-deprived H. velox. On a marginal diet of prey (one Drosophila adult on alternate days) offered to spiders as soon as they emerged, 91% of C. mildei underwent their first molt if they also received nectar, compared to 1% of controls without nectar. On a marginal diet of prey (one Drosophila adult on alternate days) offered to spiders starting two days after their emergence, 78% of the spiders also receiving nectar molted, compared to 0% of controls without nectar. Video recordings of activity showed that prey-deprived groups of C. mildei maintained their active nocturnal foraging for many days on nectar, whereas controls became increasingly quiescent until they died. Non-web-building spiders that feed on nectar may utilize its energy for foraging and thereby allocate the nutrients of prey to maintenance and growth. Keywords: Diet, extrafloral, fitness, nutritional allocation, Cheiracarnhium mildei, Hihana velox As obligate carnivores, spiders are presumed to acquire their energy for maintenance, growth, and reproduction from captured prey. Attesting to their levels of activity (and resulting energetic needs), some wandering spiders encounter and eat insect eggs (Buschman et al. 1977; Nyffeler et al. 1990; Miliczky & Calkins 2002; Pfannenstiel 2004) and the eggs of other spiders (Willey & Adler 1989). As generalist predators, spiders are useful models for investigating invertebrate nutrient-specific selective foraging (Mayntz et al. 2005), but not all spiders should be presumed to be exclusively carnivorous or to get all of their energy from prey lipids. Coll & Guershon (2002) identify “true omnivory” (i.e., feeding on both plants and prey) in spiders, citing members of two families in particular; an araneid that feeds on pollen grains in the juvenile stage, which the spiderlings trap and eat incidentally when they eat and recycle their webs (Smith & Mommsen 1984), and an anyphaenid that feeds on plant nectar (Taylor & Foster 1996). Compared to feeding on pollen grains, nectar feeding is a more directed behavior, which has been reported among all ages of spiders and among a number of different families. Independent observations of members of Thomisidae (crab spiders), Salticidae (jumping spiders), and the active, fast- moving Anyphaenidae, Miturgidae, and Corinnidae — all wanderers in foliage — suggest that all feed at the floral and extralloral nectaries (EFNs) of plants (Edmunds 1978; Vogelei & Greissl 1989; Pollard et al. 1995; Ruhren & Handel 1999; Jackson et al. 2001). Applying what Singer & Bernays (2003) might call a “behavioral perspective,” Taylor & Pfannenstiel (2008) sampled spiders they deemed most likely to feed regularly on nectar from the EFNs of cotton plants and determined that one out of four were positive for ingested fructose, a plant-derived sugar. The survey also added members of Oxyopidae to the list of families that nectar feed. Considering that the hunting success rate for some wandering spiders is thought to be low (Miyashita 1968; Anderson 1974; Nentwig 1987; Nyffeler et al. 1987; Nyffeler & Sterling 1994), we propose that plant sugars may be of direct benefit and help fuel the cursorial life of these spiders, allowing the valuable nutrients of prey to be allocated to the more complex metabolic processes of maintenance, growth, and reproduction. Plant nectars contain primarily carbohydrates and water (Percival 1961), but also amino acids, lipids, vitamins, and minerals (Baker & Baker 1975, 1983; Koptur 1992). Nectar is exuded at floral nectaries, but unless a flower’s corolla is shallow, nectar is more accessible to spiders’ small mouthparts by way of extrafloral nectaries (EFNs), nectar-bearing tissues or structures that reside anywhere on a plant outside of a flower. EFNs often occur on leaves or leaf petioles, and take many forms, such as slits, cups, bowls, or undifferentiated tissue. Arthropods, particularly ants, often visit these open, accessible EFNs (Bentley 1977). Spiders observed at nectaries are non-web-building wanderers that inhabit vegetation, and their degree of activity and nectar feeding may be correlated. Searching for prey requires wandering, and frequent wander- ing means a greater likelihood of encountering EFNs and plant nectar. Plant nectar, which contains mostly sugar, could repay the energetic costs of wandering. The active foragers, Cheiracanthium mildei L. Koch 1864 (Miturgidae) and Hihana velox (Becker 1879) (Anyphaenidae), run throughout the vegetation at night, making them good candidates to investigate the energetic contributions of nectar. Both of these spiders have been observed at plant nectaries (Taylor & Foster 1996). Three laboratory experiments tested the effects of extra- floral nectar on the survival, molting, and activity of newly emerged spiders. Hihana velox was the subject of initial survival tests. Cheiracanthium mildei, which is ecologically similar, was more easily obtained and the subject of later experiments. The experiments tested 1) the effects of nectar 232 TAYLOR & BRADLEY— PLANT NECTAR BENEEITS FOLIAGE WANDERERS 233 and two concentrations of sucrose on the survival of individually housed H. velox, 2) the effect on molting in C. milclei by adding nectar to a marginal diet of prey (Drosophila melanogaster), and 3) the effects of nectar on the nocturnal running activity of small groups of prey-deprived C. mildei. METHODS Spiders. — Experimental H. velox were offspring of adults collected in 1994 in Alachua County, Gainesville, FL, USA; experimental C. mildei were offspring of adults collected in Franklin County, Columbus, Ohio, USA. Egg sacs were either collected in the field with adult females (which guard them) or produced by females maintained in the laboratory on a varied insect diet (mainly house flies and mosquitoes). Adults lived in 7-liter clear acrylic cages (15 X 21 X 27 cm) with a screened opening at one end and a sleeved opening at the other. Experiments were conducted in a laboratory rearing room maintained on a 16:8 h light:dark diel cycle at ca 27° C and 80% relative humidity. Spiders were checked daily for molting and mortality. Each experiment or trial began within 12 h of spiderlings’ emergence from their egg sacs, which was considered Day 0. “First molt,” therefore, refers to a spider’s first molt post-emergence. Spiders were housed individually in clear, lidded, plastic containers, 5.2 cm diam. X 3.6 cm. Each container had four holes: two 12-mm, mesh-covered holes top and bottom; and two opposing 17-mm holes in the side wall, one mesh-covered and the other corked for introduction of prey and for changing the fluid wells of feeders. Feeders were small rectangles of plastic (1 X 2.5 cm) with a dimple (i.e., fluid well) drilled near each end (large dimple for water, small for nectar). Twenty of these small containers, composing an even mix of controls and treatment individuals, filled a large, clear, plastic 30 X 25 cm lidded box. Two boxes fit on a large plastic tray, lightly dusted with sifted sulfur to repel mites. Boxes were rotated daily. Spiders in the activity trials were housed in small 7-cm- square plastic lidded boxes, each with four 17-mm holes, one on each side, three mesh-covered and one corked for introduction of spiders and for refilling fluid wells and changing feeders. Each box held four feeders, totaling eight fluid wells. For the control, all eight wells contained water; for the treatment, four wells contained water, and four contained nectar. Control and treatment boxes were placed side-by-side in a lidded clear plastic box, 35 X 24 cm and filmed with an RCA closed circuit TC701 1 infrared-sensitive camera under continuous red light illumination, which does not disturb the spiders (Peck & Whitcomb 1970). Diet. — In all experiments, water was available ad libitum. All containers that held spiders also held at least one feeder. In controls, both fluid wells of the feeder contained water. In treatment groups, the large well of the feeder contained water, and the small well contained either nectar or sucrose. Water also was available from soaked No.l (9 mm) cotton dental balls. Ambient relative humidity was high, and smaller containers were kept in large boxes to keep water wells from drying out. The constant availability of free water ensured that spiders did not take nectar solely to obtain water. Sucrose and nectar, because of their viscosity, were delivered with a micro spatula in the smallest transferable