[Show abstract][Hide abstract] ABSTRACT: Skipper butterflies (Hesperiidae) are a relatively well-studied family of Lepidoptera. However, a combination of DNA barcodes, morphology, and natural history data has revealed several cryptic species complexes within them. Here, we investigate three DNA barcode lineages of what has been identified as Urbanus belli (Hesperiidae, Eudaminae) in Area de Conservacion Guanacaste (ACG), northwestern Costa Rica.
[Show abstract][Hide abstract] ABSTRACT: More than half a million specimens of wild-caught Lepidoptera caterpillars have been reared for their parasitoids, identified, and DNA barcoded over a period of 34 years (and ongoing) from Area de Conservación de Guanacaste (ACG), northwestern Costa Rica. This provides the world's best location-based dataset for studying the taxonomy and host relationships of caterpillar parasitoids. Among Hymenoptera, Microgastrinae (Braconidae) is the most diverse and commonly encountered parasitoid subfamily, with many hundreds of species delineated to date, almost all undescribed. Here, we reassess the limits of the genus Apanteles sensu stricto, describe 186 new species from 3,200+ parasitized caterpillars of hundreds of ACG Lepidoptera species, and provide keys to all 205 described Apanteles from Mesoamerica - including 19 previously described species in addition to the new species. The Mesoamerican Apanteles are assigned to 32 species-groups, all but two of which are newly defined. Taxonomic keys are presented in two formats: traditional dichotomous print versions and links to electronic interactive versions (software Lucid 3.5). Numerous illustrations, computer-generated descriptions, distributional information, wasp biology, and DNA barcodes (where available) are presented for every species. All morphological terms are detailed and linked to the Hymenoptera Anatomy Ontology website. DNA barcodes (a standard fragment of the cytochrome c oxidase I (COI) mitochondrial gene), information on wasp biology (host records, solitary/gregariousness of wasp larvae), ratios of morphological features, and wasp microecological distributions were used to help clarify boundaries between morphologically cryptic species within species-complexes. Because of the high accuracy of host identification for about 80% of the wasp species studied, it was possible to analyze host relationships at a regional level. The ACG species of Apanteles attack mainly species of Hesperiidae, Elachistidae and Crambidae (Lepidoptera). About 90% of the wasp species with known host records seem to be monophagous or oligophagous at some level, parasitizing just one host family and commonly, just one species of caterpillar. Only 15 species (9%) parasitize species in more than one family, and some of these cases are likely to be found to be species complexes. We have used several information sources and techniques (traditional taxonomy, molecular, software-based, biology, and geography) to accelerate the process of finding and describing these new species in a hyperdiverse group such as Apanteles. The following new taxonomic and nomenclatural acts are proposed. Four species previously considered to be Apanteles are transferred to other microgastrine genera: Dolichogenidea hedyleptae (Muesebeck, 1958), comb. n., Dolichogenidea politiventris (Muesebeck, 1958), comb. n., Rhygoplitis sanctivincenti (Ashmead, 1900), comb. n., and Illidops scutellaris (Muesebeck, 1921), comb. rev. One European species that is a secondary homonym to a Mesoamerican species is removed from Apanteles and transferred to another genus: Iconella albinervis (Tobias, 1964), stat. rev. The name Apanteles albinervican Shenefelt, 1972, is an invalid replacement name for Apanteles albinervis (Cameron, 1904), stat. rev., and thus the later name is reinstated as valid. The following 186 species, all in Apanteles and all authored by Fernández-Triana, are described as species nova: adelinamoralesae, adrianachavarriae, adrianaguilarae, adrianguadamuzi, aichagirardae, aidalopezae, albanjimenezi, alejandromasisi, alejandromorai, minorcarmonai, alvarougaldei, federicomatarritai, anabellecordobae, rostermoragai, anamarencoae, anamartinesae, anapiedrae, anariasae, andreacalvoae, angelsolisi, arielopezi, bernardoespinozai, bernyapui, bettymarchenae, bienvenidachavarriae, calixtomoragai, carloscastilloi, carlosguadamuzi, eliethcantillanoae, carlosrodriguezi, carlosviquezi, carloszunigai, carolinacanoae, christianzunigai, cinthiabarrantesae, ciriloumanai, cristianalemani, cynthiacorderoae, deifiliadavilae, dickyui, didiguadamuzi, diegoalpizari, diegotorresi, diniamartinezae, duniagarciae, duvalierbricenoi, edgarjimenezi, edithlopezae, eduardoramirezi, edwinapui, eldarayae, erickduartei