The move to open access and growth: experience from Journal of Hymenoptera Research

Seven microsatellite loci were isolated and characterized from a microsatellite enriched genomic library for the stingless bee Scaura latitarsis. Primers were tested in 12 individuals. The number of alleles per locus ranged from 3 to 6 (mean = 4.29) and observed heterozygosity ranged from 0.00 to 0.75 (mean = 0.40). Cross-species tests showed successful amplification for Scaura atlantica, Scaura longula, Scaura tenuis, Schwarzula timida, Schwarziana quadripunctata, Plebeia droryana, and Plebeia remota.

Two groups of Hymenoptera stand out as receiving considerable attention in terms of published articles: Braconidae and aculeate wasps. In 2011 and 2012, a total of a dozen articles were published about Braconidae and they make up for over half of the top ten most viewed articles (Table 1). The majority of the most accessed articles comprise revisionary studies, including the large-scale revision of the genus Quartinia in Southern Africa (Gess 2011a, b;2012a, b), with part IV being the most viewed article (Table 1). Eleven,or 19%, of all published articles in 2011/2012 had behavioural ecology as their main subject, with a prevalence of articles concerning aculeate wasps (Alcock 2011, Buys 2012, Nalepa 2012, Dalmazzo and Roig Alsina 2012, Barthelemy and Pitts 2012, Sarzetti et al. 2012, Wcislo et al. 2012. Although there is a predominance of articles that have systematics, phylogeny, and ecology as their prime subjects, there have been several articles dealing with important, more general subjects. Gates and Buffington (2011) describe innovative techniques for processing the increasing number of parasitoid specimens that are obtained through biodiversity studies. The tools developed and provided by Seltmann et al. (2012) in their article describing the Hymenoptera Anatomy Ontology represent an important milestone for the future of sustainable digital publication, allowing authors to reference their hymenopteran anatomical terminology to well-defined and illustrated anatomical concepts.
Authors benefit from the move to the Pensoft online editorial platform in that they can expect a faster turnaround time for their manuscripts, usually ranging from one to a few months from submission to publication. The open access model guarantees a better dissemination and visibility of their articles through the simple but important fact of accessibility, augmented by tailored use of cutting-edge Web 2.0 technologies. Manuscripts are submitted through a highly automated online editorial management system that provides authors and editors with a more streamlined editorial pathway. The move to electronic publication also means that there is no page limit for manuscript length, although for large revisionary works, checklists, catalogues, monographs, etc., there is the option of separate publication as a supplement. Nor is there a limit (or extra charge) for colour illustrations or the integration with multimedia (e.g. as in Wcislo et al. 2012), and authors are encouraged to submit non-printable matter like movie clips, sound recordings, or 3D animations. At the same time, we continue to produce a high resolution full-color printed version with a subscription option for individuals and institutions, and Society members benefit from a discount on printed copies and open access fees.
The technological development of JHR continues and the next step will not delay too long. JHR will be transferred in 2013 to the brand new Pensoft Journal System (PJS 2.0), a novel editorial management system that will greatly facilitate the editorial process for our authors, reviewers and editors through improving usability and navigation. PJS 2.0 will offer a number of additional tools, that can be used optionally by the JHR authors, such as community and public peer review, and an online, collaborative, article-authoring tool (Pensoft Writing Tool, PWT) that provides a large set of predefined, but flexible, templates of different types of article. In the PWT environment, authors can work collaboratively on their manuscripts online and may also invite external contributors, such as mentors, potential reviewers, linguistic and copy editors, colleagues, etc., who may watch and comment on the text during the manuscript preparation. The PWT will also facilitate upload of occurrence data in DarwinCore, provide automated registration of new taxa in ZooBank, import of references, embed multimedia, compose plates from single images, and many other features. Guyana, Suriname, Peru, Bolivia, and Brazil (Camargo and Pedro 2012). The large geographic distribution and the well reported female philopatric behavior in stingless bees (Nogueira-Neto 1954, Brito and Arias 2010, Francisco and Arias 2010 suggest that S. latitarsis populations should be highly differentiated. In general, population studies apply combined molecular markers of nuclear and cytoplasm origins. Mitochondrial polymorphism is easily assessed due to universal primers already described (Simon et al. 1994). However, the same is not true for nuclear markers. Microsatellites have been the nuclear marker most used in population genetic studies. As they present Mendelian and biparental inheritance, it is possible to evaluate the parental contribution, especially from males when mitochondrial data are also available (e.g. Estoup et al. 1996, Franck et al. 2001, Quezada-Euán et al. 2011. For future population studies, our first goal was to construct a microsatellite-enriched genomic library to isolate and characterize microsatellite loci for S. latitarsis. Total DNA of 12 workers was extracted using a phenol/chloroform protocol. The microsatellite-enriched genomic library was constructed according to Billotte et al. (1999) with some modifications (Brito et al. 2009). From 96 selected colonies of transformed Escherichia coli DH5α lineage, 48 were sequenced and 25 contained microsatellite sequences. Fifteen of these were chosen for primers design based on sequence quality. The software Primer3 was used (Rozen and Skaletsky 2000) to indicate the best primer sequences. Primers were tested on 12 workers of S. latitarsis from seven different locations in Brazil: Ribeirão Preto, SP (n = 2), Cajuru, SP (n = 2), Pedregulho, SP (n = 2), Ribeirão Cascalheira, MT (n = 2), Nova Xavantina, MT (n = 1), Manaus, AM (n = 2), and Londrina, PR (n = 1). Amplification reactions were conducted in a volume of 20 µL containing 2 µL of the PCR reaction buffer (Invitrogen), 1 µL of dNTP 2 mM each, 0.6 µL of MgCl 2 50 mM, 0.4 µL of each primer 20 µM, 15 ng of template DNA and 0.3 µL of Taq DNA polymerase 5U/µL (Invitrogen). PCR conditions were the following: 93 °C for 4 min, 35 cycles of denaturation at 93 °C for 40 s, annealing at 60 °C for 50 s and elongation at 72 °C for 40 s. A final extension step of 72 °C for 5 min was performed. PCR products were separated on 5.6% polyacrylamide gels and stained with silver nitrate. Allelic richness (k), observed (H O ) and expected (H E ) heterozygosity were calculated in FOFPOP 2.0 (Francisco 2009). GENEPOP 4.1.4 (Rousset 2008) was used for Hardy-Weinberg equilibrium (HWE) and linkage disequilibrium tests. Bonferroni corrections were used for multiple comparisons.
Seven out of 15 primer pairs generated fragments of expected size, and they were polymorphic (Table 1). The number of alleles ranged from 3 to 6, with an average of 4.29. Values of H O ranged from 0.000 to 0.750 (mean = 0.405) and H E from 0.531 to 0.736 (mean = 0.664). After Bonferroni correction, only the loci SLAT 16 and SLAT 40 showed significant deviation from Hardy-Weinberg-expectations. Linkage disequilibrium was detected after Bonferroni correction between SLAT 16/SLAT 24, SLAT 24/SLAT 43, and SLAT 40/SLAT 43. Deviations from HWE and linkage disequilibrium are expected when individuals are sampled from genetically differentiated populations (Brito et al. 2009). Using the same conditions aforementioned we tested the primers for the polymorphic loci in seven stingless bee species: Scaura atlantica, Scaura longula, Scaura tenuis, Schwarzula timida, Schwarziana quadripunctata, Plebeia droryana, and Plebeia remota. Cross-species amplification was 100% efficient for SLAT 10 and SLAT 44 (Table 2). However those loci were the only ones amplified in S. longula and S. tenuis. At least five out of seven primer pairs were successfully amplified in species from three other genera.
The microsatellite loci described here will be very helpful to gather molecular data of S. latitarsis to understand the population structure of this widely distributed species. If queen philopatry is confirmed for this species, these microsatellite loci will allow an evaluation of male's contributions as gene flow mediators.

acknowledgements
We thank Susy Coelho for technical support, Dr Anete P. de Souza for helping with the genomic library protocol, Fundação de Amparo à Pesquisa do Estado de São Paulo for PhD scholarships to FOF (2008/08546-4) and FCP, and for financial support (2004/15801-0; 10/50597-5) ter group of Megachilinae (Roig-Alsina and Michener 1993, Engel 2001, Michener 2007, Gonzalez et al. 2012 Moure and Melo 2007). Snelling (1983) suggested that L. huberi was adventive to South America and probably a synonym of the Indo-Australian species L. atratus Smith given the morphological similarity between the two. Such a hypothesis is also supported by the wood-nesting habits of Lithurgus, which facilitates dispersion across great distances. We confirmed the close morphological similarity between L. atratus and L. huberi, as noted by Snelling (1983). However, we also noted some morphological features in both sexes of L. huberi that seem to be consistent and that may prove to be useful in species recognition. Accordingly, the purpose of this note is to document and illustrate those characters, as well as record L. huberi for the first time for Paraguay and supplementing this with new distribution records for Argentina (vide infra).