amount, between 1-2 pi, smeared into the smaller fluid well of the plastic feeders. Water, sucrose, and nectar were changed daily. Prey consisted of live, vestigial-winged Drosophila melanogaster maintained on instant (blue) Drosophila medium (Carolina Biological Supply). Diets combining prey and nectar were offered separately on alternate days to ensure that spiders were willing and able to consume nectar directly, rather than by way of prey that had ingested nectar. All nectar was extrafloral to avoid introduction of pollen as a possible source of protein (Smith & Mommsen 1984). For the first trial of the first experiment with H. velox, extrafloral nectars were collected and combined from various greenhouse plants, such as Hibiscus and orchids. The nectar was slightly diluted to an unknown concentration to ease handling. For the second trial and all of the following experiments, nectar was undiluted and came solely from Terminalia cattapa (Indian almond, also growing in the university greenhouse), which produces copious nectar at EFNs on the base of the leaf near the petiole. Nectar from T. cattapa EFNs was 87.5% sugar constituents (variety unknown) determined from serial dilu- tions and a Reichert-Jung refractometer. The nectar was collected with a microspatula and stored at —45° C. Experiments. — 1. Survival: Two trials compared survival in individually housed H. velox on diets of water only, sucrose, or extrafloral plant nectar. For each trial, spiders from a single egg sac were divided among the control and two treatments. Both trials included a sucrose treatment to distinguish contributions of carbohydrates from possible contributions of other nectar components, such as amino acids or lipids. In the first trial, sucrose was relatively “low” (25%), in the second trial, “high” (69%), to more closely imitate the high sugar concentration of extrafloral nectars. Individuals were checked daily for mortality. 2. Molting: Two trials compared molting in individually housed C. mildei receiving marginal diets of prey (Drosophila) with and without nectar from T. cattapa. For each trial, spiders from a single egg sac were divided between the control and the treatment. In both trials, spiders were fed a single Drosophila adult on alternate days until the spider molted. On days without Drosophila, spiders received water (controls) or nectar. In the first trial. Drosophila were introduced on Day 1 . In the second trial, introduction of Drosophila was delayed until Day 3. Nectar-fed spiders on the delayed Drosophila diet received nectar for the first two days. Spiders were part of the trial until they molted once. 3. Activity: Because both H. velox and C. mildei wander energetically in vegetation at night and are inactive during the day, we filmed two groups of cohabiting spiders at night in the laboratory, one with and one without access to nectar. Both had access to water ad libitum. For both replicates spiders from a single egg sac were divided between the control and treatment. From tapes, we quantified nightly activity as the number of spiders simultaneously running during a one- minute period at 10-min intervals, for 54 periods covering crepuscular light and the eight hours of scotophase. The mean of these 54 periods represented that night’s activity. Analysis: We analyzed data with Statistica for Windows (2000), StatSoft, Inc. Survival analysis (Kaplan-Meier) em- ployed log-rank tests, which were adjusted for multiple comparisons, for which the calculated comparison-wise error rate of 0.008 is based on = 3 treatments (Hardin et al. 1996). 234 THE JOURNAL OF ARACHNOLOGY Figure 1 . — Survival estimates for Hihcma velox fed water only (/; = 24), 25% sucrose (/; = 25), or extrailoral nectar (n = 24). Curves with different letters are significantly different (adjusted pair-wise com- parison). We used chi-square tests to compare molting and the Mann- Whitney t/-test to compare nocturnal activity. RESULTS 1. Survival. — In the first trial, prey-deprived spiders survived significantly longer than water-only controls if they received 25% sucrose (log-rank test statistic = 3.71, L* = 0.0002), or if they received nectar (log-rank test statistic = 4.39, P = 0.0001). The 25% sucrose and nectar treatments were not significantly different (log-rank test statistic = —1.43, P = 0.1566) (Fig. 1). The second trial produced similar results. Spiders survived longer than water-only controls if they received 69% sucrose (log-rank test statistic = 3.36, P = 0.0008), or if they received nectar (log-rank test statistic = 3.66, P — 0.0003) (Fig 2). The 69% sucrose and nectar treatments were not significantly different (log-rank test statistic = 0.770, P = 0.4412) (Fig. 2). Molting occurred in all of the groups except the controls of Trial 2 (Table 1). 2. Molting. — Both trials ended when all of the spiders in the nectarless control died. Day 15 for the first trial and Day 16 for the second. Nectar added to a marginal diet of prey (one Drosophila adult on alternate days) significantly increased the numbers of spiders that underwent their first molt whether the Drosophila diet began on Day 1 (97% vs. 7%) or Day 3 (78% vs. 0%). Delaying the introduction of prey, however, had a significant effect on the ability to survive the process of molting. Among the 97% (29/30) of spiders that molted receiving nectar and Drosophila on Day 1, 100% survived the molting process. Among spiders receiving nectar from Day 1 and Drosophila first on Day 3, 78% (38 /49) initiated molting. Figure 2. — Survival estimates of Hihami velox fed water only (« = 20), 69% sucrose (n = 30), or extrafloral nectar (/? = 30). Curves with different letters are significantly different (adjusted pair-wise com- parison). but only 47% of the sample survived the molting process, a significant decrease in survival (multiple comparison x^- prey on Day \, n = 30; prey on Day 3, ii = 49, P < 0.001). Whether they received nectar or not, individual spiders on average consumed the same number of prey daily before an individual died or molted, calculated from the total number of Drosophila consumed in the experiment/total spider days survived. In the first trial, spiders with nectar ate 0.32 Drosophila daily and those without, 0.30 Drosophila (94 prey/294 d; 74 prey/243 d, respectively). In the second trial, spiders with nectar ate 0.26 Drosophila daily, and those without, 0.23 Drosophila (122 prey/466 d; 62 prey/265 d, respectively). 3. Activity. — The trials ended when any of the spiders died, which occurred in the nectarless control on Night 5 in the first replicate and on Night 4 in the second replicate. Comparisons of the total number of intervals of activity (270 for Replicate 1, 216 for Replicate 2) between the control and the nectar treatment show that nectar contributes significantly to the spider’s running, in absence of prey (Mann-Whitney U\ Replicate 1, n = 270 for both treatments, Z = —12.709, P < 0.001; Replicate 2, n = 216 for both treatments, Z = —13.377, P < 0.001). On Day 1, there was no significant difference in activity between spiders with and without nectar. On successive nights, spiders without nectar became increasingly quiescent until they died (Fig. 3). Individuals could not be distinguished from one another, and seven individuals at most could be distinguished running simultaneously, making the estimate of spider activity conservative. Table 1 . — Survival (mean ± 1 SE) and molting of Hihana velox in two trials of survival on diets of water only, sucrose, or nectar. Significantly more spiders molted than their water-only controls if they received nectar or 69% sucrose (y}, *P < 0.05, **P < 0.001). Trial Diet Survival (d) Range (d) f molt 11 1 Water only 9.8 ± .7 4-17 17% 24 1 25% sucrose 22.0 ± 3.4 6-73 28% 25 1 Nectar 32.6 ± 3.6 5-66 50%* 24 2 Water only 10.4 ± .7 8-20 0% 20 2 69% sucrose 28.4 ± 3.4 4-67 63%** 30 2 Nectar 28.1 ± 3.5 8-73 52%** 30 TAYLOR & BRADLEY— PLANT NECTAR BENEFITS FOLIAGE WANDERERS 235 Figure 3. — Nocturnal activity of eight or nine cohabiting, newly emerged, prey-deprived Cheiraccmthiiim iiiilclei, with or without nectar. Replicate 1 (solid circles) without nectar, n = 8; with nectar, n = 8. Replicate 2 (open circles) without nectar, /; = 9, with nectar, ii = 9. “Activity” is the mean number of spiders simultaneously running during a one-minute period at 10-minute intervals for 54 periods. Points are means ± SE. DISCUSSION In the lives of spiders, vegetation