, esthercentenoae, eugeniaphilipsae, eulogiosequeira, felipechavarriai, felixcarmonai, fernandochavarriai, flormoralesae, franciscopizarroi, franciscoramirezi, freddyquesadai, freddysalazari, gabrielagutierrezae, garygibsoni, gerardobandoi, gerardosandovali, gladysrojasae, glenriverai, gloriasihezarae, guadaluperodriguezae, guillermopereirai, juanmatai, harryramirezi, hectorsolisi, humbertolopezi, inesolisae, irenecarrilloae, isaacbermudezi, isidrochaconi, isidrovillegasi, ivonnetranae, jairomoyai, javiercontrerasi, javierobandoi, javiersihezari, jesusbrenesi, jesusugaldei, jimmychevezi, johanvargasi, jorgecortesi, jorgehernandezi, josecalvoi, josecortesi, josediazi, josejaramilloi, josemonteroi, joseperezi, joserasi, juanapui, juancarrilloi, juangazoi, juanhernandezi, juanlopezi, juanvictori, juliodiazi, juniorlopezi, keineraragoni, laurahuberae, laurenmoralesae, leninguadamuzi, leonelgarayi, lilliammenae, lisabearssae, luciariosae, luisbrizuelai, luiscanalesi, luiscantillanoi, luisgarciai, luisgaritai, luishernandezi, luislopezi, luisvargasi, manuelarayai, manuelpereirai, manuelriosi, manuelzumbadoi, marcobustosi, marcogonzalezi, marcovenicioi, mariachavarriae mariaguevarae, marialuisariasae, mariamendezae, marianopereirai, mariatorrentesae, sigifredomarini, marisolarroyoae, marisolnavarroae, marvinmendozai, mauriciogurdiani, milenagutierrezae, monicachavarriae, oscarchavesi, osvaldoespinozai, pablotranai, pabloumanai, pablovasquezi, paulaixcamparijae, luzmariaromeroae, petronariosae, randallgarciai, randallmartinezi, raulacevedoi, raulsolorsanoi, wadyobandoi, ricardocaleroi, robertmontanoi, robertoespinozai, robertovargasi, rodrigogamezi, rogerblancoi, rolandoramosi, rolandovegai, ronaldcastroi, ronaldgutierrezi, ronaldmurilloi, ronaldnavarroi, ronaldquirosi, ronaldzunigai, rosibelelizondoae, ruthfrancoae, sergiocascantei, sergioriosi, tiboshartae, vannesabrenesae, minornavarroi, victorbarrantesi, waldymedinai, wilbertharayai, williamcamposi, yeissonchavesi, yilbertalvaradoi, yolandarojasae, hazelcambroneroae, zeneidabolanosae.
[Show abstract][Hide abstract] ABSTRACT: Background/Question/Methods: A biodiversity inventory of the 120,000 terrestrial ha Area de Conservacion Guanacaste (ACG) in northwestern Costa Rica has been running for 33 years and has documented at least 9,000 species of Lepidoptera (and their parasitoids and food plants) of the estimated 15,000 species. In 2003 the inventory began routinely DNA barcoding all voucher specimens for identification and cryptic species discovery. How has this additional identification and species discovery technique changed the inventory results, and what are the implications of this change for community structure, specialization, and trophic web characterizations of diverse species arrays?
Results/Conclusions: DNA barcoding of every vouchered specimen in the inventory has
1) allowed a far faster and more certain field identification of inventory specimens, irrespective of whether to an interim name or a “final” scientific name;
2) found that 10 to 50 percent of morphologically-named OTUs in ACG are in fact pooled specimens (and hence biologies) of two or more species if not segregated according to their barcodes or other traits;
3) found that this species-level confusion becomes more frequent the smaller or more inconspicuous, or less studied, the specimens of the species being inventoried, with the exception that sometimes the most prominent and well-known species also turn out to be complexes, in part because “everyone knows the name of that one”;
4) found that within some higher taxa of parasitoids of caterpillars, the concept of “generalist” is rendered close to non-existent due to finding that many, though not all, “generalists” under one morphology-based name are a sympatric complex of highly host-specific species;
5) found that, as a byproduct of 4) above, three-layered trophic webs (plant-caterpillar-parasitoid) are much more specialist structured than “expected”; and
6) decided that
a) there are significantly more species in ACG than anticipated by an estimate based on centuries of taxonomists’ morphology-based intuition,
b) climate changes will impact these specialist-based arrays more severely than imagined, and
c) tropical ecology is perhaps somewhat shakily based largely on the biology of those species that have been separate long enough, or directedly selected long enough in directions readily visible to 2 m tall diurnal large mammals, to be morphologically distinguishable, while DNA barcoding all “species” allows summary studies to also include (yet species-level distinguish) those species that are selected to be or remain similar, or not separate long enough to be morphologically distinguishable.
