Nesting Biologies and Immature Stages of the Tapinotaspidine Bee Genera Monoeca and Lanthanomelissa and of Their Osirine Cleptoparasites Protosiris and Parepeolus (Hymenoptera: Apidae: Apinae)

Copyright q American Museum of Natural History 2006 ISSN 0003-0082 PUBLISHED BY THE AMERICAN MUSEUM OF NATURAL HISTORY CENTRAL PARK WEST AT 79TH STREET, NEW YORK, NY 10024 Number 3501, 60 pp., 111 1 21 figures, 3 tables January 12, 2006 Nesting Biologies and Immature Stages of the Tapinotaspidine Bee Genera Monoeca and Lanthanomelissa and of Their Osirine Cleptoparasites Protosiris and Parepeolus (Hymenoptera: Apidae: Apinae) JEROME G. ROZEN, JR., 1 GABRIEL A. R. MELO, 2 ANTO ˆNIO JOSE ´CAMILLO AGUIAR 3 AND ISABEL ALVES-DOS-SANTOS 4 Appendix: Taxonomic Notes on Monoeca and Description of a New Species of Protosiris, by Gabriel A. R. Melo CONTENTS Abstract ...................................................................... 2 Introduction ................................................................... 3 Overview of Nesting Sites .................................................... 4 Methods and Terminology ...................................................... 5 Biology of Monoeca haemorrhoidalis (Smith) ..................................... 6 Biology of Protosiris gigas Melo ................................................ 15 Biology of Lanthanomelissa betinae Urban ....................................... 20 Biology of Parepeolus minutus Roig-Alsina ....................................... 24 Ovarian Statistics .............................................................. 24 Immature Stages ............................................................... 25 1 Division of Invertebrate Zoology, American Museum of Natural History ([email protected]). 2 Departamento de Zoologia, Universidade Federal do Parana´, Curitiba, Brazil ([email protected]). 3 Departamento de Zoologia, Universidade Federal do Parana´, Curitiba, Brazil ([email protected]). 4 Universidade do Extremo Sul Catarinense, Criciu´ma, Brazil ([email protected]). 2 NO. 3501AMERICAN MUSEUM NOVITATES Egg of Monoeca haemorrhoidalis (Smith) ...................................... 25 First Instar of Monoeca haemorrhoidalis (Smith) ................................ 26 Other Larval Instars of Monoeca haemorrhoidalis (Smith) ........................ 28 Postdefecating Larva of Monoeca haemorrhoidalis (Smith) ....................... 30 Pupa of Monoeca haemorrhoidalis (Smith) ..................................... 32 Egg of Lanthanomelissa betinae Urban ......................................... 33 Postdefecating Larva of Lanthanomelissa betinae Urban ......................... 34 Egg of Protosiris gigas Melo ................................................. 37 First Instar of Protosiris gigas Melo ........................................... 38 Other Larval Instars of Protosiris gigas Melo ................................... 40 Postdefecating Larva of Protosiris gigas Melo .................................. 42 Pupa of Protosiris gigas Melo ................................................ 45 Postdefecating Larva of Parepeolus minutus Roig-Alsina ......................... 46 Discussion .................................................................... 48 Acknowledgments ............................................................. 49 References .................................................................... 52 Appendix: Taxonomic Notes on Monoeca and Description of a New Species of Protosiris (Hymenoptera: Apidae), by Gabriel A.R. Melo ........................ 54 Introduction ................................................................. 54 Monoeca haemorrhoidalis (Smith) ............................................. 54 Monoeca schrottkyi (Friese) ................................................... 56 Key to Large Species of Monoeca from Southeastern Brazil ...................... 56 Protosiris gigas Melo, new species ............................................ 57 Acknowledgments to Appendix ............................................... 60 References to Appendix ...................................................... 60 ABSTRACT The nesting biologies of Monoeca haemorrhoidalis (Smith) and Lanthanomelissa betinae Urban (Tapinotaspidini) are described from southeastern Brazil. Both are ground nesting; the nests of the former are attacked by the cleptoparasite Protosiris gigas Melo (Osirini), and those of the latter are attacked by Parepeolus minutus Roig-Alsina (Osirini). Egg eclosion, larval feeding behavior, and cocoon spinning of M. haemorrhoidalis are detailed. A female of P. gigas opens the closed cell of M. haemorrhoidalis by making a large opening in the cell cap (which is plugged after ovipositioning) through which she apparently extends her metasoma. Indirect evidence suggests that she uses her metasomal apex, and perhaps even the sting, to kill the host egg or early instar. Protosiris eggs are either attached to the cell-wall surface of the nearly vertical host cells or dropped onto the surface of the provisions. First instars of P. gigas, with strongly curved, sharply pointed mandibles, are also capable of killing host immatures or competing cleptoparasites. Cocoons of all four species are compared and contrasted. The egg, all larval instars, and pupa of Monoeca haemorrhoidalis are described, as are the egg and postdefecating larva of Lanthanomelissa betinae. The egg, all larval instars, and pupa of Protosiris gigas are described, as is the postdefecating larva of Parepeolus minutus. Both Monoeca haemorrhoidalis and Protosiris gigas have four ovarioles per ovary. The egg indices and other ovarian features of both species are identified and discussed. The possible phylogenetic relationship of the Tapinotaspidini with the Osirini is briefly explored on the basis of data from this study. Possible phylogenetic relationships of the Osirini with other cleptoparasitic apids are analyzed. In the appendix, the identity of the species of Monoeca, whose nesting biology is presented in the main paper, is discussed. The species is M. haemorrhoidalis (Smith, 1854), a species closely related to M. schrottkyi (Friese, 1902) and M. xanthopyga Harter-Marques, Cunha, and Moure, 2001. An identification key for distinguishing these three species is presented. Tetra- 2006 3ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES pedia piliventris Friese is placed as a junior synonym of M. haemorrhoidalis (new synonymy). A lectotype is designated for Pachycentris schrottkyi Friese. The species of Protosiris found attacking M. haemorrhoidalis is here described as new, P. gigas Melo sp. nov. It is structurally most similar to P. caligneus (Shanks), from which it differs by its abundant yellow marks, plumose pubescence on the lower paraocular area, protruding anterior mesoscutum, and sparser punctation on the metasomal terga. RESUMO A biologia da nidificac¸a˜o de Monoeca haemorrhoidalis (Smith) e Lanthanomelissa betinae Urban (Tapinotaspidini) e´ descrita a partir de estudos conduzidos no sul do Brasil. Ambas as espe´cies de abelhas nidificam no solo, sendo atacadas por duas espe´cies de abelhas cleptoparasitas da tribo Osirini; os ninhos da primeira sa˜o atacados pelo cleptoparasita Protosiris gigas Melo (Osirini) e os da segunda, por Parepeolus minutus Roig-Alsina (Osirini). Eclosa˜o dos ovos, comportamento de alimentac¸a˜o das larvas e de construc¸a˜o dos casulos de M. haemorrhoidalis sa˜o descritos em detalhe. As feˆmeas de P. gigas teˆm acesso a`s ce´lulas rece´maprovisionadas e fechadas de M. haemorrhoidalis abrindo um buraco no tampa˜o de fechamento (o buraco e´ fechado apo´s a oviposic¸a˜o), pelo qual elas aparentemente inserem o metassoma. Evideˆncias indiretas sugerem que as feˆmeas do cleptoparasita usam o a´pice do metassoma, e talvez o pro´prio ferra˜o, para matar o ovo ou a larva jovem do hospedeiro. Os ovos de Protosiris sa˜o colocados presos a` parede vertical das ce´lulas ou jogados sobre a massa de alimento. Larvas dos dois primeiros ı´nstares de P. gigas apresentam mandı´bulas fortemente curvas e pontiagudas e sa˜o tambe´m capazes de matar os imaturos do hospedeiro ou de potenciais competidores cleptoparasitas. Os casulos das quatro espe´cies sa˜o comparados. O ovo, todos os ı´nstares larvais e a pupa de Monoeca haemorrhoidalis edeProtosiris gigas sa˜o descritos, bem como o ovo e a larva po´s-defecante de Lanthanomelissa betinae e a larva po´s-defecante de Parepeolus minutus. Tanto Monoeca haemorrhoidalis quanto Protosiris gigas apresentam quatro ovarı´olos por ova´rio. I ´ndices para os ovos e outras caracterı´sticas dos ova´rios de ambas as espe´cies sa˜o apresentados e discutidos. A possı´vel proximidade filogene´tica entre Tapinotaspidini e Osirini e´ brevemente explorada com base nos dados do presente estudo. Possı´veis relac¸o˜es filogene´ticas de Osirini com outros grupos cleptoparasitas de Apidae sa˜o tambe´m analisadas. No Apeˆndice, e´ discutida a identidade da espe´cie de Monoeca, cuja biologia e´ apresentada no artigo principal. A espe´cie corresponde a M. haemorrhoidalis (Smith, 1854), uma espe´cie pro´xima a M. schrottkyi (Friese, 1902) e M. xanthopyga Harter-Marques, Cunha & Moure, 2001. Uma chave de identificac¸a˜o para estas treˆs espe´cies e´ apresentada. Tetrapedia piliventris Friese e´ colocada como sinoˆnimo ju´nior de M. haemorrhoidalis (sinoˆnimo novo). E ´designado um lecto´tipo para Pachycentris schrottkyi Friese. A espe´cie de Protosiris encontrada atacando M. haemorrhoidalis e´ aqui descrita como P. gigas Melo, n.sp. Estruturalmente, esta espe´cie assemelha-se mais a P. caligneus (Shanks), da qual difere pelas manchas amarelas abundantes, pela pilosidade plumosa na a´rea parocular inferior, pelo mesoscuto projetado anteriormente e pela pontuac¸a˜o mais esparsa nos tergos metassomais. INTRODUCTION We present here information on the nesting biology of two species belonging to separate genera of the Tapinotaspidini, Monoeca and Lanthanomelissa. 5 Both genera and, indeed, all members of the tribe are thought to use floral and other plant oils in nest construc5 Michener (2000) regarded Lanthanomelissa as a subgenus of Chalepogenus. For simplicity of presentation, we follow the usage adopted by Urban (1995). tion, nest provisioning, or both (Michener, 2000). The two species, Monoeca haemorrhoidalis (Smith) and Lanthanomelissa betinae Urban, nest in the ground. We found the nests of M. haemorrhoidalis attacked by an unnamed species of Protosiris, described and named P. gigas in the appendix. This is the first association of any species of Protosiris with a host. Nests of L. betinae were parasitized by Parepeolus minutus Roig-Alsina, mature larvae of which were recovered from 4 NO. 3501AMERICAN MUSEUM NOVITATES the nests. This is the first confirmed host/parasite association of a member of Parepeolus, although Rozen (1984b) found an adult of Parepeolus niger Roig-Alsina in a nest of Tapinotaspoides serraticornis (Friese) (cited as Tapinotaspis tucumana (Vachal)), and Roig-Alsina (1989) tentatively associated Ecclitodes stuardi (Ruiz) with Chalepogenus caeruleus (Friese). In addition to describing various aspects of the nesting biology of the two host species, we present biological information on their two cleptoparasites. The mode of cleptoparasitism of Protosiris gigas provides the first understanding of this matter for any member of the cleptoparasitic tribe Osirini. Because the immature stages of the Tapinotaspidini and Osirini are poorly known, we include an extensive treatment of eggs, larvae, and pupae of species whose nesting we investigated. Data resulting in this report were accumulated over period of more than 3 years. The nesting biologies of Monoeca haemorrhoidalis and Protosiris gigas were studied at Mananciais da Serra, Piraquara, Parana´, Brazil. Melo (G.A.R.M.), Rozen (J.G.R.), and Aguiar (A.A.) gathered information for several days starting November 20, 2002, and G.A.R.M. and A.A. subsequently visited the nesting site on January 24, 2003. Starting on December 3, 2003, all authors, including Alves-dos-Santos (I.A.S.), returned to the same nesting site, which was again active, and examined it on and off over a 9-day study period. G.A.R.M. and A.A. carried out further observation on the site January 17, 2004, December 5, 2004, and January 30, 2005. I.A.S. discovered one of the nesting sites of Lanthanomelissa betinae on the campus of Universidade do Extremo Sul Catarinense (UNESC), Criciu´ma, Santa Catarina, Brazil, in October 2002. All authors carried out excavations on it on November 15 and 16 of that year. Further studies were made there by I.A.S. in November 23–26, 2003. The other Lanthanomelissa site was on the campus of Universidade Federal do Parana´, Curitiba, Brazil. It was initially discovered by G.A.R.M. in late October 2001; J.G.R. excavated nests there in November 25, 2002. Thus, all authors contributed to the biological information presented herein and to collecting and preserving immature stages. J.G.R. described the immature stages herein, and G.A.R.M. identified the adults and prepared the taxonomic description of the new species presented in the appendix. O VERVIEW OF N ESTING S ITES Three nesting aggregations of tapinotaspidine bees were involved in this study at the following localities, all in Brazil. Monoeca site: The study site of Monoeca haemorrhoidalis is located in an area known as ‘‘Mananciais da Serra’’, in Piraquara, Parana´, which is administered by the state water company SANEPAR. This area has a high annual rainfall and is covered with dense, well-preserved Atlantic forest. The approximate coordinates and altitude of the study site are 25 8 28 9 40 0 S, 48 8 58 9 04 0 W and 1140 m. The nesting area itself is relatively flat, but its surroundings are very steep. Although the area is located on the western slopes of the Serra do Mar, the study site is within a long valley that opens to the east side of this mountain range. The soils there are derived from granitic rocks, which in some parts of the nesting area were still too consolidated to allow bees to burrow. Lanthanomelissa (Curitiba) site: This nesting site of Lanthanomelissa betinae is located on the campus of the Universidade Federal do Parana´ (Centro Polite´cnico). The approximate coordinates and altitude of the site are 25 8 26 9 55 0 S, 49 8 13 9 54 0 W and 940 m. The region is mostly urbanized, with only a small patch of forest and wetlands about 500 m away. Most of the area around the bank containing the nests is maintained as a grass lawn. The soil and vegetation of the area suggests that it was originally a patch of native grass. These grass fields were relatively common in the region in areas with shallow soils in which Araucaria forest would not develop. Lanthanomelissa (Criciu´ma) site: This nesting site of Lanthanomelissa betinae is located at the edge of the campus of the Universidade do Extremo Sul Catarinense (UNESC), Criciu´ma, Santa Catarina. The coordinates and altitude of the town of Criciu´- ma are 28 8 40 9 S, 49 8 22 9 W and 50 m. The site is bordered on one side by a small fragment 2006 5ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES of secondary forest and by open areas covered by weeds and cultivated crops. The region is within the southern limit of the lowland Atlantic forest. METHODS AND TERMINOLOGY Preserved larvae were first illustrated while in ethanol, resulting in pencil illustrations of the entire specimen and enlarged frontal and lateral views of the head. Heads were then separated from the bodies, and both were cleared in a heated aqueous solution of sodium hydroxide until all tissue was removed. If large, specimens