is considered important as a support for webs, as refugia, or as a food source for the insects that spiders catch (Turnbull 1973; Hatley & MacMa- hon 1980; Greenstone 1984; Uetz et al. 1999). Only recently have researchers considered the possibility of vegetation as a direct food source, and some spiders as true omnivores among terrestrial invertebrates. Taking this recognition one step further, we investigate why nectar feeding should be a likely activity among some spiders and for the first time measure the direct biological benefits to spiders that nectar-feed. Why nectar feeding is a likely activity. — The likelihood that nectar can and does play a role in the energy budget of some spiders is unsurprising. Nectar’s value as a dietary source of energy has been well established for nectarivorous insects, such as bees and butterflies; and predaceous arthropods other than spiders have been shown to survive periods of prey deprivation by feeding on plant nectars (Yokoyama 1978; Hagen 1987; van Rijn & Tanigoshi 1999; Limburg & Rosenheim 2001). Cursorial spiders that wander in vegetation with EFNs are likely to encounter nectar, which they have the potential to detect with “gustatory” hairs on their tarsi (Barth 2002). Cheiracanthium mildei, for example, oriented immedi- ately to sugar and inserted its mouthparts as soon as a fore- tarsus touched it (RMT personal observation). Encountering nectar, spiders are predisposed to ingesting their food in liquid form, given their form of extra-oral digestion (Cohen 1998), which may also help them ingest nectars that can be too viscous for other nectar feeders to handle (Wackers et al. 2001). Spiders respond positively to nectar, shown by preference tests (Jackson et al. 2001) and by their willingness to ingest chemicals, such as LSD, caffeine, and strychnine, if they are delivered in a sucrose solution (Christiansen et al. 1962; Witt 1971). And, both spiders that have been analyzed for digestive enzymes (a tarantula and an agelenid) possess the enzyme sucrase (Pickford 1942; Mommsen 1977), which can digest nectar Concentration of sugars at EFNs. — Our experiments show that even when water was available, spiders still drank nectar when offered. Hibana relax without prey survived significantly longer and had a significantly higher incidence of molting than water-only controls if they had access to nectar or to the high (69%) concentration of sucrose (Table 1). Such high concen- trations of sugar are not unusual in EFNs. The sugar concentration of T. cattapa extrafloral nectar that we determined to be 87.5% is nearly identical to the concentration of sugars (872 mg/ml) from the EFNs of castor bean (Riciinis communis) (Baker et al. 1978), and is similar to the concentration of sugar {11 .1%) exuded at the EFNs of cashew (Anacardium occidentale) (Wunnachit et al. 1992). Hibana relax has been observed feeding at both of these species (Taylor & Foster 1996). Other Hibana spp. and C. inclusum have been observed at the EFNs of cotton (Taylor & Pfannenstiel 2008), which produce nectars with a sugar concentration between 62% (Wackers et al. 2001) and 86% (Butler et al. 1972). Nectar fulfills energy requirements. — In experiments provid- ing C. mildei with Drosaplnla on Day 1, 97% of the spiders molted if they also had access to nectar, compared to 7% of controls without nectar. In experiments measuring activity, nectar contributed significantly to the energetic needs of C. mildei, conferring not only survival but also allowing them to keep up their frenetic running all night, every night that they were filmed. These results offer an opportunity to tease apart how these spiders are allocating nectar and prey-derived nutrients and can begin to address Uetz’s (1992) question, “Is energy the sole currency involved in spider foraging, or do nutrients play a critical role?” Both nectar and pure sucrose contributed to a higher incidence of molting in prey-deprived H. relax (Table 1), suggesting that it was the sugar component of nectar that contributed most to molting. Molting is an energy-depleting event that can increase respiration three-fold (Stranzy & Perry 1987). Nearly half of the components of a spider’s cuticle, however, consist of proteins (Dalingwater 1987). Because sucrose contributed to the same incidence of molting as nectar, but molting requires not only carbohydrates but also protein for new cuticle, it appears that sugars fulfilled much of the energetic demand of sustained nocturnal locomotion (i.e., foraging), survival, and ecdysis (the molting event), allowing the protein contained in yolk reserves and prey to be allocated primarily to growth and/or new cuticle deposition. This may explain why H. relax provided with nectar but no prey survived long but did not grow (only some undergoing a single molt: Table 1), and why C. mildei — a larger spider at emergence with perhaps fewer reserves — provided with a marginal amount of prey but deprived of nectar, died early without molting. That is, a marginal amount of prey divided between activity and growth could not support both. The addition of nectar substantially changed the outcome: on average, both control and treatment C. mildei ingested nearly identical amounts of prey (0.30 vs. 0.32, and 0.23 vs. 0.26 Drosophilal^'pidQrlAay in trials 1 and 2, respectively), but molted only if their diet was supplemented with nectar. 236 THE JOURNAL OF ARACHNOLOGY suggesting that they were at the margins of their nutritional requirements. Nectar feeding, by providing the energy for activity, may allow spiders to subsist on marginal amounts of prey, and, depending on the minimum amount required to reach functional maturity, might substantially reduce a spider’s prey requirements. Spiders that can reduce their prey intake also are likely to reduce the energy and risk associated with attacking and subduing prey. It is not clear why C. mildei receiving their initial Drosophila on Day 3 underwent a first post-emergent molt after consuming fewer prey than when Drosophila were introduced on Day 1. The consequences of delaying the introduction of prey by two days are dire: a 53% reduction in first-molt survival. This hints at some possible protein requirement for normal development within the first two days of spiderling emergence, or perhaps some developmental timeline triggered by the presence of protein in the diet. Fulfilling either of these requirements would make nectar-fueled survival and hunting all the more valuable. ACKNOWLEDGMENTS We thank Woodbridge Foster and all members of his laboratory for controlled-environment working space, equip- ment, materials, and discussion; Traci Solli and the (former) Introductory Biology Program for materials; George Keeney for access to the OSU insectary and advice; Joan Leonard for access to the OSU greenhouse; OSU Physical Facilities for use of their Genie (electric ladder) for collecting nectar at all heights of the T. cattapa tree in the OSU greenhouse; and two anonymous reviewers for improving the manuscript. 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On the association between Myrmarachne spp. j (Salticidae) and ants. Bulletin of British Arachnological Society 4:149-160. : Greenstone, M.H. 1984. Determinants of web spider species diversity: | vegetation structural diversity vs. prey availability. Oecologia | 62:299-304. | Hagen, K.S. 1987. Nutritional ecology of terrestrial insect predators. ( Pp. 533-577. In Nutritional Ecology of Insects, Mites, Spiders, and Related Invertebrates. (F.J. Slansky & J.G. Rodriguez, eds.). John Wiley & Sons, New York. Hardin, M.J., J.L. Willers & T.L. Wagner. 1996. Nonparametric multiple comparisons of survivorship distributions. Journal of Economic Entomology 89:715-721. ' Hatley, C.L. & J.A. MacMahon. 1980. Spider community organiza- i tion: seasonal variation and the role of vegetation architecture. Environmental Entomology 9:632-639. Jackson, R., S.D. Pollard, X.J. Nelson, G.B. Edwards & A.T. Barrion. 2001. Jumping spiders (Araneae: Salticidae) that feed on nectar. Journal of Zoology (London) 255:25-29. Koptur, S. 1992. Extrafloral nectar-mediated interactions between ! insects and plants. Pp. 81-129. In Insect-Plant Interactions. Volume IV. (E.A. Bernays, ed.). CRC Press, Boca Raton, I Florida. Miliczky, E.R. & C.O. Calkins. 2002. Spiders (Araneae) as potential ; predators of leafroller larvae and egg masses (Lepidoptera;Tor- ,, tricidae) in Central Washington apple and pear orchards. Pan- ! Pacific Entomologist 78:140-150. Limburg, D.D. & J.A. Rosenheim. 