Materials and methods
We examined (V.H.G. & M.S.E) the type series of L. atratus as well as of L. dentipes Smith, a species that has been considered a synonym of L. atratus, deposited in The Natural History Museum, London (NHML). Additionally, we examined 42 specimens of L. atratus from India, Celebes, Borneo, Solomon Islands, and Australia deposited in the Snow Entomological Collection (SEMC), Division of Entomology, University of Kansas Natural History Museum, Lawrence, Kansas, USA, and the U.S. National Pollinating Insects Collection, Bee Biology and Systematics Laboratory (BBSL), Utah State University, Logan, Utah.
Photomicrographs were prepared using a Canon 7D digital camera attached to an Infinity K-2 long-distance microscope lens, and were assembled with the Com-bineZM TM software package. Specimens are deposited in SEMC and the entomological collection of the Museo de La Plata, Argentina (MLP). Occurrence data for L. huberi were plotted using CorelDRAW ® X5 and were taken from the literature (i.e., Ducke 1907, 1908, 1910, Camillo et al. 1983, 1994, Wittmann and Hoffmann 1990, Roig-Alsina 2006, Pick and Schlindwein 2011

Results
Females of L. huberi primarily differ from those of L. atratus by the facial prominence. In L. huberi it is more strongly punctate and more depressed along the epistomal sulcus than in L. atratus (cf. Figs 1,3 vs. 2,4). Likewise, males of L. huberi differ from those of L. atratus in the inner glabrous surface of the metabasitarsus, which is broader and more projected (cf. Figs 5 vs. 6). Although subtle, such morphological differences appear to be consistent across the specimens examined. However, we have had access to a limited number of individuals from a restricted set of localities and cannot rule out that such differences represent mere geographic variations.
Despite the small number of species in Lithurgus s.str., the status of most of them remains questionable, particularly those from Southeast Asia. For example, at least eight 'species' that are closely related to L. atratus have been suggested to represent a single taxonomic unit (Michener 1965), although they could be a complex of rather cryptic species. Some of these are practically indistinguishable from L. atratus, differing mostly in body size (e.g., L. atratiformis Cockerell). Undoubtedly, a revision of the group that includes a great number of specimens from multiple locations (thereby necessitating extensive fieldwork as existing collections have significant gaps for Lithurgus across its distribution) is needed before any taxonomic action can be taken with certainty. Such a work would ideally be accompanied by molecular data to further test putative species boundaries as well as to elucidate possible routes and times of dispersion.
In conclusion, we support the view that L. huberi is closely related to the Indo-Australian L. atratus species or species group and, if dispersed by human activity, it might have reached South America at least 100 years ago when it was described (Ducke 1907). However, we show here consistent, albeit somewhat subtle, morphological features to distinguish L. huberi from L. atratus. For the time being we recommend that they should be treated as separate species.

acknowledgements
We are grateful to David Notton for his constant and valuable support and who kindly arranged the loan of the syntypes of L. atratus and to two anonymous reviewers for comments and suggestions that improved this note. This project was supported in part by US National Science Foundation grant DBI-1057366 (to M.S.E) and by the Department of Biological Sciences, Southwestern Oklahoma State University. This is a contribution of the Division of Entomology, University of Kansas Natural History Museum.  insects can be seen in Fig. 3. Eggs from this stick insect were regularly collected during these infestations and two new species of chrysidid egg parasites, Exova tunana sp. n. and Cladobethylus insularis sp. n. described below were reared from the eggs. There is no information on the ecology of these parasitoids except for the rearing information.

References
There are seven named species of Cladobethylus and two species of Exova described prior to those described below (Kimsey andBohart 1991, Kimsey 2011). The new species of Cladobethylus described below is the second one recorded from New Guinea. Exova is known from northern Australia and Fiji, so it is not surprising to find a species in Papua New Guinea.  Diagnosis. This is the largest bodied species of Cladobethylus, with males averaging twice the length of the known other species. Male insularis can be distinguished from aquilus, the only other species known from New Guinea by the longer malar

5.
space (4 midocellus diameters versus 3.5 in aquilus), pronotum not blue-tinted, much broader zone of cross-ridging in the scapal basin, legs brown instead of yellow (aquilus) flagellomere I shorter (4.5× as long as broad versus 5× in aquilus) and flagellomere XI shorter (5× as long as broad versus 6× in aquilus). Features of female insularis not shared with other Cladobethylus include the bicolored antenna and legs, long clypeus (0.8 midocellus diameter long versus 0.3-0.6 midocellus diameters in other species), and narrow distance between the midocellus and nearest eye margin (up to 2 midocellus diameters versus 2.6-2.7 midocellus diameters in the others).
Metasoma. Tergum I polished with few tiny highly scattered punctures; tergum II with two large ovoid patches of small punctures, 1 puncture diameter apart, separated medially by impunctate band and with broad apical impunctate band; terga III-IV with tiny punctures 1 puncture diameter apart, becoming sparser toward apical margin.
Color. Head, meso-and metasoma black with metallic bluish green highlights dorsally on head, pronotum and scutellum, sometimes faint on metasomal terga; mandible and antenna dark brown; legs light reddish brown, except coxae dark brown to black basally; wing membrane brown-tinted, darkest in vicinity of stigma; wing veins dark brown.
Etymology. The species is named after the host species. Diagnosis. This is the largest bodied species of Exova. Female tunana can be distinguished from fijiensis Kimsey by the long spine-like propodeal angle (a feature shared  with tetraspina), which is conical in fijiensis. Female fijiensis can be distinguished from tetraspina by the more weakly produced medial propodeal projections and the highly polished and smooth metapleuron and propodeum. Male tunana can be distinguished from fijiensis by the shorter distance from the midocellus to ocular margin (2 midocellus diameters or less in tunana versus 2.5 midocellus diameters in fijiensis), flagel-lomeres I and II equivalent versus I much longer than broad than II in fijiensis and the body with bluish green tints versus brassy in fijiensis.
Color. Head, meso-and metasoma black with metallic bluish green tints dorsally; antenna black, legs pale yellow; wing membrane brown-tinted, veins dark brown.
Metasoma. Polished; tergum I with fine dense scratches in two touching posteromedial patches, scratches parallel and curving laterally; tergum II with two anteromedial patches of dense curved scratches subtended by large medial patch of dense posteriorly curved fine scratches; terga III and IV smooth with scattered tiny punctures.
Etymology. The species is named after the collection site in Northern Province, PNG.

acknowledgements
This study was made possible by the efforts of Entomology Section staff at the PNGO-PRA Higaturu Research Centre, Northern Province. CFD acknowledges The Director of Research, PNGOPRA for permission to publish this article. abstract The genitalia of male ants are interpreted in the context of the broader Hymenoptera. For the first time muscle homologies are established for twenty six species of ants in nine subfamilies: Amblyoponinae, Cerapachyinae, Dolichoderinae, Ecitoninae, Ectatomminae, Formicinae, Leptanilloidinae, Myrmicinae and Ponerinae. Fifteen muscles in total are found in the external genitalia of male ants and all are homologous with the musculature of basal Hymenoptera (Schulmeister 2001). Secondary fusion, reduction or losses of muscles have occurred in different lineages. From generalized to derived, the functional morphology and qualitative mechanics of three taxa, Formica obscuripes Forel 1886, Messor andrei Mayr (1886) and Labidus coecus Latreille (1802) are described and compared. Special reference is made to the Ecitoninae, where the work of Birket-Smith (1981) is reinterpreted and volsellar homology is clarified. The true digitus of Labidus is revealed to be a small sclerite at the base of the distal volsellar element which is re-identified as the cuspis.