were examined in ethanol, or if small (usually), they were placed in glycerin. Details of anatomy such as setae and sensilla were added to the illustrations, and mandibles and spiracles were dissected and illustrated. In estimating the approximate length of a strongly curved larva, four or five relatively straight sections of it were measured in lateral view at the level of the spiracles; these measurements were then summed, as illustrated in figure 52. The term mature larva refers to the fifth instar after it has finished feeding. We use the term postdefecating larva (often called the prepupa by others) for the mature larva after it voids its feces. The term predefecating larva refers to the mature larva before it defecates. These two stages, although of the same instar and therefore usually identical in such structures as mandibles and spiracles, are quite different in overall shape, integumental texture, and behavior. The predefecating form is more robust, its integument is thinner and far less wrinkled, often of a different hue, and it is active (i.e., its head and body can move and its mandibles open and close). The postdefecating larva is more slender, often with body tubercles more pronounced, its integument is thicker and more wrinkled, and, when in full diapause, the larva is completely inactive. For most univoltine species, the postdefecating larva is the stage in which the bee diapauses until the next nesting season. Because postdefecating larvae are the forms most frequently collected, descriptions of bee larvae are usually based on that stage. Descriptions of the mature larvae presented here are based on postdefecating forms. However, in cases where we also have been able to study mature predefecating larvae, we described significant differences with respect to body tubercles and the shape of the terminal abdominal segments, following the description of the postdefecating form. Other differences, such as body color of predefecating forms, are not mentioned, and similarly the more extensive expression of internal head ridges of predefecating larvae are not described, except in the case of Lanthanomelissa betinae, which will serve as an example. It seems likely that the less prominent internal ridges in the head of postdefecating forms result from the thickening of the cuticle elsewhere as the larva enters diapause, so that there is less contrast between ridges and nonridged areas. In the frontal-view diagrams of larval heads, sensilla and internal ridges are indicated on only their left side. Larvae and eggs were examined with a Hitachi S-5700 scanning electron microscope (SEM) in the Microscopy and Imaging Facility of the American Museum of Natural History after they were critical-point dried and coated with gold/palladium. Mature larvae so examined unexpectedly revealed elongate, setiform papillae associated with the salivary openings (figs. 60, 68, 96, 99). These structures are termed salivary papillae rather than spicules because they appear to be nonsclerotized. Their patterning differed from one species to another but suggests that they may have some function associated with the application of the silk in cocoon construction, such as serving like the end of a paint brush in distributing liquid over a broad surface. The papillae were scarcely visible when viewed with a light microscope and thus may have been overlooked in previous studies of other taxa. The extent to which other cocoon-spinning mature larvae have them should be investigated. The egg index, referred to in the descriptions of eggs and in the section on Ovarian Statistics, is a method of defining the size of a bee’s egg relative to the size of the female’s body, as developed by Iwata and Sakagami (1966). It is calculated by dividing the length of the egg or mature oocyte by the distance between the outer rims of the tegulae. We 6 NO. 3501AMERICAN MUSEUM NOVITATES gathered these data two ways. For the section on Ovarian Statistics, we measured the length of the longest mature oocyte in a female and divided that figure by her intertegular distance. For the descriptions of eggs of the taxa, we averaged the lengths of eggs recovered from nests and then divided the resulting figure by the average intertegular distance of a sampling of females. Iwata and Sakagami (1966) proposed a five-category classification of bee eggs (mature oocytes) based on their indices; that is, length of the egg (oocyte) (E) divided by the distance between the outer rims of the tegulae, or metasomal width (M): dwarf (E/M # 0.50); small (0.50 , E/M # 0.75); medium (0.75 , E/M # 1.00); large (1.00 , E/M # 1.10); giant (0.10 , E/M). The abbreviations used for adult structures are: T for metasomal tergum and S for metasomal sternum, followed by an Arabic number denoting the segment (e.g., T6 refers to sixth metasomal tergum). BIOLOGY OF MONOECA HAEMORRHOIDALIS (SMITH) Several nesting sites of Monoeca haemorrhoidalis were discovered in 2002, 2003 and 2004 along a seldom-traversed, unpaved road extending more than 3 km through the heavily forested mountainous area. The largest site (fig. 1) was excavated during these 3 years to provide the information presented here. It occupied about a 12-m-long, horizontal section of the roadway. Being about 3 m wide at either end, its midsection widened to about 6 m and contained the most nests. The site was shielded marginally by the forest canopy so that much of it received considerable sunlight on clear days. Ground cover, mostly grasses, obscured much of the surface, so that many nest entrances were partly or completely hidden (fig. 2). In 2002, several smaller nesting sites along the forested roadway were discovered, but none of these showed activity of Protosiris gigas. However, in 2003 we discovered this cleptoparasite at a less populous site about 1 km from the study site. In early December 2003 the main site was extremely active midmorning to late afternoon, with flying Monoeca haemorrhoidalis and Protosiris gigas producing a constant hum. Both species tended to fly no more than 0.5 m above the ground, although occasionally individuals briefly landed a meter or more above the surface on surrounding vegetation. Because of the dense, low groundcover, nest density could not be measured. Rough estimates of the number of individuals of both species combined ranged from approximately 50 per m 2 on December 4, 2003, during a very active period, to 25 per m 2 the following day, a reduction presumably resulting from different weather conditions. At these times, most of the Protosiris were males, whereas the Monoeca population was of both sexes, though males seemed more abundant. Although hosts and cleptoparasites were generally scattered over the entire nesting area, we frequently saw small flying aggregations of mostly male Protosiris, suggesting that they may have been pursuing a receptive female. In December 2003, females of Monoeca haemorrhoidalis were seen near the nest aggregation, visiting flowers and flower buds of Tetrapterys guilleminiana A. Juss. (Malpighiaceae), a tree-climbing woody vine that presents yellow flowers arranged in large inflorescences. Most plants were in the tops of trees, but some branches had fallen onto the forest edge. One female collected on these Tetrapterys flowers carried only oil in the scopae of the hindlegs. On one plant of T. guilleminiana with nonglandular flowers, we saw one Monoeca female collecting pollen and another female with barren scopae approaching three flowers but leaving without landing. The scopal loads of five females carrying pollen consisted of Tetrapterys pollen. Also, all samples of pollen from seven broad cells contained Tetrapterys pollen, with most of them composed solely of Tetrapterys pollen. Only two of the cells analyzed revealed a half-and-half mixture of Tetrapterys pollen and unidentified pollen grains (triangular pollen grains with three furrows, each one crossed by a transverse furrow). Adult activity at the site was greatest on 2006 7ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES December 4, 2003, and declined gradually afterward although both parasites and hosts were still moderately abundant when we last visited the site on December 11, 2003, at the end of the joint study period. When G.A.R.M. and A.A. revisited the site on January 17, 2004, they discovered many flying males and nesting females of Monoeca haemorrhoidalis, which seemed about as abundant as in early December, but adults of Protosiris gigas were completely absent. The start of the nesting activities of the next generation in 2004 seemed to have been delayed at the site. The adult activity on December 5, 2004, was only about one-fifth to one-sixth of that observed in December 2003, despite the favorable weather conditions. Also, no fresh cocoons or full-grown larvae were found. Copulating Monoeca haemorrhoidalis were occasionally observed on the low vegetation covering the nesting site, with one or more other males often attempting to intercede with the copulating pairs. Females of Monoeca haemorrhoidalis presumably spent the night in nest burrows. Sleeping places of males were not found. Only a single female occupied each nest. Nest entrances of Monoeca haemorrhoidalis were surrounded by abundant moist tumuli of excavated soil, and they lacked distinct turrets, although the worked wall of the main burrow extend through the tumulus at least on some occasions. Entrances were randomly scattered, with some being 5 cm apart and others more distant. Many could be seen at one time over a broad area, but most were partly to nearly completely hidden by ground cover (fig. 2). Main burrows descended vertically with limited turning through the moist, nearly rock-free substrate and were open their entire length. Because of on-going rainy spells, the substrate was always moist, but heavy downpours did not flood our large study excavations, attesting to the drainage capabilities of the nesting substrate. Burrow diameters ranged from 7.0 to 9.0 mm, and burrow walls were smooth and moderately shiny (fig. 3). At least in one area, the burrow walls were lined with extremely fine, reflective, claylike material about 1 mm thick that was red and contrasted with the browner surrounding substrate. Their color matched that of the lower substrate and no doubt resulted from the lower substrate material being brought to the surface during nest building. However, whether the tunnel linings were an accidental byproduct of nest excavation or a special construction by the female M. haemorrhoidalis is unknown. The surface of the burrow lining showed distinct, obviously repetitive, tamping impressions only in some areas (fig. 3). These impressions are thought to have been from the female using her pygidial plate, an activity known in other bees. A water droplet applied to the wall remained beaded on the surface for at least a minute. The surface was not coated with the same waterproof material used on the cell walls, so that its shiny, hydrophobic surface may have resulted from the mechanical compression of soil particles through the tamping process. Water-retardant burrow walls are an uncommon feature among solitary, groundnesting bees. Perhaps water leaking through the walls of deep burrows with large diameters is hazardous in wet environments. Overall nest configuration of this species could not be determined, although nests were obviously deep and almost certainly consisted of numerous cells. Too many main tunnels from the current and previous generations penetrated the soil to permit any one to be followed to reveal the branching pattern, and too many cells, both current and old, were encountered to assign them to a single main burrow. In one area fresh cells were first encountered at a depth of about 70 cm, and others may have extended below the 1 m level. In other areas a few cells were found as shallowly as 30 cm. All cells tended to be vertical but were tipped slightly, although we noticed one inclined as much as 45 8 . Cells were bilaterally symmetrical around their long axes. One side was more outcurved, and the opposite side was more or less incurved above and straighter below, so that the long axis of the cell tended to be curved (figs. 7, 9). This feature is uncommon in bees, but interestingly it is characteristic of the distantly related Diphaglossinae (Rozen, 1984a), where it is often more accentuated. (Andrenidae and Halictidae commonly have one side more outcurved and the opposite side nearly flat, but the opposite side 8 NO. 3501AMERICAN MUSEUM NOVITATES Fig. 1. Nesting site of Monoeca haemorrhoidalis, showing dense ground cover in foreground. Fig. 2. Close-up of two nest entrances of same. Fig. 3. Main burrow of Monoeca haemorrhoidalis show- 2006 9ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES Figs. 7–10. Cells of Monoeca haemorrhoidalis, diagrammatically represented. 7. Fully provisioned with Monoeca egg on provisions, and egg of Protosiris gigas attached to cell wall. 8. In early stages of being provisioned. 9. Depicting partially eaten provisions. 10. Containing cocoon of Monoeca haemorrhoidalis, with empty space between top of provisions and cell closure. Scale line 5 5.0 mm. ← ing repetitive tamping impressions on shiny burrow wall. Fig. 4. Monoeca haemorrhoidalis, spiral inner surface of cell closure. Figs. 5, 6. Closure ends of cells of Monoeca haemorrhoidalis, showing plugged entrance holes made by females of Protosiris gigas; holes are filled by them as they depart. is not incurved as in the Diphaglossinae and Monoeca haemorrhoidalis.) The cell lining was reflective, waterproof, and faintly milky. It not only coated the cell wall but also extended partly into the connecting lateral. When examined with a stereomicroscope, the material appeared brittle, fracturing rather than bending under stress. Its surface was patterned with faint curved parallel lines, one patch adjoining another, as if swept by a series of brushstrokes. A sweeping by the female’s much enlarged pygidial plate (see fig. A7) will likely be found to account for these marks as she applies the coating to the cell surface. When cells were allowed to dry over time, their lining tended to fracture and pull away from the soil, so that pieces could be removed from the cell wall with forceps. When we placed a flake on a microscope slide that was then heated on a hotplate, the flake melted into a nearly clear liquid, which solidified when cooled to room temperature. The cooled material became less transparent, and, when scraped with forceps, the scrape mark was shiny. Thus, the material had the physical characteristics of wax. We would be surprised if this substance did not come from the large, pale dorsal wax gland on the anterior part of the female’s sixth metasomal tergum (T6) that is normally hidden under the previous tergum (see also appendix, figs. A7–A8). Santos et al. (2004) studied the histology of integumental glands in the metasoma of Monoeca xanthopyga Harter-Marques, Cunha, and Moure. The large gland present in the female’s T6 was found to be a class I gland that histochemically produced lipids. We assume that the gland in M. haemorrhoidalis is homologous to that present in M. xanthopyga. We examined additional species of Tapinotaspidini and found that the external evidence of the gland (a transverse depression at the base of the female’s T6) was 16 NO. 3501AMERICAN MUSEUM NOVITATES Fig. 24. Sleeping male Protosiris gigas with wings plated, holding onto edge of leaf with mandibles; forelegs not normally involved with grasping substrate. Figs. 25–27. Eggs of Protosiris gigas.25. On cell wall; note dead second instar of Monoeca haemorrhoidalis below. 