2001. Extrafloral nectar con- |: sumption and its influence on survival and development of an omnivorous predator, larval Chrysoperla plorabunda (Neuroptera: Chrysopidae). Environmental Entomology 30:595-604. Mayntz, D., D. Raubenheimer, M. Salomon, S. Toft & S.J. Simpson. 2005. Nutrient-specific foraging in invertebrate predators. Science 307:111-112. Miyashita, K. 1968. Growth and development of Lycosa T-insignata Boes. et Str. (Araneae: Lycosidae) under different feeding conditions. Applied Entomology and Zoology 3:81-88. i Mommsen, T.P. 1977. Digestive enzymes of a spider {Tegenaria acrica Koch). II. Carbohydrases. Comparative Biochemistry and Physi- ology 60A:37 1-375. | Nentwig, W. 1987. The prey of spiders. Pp. 249-263. In Ecophysi- f ology of spiders. (W. Nentwig, ed.). Springer, Berlin. Nyffeler, M., R.G. Breene, D.A. Dean & W.L. Sterling. 1990. Spiders :: as predators of arthropod eggs. Journal of Applied Entomology t 109:490-501. | Nyffeler, M., D.A. Dean & W.L. Sterling. 1987. Predation by green ' lynx spider, Peucetia viridans (Araneae: Oxyopidae), inhabiting , cotton and woolly croton plants in East Texas. Environmental Entomology 16:356-359. | Nyffeler, M. & W.L. Sterling. 1994. Comparison of the feeding niche of polyphagous insectivores (Araneae) in a Texas cotton planta- tion: estimates of niche breadth and overlap. Environmental ; Entomology 23:1294-1303. j' Peck, W.B. & W.H. Whitcomb. 1970. Studies on the biology f of a spider, Chiracanthium inclusum (Hentz). University of ( Arkansas, Agricultural Experiment Station Bulletin 753. 76 pp. ■ Percival, M.S. 1961 . Types of nectar in angiospenns. New Phytologist 60:235-281. Pfannenstiel, R.S. 2004. Nocturnal predation of lepidopteran eggs in ; south Texas cotton-2002. Pp. 1594-1600. In Proceedings of the ' TAYLOR & BRADLEY— PLANT NECTAR BENEFITS FOLIAGE WANDERERS 237 Beltwide Cotton Conference. National Cotton Council, Memphis, Tennessee. Pickford, G.E. 1942. Studies on the digestive enzymes of spiders. Transactions of the Connecticut Academy of Arts and Sciences 35:33-72. Pollard, S.D., M.W. Beck & G.N. Dodson. 1995. Why do male crab spiders drink nectar? Animal Behaviour 49:1443-1448. Ruhren, S. & S.N. Handel. 1999. Jumping spiders (Salticidae) enhance the seed production of a plant with extrafloral nectaries. Oecologia 119:227-230. Singer, M.S. & E.A. Bernays. 2003. Understanding omnivory needs a behavioral perspective. Ecology 84:2532-2537. Smith, R.B. & T.P. Mommsen. 1984. Pollen feeding in an orb- weaving spider. Science 226:1330-1333. Stranzy, F. & S.F. Perry. 1987. Respiratory system: structure and function. Pp. 78-94. In Ecophysiology of Spiders. (W. Nentwig, ed.). Springer- Verlag, Berlin. Taylor, R.M. & W.A. Foster. 1996. Spider nectarivory. American Entomologist 42:82-86. Taylor, R.M. & R.S. Pfannenstiel. 2008. Nectar feeding by wandering spiders on cotton plants. Environmental Entomology 37:996-1002. Turnbull, A.L. 1973. Ecology of the true spiders (Araneomorphae). Annual Review of Entomology 18:305-348. Uetz, G.W. 1992. Foraging strategies of spiders. Trends in Ecology & Evolution 7:155-158. Uetz, G.W., J. Halaj & A.B. Cady. 1999. Guild structure of spiders in major crops. Journal of Arachnology 27:270-280. van Rijn, P.C.J. & L.K. Tanigoshi. 1999. The contribution of extrafloral nectar to survival and reproduction of the predatory mite Ipliiseius degenerans on Ricinus coninnmis. Experimental & Applied Acarology 23:281-296. Vogelei, A. & R. Greissl. 1989. Survival strategies of the crab spider Thomisiis omistus Walckenaer 1806 (Chelicerata, Arachnida, Thomisidae). Oecologia 80:513-515. Wackers, F.L., D. Zuber, R. Wunderlin & F. Keller. 2001. The effect of herbivory on temporal and spatial dynamics of foliar nectar production in cotton and castor. Annals of Botany 87:365-370. Willey, M.B. & P.H. Adler. 1989. Biology of Peucetui viridanx (Araneae, Oxyopidae) in South Carolina, with special reference to predation and maternal care. Journal of Arachnology 17:275-284. Witt, P.N. 1971. Drugs alter web-building of spiders. A review and evaluation. Behavioral Science 16:98-113. Wunnachit, W.C., C.F. Jenner & M. Sedgley. 1992. Floral and extrafloral nectar production in Amicardium occidentale L. (Anacardiaceae): an andromonoecious species. International Jour- nal of Plant Science 153:413^20. Yokoyama, V.Y. 1978. Relation of seasonal changes in extrafloral nectar and foliar protein and arthropod populations in cotton. Environmental Entomology 7:799-802. Manuscript received 28 September 2007, revised 20 March 2009. 2009. The Journal of Arachnology 37:238-240 SHORT COMMUNICATION Caddo agilis and C. pepperella (Opiliones, Caddidae) diverged phylogenetically before acquiring their disjunct, sympatric distributions in Japan and North America Jeffrey W. Shultz: Department of Entomology, University of Maryland, College Park, Maryland 20742, USA. E-mail: jshultz@umd.edu Jerome C. Regier: Center for Biosystems Research, University of Maryland Biotechnology Institute, College Park, Maryland 20742, USA Abstract. The harvestmen Caddo agilis Banks 1892 and C. pepperella Shear 1975 (Caddidae, Caddinae) share a disjunct distribution in eastern Asia and eastern North America that has been attributed to either recent (Pleistocene) evolution of a C. pepperella morph from C. agilis in each region or to a pre-glacial separation within each of two established species. The present study used 2,130-base sequences from two nuclear protein-coding genes (EFla, Pol II) to test the phylogenetic predictions of both hypotheses using representatives from the two Caddo species from both regions and two acropsopilionine outgroup species. The results supported the hypothesis that the two Caddo species were distinct prior to their respective biogeographic disjunctions; C. agilis and C. pepperella were each recovered as monophyletic and each appears to have undergone separation into Asian and North American groups. Keywords: Harvestmen, phylogeny, elongation factor-la, RNA polymerase II This study focuses on the phylogeny and biogeography of the two extant species of Caddo, C. agilis Banks 1892 and C pepperella Shear 1975 (Caddidae, Caddinae), both of which occur in eastern North America and Japan. Two scenarios have been offered in the arachnological literature to explain the disjunct sympatric distribu- tion of the two species: the parallel-evolution hypothesis (Shear 1975, 1996, 2004) and the habitat-fragmentation hypothesis (Suzuki 1976). Shear (1975) originally suggested that the smaller C. pepperella evolved in North America as a paedomorphic (progenetic) variant of C. agilis, perhaps as an adaptation to shorter growing seasons associated with glacial conditions during the Pleistocene. The subsequent discovery of C. pepperella in Japan (Suzuki 1976) was inconsistent with Shear’s hypothesis, as it seemed to require an improbable recent dispersal between eastern North America and Japan. Shear (1996, 2004) countered that a C. pepperella morph may have evolved independently in North America and Japan during the Pleistocene. In contrast, Suzuki (1976) proposed that both species originally inhabited an ancient circumboreal ecosystem that now consists of isolated postglacial fragments in eastern Asia and eastern North America. The parallel-evolution and habitat-fragmentation hypotheses can be tested by means of molecule-based phylogenetic analysis, with the former predicting diphyly within C. pepperella and the latter predicting monophyly of both C. agilis and C. pepperella across their ranges. Here we test these hypotheses using 2,130 base pairs of the nuclear protein-coding genes elongation factor- la) (EF-la) and RNA polymerase 11 (Pol II) from representative C agilis and C. pepperella from North America and Japan as well as two acropsopilionine outgroup species, AustropsopUio sudamericantis Shultz & Cekalovic 2003 and Acropsopilio chileiisis Silvestri 1904. Our results corroborate Suzuki’s habitat-fragmentation hypothesis. METHODS Terminal taxa and sequences. — Specimens were collected alive and preserved in > 95% ethanol. They were stored in > 95% ethanol at —20° C up to 2 yr prior to RNA extraction. The analysis was based on sequences from six specimens, with collection data and GenBank accession numbers as follows: 1. Caddo agilis Banks 1892. USA: New Hampshire: Cheshire County, Pisgah State Park, 42.868°N, 72.448°W, 7-1 1 July 2001, J.W. Shultz (EF-la: FJ361272; Pol II: FJ476262- FJ476264). 