Keywords
Anatomy, army ants, kinesiology, morphology, reproductive biology introduction Insect male genitalia are spectacular mechanical adaptations with diverse morphology and function. Examples of this diversity include the twin hypodermic intromittent organs of the Drosophila bipectinata complex (Kamimura 2007), the sperm scoopulas of Odonata (Córdoba-Aguilar et al. 2003), 'Symphytan' suction cups (Schulmeister 2001), the copu-latory hooks of Metrioidea (Chrysomeloidea; Flowers and Eberhard 2006), and genitalia modified as stridulatory instruments for courtship (Micronecta, Corixidae; Sueur et al. 2011) or for stridulation in copulo (Tipula, Tipulidae; Eberhard and Gelhaus 2009). The diversity of male genitalia may also be cryptic and in the realm of functional morphology. For example, male Drosophila melanogaster with ablated genital clasper mechanosensory sensillae were found to copulate less frequently, and unilaterally ablated males mounted females skewed contralaterally from the typical position (Acebes et al. 2003).
The morphology and mechanics of male ant genitalic musculature remain undescribed for all ~12,000 species of ants (Bolton 1995), with the exception of Labidus coecus, Dorylus (Anomma) funereus (Birket-Smith 1981), and Myrmica kotokui Forel 1911 (Ogata 1991). The function of ant genitalic valves has been described from primary observations for few taxa (Allard et al. 2002(Allard et al. , 2006Baer and Boomsma 2006), while the entire sequence of copulation has never been reported; a composite description is as follows. Copulation of all non-'Symphyta' Hymenoptera is orthandrous (i.e. male atop female, facing same direction; Schulmeister 2001), which holds true for ants. Males of Carebara vidua F. Smith 1858 and Plectroctena mandibularis F. Smith 1858 were observed to use their genitalia to probe the apex of the female's gaster while grasping her with his legs (Robertson and Villet 1989;Villet 1999). Probing is likely a general requisite of ant copulatory behavior (Ross 1982;Schulmeister 2001) followed by penetration which may be via the penisvalvae apices, the digiti or both. The penisvalvae of the aedeagus anchor the male inside of the female's bursa copulatrix or the mussel organ of Atta (Baer and Boomsma 2006), sometimes causing female scarring (Kamimura 2008) while the endophallus is putatively everted (Allard et al. 2002; although see Schulmeister 2001). The typically opposable digitus and cuspis have been observed to clasp the terminal female sternite in a ponerine Diacamma sp. (Allard et al. 2002). The parameres serve as a clamp-type attachment mechanism (Gorb 2008;Ogata 1991;Schulmeister 2001), potentially enhanced by wet adhesion from a putative telomeral gland (Francoeur 1988). The male will often release grasp the female and hang on to her by his genitals only (Robertson and Villet 1989) and will remain anchored within the female in this position during a variable period of time until copulation is terminated (Hölldobler and Wilson 1990;Foitzik et al. 2002).
Knowledge of genitalic functional morphology contributes to our understanding of the sexual selection pressures which act on these organs (Eberhard 2010) and may possibly be used to make predictions about the reproductive biology of ants. This is of significance given the effects of mating systems (Boomsma et al. 2005), paternity assurance (Baer 2011;Baer and Boomsma 2006) and polyandry (Hughes and Boomsma 2007;Robinson and Page Jr. 1989) on social dynamics. Furthermore, understanding male ant morphology is important for taxonomy (Fisher 2009;LaPolla et al. 2012;Yoshimura and Fisher 2012) and systematics (LaPolla 2004;Brady and Ward 2005). Thus the musculature and gross morphology of male ant genitalia represents an appealing and untapped source for new, potentially useful morphological characters (Keller 2011;Schulmeister 2003;Song and Bucheli 2010), which may also serve as synapomorphies for clades identified via molecular techniques.
The goal of this paper is to discuss the homologies of male ant genitalic musculature in the context of the broader Hymenoptera and to provide descriptions of the functional mechanics of three distinct forms. The work of Schulmeister (2001Schulmeister ( , 2003 has identified and clarified the muscular homologies within the basal Hymenoptera, or the paraphyletic 'Symphyta' (Sharkey 2007). While the external genitalia of male ants have been characterized for several subfamilies (Ogata 1991;Yoshimura and Fisher 2009, 2012Yoshimura and Onoyama 2002), musculature has been completely overlooked excepting Birket-Smith (1981) and Ogata (1991), the former of which is here reassessed.

Materials and methods
All specimens examined in this study were from the author's personal collection (BEBC) or the Longino collection (JTLC, University of Utah). Collections in 95% ethanol were made in Central America, and in the states of Arizona, California, Tennessee and Washington. Specimens were softened by boiling in water for 2-5 minutes. Dissections were carried out in petri dishes, with or without wax and minuten pins. A Leica MZ16 microscope with maximum magnification of 115× and mounted with a JVC Digital Camera KY-F708 3-CCD was used for dissections. Micrographs were generated for illustration using Auto-Montage Pro Syncroscopy (Synoptics Ltd., Frederick, MD USA) software. For especially small males, temporary mounts of minute sclerites and musculature were prepared for observation with a Leica DME compound microscope using glycerin as the mounting medium.
General hymenopteran terms preferred by the Hymenoptera Anatomy Ontology (Yoder et al. 2010) are used when homology is clear (Table 1). For an overview of terms applied to the genitalia of ants and other Hymenoptera, refer to Fisher (2011) andSchulmeister (2001) . Walker Jr. and Liem's (1994) concepts of origin and insertion are used when discussing musculature: origins are broad anchoring attachments; while insertions are the narrow, usually distal or posterior, ends of muscles which transfer force to a movable element. The muscles of hymenopteran male genitalia are named alphabetically (Boulangé 1924;modified by Schulmeister 2001): anterior to posterior, ventral to dorsal, medial to lateral; starting with sternum IX and ending with the muscles of the parameres. Thus lettering begins on the anteroventral apex of sternum IX with muscle a and ends, in the Formicidae, with the intrinsic muscle t of the parameres.
The mechanics of sclerite movement were assessed qualitatively by examining the angles of muscular origin and insertion, and the attachment or position of the sclerite relative to the local components of the genital capsule. Simple physical models were used to predict the mechanics of muscular contraction and element movement. Formica obscuripes Forel 1886, Messor andrei Mayr (1886) and Labidus coecus Latreille (1802) were chosen as models for functional morphology because their genitalia are well developed, represent cases of relatively generalized to specialized morphology, and Table 1. Anatomical terms of the external genitalia used herein, excluding specific membrane and apodeme names. Hymenoptera Anatomy Ontology (HAO) Uniform Resource Identifiers (URIs) link to concept definitions. Synonyms discussed or used are indicated.

Term HAO URI Synonyms
among these three taxa all the observed genitalic muscles of male ants are represented. The more complicated movements described here should be treated as predictions to be tested either via direct observation of copulation or via neuroethological studies.