26. On summit of provisions; note egg of Monoeca haemorrhoidalis on provisions. 27. Attached by posterior end to cell wall; note (arrow) droplet of liquid on dorsal anterior surface of host egg (for possible explanation, see text). Fig. 28. SEM micrograph of first instar of Protosiris gigas preserved as it was emerging from chorion; two arrows on side of chorion point to presumed tear in chorion. 2006 17ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES with her mandibles (Bennett, 1966, for Hoplostelis bilineolata (Spinola); Bennett, 1972, for Exaerete dentata (Linnaeus)); and (2) the cleptoparasite female makes a small opening in the side of the host cells using her mandibles, reverses her position, and kills the host egg with her metasomal apex (Garo´falo and Rozen, 2001, for Exaerete smaragdina (Gue´rin-Me´neville)). All other known adult cleptoparasitic Apidae that introduce their eggs into sealed host cells do not kill the host offspring, but rather they have larvae that are hospicidal at one stage or another (Rozen, 2003: table 1 and references therein). In the case of Protosiris gigas, indirect evidence supports the hypothesis that the female uses her metasomal apex, and perhaps the sting, to kill the Monoeca egg or early instar. A total of 5 eggs (and one shed chorion) of Protosiris gigas were found attached to cell walls approximately halfway between the cell closure and the top of the provisions (figs. 7, 25, 27), proof that the female must be able to extend her metasoma through the hole in the cell closure. In all of these instances, the host immatures (4 eggs, 1 second instar) were clearly dead, showing loss of turgor or even being misshapen, but none was removed from its normal position on the food mass, as might have been the case if it had been killed by the mandibles of the Protosiris female. Also, none was missing, as if it might have been eaten by the cleptoparasite. Because we found another 5 live Protosiris eggs at various locations on the provisions (e.g., on the summit, fig. 26; on the periphery), egg placement by Protosiris gigas may be variable, or the eggs may have originally been attached to the wall and the attachments failed, sending them onto the surface of the provisions. On two other occasions a seemingly live host egg with an apparent fine puncture exuding a droplet of yellowish fluid on its anterior dorsal surface was noticed in a cell with a live Protosiris egg (fig. 27). This may have resulted from a puncture created by the parasite female’s sting. We perhaps encountered these cells shortly after they had been attacked, so that the host eggs had not yet lost their turgor. We reject the hypothesis that the female Protosiris used her mandibles to kill the host immature, both because there is no evidence that the Monoeca immatures were dislodged or missing from their normal positions and because it is doubtful that the opening of the cell closure made by the female would have been sufficiently large to allow her to reach into the cell. Furthermore, it seems unlikely that the Protosiris female would then reposition herself to attach her egg to the cell wall. However, is it reasonable to think that the sting (and metasomal apex) of the Protosiris female can reach the host egg, that is, to the level of the top of the central mound of the provisions? In the laboratory, we measured the maximum metasomal diameter and the length of the female metasoma from the petiole to the apex of the sting on two dead females with the metasomas fully extended. In both cases, the maximum metasomal diameter was 3.0 mm, and the distance from petiole to sting apex was 15 mm. The size of the actual holes in the cell closures (given above) would certainly permit the metasomas to extend into the lumen of the cell. Three measurements of the distance between the cell closure and the top of the provisions were 12, 14, and 15 mm. We conclude partly on the basis of these distances that the female Protosiris probably inserts her entire metasoma through the hole in the cell closure so that the sting can damage the host egg or larva. At this time the cleptoparasite presumably places her egg on the cell wall (or drops it onto the provisions). This conclusion is also partly based on the fact that the evidence does not point to any other explanation. The sting apparatus of Protosiris gigas is elongate relative to body size, presumably an adaptation for reaching the host egg and depositing its own, with the sting shaft, gonostyli, first and second rami, and furcula being longer than those of most of other bees. However, these structures are not as long as those of the related Osiris (Packer, 2003). Certain matters, however, remain unclear. Might the posterior part of a female’s mesosoma also be inserted through the cell closure, enhancing her ability to reach the host immature? How does the female manage to penetrate the thick soil-filled closure, or does she only reach the closure prior to the lateral 18 NO. 3501AMERICAN MUSEUM NOVITATES being filled? How does she position her legs when she extends her metasoma? A host cell containing a dead Monoeca second instar and a live Protosiris egg (fig. 25) indicated that a parasite female can gain access to a cell that has been closed for some time after cell closure. Whether she is able to penetrate a soil-filled lateral is unknown. The presence of a live Protosiris first instar with a dead Protosiris egg in another cell indicated either that a cell can be attacked sequentially by two parasite females or (less likely) that one female parasite may places two eggs in a cell at a time. In two cells in which the Protosiris egg was attached to the wall, the attachment was clearly by its posterior end (one egg so attached projected into the cell lumen at nearly a right angle to the cell wall; fig. 27). In all other cases cleptoparasite eggs seemed attached by their full lengths to the walls. SEM examination of an egg (fig. 76) did not reveal any special modification of the sculpturing of either the posterior end or other surfaces that might function to hold the egg to the wall. In the distantly related Melectini, which attach their eggs to the cell closures or upper cell walls by their posterior ends, the surface sculpturing of this end is quite different from the rest of the chorion (Rozen and O ¨zbek, 2003: figs. 63, 65; 2005: fig. 11), being possible adaptations to enhance attachments to cell surfaces. The chorion left behind by a newly emerged Protosiris larva was carefully examined for remnants of a cast first-instar exoskeleton. Since none was detected, we concluded that the active larva next to the chorion was the first instar (the five larval instars of this species are differentiated and described below). All first instars found were active and agile. One was observed crawling over the surface of the provisions and returning several times to chew at the remnant of the host egg. This larva was observed clinging to the substrate with the venter of the last three abdominal segments and raising the anterior part of its body until its body looped backward so that its head was posterior to its abdominal apex. The narrow, transverse anterior folds on the venter of abdominal segments 3–7 (see fig. 78) may assist the larva in this maneuver by providing extra ventral integument to elongate the larva’s backward reach. With a microscope, we observed a larva crawling upside down on a glass surface. We saw that these folds also permitted the larva to crawl forward; as the abdominal apex pushed against the substrate, the larva’s preceding segments stretched, one after another, so that the anterior end of the larva moved forward. The larva evinced no special structure (pygopod) at its abdominal apex to accomplish this, although the ventral surface of the larva including that of the abdominal apex was spiculate. Although the female Protosiris presumably usually kills the host egg (or larva), the first-instar Protosiris seems perfectly capable of killing a host immature. We observed a first instar repeatedly attacking a dead host immature. Another attempted to bite the tip of a pin when teased, and it repeatedly opened and closed its mandibles as it searched the air for the pin. The anatomy of the head with its sharply pointed, strongly curved mandibles (figs. 77, 79–81) also indicates that the larva is capable of killing the host egg or larva. It is likely that the evolutionary driving force for these behavioral and anatomical adaptations is the need to be able to battle other cleptoparasites in the same cell rather than for host killing. In the case where a dead Protosiris egg and a live first-instar Protosiris were found together in a cell, we cannot be certain whether the egg was killed by the first instar or by the second Protosiris female to find the cell. Both scenarios are possible. We also observed a third-instar Protosiris responding to our probing forceps by turning its head in the direction of the attack and repeatedly opening and closing its sharply pointed mandibles (fig. 83) (see Other Larval Instars of Protosiris, below). Thus, this aggressive behavior appears to continue beyond the first stadium and indirectly implies that the successful ownership of the provisions may go undecided for the duration of several stadia. Interestingly, Garo´falo and Rozen (2001) reported that the female of the distantly related cleptoparasite Exaerete smaragdina also normally destroys the host egg and that her offspring (but the second and not first instar) also are capable of battling with other cleptoparasites or with a host immature that might have survived. 2006 19ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES Cocoons of Protosiris gigas (fig. 19) resemble closely those of the host externally. They are tan and composed of fine silk strands, and they have the same shape and adhered closely to the lower part of the cell. They exhibited the same planar, truncated top surface of the Monoeca cocoon. They have the following dimensions (N 5 5): length, 14.5–18 mm (x¯ 5 16.3 mm); maximum diameter, 8.0–9.0 mm (x¯ 5 8.6 mm); diameter of top, 6.0–8.0 mm (x¯ 5 6.9 mm). The cocoon wall (i.e., except for the top) in cross section appeared thicker (at least on fresh, moist specimens) than that of Monoeca haemorrhoidalis. This was probably due to the fact that the cocoon consisted of three layers: an outer silken layer, an inner silken layer, and in between a layer composed of numerous fine sheets of silk sandwiching the entire meconial mass. Consequently, the inner surface of the cocoon exhibited no feces, contrasting with the cocoon of the host. Another noticeable difference between the host and parasite was in the tops of their cocoons. The cocoon top of Protosiris consisted of numerous densely layered fibrous sheets of silk (fig. 20), so that when held in front of a light source, no light penetrated through the top. Thus, the top is thicker, denser, and stronger than that of Monoeca. The open, meshlike outer surface of the Monoeca cocoon that loosely attaches to the inner surface is replaced in Protosiris with a sturdier fabric, the top surface of which is not easily compressed. On drying, the cocoons of both species shrink; the strong, dense disc of the top of the Protosiris cocoon shrinks less, giving the upper part of the cocoon a slight hourglass shape. On eclosion, the Protosiris adult does not demolish the top (front) end of the cocoons, as is characteristic of Monoeca; instead, the emerging adult chews its way out of the cocoon by burrowing through the middle of the top, leaving the periphery in place (fig. 22). Although the ratio of Monoeca haemorrhoidalis to Protosiris gigas flying over the nesting site on December 4, 2003, appeared to be roughly 1:1, G.A.R.M. and A.A. inventoried the cells on January 24, 2003, when the site was quiescent. They recovered cells of 87 Monoeca,8Protosiris, and 28 containing Tetraonyx (Paratetraonyx)distincticollis Pic (Meloidae) (adults of which were tentatively identified by Dr. John Pinto). 6 When G.A.R.M. and A.A. sampled the nesting site on January 17, 2004, 96 cells contained Monoeca (36 eggs and active immatures 1 60 postdefecating larvae in cocoons), 8 Protosiris postdefecating larvae in cocoons, and 25 Tetraonyx distincticollis and yielded a host/cleptoparasite ratio of approximately 11:1. The discrepancy between this ratio and the estimated ratio of 1:1 based on flying adults on December 4, 2003, was caused by the earlier emergence of the cleptoparasite compared with that of the host, by a briefer period of emergence of the parasite compared with a longer emergence period of the host, by host females being away foraging while the parasites were concentrated at the nesting site, or by some combination of these phenomena. There is a selective advantage for a host to prolong its emergence so that it outlasts the seasonal activity of the cleptoparasite. Data obtained in January 30, 2005, revealed an even lower host/cleptoparasite ratio: 63 cells contained Monoeca (17 predefecating larvae 1 46 postdefecating larvae in cocoons, 6 of them dead), 2 Protosiris (1 dead larva and 1 dead male imago in their cocoons, both apparently from the previous season), and 2 T. distincticollis. The few cells containing young immatures had no indication of parasitism by Protosiris (only one cell had a young meloid larva). 6 John Pinto, in an e-message sent after he had received adults of the beetle for identification, wrote: ‘‘They are of the genus Tetraonyx. Using the Selander and Martinez (1984) key to the Argentinian species, they fall at T. distincticollis Pic. It currently is the only species in the subgenus Paratetraonyx Kaszab, a poorly differentiated subgenus of questionable validity. S & M apparently never saw this species and simply used the literature to incorporate it into their key. It is not well known at all—recorded from Argentina (no localities specified) and, originally, from the state of Santa Catarina, Brazil. Tetraonyx is an enormously complex genus; with about 101 named species, almost all of them from South America. Except for the faunal review of Selander and Martinez for Argentina, all we have is an 1879 publication of Haag-Rutenberg. I mention this because the taxonomy of the genus is in dire need of revision, and I don’t know the group particularly well. Thus, the species identification has to be considered tentative.’’ 