2. Caddo agilis Banks 1892. JAPAN: Tot tori Prefecture: Chizu- cho, Ashizu Tunnel, 660 m, 20 June 1998, N. Tsurusaki (EF-la: AF240838; Pol II: AHO 10430). 3. Caddo pepperella Shear 1975. USA: New Hampshire: Cheshire County, Pisgah State Park, 42.868°N, 72.448°W, 7-11 July 2001, J.W. Shultz (EF-la: FJ361272; Pol II: FJ476265- FJ476267). 4. Caddo pepperella. JAPAN: Tot tori Prefecture: Mt. Nagi, 630 m elev., 20 June 1998, N. Tsurusaki (EF-la: AF240863; Pol II: AH0I0457). 5. Acropsopilio chUensis. CHILE: Provinica de Concepcion: Cerro Caracol, 5 October 2003, T. Cekalovic (EF-la: FJ361275; Pol II: FJ476256 - FJ476258). 6. AustropsopUio sudamericanus. CHILE: Provincia de Valdivia: Cerro Oncol, April 2001, T. Cekalovic (EF-la: FJ361274; Pol II: FJ476259-FJ476261). A voucher specimen of each species is deposited in the National Museum of Natural History (Smithsonian Institution) except for C. pepperella from Japan, because the only specimen available was consumed in genomic extraction. Molecular methods. — Detailed procedures for generating sequence data, including primer sequences, have been published elsewhere (Regier & Shultz 1997). In brief, total nucleic acids were isolated; complementary DNA of EF-la and Pol II mRNA was made by reverse transcription; ds-DNA copies were amplified by PCR and subsequently gel isolated; the resulting PCR fragments were used as templates for another round of PCR amplification with nested primers; and the resulting fragments were gel isolated and sequenced. When the resulting fragment concentration was too low to sequence directly, it was either concentrated or reamplified using the M13 sequences present at the 5' ends of all primers. The same M13 238 SHULTZ & REGIER— PHYTOGENY & BIOGEOGRAPHY OF CADDO 239 sequences were also used as primers for thermal cycle/dideoxy sequencing. Sequencing reactions were fractionated and preliminary analyses were performed with Perkin-Elmer/ABI automated DNA sequencers. Automated DNA sequencer chromatograms were edited and contigs were assembled using the pregap and gap4 programs within the Staden software package (Staden et al. 1999). Sequences were aligned and Nexus-formatted nucleotide data sets were constructed using the Genetic Data Environment, version 2.2 (Smith et al. 1994). All sequences lacked indels. Amino acid data were inferred from nucleotide sequences using the universal nuclear genetic code option in MacClade, ver. 3.08 (Maddison & Maddison 1992). Phylogenetic analysis. — Parsimony analyses of three data sets [all nucleotides (ntI-3), third codon positions (nt3) and inferred amino acids (aa)] were performed in PAUP4.0 (Swofford 1998) using unordered, equally weighted characters. Analyses consisted of exhaustive searches followed by bootstrap analyses (Felsenstein 1985) based on branch-and-bound searches of 1,000 pseudoreplicates. In conducting maximum-likelihood (ML) analysis, the program Modeltest, ver. 3.7 (Posada & Crandall 1998) was used to choose a model for the ntl-3 and nt3 data sets using AIC (Posada & Buckley 2004), with specific parameter values being estimated during subsequent phylogenetic analysis. The ML analyses were conducted in PAUP* using exhaustive searches and nonparametric bootstrap analyses were performed using branch-and-bound searches of 1,000 pseudoreplicates. RESULTS Parsimony analyses of all nucleotides intl-3), 3rd codon positions (nt3) and inferred amino acids {aa) produced identical, fully resolved topologies with bootstrap percentages (BP) of 98-100 for all internal nodes (Fig. 1) (utl-3: 2,130 characters, 398 informative, length = 897, Cl = 0.8305; nt3\ 710 characters, 317 informative; length = 756, Cl = 0.816; aa-. 710 characters, 55 informative; length = 94, Cl = 0.9492). Caddo agilis and C. pepperella were each recovered as monophyletic and reconstructed as sister groups with respect to the acropsopilio- nines, Acropsopilio chilensis and AustropsopiUo sudamericanus. Comparison of alternative likelihood models in Modeltest indicat- ed that the ntI-3 data should be analyzed using a GTR-rr4-i-I model and that the nt3 matrix should be analyzed under the TVM+r4 model. Exhaustive likelihood searches using these models recovered topol- ogies identical to those derived from parsimony-based analyses (ntl-3-. -In likelihood = 6649.2923; }U3, -In likelihood = 3391.9731). Specifically, the clades C agilis, C. pepperella, and Caddo were each recovered as monophyletic with strong support (BP 99-100%) (Figs. 2, 3), a result predicted by Suzuki’s hypothesis. The two data sets were reanalyzed under their respective models with the analyses constrained to yield Shear’s hypothesis of independent evolution of C. pepperella from C. agilis in Asia and North America and Suzuki’s hypothesis of monophyly of both C. agilis and C. pepperella throughout their ranges. Kishino-Hasegawa tests (Kishino & Hasegawa 1989) showed the trees constrained to the prediction of Shear’s hypothesis to be significantly less likely than those con- strained to Suzuki’s hypothesis (P < 0.001). DISCUSSION Our results indicate that Caddo agilis and Caddo pepperella are monophyletic species that diverged phylogenetically before each acquired a disjunct geographic distribution in Japan and eastern North America. This supports Suzuki’s (1976) habitat-fragmentation hypothesis and is inconsistent with Shear’s (1975, 1996, 2004) parallel- evolution hypothesis. These findings are consistent with current understanding of climatic and biogeographic events during the late Tertiary (Sanmartin et al. 2001). The eastern Asia-eastern North America disjunction exemplified by Caddo parallels a long-known biogeographic pattern among flowering plants (Wen 1999; Xiang et al. 2000). During the mid-Cenozoic, eastern Asia and eastern North 1 1UU/1UU/1UU i4/ia/n -|P7/-|rin/Q I 100/100/100 19/18/0 219/152/46 100/100/98 1 91/91/n 94/84/4 I — 21/21/1 197/163/13 205/183/21 Caddo agilis NA Caddo agilis Japan Caddo pepperella NA Caddo pepperella Japan AustropsopiUo Acropsopilio 100 100 0.1 substitutions/site 1 99 { Caddo agilis NA Caddo agilis Japan Caddo pepperella NA Caddo pepperella Japan - AustropsopiUo — Acropsopilio 100 100 0.1 substitutions/site I 100 Caddo agilis NA Caddo agilis Japan Caddo pepperella NA Caddo pepperella Japan AustropsopiUo — Acropsopilio Figures 1-3. — Results of phylogenetic analysis. 1. Parsimony tree based on separate analysis of all nucleotides, third codon positions and inferred amino acids, respectively. Numbers above branches are non-parametric bootstrap percentages based on 1000 pseudorepli- cates. Numbers below branches are estimated branch lengths under acctran optimization. 2. Maximum-likelihood tree based on all nucleotides using the GTR -i- r4 + I model. 3. Maximum-likelihood tree based on third codon positions using the TVM + r4 model. America were spanned by mesophytic forests that were eventually separated into Asian and American components during the early Pliocene, a culmination of long-term trends in the cooling and drying of central and northern North America. As a consequence, many plant genera have representative species in both eastern Asia and eastern North America, and relative rates tests conducted on 12 species pairs using the rhcL gene indicated a divergence time of 5.4 ± 2.6 million years ago (Xiang et al. 2000). Zoologists have not explored the Asian-North American disjunction to the same extent as botanists, but several examples are known among animals (Sanmartin et al. 2001), including the non-caddine harvestmen Acropsopilio hoopis (Crosby 1904) and Crosbycus dasycnemus (Crosby 1911), Okeantohates millipedes (Enghoff 1993), plethodontid salamanders (Min et al. 2005) and, among late Tertiary fossils, lesser pandas and meline badgers (Tedford & Harington 2003; Wallace & Wang 2004). In contrast, because the two Caddo species are roughly sympatric and often syntopic in both North America and Japan, vicariant or climatic events cannot readily explain their phylogenetic divergence or morphological differences. It is possible that the two extant Caddo species diverged due to resource or habitat partitioning, as C. agilis tends to occupy exposed surfaces (e.g., tree trunks, logs, stones) and C. pepperella occurs on the ground in the leaf litter and under fallen objects (Suzuki 1976; Shultz, unpubl. obs.). Given the similarity between