Genitalic morphology (Figs 1-17)
External genitalia (Figs 1-16) The external genitalia of ants are composed of five elements: sternum IX, the cupula, and the paired valves of the penisvalvae, volsellae, and parameres (Figs 1-4). Sternum IX (Fig. 4) is produced anteromedially into a process called the spiculum (Schulmeister 2001) and has a cranial apodeme traversing the mesal face (Forbes 1967;Forbes and Hagopian 1965), which may be a modified sternal antecosta. The term cupula is preferred over basal ring, as the latter refers to non-homologous structures in the Protura and Diptera (Tuxen 1970). The cupula (Figs 1, 2, 5-7) is a hymenopteran synapomorphy (Sharkey 2007) and is variable in form for ants. The anterior opening of the cupula is the foramen genitale (Fg, Figs 5-7;Schulmeister 2001), and is constricted to varying degrees across different lineages. In the ecitonines, the foramen genitale is reduced to a miniscule aperture through which only the ductus ejaculatorius and tracheae pass (Fig. 6). In all ant taxa I examined, the ventral gonocondyle (Schulmeister 2003) of the cupula was absent. Lateral to medial, the three paired valves are the parameres, volsellae, and penisvalvae (Figs 2 and 3). Homology of the parameres of the Formicidae with those of other Hymenoptera is reasonable, as the following complex characters are shared: 1) at least four of the five penisvalvae muscles attach to the basimeres; 2) an intrinsic muscle has an origin in the basimere and inserts in the telomere; and 3) a membrane is present at the telomere articulation with the basimere. It should be noted that this telomeral membrane is variable in development for the Formicidae, from well-developed to ab- sent across numerous clades (Yoshimura and Fisher 2011). Thus the specific homology of the telomere with the basal hymenopteran harpe is dubious; therefore the application of "basimere" and "telomere" is preferable over "gonostipes" and "harpe" for ant genitalia. The parameres of ants are thus composed of a dish-like basimere and distal telomere (Figs 1 and 3). A ventromedial extension of the basimere is the origin site of several volsellar and penisvalvar muscles, and is termed the gonostipital arm (Ga, Fig. 8); this structure is not a synapomorphy of the Myrmicinae as claimed by Ogata (1991) because the gonostipital arm is present in other ant subfamilies (this study) and in many other hymenopteran lineages (Schulmeister 2001(Schulmeister , 2003. The volsella, another hymenopteran synapomorphy along with the cupula (Sharkey 2007), is composed of the digitus which is basolaterally articulated with the parossiculus (Figs 8-12). The parossiculus is generally divided into the basivolsella and cuspis (= distivolsella, Figs 9-11). As the volsella is unique to the Hymenoptera, term preference is ambiguous; however, given that distivolsella has never been applied to Formicidae, I propose that "cuspis" be maintained. The cuspis is usually setose (Schulmeister 2001); this is indeed the case for all ant taxa examined (Cs , Figs 9-12).
Although the term aedeagus has a long history and is frequently used in the myrmecological literature to describe the middle-most valves (Brown Jr. 1960;Serna and Mackay 2009;Smith 1942;Taylor 1965;Yoshimura and Fisher 2012), this term is correctly applied to the whole intromittent apparatus which includes the sclerites, or penisvalvae, and the endophallus. The term "penisvalva" is preferred over "aedeagal plate" as the former term is virtually restricted to and broadly used for the Hymenoptera and represents the homology of the sclerite in question between ants and other hymenopterans, while the latter term is used for non-homologous structures among several insect orders ("penisvalva" e.g. Apoidea, Pesenko and Astafurova 2006;"Parasitica", Karlsson and Ronquist 2012;Vespoidea and 'Symphyta', Schulmeister 2003; "aedeagal plate" e.g. Hemiptera : Rivers 1948;Diptera: Amorim 2007; Hymenoptera: Ogata 1987). The homologies of the cranially-directed apophysis and the distal component of the ant penisvalva with the valvura and valviceps are highly likely; the location and musculature of the apophysis are equivalent to those in the basal Hymenoptera and Aculeata (Schulmeister 2003). I thus adopt the terms valvura for the arm-like apophysis and valviceps for the distal component of the penisvalva (Figs 13-15; Schulmeister 2001). The penisvalvae are joined mediodorsally via a tough membrane (Pm ,  Figs 2 and 3), and an aedeagal gland is present in various forms between the valves (not figured; Hölldobler and Engel-Siegel 1982). A lower oblique carina (Ward 2001) is present in some taxa along the ectal face of the valviceps (Lp , Figs 13-15), forming a channel-like groove in formicines. Forbes (1952) hypothesized that the channel functions as a "sperm gutter"; this seems unlikely as the channel is separated from the phallotrema by the blade of the valviceps. I hypothesize that the lower oblique carina functions as an arch, transferring the force of the pensivalvar muscles to the valviceps apex.
Internal genitalia (Figs 16 and 17) As the aedeagus is the interface between the internal and external genitalia, a general description of the internal genitalia is warranted (Fig. 16). Spermatozoa are produced in the follicles of the paired testes and pass through the vas deferens (Snodgrass 1935;Schulmeister 2001) into the vasicula seminalis where they are kept until copulation. The two vasiculae seminalae join together into the ductus ejaculatorius which opens into the endophallic bladder via the gonopore ( Fig. 17; Snodgrass 1935). The endophallic bladder was formerly known as the aedeagal bladder, but it is definitely a component of the endophallus (Snodgrass 1935); its function remains unknown . A sclerite is present in the ductus ejaculatorius which has been identified as the fibula ducti in 'Symphyta' (Schulmeister 2003), the wedge in Neivamyrmex (We, Fig. 16; Forbes and Do-Van-Quy 1965) and sperrkeil in Formica rufa (translates contextually to keel-shaped valve, Clausen 1938). Although this sclerite was found in most taxa examined, it was not homologized across the Hymenoptera or among ant subfamilies as its morphology was not explicitly examined. The endophallus opens externally via the phallotrema at the bases or the dorsal apices of the penisvalvae (Pt, Fig. 3).

Musculature
The homology of genitalic musculature for each species examined is summarized in Table 2. Fifteen total muscles were observed among the examined ant taxa. Considerable variation exists in the morphology of tergites IX and X, but as these are not primary components of the genitalia these were excluded from this investigation. Three muscles, a, b and c, have their origins on sternum IX and insert on the cupula (Figs 4, 5-7). Muscle a is paired and originates on the spiculum from which it diverges to the ventrolateral margin of the cupula, except in the ecitonines where a is inserted in a median groove ventrad to the reduced foramen genitale. The origin of the unpaired muscle b is along the cranial apodeme; the muscle converges anteriorly to its ectal insertion on the medioventral margin of the cupula. Each element of the paired muscle c originates on an anterolateral lobe of the sternum and inserts on the ventrolateral margin of the cupula.
Four muscles, d, e, f, and g, have their origins on the cupula (Figs 1-3, 5-8). Muscles e, f, and g are paired, while d is unpaired. The origins of the cupula muscles are usually associated with an apodeme: muscle d with the ventral, e with the lateral, f with the dorsolateral and g with the dorsal apodeme (Figs 1, 2, 6). In general the gonostipital arms, or anteromedian projections of the basimeres, receive the insertions of d on the ventral margin and e ventrolaterally (Figs 1, 5, 7, 8). Both f and g insert ectally on the basimere, f on the lateral margin and g on the dorsal margin (Figs 1-3, 5-8). Loss of cupula muscles was observed for three species: Atta cephalotes L. (1758), Crematogaster nigropilosa Mayr 1870, and Leptanilloides sp. in which the cupula is reduced to a thin strip with only muscle e present. Muscles f and g are fused in Pheidole californica Mayr 1870, and muscle d could not be confirmed for Hypoponera opacior Forel (1893), Odontomachus chelifer Latreille (1802), and Cerapachys nr. augustae.
In total, five penisvalva muscles (Figs 13-15) were found; h, i, j, and k were consistently present except in the reduced genitalia of Leptanilloides sp. (Table 2), while l was lost in disparate taxa. These muscles are paired, with one element of each associated with a left or right penisvalva. Three of the penisvalva muscle origins are on the mesal face of the basimere: i on the ventral, j on the dorsolateral, and k on the dorsomedian face (j, k Figs 1-3; i Fig. 8). The origin of muscle h is variable, in some cases spanning the apices gonostipitum to the ventral apodeme of the cupula. Although the dorsome- dial basimeral area where k originates has been termed the parapenis in basal Hymenoptera (Schulmeister 2001), this term is not used for the Formicidae due to uncertain homology. Muscle l has its origin on the basal dorsomedian area of the telomere (Figs  3 and 8). The insertions of h and j are on the ventral and dorsal faces of the valvura apices, respectively. Muscles i and l are inserted on the penisvalva lateral apodeme (Figs 13-15;Ward 2001), an equivalent structure to the ergot of basal hymenoptera (Schulmeister 2001). Finally, k is mesally inserted on the base of the valvura or the valviceps (Fig. 15).  Ogata (1991) in which the following muscles were mislabeled: f, i, j, k, qr, t.
Muscle p has its origin on the internal lateral face of the basimere. Reduction of the cuspis and loss of p was observed in Aphaenogaster nr. rudis, Messor andrei, Pheidole californica and Hypoponera opacior. This contrasts with the intrinsic volsellar muscle qr which was present in all but two taxa examined, and could not be confirmed for a third. The origin on qr is on the basivolsellar apodeme.
A single intrinsic muscle of the paramere, t, was found in several of the taxa examined. Muscles u and v (Schulmeister 2001) were never present. Muscle t is paired, with the left or right element originating anterolaterally on the respective basimere ( Fig.  8) and inserting on the telomeral corium (Fig. 1). Presence of t was inconsistent; in the present dataset, loss is associated with telomere fusion to the basimere, in reduced genitalia, and with change in telomere function. In these circumstances, other muscles including the volsellar p and penisvalvar j are hypothesized to affect the grasping capacity of the parameres via elastic forces.