20 NO. 3501AMERICAN MUSEUM NOVITATES Fig. 29. Nesting area (bank, middle background) of Lanthanomelissa betinae on campus of Universidade do Extremo Sul Catarinense, Criciu´ma. Fig. 30. A.A. looking at sloping nesting area of Lanthanomelissa betinae on the campus of Universidade Federal do Parana´, Curitiba. BIOLOGY OF LANTHANOMELISSA BETINAE URBAN The nesting biology of this species was original described by Sakagami and Laroca (1988) under the name of Lanthanomelissa goeldiana (Friese) from a nesting site in Castro, Parana´, Brazil. Urban (1995) assigned two females from that nesting site as paratypes of L. betinae when she described the species. Our observations are from notes we gathered primarily on the campus of the Universidade do Extremo Sul Catarinense, Criciu´ma, in 2002 and 2003, although information observed at the Curitiba nesting site was similar. At both localities, burrow entrances were discovered on sloping surfaces (figs. 29, 30) close to grassy areas where the floral oil/pollen plant Sisyrinchium micranthum Cav. (Iridaceae) grew. Although both sites were ecologically degraded because of campus landscaping and lawn maintenance, adult bees also occurred at other localities with relatively unaltered landscapes. The presence of this bee on these campuses was possible because of the persistence of Sisyrinchium under such situations. Early in the 2002 season in Curitiba, we observed males of Lanthanomelissa betinae patrolling flowers of Sisyrinchium and Cuphea gracilis Koehne (Lythraceae) and sleeping in aggregations when females were not yet present (thus indicating protandrous emergence). Fifty-eight sleeping male aggregations of L. betinae were sighted between October 13 and December 18, 2002, all of them along the bank where we found the nests (fig. 30). Most sleeping male aggregations were 10–80 cm above the ground, on the top leaves of four herbaceous plant species: Chromolaena hirsuta (Hooker and Arnott) R.M. King and H. Robinson (N 5 27), Eupatorium laevigatum Lamarck (N 5 3), Chromolaena pedunculosa (Hooker and Arnott) R.M. King and H. Robinson (N 5 18), and Solidago microglossa de Candolle (N 5 4) (Asteraceae). Few other males were found between the inflorescences of Cuphea gracilis (N 5 3) and Taraxacum officinale G.H. Weber ex Wiggers (Asteraceae) (N 5 2). The largest number of the males was observed on October 26, 2002, when 166 males were counted, distributed in 24 aggregations ( m5 6.91 6 13.2). The largest sleeping aggregations were observed in Chromolaena hirsuta (N 5 3; maximum numbers, 53, 39, 39) and Solidago microglossa (N 5 1; maximum number, 18). Male activity started to drop by early November and ceased after mid-December. Females presumably slept in their nests. Males were also previously found by I.A.S. in Criciu´ma sleeping on flowers of Asteraceae and in flowers of Petunia integrifolia (Hook.) Schinz and Thellung (Solanaceae). On Asteraceae, they were just resting 2006 21ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES Figs. 31, 32. Cells of Lanthanomelissa betinae, diagrammatically represented. 31. Early load of provisions. 32. Egg on completed provisions in closed cell. Scale bar 5 5.0 mm. on the flowers with their head partly inserted into the flower. In Rio Grande do Sul, she once observed five males sleeping end-toend around the ovary at the bottom of the corolla of P. integrifolia. Nests, each occupied by a single female, were compact (maximum lateral spread of cells at most ca. 10 cm) and shallow (maximum depth ca. 15 cm) with cells close to one another. Dry tumuli (ca. 3.5 mm in diameter) were on the downhill side of entrances. Open entrance tunnels descended obliquely with some turning. Burrow walls were nonreflective and had no special lining. Cells were elongate ovals, 7.0–8.0 mm in maximum length and 4.5–5.0 mm in maximum diameter (based on actual measurements and on cocoon dimensions); thus, their lengths were somewhat more than 1.5 times their maximum diameters, contrasting with the elongate cells of Monoeca haemorrhoidalis. All were arranged singly (i.e., not in linear series). They were broadly rounded at the rear end, and their maximum diameter was onethird the distance from the rear end, forward from which they narrowed gradually to the front entrance, which was ca. 3.0 mm in diameter. Cells seemed to be radially symmetrical around their straight long axis, with one side not being straighter than the opposite one, contrary to the cells of Monoeca as described above. However, the true symmetry of the cells of Lanthanomelissa may have been obscured by their small size. Cell closures on the inside were a deep concave spiral of 3–4 coils. The long axes of cells ranged from 35 to 80 8 from horizontal, with the front of the cell always higher than the rear. Cells possessed a conspicuous, waterproof, grayish lining beneath which the cell wall appeared dark, but whether the color resulted from mechanical compression of the substrate or from some applied substance (e.g., floral oils, glandular secretions) is unknown. When cell fragments dried, the lining tended to crack, but flakes could not be easily removed from the wall. When the lining was heated on a glass slide on a hotplate, it did not melt, as did the lining of the Monoeca cells. The source of the lining material is unknown. Laterals were of various lengths, with the shortest being ca. 1 cm long; all were filled with soil after cell closure. The first provisions brought into a cell were placed as an unshaped mass occupying the bottom of cell (fig. 31). The final provisions were shaped as an elongate, mealymoist loaf, in one case 4.2 mm long and 2.8 mm high (fig. 32). When newly formed, the loaf was, at least in a few cases, attached by its ventral or posteroventral surface to the cell wall, so that the rear surface was separated from the rear of the cell, and sometimes the anteroventral surface was also separated from the cell wall (fig. 32). The female deposited her egg on the top surface of the provisions near the front end (fig. 32). Young larvae crawled over the loaf while feeding, and intermediate-stage larvae circled the food mass, which had by this time slumped in the cell so that its posterior end became attached to the rear end of the cell. 22 NO. 3501AMERICAN MUSEUM NOVITATES Fig. 33. Cocoon of Lanthanomelissa betinae from which larva had been removed, lateral view. Fig. 34. Cocoon of Parepeolus minutus from which larva had bee removed, lateral view. Fig. 35. Inner surface of front end of cocoon of Lan- ← thanomelissa betinae showing complete covering by pale feces. Fig. 36. Inner surface of front end of cocoon of Parepeolus minutus showing central area that is not coated by feces. Provisions at this time developed a central peak as the feeding larva circled the periphery with its dorsum against the cell wall. Information on the larva’s final feeding position was not noted. Externally brown, the cocoon of Lanthanomelissa betinae was 7.0–7.4 mm long (x¯ 5 7.3 mm) and 4.5–5.0 mm in maximum diameter (x¯ 5 4.8 mm) (N 5 7). Each filled the entire cell and had a shape (fig. 33) identical to the inner topology of the cell (i.e., broadly round at the rear end), tapering forward from its maximum diameter to the narrowly rounded or faintly truncated front end. All but the front end of the cocoon was composed externally of fine, densely appressed silk strands. The anterior end consisted of coarser silk strands, openly and loosely arranged (thus, far less dense than elsewhere). The entire inner surface of the cocoon including the front end (fig. 35) was coated uniformly with a smooth layer of pale, almost white feces. Thus, in cross section, the cocoon consisted of two layers, a brown, silken outer layer (with the front end more open and less dense than elsewhere) and a somewhat thinner inner layer of feces. The cocoons of Lanthanomelissa betinae and Monoeca haemorrhoidalis were similar in that their walls conformed to the similarly shaped cells, and both were composed of dense, fine, brown silk on the outside. However, they differed in a number of distinctive ways irrespective of dimensions: (1) Cocoon symmetry reflected the shape of the cell; thus, the cocoons of L. betinae appeared roughly radially symmetrical around their long axes whereas those of M. haemorrhoidalis were bilaterally symmetrical. (2) Whereas the front end of the cocoon of M. haemorrhoidalis was sharply truncated, with the periphery usually being accentuated by a slightly elevated rim, the front end of the cocoon of L. betinae was narrowly rounded, although on some specimens a small, flattened anterior surface was discernable. (3) Al- 2006 23ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES though both taxa start spinning their cocoons before the onset of defecation, L. betinae spins its entire cocoon before applying feces to the inner surface. Monoeca haemorrhoidalis continues to spin layers of fiber while defecating, so that the feces are incorporated into the fabric of the cocoon as well as on the inner surface of the completed silken cocoon. (4) Feces of L. betinae completely cover the front interior surface of the cocoon as well as the rest of the cocoon, whereas the feces of M. haemorrhoidalis are incorporated throughout the cocoon except for the front surface. (5) The front end of the cocoon of M. haemorrhoidalis was far removed from the cell closure, whereas the front end of the cocoon of L. betinae approached the cell closure closely. This feature is the result of the more elongate cells of M. haemorrhoidalis. Although several nests of Lanthanomelissa betinae with only a few cells were excavated, one nest seemed to contain 21 active cells, another had 11 cells, and a third had 9 cells. However, the first two nests mentioned included many other cells from which the adults had emerged, leaving behind vacated cocoons. This strongly suggests that these main burrows may have been used by more than one generation, a possibility supported by the fact that a single bee pupa 7 was encountered in one nest. Either the pupa was a member of the generation from the previous year or some members of the current generation may be bivoltine. So many cells in the nest favor the first alternative. Large nests contained offspring that had already constructed cocoons, freshly deposited eggs, and even open cells still being provisioned. The large range of developmental stages might be explained (assuming that only a single generation comprises the nest contents) either by a rapid development of immatures or, conversely, by a slow rate of emergence, nest 7 This pupa, a female, was badly damaged while being excavated and may have been either Lanthanomelissa betinae or Parepeolus minutus. However, the fact that the hindbasitarsus was not greatly enlarged suggests that it was the latter species. The pupa lacked mesoscutal tubercles, possessed low but distinct, paired, mesoscutellar tubercles, and an apical rows of rather large, sharply pointed tubercles on each of T2–T5, but not on T1. These are also features shared by Monoeca haemorrhoidalis and Protosiris gigas. construction, and provisioning. These matters cannot be determined by available data. The observations on the nesting biology of Lanthanomelissa betinae (cited as goeldiana (Friese)) reported by Sakagami and Laroca (1988) concur with ours in that the species seems to prefer a sloping nesting surface, although the slope at their site appears to have been steeper than ours. In both cases, entrance turrets were absent, main tunnels tended to be short and remained open, and a single female inhabited each nest. Laterals tended to be short and were soil-filled after cell closure for both. Whereas cells we observed were arranged singly, they stated that cells were often in series of twos. Furthermore, they indicated that cells tended to be more vertical, whereas we found their orientation more variable, with their long axes ranging from 35 to 80 8 from horizontal. With both sets of observations, cell lengths were substantially less than twice their maximum diameters, and nests were quite compact and shallow because of short main tunnels and laterals. The provisions in both cases were arranged as loaves attached to the side of the cell, with eggs on the opposite surface from the attachment. They reported that intermediate larvae circled the provisions, so that they did not contact the cell wall, whereas we found such larvae circling the provisions while the food was still attached to the rear of the cell; this difference, however, may be a discrepancy in the ages of the two larvae observed. Sakagami and Laroca found no cleptoparasites associated with their nests. The floral relationships of Lanthanomelissa betinae were reported by Cocucci and Vogel (2001) and Truylio et al. (2002). The latter found females of L. betinae to be the main visitors of Sisyrinchium micranthum in Rio Grande do Sul, Brazil. Roig-Alsina (1997) described the oil-collecting modifications of the foretarsus found in the genus Lanthanomelissa. In an unpublished study in 1994, I.A.S. analyzed the provisions from 10 nests of Lanthanomelissa betinae in Rio Grande do Sul and found that all were provisioned with pollen only from Sisyrinchium. Thus, this bee species obtains both pollen and oil from the same plant. 24 NO. 3501AMERICAN MUSEUM NOVITATES TABLE 1 Comparative Data on Number of Ovarioles and Number and Sizes of Mature Oocytes of Taxa in Current Study Figures in the first three columns of figures are averages because more than one female of each species was measured. For further explanation, see section on Ovarian Statistics. Taxon Egg index Total mature oocytes Mature oocytes per ovariole Ovariole formula No. of specimens Monoeca haemorrhoidalis Protosiris gigas 0.65 0.82 1 2.33 0.125 0.29 4:4 4:4 2 3 BIOLOGY OF PAREPEOLUS MINUTUS ROIG-ALSINA Adults of Parepeolus minutus were observed apparently searching for and entering nests of Lanthanomelissa betinae at the nesting areas at both Criciu´ma and Curitiba. Four postdefecating Parepeolus larvae, three of which came from one nest, were recovered from nests at Criciu´ma, confirming the host/ parasite association. In external appearance their cocoons (fig. 34) were indistinguishable from those of the host, except that in two examples (the third was not noted) the front end of the cocoon contained no dark, slightly thicker strands of silk. However, from the inside, the cocoons of the two taxa could be immediately recognized because the fecal linings at the front ends of the cocoons of P. minutus were incomplete, leaving a hole through which the open silk fibers of the cocoon were easily visible (fig. 36) and light could be transmitted. Information on the mode of parasitism of Parepeolus minutus is not available. Other parasites were not observed in the host nests. OVARIAN STATISTICS Table 1 provides comparative data on the number of ovarioles and number and sizes of mature oocytes of Monoeca haemorrhoidalis and Protosiris gigas. Although we attempted to dissect the ovaries of Lanthanomelissa betinae from four females, we were unable to retrieve information concerning ovarian anatomy or oocyte development, either because of poor preservation or because of unusual morphology. Females of Parepeolus minutus were not available for dissection. The egg index of Monoeca haemorrhoidalis was 0.65, whether calculated on the basis of the length of its mature oocyte or of its egg (see Methods and Terminology for methods of calculating the egg index). This value falls in the category of small. The only other tapinotaspidine for which we have information is Lanthanomelissa betinae, reported below. This egg index, based on a single female and her egg, was 0.99, close to the upper limit of category medium. It is unknown if such a range in values within a single tribe of solitary bees is unusual. The surprising statistics in table 1 are the range in values of the egg indices between Protosiris gigas and Monoeca haemorrhoidalis. Cleptoparasitic bees tend to have lower egg indices than do those of solitary bees (Rozen, 2003), but here we find that the egg length of the cleptoparasitic bees is nearly as long as that of its solitary host (compare figs. 37 and 38). The egg index of P. gigas is 0.82 based on the mature oocyte (table 1) and 0.86 based on a sampling of eggs from females (see Egg of Protosiris gigas Melo), with both values being well within the medium category. We can partly account for this apparently anomalous situation in a number of ways. First, the egg of Monoeca haemorrhoidalis is curved and hence measures shorter than its curved length, in contrast to straighter egg of Protosiris. Second, the egg of M. haemorrhoidalis is more uniformly thick; it does not taper posteriorly, as does the egg of P. gigas. Third, the host, a robust bee, has a very wide mesosoma compared to its body length, whereas the cleptoparasite is slender, with a narrow mesosoma. Hence, the denominators in both calculations of the egg indices are skewed. Finally, the egg of P. gigas is obviously much thinner than that of its host. When the two eggs are viewed side by side 2006 25ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES Figs. 37–39. Comparative diagrams of eggs, lateral views, anterior ends up, all drawn to same scale with camera lucida. 