C. pepperella and juvenile C. agilis, it is possible that such habitat specialization produced morphological differences between the 240 THE JOURNAL OF ARACHNOLOGY two species via heterochrony, in a manner similar to that proposed by Shear ( 1975, 1996, 2004). Still, there is no clear evidence as to whether C agilis is peramorphic with respect to its ancestor, whether C pepperella is paedomorphic with respect to its ancestor, both, or neither. Outgroup comparison with the small soil- or litter-dwelling acropsopilionines (Caddidae) would seem to favor C. pepperella as the better model for the common ancestor of extant Caddo and thus evolution of C. agilis via hyperniorphosis. However, without relevant information about the morphology and development of ancestral and extant Caddo, this matter will remain an exercise in speculation. ACKNOWLEDGMENTS We thank Tomas Cekalovic and Nobuo Tsurusaki for specimens and two anonymous reviewers for comments. This research was supported by NSF grants 9981970 and 0640179. JWS was supported by the Maryland Agricultural Experiment Station. LITERATURE CITED Enghoff, H. 1993. Phylogenetic biogeography of a Holarctic group: the julidan millipedes. Cladistic subordinateness as an indicator of dispersal. Journal of Biogeography 20:525-536. Felsenstein, J. 1985. Confidence limits on phytogenies: an approach using the bootstrap. Evolution 39:783-791. Kishino, H. & M. Hasegawa. 1989. Evaluation of the maximum likelihood estimate of the evolutionary tree topologies from DNA sequence data, and the branching order in Hominoidea. Journal of Molecular Evolution 29:170-179. Maddison, W.P. & D.R. Maddison. 1992. MacClade: Analysis of Phytogeny and Character Evolution. Version 3. Sinauer Associ- ates, Sunderland, Massachusetts. 398 pp. Min, M.S., S.Y. Yang, R.M. Bonett, D.R. Vieites, R.A. Brandon & D.B. Wake. 2005. Discovery of the first Asian plethodontid salamander. Nature 435:87-90. Posada, D. & T.R. Buckley. 2004. Model selection and model averaging in phylogenetics: advantages of Akaike Information Criterion and Bayesian approaches over likelihood ratio tests. Systematic Biology 53:793-808. Posada, D. & K.A. Crandall. 1998. Modeltest: testing the model of DNA substitution. Bioinformatics 14:917-818. Regier, J.C. & J.W. Shultz. 1997. Molecular phylogeny of the major arthropod groups indicates polyphyly of crustaceans and a new hypothesis for the origin of hexapods. 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Computer Applications in the Biosci- I ences 10:671-675. Staden, R., K.F. Beal & J.K. Bonfield. 1999. The Staden Package, 1998. Pp. 115-130. In Bioinformatics Methods and Protocols. (S. Misener & S. Krawetz, eds.). The Humana Press, Totowa, New | Jersey. j Suzuki, S. 1976. The harvestmen of the family Caddidae in Japan ! (Opiliones, Palpatores, Caddoidea). Journal of Science, Hiroshima | University, Series B, Division 1 26:261-273. j Swofford, D.L. 1998. PAUP* 4.0 Beta Version. Sinauer Associates: Sunderland, Massachusetts. ji Tedford, R.H. & C.R. Harington. 2003. An arctic mammal fauna ’i from the early Pliocene of North America. Nature 425:388-390. ( Wallace, S.C. & X. Wang. 2004. Two new carnivores from an unusual i late Tertiary forest biota in eastern North America. Nature i; 431:556-559. I Wen, J. 1999. Evolution of eastern Asian and eastern North American disjunct distributions in flowering plants. Annual j Review of Ecology and Systematics 30:421-455. i Xiang, Q-Y., D.E. Soltis, P.S. Soltis, S.R. Manchester & D.J. ;i Crawford. 2000. Timing of the eastern Asian-eastern North American floristic disjunction: molecular clock corroborates paleontological estimates. Molecular Phylogenetics and Evolution 15:462^72. f Manuscript received 1 August 2008, revised 5 December 2008. 2009. The Journal of Arachnology 37:241-242 SHORT COMMUNICATION New spider host associations for three acrocerid fly species (Diptera, Acroceridae) Maxim Larrivee and Christopher J. Borkent: Department of Natural Resource Sciences, McGill University, Macdonald Campus, 21111 Lakeshore Road, Ste. Anne de Bellevue, Quebec, H9X 3V9. Canada. E-mail: maxim. larrivee@ mail.mcgill.ca Abstract. Acrocerid flies are endoparasitoids of spiders. New host associations are reported for Ogcodes mekimpiis Loew 1872, O. eugonatus Loew 1872, and Acrocera sp. (Group IV; semu Sabrosky 1944) from reared individuals of two Salticidae species, Pelegrina proterva (Walckenaer 1837) (both Ogcodes species), and Eris militaris (Hentz 1845) (the Acrocera sp.) (Group IV; sensu Sabrosky 1944). The spiders were sampled in the canopy and understorey of a mature north-temperate hardwood forest at the Morgan Arboretum, Quebec, Canada. Keywords: Endoparasitoids, Salticidae, canopy, maple, beech Acrocerid flies (Diptera, Brachycera) are endoparasitoids of spiders. Each larval instar is morphologically unique and has a distinctive lifestyle (hypermetamorphosis: Schlinger 1987). Their planidial first instar larvae actively seek their spider host or, only in the genus Acrocera, attach themselves to the substrate where they have hatched, waiting for a host spider to pass by (Schlinger 1987, 2003; Nielsen et al. 1999). Once a host is found, the planidium climbs on to the spider, migrates to the spider’s abdomen, and cuts a small hole to enter the spider en route to the booklungs (Schlinger 1987; see Nielsen et al. 1999, for an alternative strategy to enter the host). In the booklungs, the larva molts again, attaches itself to a booklung, and enters a resting stage. After molting, the fourth instar larva feeds actively inside the spider and causes the parasitized spider to spin a molting-web like retreat. The acrocerid larva then emerges from the spider, finishes feeding, fixes itself to the web and pupates (Schlinger 1987). Acrocerid flies show a preference for wandering, fossorial, and web-building spiders that live close to the ground and wander in adjacent vegetation (Cady et al. 1993). We report new spider (Araneae) host associations for Ogcodes melampus Loew 1872, O. eugonatus Loew 1872, and Acrocera sp. (Group IV; sensu Sabrosky 1944) (Diptera: Acroceridae). Foliage spiders were sampled by beating live and dead branches between 10 May and 24 September 2007 in the canopy of mature trees and understorey saplings of sugar maple (Acer saccharum Marsh.) and American beech (Fagus grandifolia Eher.) at the Morgan Arboretum, Sainte-Anne-de-Bellevue, Quebec, Canada (45°25'55"N; 73°56'58"W). In the laboratory, the spiders were housed individually in small plastic containers and kept alive in preparation for a ballooning dispersal experiment. During this experiment, cream yellow pupae were noticed inside containers of three, dead, sub-adult individuals of Pelegrina proterva (Walckenaer 1837) (Araneae: Salticidae, body size = 3.9 mm, n = 4). Two of these individuals of P. proterva were sampled in the canopy of mature American beech trees and one in the canopy of a mature sugar maple on 7 June 2007. Adult flies emerged in the plastic containers approximately 28 days later, in early July 2007. The adults were determined to be two females of O. eugonatus (one from American beech and the other from sugar maple) and one female of O. melampus (from American beech). In similar fashion, a female Acrocera sp. (Group IV, near female 1 sensu Sabrosky 1948) emerged from a sub adult individual of Eris militaris (Hentz 1845) (Araneae: Salticidae, body size = 5.2 mm, n = 6). This individual of E. militaris was sampled on an American beech sapling in the understorey on 3 July 2007, the acrocerid larva had pupated a week later, and the adult fly emerged approximately 2 wk later. Overall, 0.88 percent of the spiders in our study were parasitized by Acroceridae. Acrocera is known to lay its eggs on grass stems (Schlinger 1987), potentially not far removed from American beech saplings. Eris militaris, the host spider, is also significantly associated with the understorey layer in this habitat type (Larrivee & Buddie 2008). In contrast, the three infected individuals of P. proterva originated from the canopy. Females from the genus Ogcodes lay their eggs on the tips of dead twigs (Schlinger 1987), common in the canopy of beech trees. Only four acrocerid parasites were found in our study but the rarity of these flies makes this an important life history observation. Ogcodes specimens were only in spiders from the canopy and the Acrocera specimen in an understory spider. Future research on