Formica obscuripes Forel, 1886
Among the taxa examined this species has relatively generalized genitalia, with all fifteen muscles present and without extreme modifications of the sclerites. The three muscles of sternum IX, a, b, and c, are present in this species (Figs 4 and 5). Simultaneous contraction of both elements of a contracts the genital capsule into the gaster to varying degrees. Antagonistic contraction of either the left or right element of a adducts the genital capsule towards the respective side, an action which is synergistic with the genital capsule protractor b. Muscle c inserts ventrolaterally on the ventral apodeme of the cupula, above the insertions of a and b (Fig. 5), and torques the genital capsule via elevation of the basal margin of the cupula toward the muscle's origin.
Muscles of the cupula rotate the genitalic valves within the cupula similarly to a ball-in-socket mechanism. Each muscle is inserted on the anterior margin of the basimere except for d, which inserts on the anteroventral face of the endophallic bladder (Figs 5 and 8). Muscle e adducts the valves ventrally, while g acts antagonistically to elevate the apices while contracting the valves. Muscle e also adducts the telomere to the mediosagittal plane by drawing the medioventral apices of the basimeres dorsolaterally. Contraction of f may affect torsion of the valves.
All five muscles of the penisvalvae are present in this species (Fig. 13). The penisvalvae can move independently along the mediosagittal plane and are capable of rotation, pivoting on their lateral apodemes. Elevation of the valviceps is caused by contraction of h, while depression is caused by j. Muscle k is inserted basally on the valviceps and may dig the basal teeth into the female's bursa copulatrix (Kamimura 2008) by drawing the valviceps anterad and adducting the apices ventrally. Rhythmic contraction of i may enhance this hypothetical function in order to gain greater purchase in the female via a motion analogous to mastication. Muscle l abducts the valviceps ventrolaterally swinging the apices apart, potentially to allow passage of ejaculate during copulation or to remove the sharply curved valviceps teeth from the female.
Two muscles, p and qr, are attached ectally and mesally to the cuspal apodeme of the volsellae, respectively, and act antagonistically to open or close the cuspides (Fig.  9). Adduction of the cuspis to the digitus is caused by contraction of qr, which bends the volsellar plate toward the penisvalvae while drawing the cuspal apex toward the digitus. The antagonistic abduction of the cuspis is induced by p, which extends laterally to the basimere from the ectal face of the cuspal apodeme (Figs 8 and 9).
The single intrinsic muscle of the paramere, t, is relatively well developed in this species. Contraction of t flexes the telomere ventrally via folding of the telomere corium. Upon relaxation of t, the elastic force of the sclerotic bridge between the basimere and telomere sclerite erects the telomere. Flexion of the telomere may also be affected by the penisvalvar muscle j which has a major portion of its origin along a mediodorsal inflection of the basimere. Intense contraction of j may roll the telomere ventrally via torsion of the inflection ectally, while also flexing the penisvalvae ventrally; this may function to strengthen the clamp-hold of the female.

Messor andrei Mayr, (1886)
With the loss of p, fourteen of the fifteen genitalic muscles are found in this species. Although the foramen genitale is somewhat constricted (Fg, Fig. 7), the muscular function of the cupula and sternum IX are not distinct enough from Formica obscuripes to warrant separate discussion. The clasping function of the paramere is maintained despite the replacement of most of the telomere corium by sclerite. In this case, there is very little telomere flexion. Rather, t adducts the apices of the telomere toward the mediosagittal plane while pulling the medioventral faces dorsally. The mesal face of the telomere has a cup-like lamina which, coupled with hypothetical secretions of the telomeral gland, may enhance the male's grasp on the female.
The valviceps is produced dorsally with a laterally-produced lamina (Lm, Fig. 2) which is divided into anterior and posterior lobes by a constriction, making the valviceps similar in shape to an anvil. The dorsal laminae may function as a passive anchor in the female's bursa copulatrix while the aedeagus is intromittent. Between the two valviceps, an ellipsoid chamber is formed with the penisvalvar membrane closing it dorsally. I hypothesize that this chamber acts as a mechanical sperm pump, forcefully pulsing ejaculate out of the anterior-directed phallotrema via contraction of l, which spreads the valviceps bases. Contraction of j may synergistically contribute to this hypothetical action via rotation around a vertical axis placed at the dorsal constriction, drawing the dorsal posterior lobes together while spreading the anterior lobes and valviceps apices ectally. Muscle j also adducts the valviceps ventrally. The basal wall of the valviceps, where k is inserted, is inflected dorsally and ventrally into a rough parabola with its vertex pointed anteriorly. This inflection divides k into dorsal levator and ventral contractor halves. Motion of l and k together may pull the valviceps anteroventrally, potentially digging the valviceps teeth into the female. Muscle h has most, if not all, of its origin along the endophallic bladder.
Clasping is not the function of this species' modified volsella (Fig. 10), since the digitus is completely fused with the setose cuspal shaft, and is in the form of a crescent with a basal flange and apical process. The apical component of the cuspis is reduced to a small ellipsoid plate that articulates with the digitus laterally (Cs, Fig. 10, inset). Muscle p has been functionally replaced by qr which is divided into dorsal and ventral halves by the flange-like basivolsellar apodeme (Ba, Fig. 10). The dorsal margin of the cuspis is produced into a lamina which twists ~180° around the basal-distal axis. The reduced cuspis apex acts as a pivot for rotation of the digital crescent, while the twisted lamina provides strength and elastic flexibility for torsion of the volsella. Ventral contraction of qr in the left volsella rotates the apex of the digitus medioventrally clockwise, while dorsal contraction rotates the digital flange dorsolaterally counterclockwise. Thus qr has a fine-tuned forward and reverse function via differential contraction of the dorsal or ventral sides coupled with the elastic torsion of the cork-screw cuspis. The copulatory function is hypothesized to be prying the terminal plates of the female open, akin to a lock-pick, via insertion of either the digitus or digital flange into the female followed by lifting or depressing torsion.