37. Monoeca haemorrhoidalis.38. Protosiris gigas.39. Lanthanomelissa betinae. (figs. 37 and 38), clearly the cleptoparasite has a considerably smaller egg volume. Part of the problem is with our having used egg length and intertegular distance, two linear statistics, to evaluate volume. While this explanation may partly explain the large size of the egg of Protosiris gigas, other Osirini tend to have egg indices higher than those of many parasitic bees: Epeoloides coecutiens (Fabricius), 0.70 (small) (Rozen, 2001); Osirinus lemniscatus Roig, 0.79 (medium) (Alexander, 1996); and Parepeolus aterrimus (Friese), 0.88 (medium) (Alexander, 1996). Thus, it appears that osirines as a group do not have egg indices that fall into the dwarf category, as do many parasitic bees. Now that we know that females of P. gigas deposit their eggs into a closed host cell, we can probably interpret these relatively high indices as indicating that all members of the tribe invade a closed host cell for ovipositioning; as Rozen (2003) pointed out, cleptoparasitic taxa that invade closed host cells tend to have larger eggs than do those that hide their eggs in cells still being visited by host females. The remaining statistics in table 1 are normal. Cleptoparasitic bees tend to have more mature oocytes than do solitary bees, presumably because of the need for cleptoparasites to lay eggs whenever appropriate host cells are discovered. Four ovarioles per ovary is plesiomorphic in the Apidae (Michener, 2000). IMMATURE STAGES In this section we describe the immature stages of Monoeca haemorrhoidalis,Lanthanomelissa betinae,Protosiris gigas, and Parepeolus minutus, respectively, to the extent that collected material permits. Most of the information presented under Methods and Terminology is applicable to this section. In the descriptions of the intermediate larval instars of both M. haemorrhoidalis and P. gigas, dates and collector names refer to exemplars that were cleared for careful study. E GG OF M ONOECA HAEMORRHOIDALIS Smith) Figures 7, 37, 40–42 D ESCRIPTION : Length 2.9–3.2 mm; maximum diameter 07.3–0.75 mm (N 5 3); egg index 0.65 (category small) (based on mean length of three eggs divided by mean intertegular width of five females selected at random). Shape approximately symmetrical along its strongly curved long axis; broadly rounded at both ends, anterior end slightly to distinctly wider than posterior end, widest about one-fourth length from anterior end; micropyle a tight cluster of small pores at anterior pole, these pores directed toward outcurved surface. Color nearly white. Chorion viewed through stereomicroscope somewhat shiny, much more so than that of Protosiris gigas. Chorion viewed by SEM without obvious patterning over most of surface but with faint polygonal sculpturing behind micropyle (fig. 41). M ATERIAL E XAMINED : Three eggs, Brazil: Parana´, Mananciais da Serra, Piraquara, 20XI-2002 (J.G. Rozen). R EMARKS : The three eggs described above were quite similar to one another, and one of these was illustrated (fig. 37). Another egg (fig. 40) collected and preserved in 2003 was used for SEM examination and measured after being subjected to critical-point drying and coating with palladium/gold. It proved to be shorter (2.5 mm) and wider at maximum width (3.0 mm) than the others, and its posterior half tapered somewhat more than that 32 NO. 3501AMERICAN MUSEUM NOVITATES Figs. 63, 64. Monoeca haemorrhoidalis, pupa. 63. Male, lateral view, with enlargement of tergal tubercle. 64. Hindleg of female, lateral view. Scale bar ( 5 1.0 mm) refers to both figures. but not meeting along midline; those of abdominal segmented 9 more nearly meeting at midline and finely spiculate, thus forming medially interrupted, narrow, transverse, spiculate band (fig. 62); abdominal segment 10 attached in approximate middle of segment 9 in lateral view (fig. 52); anus appearing to be positioned near middle of segment 10 in posterior view (fig. 62) but actually dorsal in position (see predefecating form, below); low transverse swelling curving above anus (fig. 62); surface of swelling distinctly verrucose; integument anterior to swelling finely spiculate laterally; anus (fig. 62) with transverse lips. Spiracles (figs. 52, 56) moderately small, pigmented, subequal in size; atrium globular, projecting above body wall, with rim; peritreme present; atrial inner surface with rows of sharp denticles concentric with primary tracheal opening; primary tracheal opening with collar; subatrium moderately short, with about 6 chambers. Predefecating Form (fig. 53): As described for postdefecating form except for following: integument smooth, not wrinkled. Intersegmental lines scarcely incised dorsally, moderately incised ventrally; intrasegmental lines scarcely evident; paired dorsal tubercles scarcely evident, although on cleared, stained specimen, certainly present, with cuticle more than twice as thick as elsewhere; pleural swellings not developed. Anus clearly dorsal in position (fig. 53). M ATERIAL S TUDIED : 12 postdefecating larvae, Brazil: Parana´: Mananciais da Serra, Piraquara, 20-XI-2002 (J.G. Rozen); 2 postdefecating larvae, same except 21-XI-2002; numerous predefecating and postdefecating larvae, same except 20-XII-2002 (G.A.R. Melo and A. Aguiar); numerous predefecating and postdefecating larvae, same except 4-11-XII-2003 (A. Aguiar, I. Alves-dos-Santos, R.B. Gonc¸alves, G.A.R. Melo, J.G. Rozen). R EMARKS : We could not detect evidence of mandibular wear in postdefecating larvae of this species, which might have resulted from eating the cell lining. Such evidence was discovered in the mandibles of Protosiris gigas (see Remarks in the treatment of its mature larva and also see Biology of Monoeca haemorrhoidalis (Smith), above). P UPA OF M ONOECA HAEMORRHOIDALIS (S MITH ) Figures 63, 64 D IAGNOSIS : Pupae of other Monoeca species have not been described. The only other pupal description of a tapinotaspidine was that of Paratetrapedia swainsonae (Cockerell) (Rozen and Michener, 1988). The undescribed pupa of Tapinotaspoides serraticornis is also represented in the collection of the American Museum of Natural History. Pupae of all three taxa are quite similar (except for size) and lack large mesoscutal tubercles while having obvious paired mesoscutellar tubercles; the very low mesoscutal tubercles reported for P. swainsonae may actually be homologs of the mesoscutal verrucose patches in front of the mesoscutellar tubercles of 2006 33ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES M. haemorrhoidalis. The very low verrucae associated with the ocellar area of the pupa of Monoeca haemorrhoidalis are replaced in the other two species with sharply defined ocellar tubercles. The females of these two species have apical median projections of S4 and S5, which are lacking in M. haemorrhoidalis. Pupae of P. swainsonae tend to have more pronounced axillae than do either of the other two, and both P. swainsonae and T. serraticornis bear a very small but welldefined tubercle at the base of their forewings, and M. haemorrhoidalis does not. There are, no doubt, other differences to be identified among the taxa in the pupal stage, but these differences are probably subtle and difficult to define. Although the pupae of Monoeca haemorrhoidalis and its cleptoparasite Protosiris gigas are found in cocoons of similar external appearance, the pupae can be immediately distinguished by body form (that of M. haemorrhoidalis is robust and that of P. gigas is slender). The basitarsus even of the male M. haemorrhoidalis is broad compared with the slender basitarsus of the pupa of P. gigas. The apex of the foretrochanter of M. haemorrhoidalis bears a pointed tubercle, whereas that of P. gigas is swollen and rounded. H EAD : Integument without setae, spicules, tubercles, but with weakly defined verrucae above ocelli. Labrum apically rounded in frontal view; pupal ocelli scarcely defined, nontuberculate. Mandible with subapical ventral swelling accommodating developing adult setae. Paraglossa long, approximately equal in length to first segment of labial palpus. M ESOSOMA : Integument without setae; mesoscutum with pair of obscurely defined, paramedian, verrucose patches (not discernable in fig. 63) in front of mesoscutellar tubercles; pronotum with lateral angles finely verrucose dorsally; tegula dorsally verrucose. Lateral angles and posterior lobes of pronotum moderately produced. Mesepisternum without tubercles; mesoscutum without tubercles; mesoscutellum with paired, moderately low, paramedian tubercles accommodating developing adult setae; metanotum swollen especially medially but not tuberculate. Tegula not produced, without tubercle; wings without tubercles. All coxae with moderate-sized, sharply pointed, ventroapical tubercles that accommodate developing adult setae; foretrochanter with moderately small ventroapical tubercle; midand hindtrochanters with ventroapical angle but without clearly define tubercles; forefemur with rounded, ventrobasal tubercle; midfemur of both sexes without tubercles; hindfemur of female with very small apical tubercle in front of basitibial plate (fig. 64), in male this tubercle obscure; foretibia with small apical tubercle; midtibia without tubercle; hindtibia with slender apical tubercle on outer surface; hindtibial spurs curved; basitarsus with pronounced apical tubercle; basitarsus of male (fig. 63) narrower than that of female, which is broader than length of other tarsomeres combined (fig. 64). M ETASOMA : Integument without spicules or setae. T1 without tubercles; T2–T6 (male), T2–T5 (female) with apical (or subapical) row of fine, sharply pointed tubercles, those of T2 less pronounced that those of other terga, which tend to have pigmented apices; sterna with apical margins unmodified, without rows of tubercles. Apex of metasoma without terminal spine, ending in rounded membranous lobe. M ATERIAL S TUDIED : 3 female, 7 male pupae, Brazil: Parana´: Mananciais da Serra, Piraquara, 10-XII-2003 (J.G. Rozen); numerous female and male pupae, same except same except 4–11-XII-2003 (A. Aguiar, I. Alves-dos-Santos, R.B. Gonc¸alves, G.A.R. Melo, J.G. Rozen). E GG OF L ANTHANOMELISSA BETINAE U RBAN Figure 32 The single specimen upon which this description is based was lost before it could be examined with an SEM. D ESCRIPTION (fig. 32): Length 2.2 mm; maximum diameter 0.58 mm (N 5 1); egg index 0.99 (category medium). Shape stout, approximately symmetrical along its strongly curved long axis; broadly rounded at both ends, anterior end presumably broader as in Monoeca haemorrhoidalis. Color nearly white. Chorion viewed through stereomicroscope somewhat shiny. M ATERIAL E XAMINED : One egg, Brazil: Pa- 34 NO. 3501AMERICAN MUSEUM NOVITATES Figs. 65–68. SEM micrographs of postdefecating larva of Lanthanomelissa betinae.65. Entire body, lateral view. 66. Close-up of antenna. 67. Labral apex. 68. Close-up of salivary lips showing elongate papillae, identified by rectangle in fig. 67. rana´, Curitiba, 25-XI-2002 (J.G. Rozen), nest no. 2. R EMARKS : The female collected from the nest provided the intertegular measurement for calculating the egg index. P OSTDEFECATING L ARVA OF L ANTHANOMELISSA BETINAE U RBAN Figures 65–73 D IAGNOSIS : See the Diagnosis of the postdefecating larva of Monoeca haemorrhoidalis to distinguish the mature larvae of these two species. Because the postdefecating larva of Lanthanomelissa betinae is nearly white, its antennal papilla (figs. 66, 71) are long and slender, and its mandible (fig. 72, 73) is apically bidentate to subtruncate and bears a denticulate cusp, it can be separated from that of its cleptoparasite Parepeolus minutus, which has a more yellowish cream-color body, antennal papillae (fig. 97) that are more robust, and a mandible (figs. 103, 104) that is apically pointed and lacks a denticulate cusp. Lucas-de-Oliveira (1966) described the predefecating larva of this species, under the name of Lanthanomelissa sp. Later Urban (1995) described and named the species L. betinae, based in part upon two adult paratypes from the nesting site from which the larva had been collected. D ESCRIPTION : Length (if straight) about 9 m. Head (figs. 70, 71): Integument of head capsule at most faintly pigmented, hence appearing white, concolorous with body, except following brownish: mandibles, especially apically, and salivary lips. Cranium with very fine, scattered, mostly nonsetiform sen- 2006 35ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES Figs. 69–73. Lanthanomelissa betinae.69. Predefecating larva, lateral view. 70, 71. Head of postdefecating larva, frontal and lateral views. 72, 73. Right mandible of same, dorsal and inner views. Scale bar ( 5 1.0 mm) refers to fig. 69. silla; inner apical surfaces of maxilla and epipharynx apparently nonspiculate: hypopharynx (fig. 67) with a few elongate spicules laterally well behind hypopharyngeal groove. Salivary opening apically with elongate papillae apparently arising from within opening (visible with SEM: figs. 67, 68); external surfaces of lips without papillae. Head moderate in size compared with body; head capsule moderately wider than long in frontal view (fig. 70). Tentorium complete, moderately robust; anterior tentorial pit distinctly closer to anterior mandibular articulation than to antenna; posterior tentorial pit well impressed, in normal position at junction of hypostomal and postoccipital ridges. Median longitudinal thickening of head capsule extending part way to level of antennae. Postoccipital, hypostomal, pleurostomal ridges well developed; epistomal ridge laterad of anterior tentorial pits short but well developed; epistomal ridge between pits scarcely evident, external sulcus shallow. Parietal band evident. Antennal prominence weak; antennal disc well differentiated from papilla, somewhat projecting; papilla long (two or more times basal diameter), longer than palpi, conical as seen in 36 NO. 3501AMERICAN MUSEUM NOVITATES lateral view, with approximately 3 apical sensilla. Front of head capsule in lateral view sloping normally so that labrum extends somewhat beyond clypeus, and clypeus somewhat beyond frons. Labrum moderately wide, apically subtruncate in frontal view (fig. 70) but in maximum profile (i.e., frontodorsal view) labrum apically weakly bilobed, these mounds bearing most of labral sensilla; labral sclerite (as found in Megachilidae) absent; epipharyngeal surface with brush of long setiform spicules on each side directed anteromesad. Mandible broad at base when viewed dorsally (fig. 72) or ventrally and with short apex beyond base; when viewed adorally, mandible gradually tapering to narrow, nearly flat apex, which may be subtruncate (fig. 73), indistinctly toothed, or indistinctly bilobed with either dorsal or ventral lobe slightly longer than other; cuspal area strongly denticulate (figs. 72, 73); its teeth regularly spaced along dorsal and ventral edges of area and larger than teeth within area; dorsal apical edge beyond cusp without teeth or with one or two; ventral apical edge with linear series of irregular small denticles. Labiomaxillary region moderately strongly projecting in lateral view (fig. 71). Maxillary apex distinct, with