[Show abstract][Hide abstract] ABSTRACT: An intense, 30-year, ongoing biodiversity inventory of Lepidoptera, together with their food plants and parasitoids, is centered on the rearing of wild-caught caterpillars in the 120,000 terrestrial hectares of dry, rain, and cloud forest of Area de Conservacion Guanacaste (ACG) in northwestern Costa Rica. Since 2003, DNA barcoding of all species has aided their identification and discovery. We summarize the process and results for a large set of the species of two speciose subfamilies of ACG skipper butterflies (Hesperiidae) and emphasize the effectiveness of barcoding these species (which are often difficult and time-consuming to identify).
Adults are DNA barcoded by the Biodiversity Institute of Ontario, Guelph, Canada; and they are identified by correlating the resulting COI barcode information with more traditional information such as food plant, facies, genitalia, microlocation within ACG, caterpillar traits, etc. This process has found about 303 morphologically defined species of eudamine and pyrgine Hesperiidae breeding in ACG (about 25% of the ACG butterfly fauna) and another 44 units indicated by distinct barcodes (n = 9,094), which may be additional species and therefore may represent as much as a 13% increase. All but the members of one complex can be identified by their DNA barcodes.
Addition of DNA barcoding to the methodology greatly improved the inventory, both through faster (hence cheaper) accurate identification of the species that are distinguishable without barcoding, as well as those that require it, and through the revelation of species "hidden" within what have long been viewed as single species. Barcoding increased the recognition of species-level specialization. It would be no more appropriate to ignore barcode data in a species inventory than it would be to ignore adult genitalia variation or caterpillar ecology.
PLoS ONE 01/2011; 6(8):e19874. · 3.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: We propose that the many different, but essentially similar, eye-like and face-like color patterns displayed by hundreds of species of tropical caterpillars and pupae-26 examples of which are displayed here from the dry, cloud, and rain forests of Area de Conservacion Guanacaste (ACG) in northwestern Costa Rica-constitute a huge and pervasive mimicry complex that is evolutionarily generated and sustained by the survival behavior of a large and multispecific array of potential predators: the insect-eating birds. We propose that these predators are variously and innately programmed to flee when abruptly confronted, at close range, with what appears to be an eye of one of their predators. Such a mimetic complex differs from various classical Batesian and Müllerian mimicry complexes of adult butterflies in that (i) the predators sustain it for the most part by innate traits rather than by avoidance behavior learned through disagreeable experiences, (ii) the more or less harmless, sessile, and largely edible mimics vastly outnumber the models, and (iii) there is no particular selection for the eye-like color pattern to closely mimic the eye or face of any particular predator of the insect-eating birds or that of any other member of this mimicry complex. Indeed, selection may not favor exact resemblance among these mimics at all. Such convergence through selection could create a superabundance of one particular false eyespot or face pattern, thereby increasing the likelihood of a bird species or guild learning to associate that pattern with harmless prey.
Proceedings of the National Academy of Sciences 06/2010; 107(26):11659-65. · 9.81 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Associating sexes in many species of the Neotropical hesperiid genus Porphyrogenes has been difficult, erroneous, or impossible, owing to their sexual dimorphism. Despite the extreme sexual dimorphism of Porphyrogenes peterwegei Burns, new species, full description of this rainforest skipper from Area de Conservación Guanacaste (ACG), northwestern Costa Rica, is no problem. The available sample amounts to nearly 100 reared adults, almost evenly divided between the sexes. Of these, 67 have been DNA barcoded and found to vary minimally in their COI haplotypes, with one haplotype predominating. Caterpillars and pupae consistently go through a distinctive set of developmental changes in their color patterns. Foodplant choice is conservative: of 227 immatures found in the wild, 94% were eating woody vines of the genus Machaerium, especially M. seemannii (Fabaceae), whereas 6% were feeding on the quite unrelated species Dichapetalum morenoi (Dichapetalaceae)—a peculiar pattern of larval foodplant selection seen in another skipper and two butterfly species in ACG. Caterpillars of P. peterwegei are occasionally attacked by a host-specific parasitoid (an undescribed tachinid of the genus Siphosturmia), which has not otherwise been encountered in 20,642 tachinid attacks on the caterpillars of >3,000 species of Lepidoptera in ACG. Morphologically, P. peterwegei differs significantly from its many congeners in both facies and genitalia.