Labidus coecus Latreille, (1802)
The genitalia of Labidus have been greatly modified in form relative to the nonecitonine taxa examined. Sternum IX is produced apically into a bifid lobe and muscle c is absent. An external longitudinal groove extends along the ventral face of the egg-shaped cupula (Cu, Fig. 8) and receives the insertions of muscles a and b. The origin of b has migrated up the anterolateral lobes of the sternum; the muscle extends mesally to its insertion in the ventral groove just posterad the insertion of a, which is ventrad the foramen and is longitudinally elongated. This elongation of a along the curvature of the cupula prevents pronounced dorsal or ventral pitch during contraction of a. Muscle b may either torque the capsule contralaterally with contraction of the left or right half, or both halves together may adduct the apices of the genital valves ventrally.
Contrary to the arguments of Birket-Smith (1981), I found the cupula muscles of Labidus to have clear homology with the pleisiomorphic muscles d, e, f, and g (Fig. 4). Birket-Smith (1981) suggests that muscles e, f and g have been lost during the evolution of Labidus to be replaced by a "vaguely differentiated muscle, d'." I disagree with this interpretation: the muscular origins are identical with respect to association with cupula apodemes (Do, Dl, Lb, Fig. 4) while the muscles themselves are distinguish-able by seams between the bundles. Moreover, the insertions on the apodemes of the basimeres are relatively distinct (Ad, Ac, Av, Fig. 6), especially that of g which is broadest medially and tapers laterally. The lateral basimere apodeme is emarginate between the insertion of f on the dorsal and e on the ventral half. Muscle d is inserted on a cranially-directed anteroventral process of the gonostipital arm.
Contraction of the cupula muscles may cause splaying and movement in the mediosagittal plane. The cupula of all ecitonine species investigated is extremely strong and well-constructed to withstand the forces applied to it internally. A tall longitudinal apodeme is present along most of the length of the ring, from the foramen genitale to the apices gonostipitum. This apodeme may function as a strut, strengthening the cupula from the antero-posterior compression forces exerted by the cupular muscles. The anteroventral processes of the basimeres are extended anteriorly to a greater magnitude than the dorsal margins of the basimeres. Thus the dorsal margins of the basimeres act as a fulcrum for a third-order lever: synergistic contraction of d and e act as in-forces and cause the apices of the genitalic valves to swing ventrally. As muscle e converges mesally from its ventrolateral origin, it may also be able to fine-tune the yaw of the adduction motion. Muscle f contributes primarily to the spreading of the genitalic valves around a vertical axis of rotation between the apices gonostipitum and the bases of the valviceps.
With respect to the penisvalva muscles, Birket-Smith (1981) discussed a muscle he termed h'. This is not a novel muscle; rather it is the well-developed posterior portion of muscle h which extends into the basimere. This portion may be seen in several other taxa, including Formica obscuripes in which h is not fully divided. Muscle k has the typical origins and insertions on the dorsal anteromedian faces of the basimeres (Fig. 3) and mesal face of the valvurae (Fig. 15). Muscle h has its origin across the length of the cupula on the cupula's anterior wall, opposite the insertion of a. The origin of i extends from the ventromedial to the anterolateral face of the basimere. This lateral projection was interpreted by Birket-Smith (1981) to be the only component of i. Comparison with Formica obscuripes again clarifies the homology, as muscle i of the formicine may be seen to extend to the lateral face of the basimere as well.
Four of the five penisvalva muscles seem to act synergistically by contributing forces to the ventral adduction of the penisvalva apex, while only h acts antagonistically. The muscles all act as in-forces for moving the penisvalva as a lever and may be conveniently grouped by the order of their lever mechanism: muscle h as first-order dorsal adductor; muscles k and j as first-order and i as a third-order ventral adductor. Besides contributing to ventral or dorsal adduction, h antagonistically closes or spreads the ventral apices of the penisvalvae.
The volsellae of Labidus are highly modified, which has led to confusion of their homology (Birket-Smith 1981;Brady and Ward 2005;Watkins II 1982). The entire organ has migrated to a ventral position beneath the penisvalvae (Fig. 3), and the stalked distal element hinges ventrally on the reduced basivolsella which is completely fused with the basimere. In the literature, the distal element of the volsella has been termed the digitus: it is clear that this is not the case. The true digitus is a small ellipsoid plate, hitherto unreported, attached to the volsellar corium in a dorsolateral position at the base of the distal element (Di, Fig. 11). Moreover the distal volsellar element is setose, a character state associated solely with the cuspis and potentially a deep autapomorphy for the Hymenoptera (Schulmeister 2001;Sharkey 2007). As a corollary, the digitus of Neivamyrmex longiscapus Borgmeier 1953 is retained as a rounded quadrangular plate fused with the shaft of the cuspis (Di, Fig. 12), while that of Nomamyrmex eisenbeckii Borgmeier 1936 is reduced and completely fused with the cuspal shaft.
Both muscles of the volsellae are present (Fig. 11), with their insertions on the cuspal apodeme. Birket-Smith (1981) identified the cuspal muscles r and s. As neither r nor s are present in any of the other taxa examined, it is most parsimonious at present to recognize them as p and qr, respectively. The origin of muscle p covers a large area within the basimere laterally to dorsolaterally, while the origin of qr has migrated from its position on the basivolsellar apodeme to the basimeral anterolateral region. The movements of the cuspis were difficult to ascertain. However, the muscles may act antagonistically to elevate or depress the cuspis. During depression of the cuspis, the cuspal condyle slides ventrally along a vertical shelf of the basimere until it strikes a medially-projected shelf. Because this shelf has a margin contour complementary to the basal swelling of the cuspis, the basal swelling may be pulled into the volsellar corium, potentially providing torque to the cuspis. The copulatory function of these modified volsellae is not known; nor is it known whether they are intromittent.
The final point of contention with Birket- Smith's (1981) description of the musculature of Labidus is the muscle of the telomere. From my dissections of Labidus, I found that when a telomere was not carefully disarticulated, a small muscle fragment could be found with its insertion on the medial face of its base, suggesting the presence of a unique muscle specific to the telomere. It became clear, however, after more care was taken that this was actually l, and that its insertion on the valvura could be observed. I thus conclude that Birket-Smith's putative muscle y is actually muscle l, and that no novel muscle has evolved in the telomeres of Labidus. The telomere is restricted to lateral movement as the medial base is fused to the basimere via sclerotization, while the rest of the telomere base is surrounded by membrane (Tm, Fig. 3). The differential flexibility of the sclerotic bridge and the corium swing the telomeres laterally when j is contracted, with the bridge acting as a pivot. Thus the grasping function of the telomeres of Labidus and other ecitonines has been lost, excepting Nomamyrmex.