palpus positioned slightly to outer side; galea possibly present, but only as sensilla mesad of palpus; palpus moderately large; cardo, stipes, and articulating arm of stipes evident but weakly developed and unpigmented. Labium divided into prementum and postmentum; premental sclerite not evident; labial palpus about as long as maxillary palpus. Salivary opening a transverse slit on moderately projecting lips; lips in frontal view (fig. 70) moderately narrow, reflecting narrowness of prementum. Hypopharynx not projecting, middle part nearly flat, extending backward toward mouth. Body (fig. 65): Integument of postdefecating larva coarsely wrinkled, without setae except for minute, scattered, slightly setiform sensilla mostly below anus on abdominal segment 10; body surface mostly without spicules except paired abdominal tubercles apically each with weak patches of fine spicules; spicules weakly present laterally on abdominal segment 10. Body form robust; intersegmental lines deeply incised; dorsal intrasegmental lines of mesoand metathorax and abdominal segments 1–9 moderately incised; pleural swelling (below spiracle) on most body segments strongly pronounced, more so than in Monoeca haemorrhoidalis; low paired dorsal tubercles on prothorax, on caudal annulets of mesoand metathorax, and on caudal annulets of segments 1–8, these tubercles tending to be transverse but not meeting along midline; those of abdominal segment 9 more nearly meeting at midline; most dorsal tubercles tending to be more pronounced that those of M. haemorrhoidalis; abdominal segment 10 attached in approximate middle of segment 9 in lateral view (fig. 65); anus positioned somewhat above middle of segment 10; low transverse swelling curving above anus; surface of swelling less distinctly verrucose than in Monoeca haemorrhoidalis; anus with indistinct transverse lips. Spiracles moderately small, unpigmented, subequal in size; atrium globular, projecting above body wall, with rim; peritreme present; atrial inner surface smooth; primary tracheal opening with collar; subatrium moderately short, with about 6 chambers. Predefecating Form (fig. 69): As described for postdefecating form except for following: integument smooth, not wrinkled. Intersegmental lines scarcely evident dorsally; intrasegmental lines not evident; pleural swelling not evident; paired dorsolateral body tubercles not evident on uncleared specimen, but visible on cleared, stained specimen as transverse, slightly roughened, dark patches. Anus almost certainly dorsal on segment 10, but ventral line between abdominal segments 9 and 10 not evident. Male with small paired cuticular scars on ventral midline of abdominal segment 9; female sex characters unknown. M ATERIAL S TUDIED : 4 postdefecating and 1 predefecating larvae, Brazil: Santa Catarina: Criciu´ma, UNESC, 16-XI-2002 (J.G. Rozen), from nest no. 1; 1 postdefecating larva, same except from nest no. 4; 15 postdefecating larvae, same except 23, 26-XI-2003 (I. Alves-dos-Santos), nest nos. 1, 2. R EMARKS : Postdefecating larvae are coated with an extremely thin, colorless, transparent material that adheres to the integument. 2006 37ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES Figs. 74–77. SEM micrographs of egg and first instar of Protosiris gigas.74. Front end of egg, anterior view, outcurved surface toward top. 75. Close-up of micropyle. 76. Posterior end of egg, lateral view. 77. Head of first instar, ventral view. E GG OF P ROTOSIRIS GIGAS M ELO Figures 38, 74–76 Length 2.8–3.0 mm, maximum diameter 0.48–0.55 mm (N 5 3); egg index 0.86 (based on average egg length divided by the average intertegular distance [3.39 mm] of five females collected at random) (category medium). Shape (fig. 38) slender, approximately symmetrical along its slightly curved long axis; rounded at both ends, with widest diameter slightly anterior to midbody; anterior end nearly parallel-sided; posterior end gradually tapering; anterior pole slightly produced; micropyle (fig. 75) a cluster of small pores on produced anterior pole; pores directed toward outcurved surface. Color nearly white except chorion of two eggs faintly tan at posterior end. Chorion viewed through stereomicroscope smooth, dull, without visible pattern except produced area at anterior pole somewhat shinier. As viewed with SEM, chorion at anterior pole showing fine elongate grooves radiating from micropylar pore directed only toward outcurved surface (fig. 75); somewhat farther away from micropyle but still at anterior end, chorion with radiating, strongly expressed polygons with raised borders that exhibit midline grooves (fig. 74); these grooves on outcurved surface extensions of grooves radiating from micropylar pores; polygonal grooves rapidly becoming less expressed posteriorly (fig. 74), fading to become faint polygonal patterning (lacking midline grooves) that extends over rest of chorion, except patterning becoming more pronounced, without midline grooves at extreme posterior end (fig. 76). M ATERIAL E XAMINED : 3 eggs, Brazil: Pa- 38 NO. 3501AMERICAN MUSEUM NOVITATES rana´, Mananciais da Serra, Piraquara, 20-XI2002 (J.G. Rozen) from nests of Monoeca haemorrhoidalis R EMARKS : Among the taxa of the Osirini, the egg (as a mature oocyte) of Epeoloides coecutiens (Fabricius) was described and illustrated by Rozen (2001), and the presumably mature oocytes of Osirinus lemniscatus Roig-Alsina and Parepeolus aterrimus (Friese) were illustrated in the ovariole by Alexander (1996). Eggs of Protosiris gigas and E. coecutiens are classified as small, and those of O. lemniscatus and Pa. aterrimus are small and medium, respectively, according to Alexander (1996: table 1). Those of E. coecutiens are somewhat more curved (Rozen, 2001: fig. 3) than those of Pr. gigas (fig. 38) but otherwise are approximately similar in shape. The egg of O. lemniscatus appears to be slender (Alexander, 1996: fig.14), like that of Pr. gigas, but the egg shape of Pa. aterrimus is difficult to interpret (Alexander, 1996: fig. 15). The eggs of Pr. gigas and E. coecutiens differ considerably in chorionic surface sculpturing, with that of E. coecutiens being nodular (Alexander, 1996: figs. 10, 11) while that of Pr. gigas consists of a network of polygons with raised margins, all as seen with an SEM. However, the micropylar area of both (Alexander, 1996: fig. 12, and fig. 74 herein) consists of polygons narrowing toward the anterior pole, the polygons with raised margins each of which is divided by a fine groove. At the extreme anterior end of the eggs, the polygons are obliterated, and there remains only the raised borders and, on the outcurved surface, the fine, channellike grooves, which continue to the micropylar array. This feature, shared by both species, may be a synapomorphy. In E. coecutiens, the micropylar pores were not identified by Rozen (2001), but were probably obscured by follicular debris. 8 The pores of Pr. gigas become quite visible only under extremely high magnification (fig. 75). 8 An early SEM study of the honey bee egg failed to detect the micropylar pores (Bronskill and Salkeld, 1978), but Erickson (1981: pl. 1.44) clearly demonstrated their presence. F IRST I NSTAR OF P ROTOSIRIS GIGAS M ELO Figures 77–81 D IAGNOSIS : Because osirine first instars have not been collected and described before, a comparison of the first instar of Protosiris gigas with close relatives cannot be made. See the Discussion and table 2 for features that distinguish the first instar of this species from those of other cleptoparasitic apids. Please see the diagnosis of the first instar of Monoeca haemorrhoidalis for ways to separate first instars of host and parasite. L ENGTH : About 3.0 mm. H EAD (figs. 77, 79–81): Integument of parietal moderately sclerotized, somewhat pigmented; mandibular apices strongly pigmented; head capsule and mandibles at base with scattered, inconspicuous, minutely setiform sensilla, except parietals with indistinct line of slightly larger sensilla occupying position of spinulae as found in the Melectini (see remarks below); following areas spiculate: epipharyngeal surface, maxillary apices, and labium posterior to salivary opening (hypopharynx nonspiculate). Head more or less prognathous (fig. 80); parietals faintly enlarged, moderately elongate in lateral view (fig. 80), somewhat constricted behind dorsally (fig. 80) but not laterally so that foramen not noticeably narrowed; sclerotization of parietal ending at hypostomal ridge, not invading labiomaxillary area; sclerotization of parietals ending posteriorly at postoccipital ridge. Tentorium complete but thin, with dorsal arms; anterior and posterior tentorial pits moderate in size; internal head ridges tending to be moderately developed except epistomal ridge weak laterad of anterior tentorial pits, absent between pits; integument near ridge not pebbled, wrinkled, or in other ways sculptured. Parietal bands absent. Antenna a low projection with the papilla fused with parietal and bearing 6–7 nonsetiform sensilla. Labrum moderate in size, apically subtruncate when seen in maximum profile (fig. 81) (but appearing rounded in frontal view, fig. 79); labrum weakly, if at all, sclerotized, with numerous nonsetiform sensilla, many on low elevations; labral tubercles absent; boundary between labrum and lower end of clypeus weakly defined. 2006 39ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES Figs. 78–86. Early instars of Protosiris gigas.78. First instar, entire larva, lateral view. 79–81. Head, first instar, frontal, lateral, and ventral views, respectively. 82. Head, second instar, frontal view. 83. Head, third instar, frontal view. 84. Abdominal segments 3 and 4, early third instar, lateral view. 85. Abdominal segments 3 and 4, late third instar, lateral view; for explanation see text. 86. Head, fourth instar, frontal view. Scale bar ( 5 1.0 mm) refers to fig. 78. Figures drawn in various scales, except 79– 82 to same scale and 84, 85 to same scale. 40 NO. 3501AMERICAN MUSEUM NOVITATES Mandibles (figs. 77, 79–81) heavily sclerotized, moderately pigmented, basally stout, apically attenuate, strongly curved, ending in sharply pointed, simple apex, without tubercles on outer aspect but with scattered fine sensilla; inner surface smooth, without teeth or projections. Each maxilla a strongly projecting, apically heavily spiculate, rounded lobe; maxillary palpus not evident except for subapical cluster of about six, nonsetiform sensilla on ventrolateral surface (figs. 77, 81) on cleared specimen (these sensilla resembling those of labial palpus); under SEM examination maxillary palpus a low, nonspiculate, padlike structure (fig. 77); palpus, cardo, stipes (except for articulating arm) shallow depression near base of maxilla; thin sclerite leading from maxilla to salivary opening presumably articulating arm of stipital sclerite because of its position relative of maxilla and apex of labium; maxillae and labium basally forming continuous surface and seemingly fused, but actually separated by fine suture (fig. 81) extending to their bases; labium greatly recessed so that maxillary apices extend much farther forward than apex of labium; labial palpus not evident as projection on cleared specimen but identified by cluster of nonsetiform sensilla behind and laterad of salivary opening; under SEM examination, labial palpus faintly produced, nonspiculate (fig. 77). Salivary opening small, circular; salivary duct evident. Hypopharynx identified as area above articulating arms of stipital sclerites, nonspiculate, bearing two irregular pits, possibly indentations formed by mandibular apices when mandibles are closed. B ODY : Integument without setae; venter of each body segment with extensive patch of spicules, dorsal surface without spicules; linear series of fine granules immediately above spiracular line on each side (as found in Monoeca haemorrhoidalis) absent. Form elongate, linear (fig. 78); intersegmental lines deeply incised; body without dorsal tubercles; abdominal segments not divided dorsally into cephalic and caudal annulets; abdominal segments 4–7 each with distinct transverse cleft shortly behind intersegmental line, so that area in front becomes median, backward-sloping transverse fold, anterior surface of which is spiculate while the posterior surface is smooth; abdominal segment 3 with similar but less distinct anterior ventral fold; prothorax and abdominal segment 9 not protruding ventrally; abdominal segment 10 rounded posteriorly; anus apical. All spiracles present, small, coequal in diameter, flush with body wall (i.e., not on tubercles). M ATERIAL S TUDIED : 2 first instars (one still partly enclosed in the chorion), Brazil: Parana´: Mananciais da Serra, Piraquara, 20-XI2002 (J.G. Rozen) from nests of Monoeca haemorrhoidalis. R EMARKS : The single specimen whose head capsule and body were cleared in an aqueous solution of sodium hydroxide had a number of pollen grains lodged in its esophagus, an indication that it had started to feed although its midintestine contained no pollen. The median, backward-sloping folds on the anterior venters of abdominal segments 3–7 are a feature unknown in other bee larvae. Their function is discussed in Biology of Protosiris gigas, above. O THER L ARVAL I NSTARS OF P ROTOSIRIS GIGAS M ELO The four following larval instars increase in size incrementally, both in body length and head width. All material discussed in this section was collected at Brazil: Parana´, Mananciais da Serra, Piraquara; dates of collection and collectors are identified parenthetically. The second instar (20–21-XI-2002 [J.G. Rozen]) is similar to the first, although it is somewhat larger (compare figs. 79 and 82), and the head is less elongate in lateral view. The antennae, composed of about the same number of sensilla, are perhaps less pronounced than those of the first instar. The mandibles continue to be sharply pointed, suggesting that this instar is also capable of killing conspecific and host immatures. Maxillary and labial palpi are still represented only by their sensilla (not examined with an SEM). The epistomal and pleurostomal ridges are less pronounced, and the spiculation of head and body is similar to the first instar. The two small pits in the hypopharynx of the first instar have given way to a single, larger, median, dimplelike pit, where the mandibular 2006 41ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES apices might reside if the mandibles are closed. The transverse folds of integument on the anterior ventral surfaces of abdominal segments 3–7 are no longer evident. The spiracular atrium can be identified because it is now somewhat larger than the subatrium, but other features are unclear because of the small size of the spiracle. With the third instars (5, 10-XII-2003 [G.A.R. Melo]; 6-XII-2003 [J.G. Rozen, G.A.R. Melo]), we see a gradual shift away from the rapacious features of the first and second instars, toward the anatomy of the fifth instar, adapted for feeding and cocoon spinning. This is most evident in the mandibles, which now start to broaden subapically where the surface takes on a faintly concave shape. This will transform into the scoop-shaped apical concavity in subsequent instars. For the first time a row of sharp teeth are visible along the dorsal and ventral apical mandibular margins. The mandible (fig. 83) is still acutely pointed and moderately strongly curved at the extreme apex. Observations recorded in the Biology of Protosiris gigas, above, suggest that this instar is behaviorally capable of attacking (or defending itself) in the event it encounters another larva in the cell. The