Proceedings- Entomological Society of Washington 02/2010; · 0.39 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Inventory of the caterpillars, their food plants and parasitoids began in 1978 for today's Area de Conservacion Guanacaste (ACG), in northwestern Costa Rica. This complex mosaic of 120 000 ha of conserved and regenerating dry, cloud and rain forest over 0-2000 m elevation contains at least 10 000 species of non-leaf-mining caterpillars used by more than 5000 species of parasitoids. Several hundred thousand specimens of ACG-reared adult Lepidoptera and parasitoids have been intensively and extensively studied morphologically by many taxonomists, including most of the co-authors. DNA barcoding - - the use of a standardized short mitochondrial DNA sequence to identify specimens and flush out undisclosed species - - was added to the taxonomic identification process in 2003. Barcoding has been found to be extremely accurate during the identification of about 100 000 specimens of about 3500 morphologically defined species of adult moths, butterflies, tachinid flies, and parasitoid wasps. Less than 1% of the species have such similar barcodes that a molecularly based taxonomic identification is impossible. No specimen with a full barcode was misidentified when its barcode was compared with the barcode library. Also as expected from early trials, barcoding a series from all morphologically defined species, and correlating the morphological, ecological and barcode traits, has revealed many hundreds of overlooked presumptive species. Many but not all of these cryptic species can now be distinguished by subtle morphological and/or ecological traits previously ascribed to 'variation' or thought to be insignificant for species-level recognition. Adding DNA barcoding to the inventory has substantially improved the quality and depth of the inventory, and greatly multiplied the number of situations requiring further taxonomic work for resolution.
[Show abstract][Hide abstract] ABSTRACT: DNA barcodes can be used to identify cryptic species of skipper butterflies previously detected by classic taxonomic methods and to provide first clues to the existence of yet other cryptic species. A striking case is the common geographically and ecologically widespread neotropical skipper butterfly Perichares philetes (Lepidoptera, Hesperiidae), described in 1775, which barcoding splits into a complex of four species in Area de Conservación Guanacaste (ACG) in northwestern Costa Rica. Three of the species are new, and all four are described. Caterpillars, pupae, and foodplants offer better distinguishing characters than do adults, whose differences are mostly average, subtle, and blurred by intraspecific variation. The caterpillars of two species are generalist grass-eaters; of the other two, specialist palm-eaters, each of which feeds on different genera. But all of these cryptic species are more specialized in their diet than was the morphospecies that held them. The four ACG taxa discovered to date belong to a panneotropical complex of at least eight species. This complex likely includes still more species, whose exposure may require barcoding. Barcoding ACG hesperiid morphospecies has increased their number by nearly 10%, an unexpectedly high figure for such relatively well known insects.
Proceedings of the National Academy of Sciences 05/2008; 105(17):6350-5. · 9.81 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Although central to much biological research, the identification of species is often difficult. The use of DNA barcodes, short DNA sequences from a standardized region of the genome, has recently been proposed as a tool to facilitate species identification and discovery. However, the effectiveness of DNA barcoding for identifying specimens in species-rich tropical biotas is unknown. Here we show that cytochrome c oxidase I DNA barcodes effectively discriminate among species in three Lepidoptera families from Area de Conservación Guanacaste in northwestern Costa Rica. We found that 97.9% of the 521 species recognized by prior taxonomic work possess distinctive cytochrome c oxidase I barcodes and that the few instances of interspecific sequence overlap involve very similar species. We also found two or more barcode clusters within each of 13 supposedly single species. Covariation between these clusters and morphological and/or ecological traits indicates overlooked species complexes. If these results are general, DNA barcoding will significantly aid species identification and discovery in tropical settings.