Conclusion
The intersubfamilial homology of male ant genitalic musculature is described for the Amblyoponinae, Cerapachyinae, Dolichoderinae, Ectatomminae, Formicinae, Leptanilloidinae, Myrmicinae and Ponerinae for the first time, while the musculature and sclerite homology of the ecitonines Labidus and Neivamyrmex are clarified. Overall, the generalized male ant has fifteen muscles: three on sternum IX, four on the cupula, five penisvalvar, two volsellar and one paramere muscle. Muscles of sternum IX act to protract, contract or torque the genitalic capsule, which may aide in probing and intromittent behavior. The cupula muscles attach to the basimeres, and are most variable in presence or absence among the myrmicine taxa examined. Muscles of the cupula exert greater control over the genitalic valves relative to the muscles of sternum IX. The penisvalva muscles have functions which vary with the lineage, and muscle l is lost in some. In general, the penisvalva muscles may torque the penisvalvae apices, while adducting the individual penisvalvae in the mediosagittal plane. Volsellar mechanics varies greatly with the morphology of the volsellar sclerites. In species with opposable digiti and cuspides, the muscles may spread or close the apices. In other species, the digiti may torque clockwise or counter-clockwise, while movement in Labidus is restricted to the mediosagittal plane. Finally, the parameres have a single muscle which is lost in some lineages to be replaced by any of the proximal muscles.
The mechanics of the sclerites and their muscles are labile over evolutionary time: the axes of rotation, the pivots, and the muscular origins and insertions shift across the sclerites. Detecting the patterns of morphological and mechanical evolution of male ant genitalia may provide useful phylogenetic characters, especially for clades currently with few or without morphological synapomorphies (e.g. the Formicoid clade, Ward 2011). Future studies should include representative taxa of each subfamily where males are known, emphasizing the differences among lineages. The hypertrophic genitalia of the Leptanillinae (Bolton 1990) provide an especially enticing opportunity, while the volsellar morphology of dorylomorphs (Brady and Ward 2005) may be valuable to reconsider, especially with musculature characters. A context and critical comparison of genitalic sclerites and muscles with the Vespoidea would be valuable and may potentially clarify the phylogenetic position of the Formicidae. Investigations of the mechanical processes involved in ant copulation will aid in understanding of the adaptations of the musculature and valves. Furthermore, we may use male genitalic musculature to predict evolutionary trends in behavior and reproductive biology. A foundation for interpreting genitalic musculature allows for neuroethological investigations with which comparative copulatory procedures may be experimentally observed (Kumashiro and Sakai 2001). The ground is set for describing the sequence and fine-scale interaction of the genitalic muscles on the male sclerites. Species of Asecodes Förster are described in the literature as parasitoids of chrysomelid beetles (Askew and Viggiani 1978, Kamijo 1986, Schauff 1991. Asecodes lucens (Nees) (as A. mento (Walker)), was reared from Galerucella nymphaeae (L. ) (Hippa and Koponen 1984) and Lochmaea suturalis (Thomson) (Golightly 1962) (Coleoptera: Chrysomelidae). In an unpublished study in Sweden the junior author and associates also reared A. lucens from five additional species of Galerucella, G. calmariensis (L.), G. lineola (Fabr.), G. pusilla (Duftschmid), G. sagittariae (Gyllenhal) and G. tenella (L.). However, parasitism rates differed considerably in different Galerucella species collected from the same locality. These patterns caused us to wonder whether a single species, A. lucens, was using multiple hosts or if there were populational differences in host range. To examine the possibility that population differentiation occurs within A. lucens based on host use we sequenced both mitochondrial and nuclear genes (Hambäck et al. unpubl data). This analysis indicated the occurrence of multiple species.
In this paper, we identify morphological traits that separate three species of Asecodes, corresponding also to molecular and biological data. We compared the morphotypes with previous descriptions and found that two correspond to described species, A. lucens (Nees) and A. parviclava (Thomson), whereas the third was not described. The new species is described under the name A. lineophagum.

Material and methods
The colour photos were made with a Nikon SMZ 1000 stereomicroscope and a Nikon DS-5M camera. To eliminate reflections from the metallic and shiny body, a domelight manufactured as described by Kerr et al. (2008), was used as the light source for photography. Photos were taken at different focus levels and Helicon Focus Pro version 4.75 was used to merge them into a single image. The photography of wing interference patterns is described in detail by Shevtsova and Hansson (2011). The SEM photos were made from uncoated specimens on their original cardboard mounts. These were taken in low vacuum mode on a JEOL ® JSM 5600LV SEM microscope.
Morphological terms follow Gibson (1997). For illustrations of the morphological terms see www.neotropicaleulophidae.com.
The genetic distances were estimated from pairwise comparisons of 784 base pairs corresponding to the three prime end of CO1 (as used in Hambäck et al., unpublished). In the analysis, 7 individuals of A. lucens, 23 individuals of A. parviclava and 3 individuals of A. lineophagum were used. The analysis was done in PAUP* ver. 4.0.a125 (Swofford 2002), and genetic distances were calculated under the Kimura-2-parameter model (K2P) with pairwise deletion of missing data. Additional mitochondrial and nuclear genes data were used for reconstructing the phylogeny (Hambäck et al., unpublished), but were not included in the estimate of genetic distances due to a less extensive data set than for CO1.

Designation of primary types
Five names were available for this complex of species prior to this investigation, with one name, A. lucens, considered as valid and four other names as synonyms under the latter name. However, none of the names were fixed to a primary type and three of the names lacked type material altogether. For nomenclatural stability names must be fixed to a type specimen, and we therefore designate primary types, neotypes or lectotypes, for all five available names.

Neotypes
Eulophus lucens was described by Nees from a single female caught on Robinia pseudoacacia in Sickershausen [Bavaria, Germany] June 22, 1812, which was placed in his own collection. Specimens in the collection of Nees no longer exist apart from specimens sent to Westwood, now in the Oxford University Museum of Natural History (Graham 1988). No material of E. lucens exists in the Oxford collection (Graham 1988) and it is thus safe to assume that the holotype of E. lucens was destroyed. A female from Hungary, Vas County, Szakonyfalu, collected 23.vi.1960, agrees well with the description of E. lucens and is here designated as neotype. The locality in Hungary is the closest to the type locality where it has been possible to find material of this species. Entedon mento was described by Walker from an unspecified number of males from near London and from Belfast. There is no material that agrees with the original description, either in the general collection or in the type collection of the Natural History Museum, London, where the material Walker based his descriptions is kept. In the type collection, the box supposed to contain some type material of E. mento is emp-ty, but with a note "mento??". Thus, it appears the material on which Walker based his description of E. mento is now lost. A female from England, Middlesex, Southgate, collected 6.vi.1972, fits the description of E. mento and is here designated as neotype for Entedon mento. The species was allegedly described from males, but males and females of this species are very similar. Further, Walker frequently misidentified the sex (Graham 1963), and a female is selected for neotype to make this species comparable to the other species of this complex, all of which are represented by females.
Derostenus parviclava was described by Thomson from an unspecified number of females he collected on Öland, an island in the Baltic Sea, and by G.F. Möller in Holmeja, a locality in Skåne, the southernmost province in Sweden. The collection of C.G. Thomson is in the Lund museum, and the collection of G.F. Möller is in the Natural History Museum in Gothenburg, both in Sweden. However, neither collection has any specimens from the type localities of D. parviclava. There is a female under the name D. parviclava in the G.F. Möller collection from Bökeberg (labeled "Bök"), which is a locality very close to Holmeja, one of the type localities. Because Thomson was very specific concerning localities from the province Skåne, where he lived and worked, it seems unlikely that he interchanged "Holmeja" with "Bökeberg". However, the female from Bökeberg agrees well with the original description of D. parviclava and it is from a locality very close to one of the original type localities. This specimen fulfills the criteria for a neotype for Derostenus parviclava and is designated as such here.

Lectotypes
The descriptions of Entedon chthonia and E. metagenes do not have information on the number of specimens used, and neither has been fixed to a primary type. The type collection of the Natural History Museum, London, has a specimen each of E. chthonia (type no. 5.2603) and E. metagenes (type no. 5.2604). These specimens fit the original descriptions and are here designated as lectotypes.

Biology of the parasitoids
The Asecodes species included here are gregarious koinobiont endoparasitoids of beetle larvae (Stenberg and Hambäck 2010). Females lay their eggs in the early larval stage and successful parasitoid development leads to a mummification of the host larva. The host larval mummies are black and morphologically resemble larvae, whereas unparasitized larvae form soft yellow pupae typical of chrysomelid beetles (photos in Hambäck 2004). After successful development the parasitoid larvae pupate inside the mummified host larva. The number of parasitoid pupae in the mummies is highly variable, from 1-14 within one mummy. The emergence of adult parasitoids typically occurs in intervals, which probably reflects separate egg-laying events and thus indicates superparasitism as a common trait. The number of parasitoids in each host affects both the sex ratio and the adult body size of emerging offspring. Single emerging parasitoids are invariably females, but at high parasitoid densities the sex ratio is male biased (up to 80%) (Stenberg and Hambäck 2010). It also seems that density dependence in sex ratio and adult body size are correlated with host species, as indicated by a comparison of parasitoids emerging from Galerucella tenella and G. calmariensis (Stenberg and Hambäck 2010).
Parasitism rates may at times be very high, close to 100%, but may at other times be very low. In Sweden, where this study was performed, there seems to be a latitudinal shift in parasitism rates, at least for some hosts. The parasitism rates for G. calmariensis and G. tenella in northern localities, close to Umeå, are typically very high, between 50% and 100%, but less than 10% in more southern localities. The genus contains both strictly monophagous species and oligophagous species and the different Galerucella species often occur in the same localities, but on different wetland plants. There are often large differences in the parasitism rates between Galerucella species within the same locality. For instance, parasitism rates may be very high on G. lineola and very low on other Galerucella species in one locality, whereas parasitism rates are high in another species in another locality. The different parasitism rates are not likely to be due to phenological differences or spatial distributions within localities because host plants colonised by different larval species may occur on neighboring plant individuals.