antennal papilla is large but shorter than its basal diameter. The maxillary and labial palpi are slightly produced, blisterlike swellings. The salivary opening is slightly transverse but without lips. Spiculation is now faintly evident on the hypopharynx and the inner apex of the maxillae, and the ventral surface of the body is strongly retrorse spiculate. Fine spicules also occur dorsally on more posterior abdominal segments. As indicated by the reshaped mandible, the third instar is primarily adapted for feeding. Because we were able to observe exemplars preserved in various stages of their feeding, we were able to record anatomical changes in their body shape. In its early stage (fig. 84) the body is quite linear, and abdominal segments at least 1–4 (but probably 1–8) are divided dorsally into cephalic and caudal annulets. Ventrally each of abdominal segments 2–8 posseses a narrow transverse fold of spiculate integument in front of the main ventral integument, similar to the folds found on segments 3–7 of the first instar. By the end of the third stadium (fig. 85), both the dorsal and ventral folds have disappeared because of the swollen midsection of the body. Thus, the greatly folded integument of the body midsection during the early part of the stadium allows the instar to ingest a large quantity of the provisions. The fully fed third instar appears quite physogastric, contrasting with its linear shape at the beginning of the stage. The spiracular atria are globular but much flattened; the primary tracheal opening is without a distinct collar and the subatrium cannot be recognized. The fourth instar (fig. 86) (11, 20-XII2002 [G.A.R. Melo]; 6-XII-2003 [J.G. Rozen, G.A.R. Melo]) in most ways is anatomically intermediate between the third and fifth instars. The mandible may no longer be effective (and presumably does not need to be) for attacking other cleptoparasites and probably serves well for ingesting provision because the scoop-shaped apex is more pronounced than in the third instar and is further broadened by the rows of apical teeth. The mandible changes little, except for size, between this instar and the final one. The hypopharynx has assumed a dorsally protruding form (which may actually have been the case in the preceding instar), its surface now distinctly spiculate, as in the last larval instars. Maxillary and labial palpi are projecting considerably more than in the previous instar but not as far as their basal diameters, which is also true for the antennal papillae. Parietal bands and the frontal depression above and mesad of each antenna are faintly present. Division of the labium into prementum and postmentum is questionably distinct in this instar. The salivary opening is narrowly transverse and has slightly projecting lips with papillae appearing on its dorsal surface and just above it. It is unknown if the abdominal segments exhibit intrasegmental lines early in this instar as they do in the third. The body integument is strongly spiculate ventrally, more weakly so dorsally. The spiracular atrium of the fourth instar is clearly globular but shallow compared with that of the last instar, possesses a peritreme, and the subatrium is sclerotized but lacks distinct chambers. 48 NO. 3501AMERICAN MUSEUM NOVITATES lips in frontal view (fig. 102) moderately narrow, reflecting narrowness of prementum. Hypopharynx a dorsally projecting lobe, truncated on top, behind articulating arms of stipites and premental sclerite. Body (fig. 101): Integument without setae except for small cluster of minute setiform sensilla mostly on abdominal segment 10, below anus; body surface spiculate dorsally and ventrally, but nonspiculate laterally in vicinity of spiracles and below them; integument without spines or sclerotized tubercles. Body form moderately robust; intersegmental lines moderately weakly incised; dorsal intrasegmental lines absent on thorax, weakly present on abdominal segments 1–9; pleural swelling (below spiracle) scarcely noticeable; very low paired dorsal tubercles on thoracic segments and on caudal annulets of abdominal segments 1–9; abdominal segment 10 attached in approximate middle of segment 9 in lateral view (fig. 100); anus somewhat dorsal on segment 10; distinct transverse swelling curving above anus; surface of swelling finely roughened but not verrucose; anus without distinct lips. Spiracles moderately small, darkly pigmented, subequal in size; atrium globular, projecting slightly above body wall, with rim; peritreme present; atrial inner surface smooth; primary tracheal opening with collar; subatrium short, usually with two larger outer chambers and one or two smaller inner chambers. M ATERIAL S TUDIED : 1 postdefecating larva, Brazil, Santa Catarina, Criciu´ma, UNESC, 16-XI-2002 (J.G. Rozen), from nest no. 1 of Lanthanomelissa betinae; 1 postdefecating larva, same except 23-XI-2003 (I. Alves-dosSantos), from nest no. 1 (different nest from previous year); 1 postdefecating larva, same except 26-XI-2003, from nest no. 2. R EMARKS : As with the other postdefecating larvae described herein, that of Parepeolus minutus, when removed from the cocoon, was found to be covered by a dry, clear, nearly colorless film that closely adhered to it body and tended to break into flakes after the larva was preserved in Kahle’s solution. DISCUSSION The reader is referred to ‘‘Synopsis of the nesting biology of the Tapinotaspidini bees’’ (Aguiar et al., 2004) for a comparison of the nesting biology of the members of this tribe based on the information from this study and from previously published accounts. As the authors pointed out, members of the tribe nest in horizontal ground, earthen banks, and rotten wood. In addition, Chalepogenus rozeni Roig-Alsina has been observed 26 km south of Vicun˜a, Elqui Prov., Chile, nesting in deep cracks in the soil, as much as 30 cm below the surface. These nests have short main tunnels extending obliquely downward and cells appearing within 5 cm of the vertical crack surfaces (Rozen, unpubl. data). Such a nesting site can probably be considered akin to nesting in an earthen bank. It would be interesting to compare the nesting biology and immatures of the Tapinotaspidini with those of the Exomalopsini because, until recently, the included taxa were placed in the same tribe, the Exomalopsini. However, such a comparison would be premature because so little has been published concerning the immatures and biologies of the various genera. Nonetheless, one biological feature seems to distinguish the Tapinotaspidini, that is, their behavior of collecting floral oils and their ability to transport and manipulate them. The nonparasitic Ctenoplectrini do collect floral oils (Michener, 2000) and the postdefecating larva of one species has been described (Rozen, 1978). While this larva is remarkably similar to those of the Tapinotaspidini, the similarities (body form, details of the head and mouthparts) appear to be plesiomorphic. To the extent known, the Ctenoplectrini are cavity nesters, unlike the Tapinotaspidini. However, a more detailed analysis of larval features is warranted to determine if synapomorphies can be identified that show a relationship between these two tribes in light of some of the analyses of Roig-Alsina and Michener (1993) that show them to be sister groups. In attempts to determine the nonparasitic ancestor that gave rise to cleptoparasites, host lineages are often identified as being likely ancestors. Is there evidence that the Osirini are derived from a tapinotaspidinelike ancestor? The fact that osirine larvae can metabolize floral oils mixed with pollen may be a synapomorphy of the two tribes. Mature 2006 49ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES larvae of the two tribes show broad resemblances, but most of the features appear plesiomorphic and therefore are of little help in evaluating relationships. The first instar of Protosiris gigas is obviously adapted to seek out and destroy host immatures and competing cleptoparasitic immatures. Its anatomy and biology therefore can be used to address the question whether Protosiris (and other Osirini) are closely related to other cleptoparasitic groups. Rozen (1991: table 1) identified 21 features of first instars that varied among the Nomadinae, Protepeolini, Melectini, Rhathymini, Isepeolini, and Ericrocidini. These same features are presented in table 2 here, to which two more columns have been added for Coelioxoides (Tetrapediini) and Protosiris (Osirini). Primitive features are scored 0, and derived features are scored 1, 2, and so on. Two features of the Isepeolini, then unknown, are now known: (1) females of Isepeolus introduce their eggs into host cells that are still open (scored 1), and (2) when deposited, the eggs are flat against the cell lining (Rozen, 2003). The latter is also scored 1, to be the same as ‘‘inserted in the cell wall’’ because of indirect evidence that the related Melectoides deposits its eggs this way in the cells of Canephorula (Michellete et al., 2000; Rozen, 2003). Data for Coelioxoides come from Alves-dos-Santos et al. (2002). It is now possible to analyze the behavioral and anatomical features of the first instar of Protosiris (the only member of the Osirini studied so far) with those of seven of the nine other cleptoparasitic lineages of the Apidae (Rozen, 2000b). Two cleptoparasitic lineages, Exaerete and Aglae (both Euglossini), are excluded, Aglae because its mode of parasitism is unknown, Exaerete because its second instar (not the first) is hospicidal (Garo´falo and Rozen, 2001). Table 3 shows the features of egg deposition and first-instar anatomy that Protosiris shares with each of the other seven lineages of cleptoparasitic apids. Based on the number of shared features, Protosiris and the Ericrocidini are the most similar, but they share no feature uniquely, a fact that suggests that these character states are not particularly strong. The complete sclerotization of the labiomaxillary region of the ericrocidines contrasts sharply with the totally membranous labiomaxillary region of Protosiris, as does the dorsoventrally flattened, large, projecting ericrocidine antenna with the small antennal papilla of Protosiris. The ericrocidine sclerotized labrum fused with the clypeus is also unlike the nearly membranous labrum of Protosiris. These strong dissimilarities do not support the idea that these two taxa are related. The only possibly unique character shared by Protosiris with any of the lineages is character 6, the presence of spinulae found also in Melectini. While spinulae have been a unique feature of melectines, present in all of them, the microscopic morphology of a spinula (Rozen, 1991: fig. 22) is not that of a simple setiform sensillum as appears to be true in Protosiris. Thus, while the spinulae of the Melectini and the band of setiform sensilla of Protosiris occupy the same position on the head capsule, they may not be homologous. The numerous differences between the first instars of these two groups also argue against a close relationship (table 2). One is left with the impression that Protosiris and thus the Osirini represent an independent origin of cleptoparasitism in the Apidae. The presence of the unique, ventral, integumental folds on the anterior edges of segments 3–7 seems to support the concept that many hospicidal bee larvae must be able to move with agility in order to find and kill host eggs (or larvae) and those of competing cleptoparasites, and that Protosiris (or its ancestor) found a new way to crawl. It is not out of the question that the Osirini arose from a tapinotaspidine-like ancestor despite the lack of evidence supporting such a conclusion at present. ACKNOWLEDGMENTS Robert G. Goelet, Chairman Emeritus, Board of Trustees, American Museum of Natural History, kindly supported the two fieldtrips to Brazil undertaken by J.G.R. to carry out this investigation. We thank the State of Parana´ water company SANEPAR for granting permission to G.A.R.M. to conduct the studies on Monoeca at the Mananciais da Serra. John Pinto, University of California, Riverside, tentatively identified meloid adults 50 NO. 3501AMERICAN MUSEUM NOVITATES TABLE 2 Comparison of the Modes of Parasitism and of Anatomical Features of First Instars of Cleptoparasitic Apidae 2006 51ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES TABLE 2 (Extended) [Data in table 2 from Rozen (1991: table 1, except as noted in Discussion), from Coelioxoides (Alves-dos-Santos et al., 2002), and from Protosiris (current study)]. 52 NO. 3501AMERICAN MUSEUM NOVITATES TABLE 3 Derived Character States of Egg Deposition Features and First Instars (Shared by Protosiris gigas with other Cleptoparasitic Apidae; for explanation, see Discussion) Taxa Shared derived character states No. of states Protosiris/Nomadinae Protosiris/Protepeolini Protosiris/Melectini Protosiris/Rhathymini Protosiris/Isepeolini Protosiris/Ericrocidini Protosiris/Coelioxoides 2, 11, 16 11 0, 1, 2, 6 a 0, 1, 3, 11, 17 2, 11, 16, 17 0, 1, 2, 11, 16, 17 0, 1, 3, 16 3 1 4 5 4 6 4 a The homology of this feature is questionable; see Discussion. taken from the nesting site of Monoeca haemorrhoidalis as Tetraonyx (Paratetraonyx) distincticollis Pic. We thank the following Scientific Assistants of the American Museum of Natural History for their contributions to this project: Valerie Giles prepared all larval specimens for SEM examination and took the micrographs; Steve Thurtson arranged and labeled the illustrations for publication. Division Secretary Joanna Bilz assisted in numerous ways, including the formatting of table 2. We also express our appreciation to John S. Ascher and two anonymous reviewers for their thoughtful suggestions and comments that have improved the manuscript. REFERENCES Aguiar, A.J.C., G.A.R. Melo, J.G. Rozen, Jr., and I. Alves-dos-Santos. 2004. Synopsis of the nesting biology of Tapinotaspidini bees (Apidae: Apinae). Proceedings of the 8th IBRA International Conference on Tropical Bees and VI Encontro sobre Abelhas, pp. 80–85. Ribeira˜o Preto, Brazil; CD-ROM. Alexander, B.A. 1996. Comparative morphology of the female reproductive system of nomadine bees (Hymenoptera: Apidae: Nomadinae). 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Comparative nesting biology of the bee tribe Exomalopsini (Apoidea, Anthophoridae). American Museum Novitates 2798: 1–37. Rozen, J.G., Jr. 1991. Evolution of cleptoparasitism in anthophorid bees as revealed by their mode of parasitism and first instars (Hymenoptera: Apoidea). American Museum Novitates 3029: 1–36. Rozen, J.G., Jr. 2000a. Pupal descriptions of some cleptoparasitic bees (Apidae), with a preliminary generic key to pupae of cleptoparasitic bees. American Museum Novitates 3289: 1–19. Rozen, J.G., Jr. 2000b. Systematic and geographic distributions of Neotropical cleptoparasitic bees, with notes on their modes of parasitism. In M.M.G. Bitondi, K. Hartfelder, et al. (editors), Anais do IV Encontro sobre Abelhas, pp. 204–210. Ribeira˜o Preto, Brazil. Rozen, J.G., Jr. 2001. Ovarioles and oocytes of two old world cleptoparasitic bees with biological notes on Ammobatoides (Apoidea: Apidae). American Museum Novitates 3326: 1–9. Rozen, J.G., Jr. 2003. Eggs, ovariole numbers, and modes of parasitism of cleptoparasitic bees, with emphasis on Neotropical species (Hymenoptera: Apoidea). American Museum Novitates 3413: 1–36. Rozen, J.G., Jr., and S.L. Buchmann. 1990. Nesting biology and immature stages of the bees Centris caesalpiniae,C. pallida, and the cleptoparasite Ericrocis lata (Hymenoptera: Apoidea: Anthophoridae). American Museum Novitates 2985: 1–30. Rozen, J.G., Jr., K.R. Eickwort, and G.C. Eickwort. 