hardwood forest Ogcodes and Acrocera species should test their potential preference for canopy and understorey spiders respectively. Ogcodes melampus is mainly found in the western part of North America with previous records placing it as far east as Minnesota (Schlinger 1960). This specimen represents a significant range extension for this species. Other northeastern specimens of O. melampus were found in the Canadian National Collection in Ottawa, from both Michigan and Ontario. There is no life history available for this species (Schlinger 1960) and it has been reared from only two spider species, a lycosid and a thomisid (Schlinger 1987). Our record adds the family Salticidae and the species P. proterva to its host list. Ogcodes eugonatus has been reared from Lycosidae, Oxyopidae, Thomisidae, and Salticidae (species are listed in Schlinger 1987) though this is the first record of this species from a P. proterva host. The genus Acrocera occurs across North America though records from Acrocera Group IV (sensu Sabrosky 1944) mostly originate from eastern North America. They are known endoparasitoids of seven spider families: Plectreuridae, Lycosidae, Agelenidae, Amaurobiidae, Clubionidae, Gnaphosidae, and Salticidae (Schlinger 1987). Acrocera bulla Westwood, a member of Group IV, is the only other known species from the genus Acrocera that is an endoparasitoid of the family Salticidae. Our observation adds E. militaris to the host list of spiders for the genus Acrocera. Specimens are deposited at the Lyman Entomological Museum, McGill University, Macdonald Campus, Ste-Anne-de-Bellevue, Que- bec, Canada. ACKNOWLEDGMENTS We thank Chris Buddie for his support of this project including the use of the DINO 260xt mobile aerial platform to access the tree canopies (Canadian Foundation for Innovation New Opportunities Grant (Project #9548). Cristina Idziak allowed us to sample in the 241 242 THE JOURNAL OF ARACHNOLOGY Morgan Arboretum. Jeff Gumming from the Diptera section at the Canadian National Collection kindly provided determined specimens of Ogcodes and Acroceni for comparison. Finally, we thank Robb Bennett and two anonymous reviewers for comments on an early draft of the manuscript. LITERATURE CITED Cady, A., R. Leech, L. Sorkin, G. Stratton & M. Caldwell. 1993. Acrocerid (Insecta: Diptera) life histories, behaviours, host spiders (Arachnida; Araneida), and distribution records. Canadian Ento- mologist 125:931-944. Larrivee, M. & C.M. Buddie. 2008. Diversity of canopy and understorey spiders in north-temperate hardwood forests. Agri- cultural and Forest Entomology. DOI; 10. 1 1 1 1/j. 1461-9563.2008. 0042 Lx Nielsen, B.O., P. Punch & S. Toft. 1999. Self-injection of a dipteran parasitoid into a spider. Naturwissenschaften 86:530-532. Sabrosky, C.W. 1944. A revision of the American spider parasites of the genera Ogcodes and Acrocera (Diptera, Acroceridae). Amer- ican Midland Naturalist 31:385-413. Sabrosky, C.W. 1948. A further contribution to the classification of the North American spider parasites of the family Acroceridae (Diptera). American Midland Naturalist 39:382-430. Schlinger, E.I. 1960. A revision of the genus Ogcodes Latreille with particular reference to species of the western hemisphere. Proceedings of the United States National Museum 111:227-336. Schlinger, E.I. 1987. The biology of Acroceridae (Diptera): true endoparasitoids of spiders. Pp. 319-327. In Ecophysiology of Spiders. (W. Nentwig, ed.). Springer-Verlag, Berlin. Schlinger, E.I. 2003. Acroceridae, spider-fly endoparasitoids. Pp. 734- 740. In The Natural History of Madagascar. (S.M. Goodman & J.P. Benstead, eds.). University of Chicago Press, Chicago. Manuscript received 18 July 2008, revised 5 November 2008. 2009. The Journal of Arachnology 37:243-245 SHORT COMMUNICATION Description of Toca^ a new neotropical spider genus (Araneae, Ctenidae, Calocteninae) Daniele Polotow: Departamento de Zoologia, Institute de Biociencias, Universidade de Sao Paulo, Sao Paulo, SP, Brazil. E-mail: danielepolotow@yahoo.com.br Antonio D. Brescovit: Laboratorio de Artropodes, Institute Butantan, Avenida Vital Brasil, 1500, CEP 05503-900, Sao Paulo, SP, Brazil. E-mail: adbresc@terra.com.br Abstract. Toca new genus is proposed to include two new species: the type species T. hossanova new species from Rio de Janeiro, Brazil, and T. samba new species from Parana and Minas Gerais, Brazil. Toca may be related to Calocleniis Keyserling and Gephyrocteiius Mello-Leitao, with which it shares the scales on the abdominal dorsum and the epigynum as a single, slightly sclerotized, fold. The genus can be distinguished among the Calocteninae genera by its unique genital structures. Keywords: Systematics, taxonomy, Brazil The subfamily Calocteninae was proposed by Simon (1897) based mainly on the shape of the labium, sternum, and carapace; and by the numerous and elongated spines on the first and second pairs of legs. Currently it contains four genera: Caloctenus Keyserling 1877 and Gephyroctenus Mello-Leitao 1936, both from South America; Diallomus Simon 1897 from Sri Lanka; and Apohmia Simon 1898 from the Seychelles Islands (Silva 2003; Platnick 2008). The subfamily is characterized by the following synapomorphies: presence of a set of elongated spines on tibia and metatarsus of the first and second pairs of legs, six thickened and elongated setae on the anal tubercle, and a reduced number of cylindrical gland spigots on the posterior median spinnerets (Silva 2003). In addition to the four Calocteninae genera already described, we propose the new genus Toca to include two new species: T. hossanova from Rio de Janeiro, Brazil, and T. samba from Parana and Minas Gerais, Brazil. Toca may be related to Caloctenus and Gephyroctenus, with which it shares scales on the abdominal dorsum and epigynum as a single, slightly sclerotized fold (Silva 2003, 2004; Polotow & Brescovit 2008). Toca can be distinguished by the unique genital structures within the subfamily, which support the proposal of a new genus. The material examined belongs to Institute Butantan, Sao Paulo (IBSP, A. D. Brescovit) and Museum National d’Histoire Naturelle, Paris (MNHN, C. Rollard). All measurements are in millimeters. Terminology follows Silva (2003). TAXONOMY Ctenidae Keyserling 1877 Calocteninae Simon 1897 Toca new genus Figs. 1-9 Type species. — Toca hossanova new species Etymology. — The generic name is an arbitrary combination of letters. The gender is feminine. Diagnosis. — Toca resembles Caloctenus and Gephyroctenus by the presence of scales on the abdominal dorsum (Silva 2003; Silva 2004). Males can be distinguished by an embolus with a rounded base (Figs. 4, 8) and large conductor (Figs. 1, 7) with a surrounding groove to accomodate the embolus (Figs. 3, 8). The female of T. hossanova resembles the female of Diallomus fuliginosus (type ' Current address: Laboratorio de Artropodes, Instituto Butantan, Avenida Vital Brasil, 1500, CEP 05503-900, Sao Paulo, SP, Brazil. specimen, deposited in MNHN, examined) with the epigynum containing a slightly sclerotized single fold and an anterior hood (Fig. 5). The female can be distinguished from the remaining genera by the elongated copulatory ducts and anterior glandular projection (Fig. 6) of the epigynum. The female of T. samba is unknown. Description. — Ecribellate ctenids. Total body length (males and females) 3.40-4.40. Carapace pale brown with longitudinal lighter stripe from eyes to posterior margin of carapace; chelicerae, labium, endites, sternum, and legs pale brown; posterior median and lateral eyes on black tubercles; legs with dorsal, transverse dark spots. Carapace flattened. Eyes: ctenoid pattern, 2-4-2. Chelicerae: three prolateral teeth and five to six small retrolateral teeth. Labium short, wider than long. Eovea short, positioned in posterior third of carapace. Legs I and II with set of numerous elongated spines on femur, tibia, and metatarsus. Trochanter slightly notched. Abdomen flattened, subpentagonal. Six erect bristles distally positioned on anal tubercle. Palp: tibia short; RTA divided into ventral and dorsal branches (Figs. 1, 2, 7, 9); cymbium with retrolateral basal projection (Figs. 3, 7); subtegulum prolateral; tegulum covered by conductor; median apophysis hook shaped (Figs. 1, 7); embolus surrounding tegulum, supported by conductor; conductor sclerotized ventrally, with retrolateral laminar projection supporting embolus tip (Figs. 2, 