Identification
For identification of the species treated here the following additions can be made to the latest key to European species of Asecodes in Askew and Viggiani (1978).
Antenna as in Fig. 10. Frons below frontal suture with weak reticulation, above suture smooth; antennal scrobes join on frontal suture. Vertex with very weak reticulation inside ocellar triangle, smooth outside triangle.
Hosts and sex ratio. The Swedish specimens were reared from Galerucella calmariensis and G. pusilla (Coleoptera: Chrysomelidae) on Lythrum salicaria, and G. tenella on Filipendula ulmaria. The number of samples is smaller than for the other two species (n = 23, 10 from G. calmariensis, 6 from G. pusilla, 7 from G. tenella

discussion
The separation of A. lucens into three species based on molecular and morphological evidence is supported by biological data. Asecodes lucens and A. lineophagum were reared from only one host species, Galerucella sagittariae and G. lineola, respectively, whereas A. parviclava was reared from three host species, G. tenella, G. calmariensis and G. pusilla. These observations have also been confirmed with independent observations of parasitoid behaviour in the laboratory, where females were found to attack the respective host species, but not other species (L. Fors, unpubl. data). The delimitation of three species is also supported by observations in the field, where one species of Galerucella larvae may be heavily parasitized and another is not attacked in the same locality. Such parasitism patterns have, however, not been observed for G. tenella, G. calmariensis and G. pusilla. In fact, earlier studies show strong correlations in parasitism rates between G. tenella and G. calmariensis among localities (Hambäck et al. 2006). Moreover, field observations suggested that parasitism rates on G. tenella were consistently higher when this species was sympatric with G. calmariensis. The reason for this pattern was not resolved (Hambäck et al. 2006), but the pattern suggest that the parasitoid population may mediate indirect interactions between its hosts, as is known for other host-parasitoid systems. The current information on the species delimitation within Asecodes was important to identify pairs of host species where such effects would be likely. Based on previous information, we could have expected similar indirect effects also for other species pairs but the novel information on population differentiation among parasitoid individuals suggest this not to be the case.  Figures 2-9. Asecodes spp.,wings, females 2-3 A. lineophagum sp. n. 2 transparent wings 3 wing interference pattern 4-5 A. parviclava (Thomson) 4 transparent wings 5 wing interference pattern 6-7 A. lucens (Nees) 6 transparent wings 7 wing interference pattern 8 A. lucens, forewing, neotype 9 A. parviclava, forewing, neotype.
In view of our findings, the previous host records of Galerucella nymphaeae and Lochmaea suturalis for A. lucens need confirmation. Investigation of Asecodes specimens reared from these hosts might quite possibly reveal additional cryptic species in this group. Figures 10-15. Asecodes spp., antennae 10-12 females 10 A. lineophagum sp. n. 11 A. parviclava (Thomson)

acknowledgements
Thanks to museum staff (names listed under the museum acronyms above) for loan of type material, to the Department of Biology, Lund University, for the use of their SEM facility, and to Elisabeth Weingartner and Johannes Bergsten for assisting in handling the material. The study was funded by a grant from the Swedish Research Council Vetenskapsrådet (contract no. 621-2009-4943).

Hybrizon Fallén (Hymenoptera, ichneumonidae, Hybrizoninae) found in Hunan (China)
and Yu and Horstmann (1997) because of the structure of the connection of the second and third metasomal tergites and the venation of the hind wing, both indicate a closer relationship with the family Ichneumonidae (Sharkey and Wahl 1987;Wahl and Sharkey 1988) than with the Braconidae. From analysis of the 28S ribosomal RNA from the genus Hybrizon Fallén, 1813, it may be concluded that the Hybrizoninae are at a basal position of the Ichneumonidae-lineage (Belshaw et al. 1998;Quicke et al. 2000;Belshaw and Quicke 2002), but Gillespie et al. (2005) documented the unusual structure of 28S in Hybrizon, which makes alignment difficult. Quicke et al. (2009) found that Hybrizon likely is a derived subfamily within the ophioniformes-group of the Ichneumonidae, which agrees with the derived morphology of the Hybrizoninae. The subfamily is known only from the Holarctic region and we report for the first time two species of the genus from the Oriental part of China. There are only two reports of the genus Hybrizon from China (He 1981) but only from Palaearctic northern China (H. buccatus (de Brébisson, 1825) from Jilin and Heilongjiang and H. ghilarovi Tobias, 1988, from Jilin). The second author collected in Hunan province two species of the genus, resulting in an enormous extension of the known distribution by 2200+ km southwards.
The biology of the Hybrizoninae has been for long time uncertain, but recently oviposition has been documented by photographing and filming two different genera (Komatsu and Konishi 2010;. It shows that the final instar ant larva is used for oviposition when the worker ants transport the larvae outside the nest. Of one species (H. buccatus) we have some host records indicating that predominantly ant larvae from the subfamily Formicinae (Formicidae) are selected, but also larvae from non-Formicinae may be used (Gómez Durán and van Achterberg 2011). It is too early to conclude a lack of specialisation, because in most cases the true nature of the associations has not really been established and the host associations are largely unknown for the other species.

Material and methods
The collecting site is at the border of the Southeast Lake near Yuanjiang (N. Hunan) in the common reed (Phragmites australis (Cav.)) zone, with Oriental "Lasius fuliginosus" (= Lasius fuyi Radchenko, 2005; see Radchenko 2005) as possible host. The collecting in this wetland habitat along the lake was rather cumbersome and done by hand netting among the common reed.
For references to genera and species of Hybrizoninae, see Yu et al. (2009) and updates, for the East Palaearctic species, see Konishi et al. (2012) and for morphological terminology, see van Achterberg (1988). The specimens are deposited in the College of Bio-Safety Science and Technology, Hunan Agriculture University (HUNAU) at Changsha and in the NCB Naturalis collection (RMNH) at Leiden. Material. Reported from North China by He (1981: Heilongjiang, Jilin) and by Konishi et al. (2012: Jilin). Unknown from Oriental China.
Notes. Up to now only known from the holotype from Far East Russia (Khabarovsk kray). The holotype is illustrated by Konishi et al. (2012). New for China and for the Oriental region. Tobias, 1988 http://species-id.net/wiki/Hybrizon_ghilarovi Figs 5-8
In the study of parasitoids that attack tree borers in three forest areas of China, a new species of Triclistus has been found. The aim of this contribution is to describe and illustrate this species and provide biological data on its habitat and hosts.

Materials and methods
In the last five years the authors have been researching the parasitoids of borers of tree branches and cones in Heilongjiang, Jilin, Liaoning, Hebei, Henan, Shanxi and Qinghai provinces, situated in the Eastern Palaearctic part of China and in Hunan and Jiangxi Provinces, situated in the northern border of the Oriental part of China.
To rear parasitoids, cones and twigs of naturally heavily infested trees of Pinus armandi Franch, P. massoniana Lamb., P. sylvestris L. var. mongolica Litv., and P. tabulaeformis Carr. were brought to the laboratory and maintained in large nylon cages at room-temperature. Water was sprayed over the cones and twigs twice a week and emerged insects collected daily.
The hosts were identified by Professor Hou-Hun Li, Nankai University, Tianjing, China.
Images of whole bodies were taken using a CANON Power Shot A650 IS. Other images were taken using a Cool SNAP 3CCD attached to a Zeiss Discovery V8 Stereomicroscope and captured with QCapture Pro version 5.1.
The morphological terminology is mostly that of Gauld (1991). Wing vein nomenclature is based on Ross (1936).
Type specimens and hosts are deposited in the Insect Museum, General Station of Forest Pest Management, State Forestry Administration, P. R. China.