1978. The bionomics and immature stages of the cleptoparasitic bee genus Protepeolus (Anthophoridae, Nomadinae). American Museum Novitates 2640: 1–24. Rozen, J.G., Jr., and C.D. Michener. 1988. Nests and immature stages of the bee Paratetrapedia swainsonae (Hymenoptera: Anthophoridae). American Museum Novitates 2909: 1–13. Rozen, J.G., Jr., and H. O ¨zbek. 2003. Oocytes, eggs, and ovarioles of some long-tongued bees (Hymenoptera: Apoidea). Appendix: Parammobatodes rozeni, a new bee species from Israel, by M. Schwarz. American Museum Novitates 3393: 1–35. Rozen, J.G., Jr., and H. O ¨zbek. 2005. Notes on the egg and egg deposition of the cleptoparasite Thyreus ramosus (Hymenoptera: Apidae: Melectini). Journal of the Kansas Entomological Society 78(1): 34–40. Sakagami, S.F., and S. Laroca. 1988. Nests of an exomalopsine bee Lanthanomelissa goeldiana (Hymenoptera: Anthophoridae). Journal of the Kansas Entomological Society 61: 347–349. Santos, C.G., J.M. Oliveira, K.S. Ramos, and B. Blochtein. 2004. Plasticidade de glaˆndulas tegumentares abdominais em Monoeca xanthopyga Harter-Marques, Cunha & Moure (Hymenoptera, Apidae, Tapinotaspidini). Revista Brasileira de Entomologia 48: 221–225. Torchio, P.F. 1989a. In-nest biologies and development of immature stages of three Osmia species (Hymenoptera: Megachilidae). Annals of the Entomological Society of America 82: 599– 615. Torchio, P.F. 1989b. Biology, immature development, and adaptive behavior of Stelis montana, a cleptoparasite of Osmia (Hymenoptera: Megachilidae). Annals of the Entomological Society of America 82: 616–632. Torchio, P.F., and V.J. Tepedino. 1982. Parsivoltinism in three species of Osmia bees. Psyche 89: 221–238. Torchio, P.F., and G.E. Trostle. 1986. Biological notes on Anthophora urbana urbana and its parasite, Xeromelecta californica (Hymenoptera: Anthophoridae), including descriptions of late embryogenesis and hatching. Annals of the Entomological Society of America 79: 434– 447. Torchio, P.F., G.E. Trostle, and D.J. Burdick. 1988. The nesting biology of Colletes kincaidii Cockerell (Hymenoptera: Colletidae) and development of its immature forms. Annals of the Entomological Society of America 81: 605–625. Truylio, B., B. Harter-Marques, and W. Engels. 54 NO. 3501AMERICAN MUSEUM NOVITATES 2002. Biologia floral e polinizac¸a˜o de Sisyrinchium micranthum (Iridaceae) na regia˜o do Planalto das Arauca´rias do Rio Grande do Sul, Brasil. Biocieˆncias 10: 11–24. Urban, D. 1995. Espe´cies de Lanthanomelissa Holmberg e Lanthanella Michener & Moure (Hymenoptera, Anthophoridae, Exomalopsinae). Revista Brasileira de Zoologia 12: 767–777. APPENDIX T AXONOMIC N OTES ON M ONOECA AND D ESCRIPTION OF A N EW S PECIES OF P ROTOSIRIS (H YMENOPTERA :A PIDAE ) by Gabriel A.R. Melo INTRODUCTION This short note deals with the identity of the species of Monoeca from southeastern Brazil and describes the new species of Protosiris associated with the nesting site of Monoeca haemorrhoidalis, described above. The acronyms used herein refer to the following collections: AMNH, American Museum of Natural History, New York; BMNH, The Natural History Museum, London; DZUP, Departamento de Zoologia, Universidade Federal do Parana´, Curitiba, Brazil; MPEG, Museu Paraense Emilio Goeldi, Bele´m, Brazil; MZSP, Museu de Zoologia, Universidade de Sa˜o Paulo, Sa˜o Paulo; NHMW, Naturhistoriches Museum Wien, Wien, Austria; ZMHB, Museum fu¨r Naturkunde der Humboldt Universita¨t zu Berlin, Berlin. Monoeca haemorrhoidalis (Smith) Melissodes haemorrhoidalis Smith, 1854: 313. Type female, South America (BMNH). Tetralonia reversa Smith, 1879: 111. Type female, Brazil: Rio de Janeiro, Tijuca (BMNH). Synonymy indicated by LaBerge and Moure (1962: 11). Tetrapedia piliventris Friese, 1899: 293. Holotype male, Brazil: Sa˜o Paulo, Santos (NHMW). NEW SYNONYMY . The specimens from the population studied in Piraquara, Parana´, agree with the type material listed above, as well as with additional specimens from Rio de Janeiro (listed below). This species is most similar to M. schrottkyi (Friese, 1902) and M. xanthopyga Harter-Marques, Cunha, and Moure, 2001. These three species, together with M. pluricincta (Vachal, 1909), represent the largest members of Monoeca (from 10 to 14 mm in length); also, they possess relatively long pubescence on the mesoscutum and scutellum (longest plumose setae about 2.5–3.1 3 as long as diameter of flagellum). Monoeca pluricincta is readily distinguished by its conspicuous yellow integumental tergal bands and by its shallower medial sulcus on the upper frons. The other three species differ mostly by details of the pubescence and structure of the metasomal terga and sterna (figs. A1–A6) and can be separated using the key presented below. Females of M. haemorrhoidalis and M. schrottkyi can be readily separated from one another based on the pattern of pubescence on the metasomal terga. Many males of M. haemorrhoidalis, however, are quite similar to M. schrottkyi in possessing broad bands of yellow pubescence on the terga. Among the six species of Monoeca examined (see Biology of Monoeca haemorrhoidalis, above), females of the four species mentioned here have a large, wax-extruding area at the base of T6 (figs. A7, A8). The specimens of M. haemorrhoidalis from Piraquara, especially the males, vary considerably in color of the body pubescence and in the distribution of the pubescence on the metasomal terga. The females are less variable; most of them have the marginal zone of T2 and entire T3–T4 covered with bright yellow setae, while only a few females have the metasomal terga covered with dark pubescence. The males vary from having very light colored (pale yellow to light orange) to entirely black pubescence on the head, thorax, and legs, including all sorts of combinations. The setation on the metasomal terga also varies from very dense, with broad marginal bands on the terga, to very sparse, with only a few scattered setae along the tergal margin. M ATERIAL E XAMINED : Female type of Melissodes haemorrhoidalis, ‘‘Type H. T.’’, ‘‘B. M. Type / Hym / 17.B. 858’’, ‘‘Melissodes / haemorrhoidalis / S. Amer. Sm.’’ and ‘‘F. Sm. coll. / 79.22’’; female type of Tetralonia reversa, ‘‘Type B. M.’’, ‘‘B. M. Type / Hym / 17.B.8111’’, ‘‘Tetralonia / reversa / Type Smith’’ and ‘‘TEJUCA / Jan 1. 1857 / J. Gray’’; male holotype of Tetrapedia piliventris, ‘‘Santos / Brasilien / Dr. Brauns. / 18.10.97’’, ‘‘Tetrapedia / piliventris / det. Friese 1898 / n. sp.’’ ( ? ) and ‘‘Holotype’’; 1 female (DZUP), ‘‘Tijuca—R.J. / 16.I.56 / Moure leg.’’ and ‘‘Monoeca / reversa / (Smith, 1879) / Det. 2006 55ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES Figs. A1, A2. Monoeca haemorrhoidalis.A1. Female metasoma, dorsal view. A2. Male metasoma, ventral view. Figs. A3, A4. Monoeca xanthopyga.A3. Female metasoma, dorsal view. A4. Male metasoma, ventral view. Figs. A5, A6. Monoeca schrottkyi.A5. Female metasoma, dorsal view. A6. Male metasoma, ventral view. Scale 5 3 mm, refers to all figures. 56 NO. 3501AMERICAN MUSEUM NOVITATES Figs. A7, A8. Sixth tergum of Monoeca haemorrhoidalis, female. A7. Dorsal view of sclerite, showing well-developed pygidial plate and broad wax extruding area at base associated with epidermal gland. A8. Close-up of wax release surface as indicated by rectangle in fig. A7. Moure 1980’’; 1 male (DZUP), ‘‘Floresta da Tijuca / D. Federal BRASIL / I-1954 / C.A.C. Seabra Coll.’’ and ‘‘Monoeca / brasiliensis / Lep. Serv. / Det. J. S. Moure 1957’’; 1 female (DZUP), ‘‘Praia Lagoinha do Leste / Floriano´polis, SC, Brazil / 27.XII.2002 / A. Zillikens leg.’’ and ‘‘In nest aggregation of / Monoeca sp.’’; plus numerous males and females collected at the nesting site, as well as additional older specimens collected in the same locality and deposited in the DZUP collection. Monoeca schrottkyi (Friese) Pachycentris schrottkyi Friese in Schrottky (1901: 215), nomen nudum. Pachycentris schrottkyi Friese, 1902:187. Lectotype female (presently designated), Brazil: Sa˜o Paulo, Jundiaı´ (ZMHB). In order to properly identify the material from Piraquara, we also studied the type material of M. schrottkyi (see Comments under M. haemorrhoidalis). Friese described this species based on females collected by Schrottky in a nest aggregation found in Jundiaı´, Sa˜o Paulo. To help stabilize the taxonomy of the genus, we designate here as lectotype one of the syntype females (see label data below). M ATERIAL E XAMINED : Lectotype female (ZMHB), ‘‘Brasil / Jundiahy / 16.11.1899 / Schrottky’’, ‘‘Pachycentris / schrottkyi / 1900 Friese det. / Fr.’’ ( / ) and ‘‘Typus’’; 1 female (MZSP), ‘‘1.374’’ and ‘‘PARATYPE / Monoeca / schrottkyi (Friese, 1902) / J. S. Moure 1993’’ (the registry book of the MZSP contains the following data under the number 1.374: ‘‘Pachycentris schrottkyi Friese (cotipo), 16.xi.1899, Jundiahy, Schrottky’’); 1 female (MPEG), ‘‘JUNDIAHY / E. S. PAULO’’, ‘‘Brasil / Jundiahy / 18.11.1899 / Schrottky’’ and ‘‘Pachycentris / schrottkyi Friese’’ (Ducke’s handwritten label); 1 male (DZUP), ‘‘Sa˜o Paulo / cidade / Melzer leg. / 1914.’’ and ‘‘Tetralonia / (Thygater.)’’; 1 male (DZUP), ‘‘Barueri / Sa˜o Paulo—Brasil / 8-IV-61 / K. Lenko col.’’ and ‘‘Monoeca / schrottkyi / (Friese, 1902) / Pe J S Moure 1993’’; 1 female (MZSP), ‘‘Est. Biol. Borace´ia / Saleso´polis, SP / W. Wilms, col. / 14.12.1992’’ and ‘‘Monoeca / piliventris / (Friese, 1899) / det. W. Wilms, 1994’’; 1 female (DZUP), ‘‘Caioba´ / XII-1942’’ and ‘‘Fiorentina / schrottkyi / P. Moure det. 1947’’; 1 female (DZUP), ‘‘CORUPA ´/ S. Catarina BRASIL / XI1953 A. Maller’’ and ‘‘Monoeca / schrottkyi / (Fr.) / Det. J. S. Moure 1957’’ (on the reverse ‘‘C. W. cotype / MZ-USP / Pe. J. S. Moure / XII1954’’). Key to the Large Species of Monoeca from Southeastern Brazil 1. Wing membrane darkly infumated. Lateral portion of tergum 2 and entire terga 3 and 4 densely covered with simple dark setae, discs not distinctly different from marginal zones, except for short lateral bands of plumose dark setae on T3–T4 (fig. A3). Simple setae on mesoscutum about as long as plumose setae. Female: medial portion of clypeus entirely punctured, except for a narrow basal stripe .... xanthopyga (Parana´, Rio Grande do Sul, Santa Catarina) — Wing membrane with a yellow tint. Marginal zones of T2–T4 with distinct bands of plumose setae, usually pale yellow (sometimes black), setae on tergal discs distinctly shorter compared to those on marginal zones (figs. A1, A5). Female: a few simple 2006 57ROZEN ET AL.: TAPINOTASPIDINE BEES AND CLEPTOPARASITES setae on the mesoscutum distinctly longer than plumose pubescence; medial portion of clypeus with a narrow longitudinal stripe without punctures ............ 2 2. Marginal zones of T2–T4 only weakly depressed in relation to discs laterally (fig. A1). Pubescence on lateral portions of discs of T3–T4 not particularly sparser than on T2, except on T4 of a few males with weakly pubescent terga. Female: last three metasomal terga usually with integument bright reddish brown (rarely dark brown); clypeus with only a medial longitudinal depression or at most with a faint ridge; punctures on disc of tergum 2 almost reaching center of sclerite. Male: sternal pads less developed and occupying less than three-fourths of the sclerite width, tips of setae of posterior fringe not meeting in the middle portion of sclerite (fig. A2) .....haemorrhoidalis (Parana´, Rio de Janeiro, Santa Catarina, Sa˜o Paulo) — Marginal zones of T2–T4 distinctly depressed in relation to discs laterally (fig. A5). Pubescence on lateral portion of discs of T3–T4 distinctly sparser than on T2. Female: metasomal terga with a dark brown to black integument; clypeus with a weak, but distinct, medial longitudinal ridge; central one-third of tergum 2 almost impunctate. Male: sternal pads strongly developed and occupying over three-fourths of the sclerite widths, setae of posterior fringe distinctly long, their tips meeting in the middle portion of sclerite (fig. A6) schrottkyi ..................(Parana´, Santa Catarina, Sa˜o Paulo) Protosiris gigas Melo, new species Figures A9–A21 C OMMENTS AND D IAGNOSIS : The genus Protosiris contains four described and a few additional undescribed species (Roig-Alsina, 1989; Shanks, 1986; Melo, unpubl.). The new species proposed here, Protosiris gigas, is most similar, both in size and structure, to P. caligneus (Shanks) (comparisons based on male and female paratypes from the type locality deposited at AMNH and MZSP). Protosiris gigas can be separated from P. caligneus by its abundant yellow marks (uniformly reddish to dark brown in P. caligneus), lower paraocular area covered by plumose pubescence (mostly simple in P. caligneus), slightly longer decumbent hairs on middle portion of upper frons, mesoscutum strongly protruding on its midanterior portion (more pronounced in the female), and with a corresponding deep sulcus along the midline (fig. A18), posterior portion of male scutellum more evenly convex, medial sulcus only weakly indicated (scutellum more bulging in P. caligneus, medial sulcus more deeply marked), basal depression of metaposnotum smooth and without rugulae (fig. A12) (finely microreticulate and with lateral rugulae in P. caligneus), tergal setal punctures relatively sparse (very dense in P. caligneus, punctures about 1–2 puncture diameters apart) and lateral portions of terga 2–4 with only erect setae (in P. caligneus, with abundant short, decumbent hairs, similar to those of tergum disc). D ESCRIPTION :Holotype male. Body length: 14 mm; maximum head width: 3.1 mm; forewing length: 11.5 mm. Color: head and thorax mostly pale yellow, with many reddish to dark brown areas, as shown in figures A9 and A10; scape, pedicel, and 1st flagellomere dark brown, remaining flagellomeres dark reddish brown. Wing membrane brownish yellow infuscated, veins brown, pterostigma reddish brown. Metasomal terga 1–5 pale yellow at base and broadly brown apically; T6–T7 entirely dark brown. Sternum 1 dark brown at base and apically, its middle portion pale yellow; S2–S4 largely pale yellow basally and with triangular-shaped, apical brown band (in middle, band occupying about half of sclerite’s length); S5–S6 mostly dark brown. Pubescence: lower paraocular area covered with plumose pubescence. Longest erect setae on lateral portion of mesepisternum about as long as width of 2nd flagellomere (0.3–0.32:0.31). Lateral ventral portions of metasomal terga with only erect setae; S5 with a distinct apical band of plumose pubescence, setae directed to the middle portion of sclerite. Integumental surface: integument very smooth and polished. Setal punctures fine and inconspicuous, except for a few relatively strong punctures laterally on the clypeus and posteriorly on lateral portion of mesepisternum (weaker than those on clypeus). Metapostnotum mostly smooth, microreticulation almost imperceptible, basal depression well developed, its surface without longitudinal rugulae and only with a few transverse, inconspicuous rugulae laterally at the base (fig. A12). Setal punctures on posterior halves of discs of terga 1–4 about 3–4 puncture diameters apart transversely and 5–10 diameters apart longitudinally, slightly denser on basal halves of sclerites. Structure (measurements in mm): head about 1.3 3 wider than long (3.10:2.40); inner orbits nearly straight and parallel (upper to lower interorbital distance, 1.69:1.70); eye 1.8 3 as long as width at eye’s midlength (1.85:1.02); mandible bidentate apically, about 2.9 3 longer than its outer basal width (1.58:0.55); clypeus about 2.4 3 wider than long (1.69:0.71), distinctly protuberant, mid-