8). Epigynum: formed by single plate, slightly sclerotized, with anterior hood (Fig. 5); spermathecae rounded; fertilization ducts short, emerging from spermathecal base (Fig. 6). Composition. — Two species: Toca hossanova new species and T. samba new species Distribution. — Southern and southeastern Brazil. Toca hossanova new species Figs. 1-6 Type material. — Male holotype from Fazenda Ranchinho Porto da Roqa, Petropolis, 22°30'39"S, 43° 1 1'4"W, Rio de Janeiro, Brazil, 12- 14 November 1999, deposited in IBSP 62920; female paratype from the same locality, 8-15 February 2000, Equipe Biota, deposited in IBSP 62919; male paratype from the same locality, 15-16 August 2001, Equipe Biota, deposited in IBSP 90669. Etymology. — The species epithet is a Portuguese noun that refers to a popular rhythm of Brazilian music. Diagnosis. — Toca hossanova can be distinguished from T. samba by the elongated cymbium and conductor, the elongated retrolateral projection of the conductor, and the slender and thin median apophysis on the male palp (Figs. IM). The females can be recognized by the presence of an anterior epigynal hood (Fig. 5) 243 244 THE JOURNAL OF ARACHNOLOGY Figures 1-6. — Toca hossanova. male palp; 1. Ventral view; 2. Retrolateral view; 3. Dorsal view; 4. Prolateral view. 5-6 Epigynum: 5. Ventral view (arrow points to left copulatory opening); 6. Dorsal view. Abbreviations: c, conductor; db, dorsal branch of retrolateral tibial apophysis; e, embolus; g, glandular projection; h, hood; Ip, laminar projection of tegulum; ma, median apophysis; s, spermathecae; vb, ventral branch of retrolateral tibial apophysis. and elongated copulatory ducts, overlayed with small glands (Fig. 6) in the epigynum. Description. — Male (IBSP 62920).- Total length 3.40. Carapace 1.50 long and 1.40 wide. Clypeus 0.07 high. Eye diameter: AME 0.10, ALE 0.08, PME 0.10, PLE 0.10. Leg measurements: I: femur 1.60/ patella 0.50/ tibia 1.80/ metatarsus 1.70/ tarsus 0.60/ total 6.20; II: 1.70/ 0.45/ 1.85/ 1.90/ 0.60/ 6.50; III: 1.80/0.40/ 1.75/ 1.85/ 0.70/ 6.50; IV; 1.90/ 0.40/ 1.60/ 2.10/ 0.90/ 6.90. Leg formula: 42=31. Leg spination: tibiae I and II with eight ventral pairs of spines; metatarsi I with six ventral pairs of spines; metatarsi II with five ventral pairs of spines. Abdomen brown with posterior area white. Palp: ventral branch of RTA laminar (Fig. 1); prolateral area of tegulum visible in ventral view, with laminar process (Fig. 4). Female (IBSP 62919).- Total length 4.40. Carapace 1.60 long and 1 .60 wide. Clypeus 0.08 high. Eye diameter: AME 0. 10, ALE 0.08, PME 0.12, PLE 0.12. Leg measurements: I: femur 1.50/ patella 0.50/ tibia 1.80/ metatarsus 1.60/ tarsus 0.50/ total 4.90; 11:1 .80/0.60/ 1.80/ 1 .60/ 0.60/ 6.40; III: 1.80/ 0.50/ 1.60/ 1.80/ 0.70/ 6.40; IV: 1.80/ 0.50/ 1.50/ 1.80/ 0.70/ 6.30. Leg formula: 2=341 . Leg spination: tibiae I and 11 with eight ventral pairs of spines; metatarsi I and II with six ventral pairs of spines. Coloration of the abdomen as in male. Epigynum as in generic description. Additional material examined. — None. Distribution. — State of Rio de Janeiro, Brazil. Toca samba new species Figs 7-9 Type material. — Male holotype from Morro do Cabral, Lagoa, Tijucas do Sul, 25°55'37"S, 49°10'44"W, Parana, Brazil, November 2000, J. Ricetti, deposited in IBSP 39239; male paratype from Parque Estadual de Vila Velha, Ponta Grossa, 25°4'60"S, 50°8'60"W, Parana, Brazil, 8 December 1986, Profaupar/CIIF, deposited in IBSP 62915; male paratype from Mata Grande, Parque Estadual do Ibitipoca, Lima Duarte, 21°5riO"S, 43°47'60''W, Minas Gerais, Brazil, 27-29 October 1997, B. M. Souza, deposited in IBSP 23812. POLOTOW & BRESCOVIT— rOC/l, A NEW CTENIDAE GENUS 245 Figures 7-9. — Toca samba. Male palp: 7. Ventral view; 8. Prolateral view; 9. Retrolateral view. Etymology. — The species epithet is a Portuguese noun which refers to a popular rhythm of Brazilian music. Diagnosis. — Toca samba can be distinguished from T. bossanova by the rounded cymbium and conductor, median laminar projection on the conductor, and robust median apophysis on male palp (Figs 7-9). Description. — Male (IBSP 39239).- Total length 4.00. Carapace 1.60 long and 1.50 wide. Clypeus 0.08 high. Eye diameter; AME 0.10, ALE 0.08, PME 0.10, PLE 0.10. Leg measurements: I: femur 1.80/ patella 0.60/ tibia 2.00/ metatarsus 1.80/ tarsus 0.70/ total 6.90; II: 1.90/ 0.60/ 2.00/ 1.80/ 0.80/ 7.10; III: 2.10/ 0.50/ 1.90/ 1.90/ 0.90/ 7.30; IV: 2.20/ 0.50/ 2.00/ 2.20/ 1.00/ 7.90. Leg formula: 4321. Leg spination: tibiae I and II with eight ventral pairs of spines; metatarsi I and II with six ventral pairs of spines. Abdomen medially pale brown, with two anterior white spots, lateral area brown and posterior area white. Palp: ventral branch of RTA elongated (Fig. 9); subtegulum reduced, not visible in ventral view. Female: Unknown. Additional material examined. — None. Distribution. — States of Parana and Minas Gerais, Brazil. DISCUSSION To date, there are only two Calocteninae genera described from South America: Caloctenus and Gephyroctenus. Caloctenus contains four valid species and can be distinguished by leg spination, carapace shape, strongly sclerotized male palpal tibia at apex, and median apophysis with an apical beak. These characters were considered apomorphic by Silva (2004). Gephyroctenus contains eight species and can be distinguished by the following synapomorphies: a cymbial retrolateral groove, retrolateral origin of embolus, long and thin embolus, median apophysis with a subdistal hook, hyaline projection close to the embolus base in the male palp, fused median and lateral fields in a single epigynal plate, copulatory opening located dorsally in an atrium, and elongated copulatory ducts surrounding the spermathecae in the female epigynum (Polotow & Brescovit 2008). The two species described in this paper cannot be assigned to these previously described genera. In addition to the unique morphological characters on the male palp and female epigynum, as described above, they lack the apomorphic features that characterize Caloctenus and Gephyroctenus. Caloctenus and Gephyroctenus are closely related by the presence of four retrolateral teeth in the chelicerae, reduced anterior lateral eye lenses, and cylindrical glands with an enlarged base on the posterior median spinnerets (Silva 2003). Toca also has reduced anterior lateral eye lenses, but five to six retrolateral teeth. The presence of the cylindrical glands with an enlarged base on the posterior median spinnerets in Toca should be confirmed in the future with scanning electronic microscopy. The males of Toca share the long and filiform embolus on the male palp with Gephyroctenus. The females of Toca resemble the type species of Diallomus, D. fulliginosus, from Sri Lanka (female type specimen depo- sited in the MNHN, examined), both by the single, slightly sclerotized epigynal fold and the anterior hood on the female epigynum. Therefore, the relationship of Toca new genus to other Calocteninae genera awaits cladistic analysis with all the genera assigned to Calocteninae and representatives of the remaining subfamilies of Ctenidae. ACKNOWLEDGMENTS We are grateful to Cristina Rheims, Gustavo Ruiz, Ingi Agnarsson, and the anonymous reviewers for helpful suggestions on the manuscript. We wish to thank Cristine Rollard, curator of MNHN, for providing the type material for this study. This study was supported by CNPq and FAPESP (grant nos. 99/05446-8 and 06/55230-7). This study is part of the BIOTA/FAPESP - The Biodiversity Virtual Institute Program (www.biotasp.org.br). LITERATURE CITED Platnick, N.I. 2008. The World Spider Catalog, Version 8.5. American Museum of Natural History, New York. Online at http://research.amnh.org/entomology/spiders/catalog/index.html Polotow, D. & A.D. Brescovit. 2008. Revision of the Neotropical spider genus Gephyroctenus (Araneae; Ctenidae: Calocteninae). Revista Brasileira de Zoologia 25:705-715. Silva, D. 2003. Higher-level relationships of the spider family Ctenidae (Araneae: Ctenoidea). Bulletin of the American Museum of Natural History 274:1-86. Silva, D. 2004. Revision of the spider genus Caloctenus Keyserling, 1877 (Araneae, Ctenidae). Revista Peruana de Biologia 11:5-26. Simon, E. 1897. Histoire naturelle des araignees. Librairie encyclo- pedique de Roret, Paris 2:1-192. Manuscript received 14 March 2008, revised 24 November 2008. 2009. 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