Proceedings of the National Academy of Sciences 02/2006; 103(4):968-71. · 9.81 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: By facilitating bioliteracy, DNA barcoding has the potential to improve the way the world relates to wild biodiversity. Here we describe the early stages of the use of cox1 barcoding to supplement and strengthen the taxonomic platform underpinning the inventory of thousands of sympatric species of caterpillars in tropical dry forest, cloud forest and rain forest in northwestern Costa Rica. The results show that barcoding a biologically complex biota unambiguously distinguishes among 97% of more than 1000 species of reared Lepidoptera. Those few species whose barcodes overlap are closely related and not confused with other species. Barcoding also has revealed a substantial number of cryptic species among morphologically defined species, associated sexes, and reinforced identification of species that are difficult to distinguish morphologically. For barcoding to achieve its full potential, (i) ability to rapidly and cheaply barcode older museum specimens is urgent, (ii) museums need to address the opportunity and responsibility for housing large numbers of barcode voucher specimens, (iii) substantial resources need be mustered to support the taxonomic side of the partnership with barcoding, and (iv) hand-held field-friendly barcorder must emerge as a mutualism with the taxasphere and the barcoding initiative, in a manner such that its use generates a resource base for the taxonomic process as well as a tool for the user.
Philosophical Transactions of The Royal Society B Biological Sciences 11/2005; 360(1462):1835-45. · 6.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Astraptes fulgerator, first described in 1775, is a common and widely distributed neotropical skipper butterfly (Lepidoptera: Hesperiidae). We combine 25 years of natural history observations in northwestern Costa Rica with morphological study and DNA barcoding of museum specimens to show that A. fulgerator is a complex of at least 10 species in this region. Largely sympatric, these taxa have mostly different caterpillar food plants, mostly distinctive caterpillars, and somewhat different ecosystem preferences but only subtly differing adults with no genitalic divergence. Our results add to the evidence that cryptic species are prevalent in tropical regions, a critical issue in efforts to document global species richness. They also illustrate the value of DNA barcoding, especially when coupled with traditional taxonomic tools, in disclosing hidden diversity.
Proceedings of the National Academy of Sciences 11/2004; 101(41):14812-7. · 9.81 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Unlike most species of Lepidoptera whose DNA barcodes have been examined, closely related taxa in each of three pairs of hesperiids (Polyctor cleta and P. polyctor, Cobalus virbius and C. fidicula, Neoxeniades luda and N. pluviasilva Burns, new species) seem in- distinguishable by their barcodes; but that is when some of the cytochrome c oxidase I (COI) sequences are short and sample sizes are small. These skipper butterflies are unquestionably distinct species, as evidenced by genitalic and facies differences and by ecologic segregation, i.e., one species of each pair in dry forest, the other in adjacent rain forest in Area de Conservación Guanacaste in northwestern Costa Rica. This national park is the source of the specimens used in this study, all of which were reared. Larval foodplants are of no or problematic value in dis- tinguishing these species. Large samples of individuals whose barcodes are acceptably long reveal slight interspecific differentiation (involving just one to three nucleotides) in all three pairs of skippers. Clearly, the chronic practice of various taxonomists of setting arbitrary levels of dif- ferentiation for delimiting species is unrealistic.
[Show abstract][Hide abstract] ABSTRACT: The black and red medium-sized parasitoid wasp Bassus brooksi Sharkey n. sp. (Braconidae) is described from wild-caught specimens from dry forest habitats ranging from northern Mexico to the northwestern Costa Rican coastal plain, and from specimens reared from the Area de Conservacion Guanacaste, Costa Rica. The wasp larvae develop in the caterpillars of a relatively unrelated array of at least 24 species of pyrgine Hesperiidae. These hosts live in the broken shade and sun of forest edges while feeding on a variety of herbs, vines, and low woody plants. Oviposition occurs in an early to middle instar caterpillar, and a single wasp larva emerges from a penultimate or ultimate instar caterpillar to spin its large elongate white cocoon in the caterpillar's shelter next to the empty skin and head capsule of the caterpillar. Bassus brooksi is distinctive in not attacking any grass-feeding Hesperiidae caterpillars (Hesperiinae) in this same habitat, and in apparently ignoring many species of sympatric pyrgine hesperiids as well as all other taxa of caterpillars. Bassus brooksi is closely related to Bassus spiracularis, which ranges over much of North America, is broadly sympatric with B. brooksi in northern Mexico, and has been reared only from pyrgine hesperiid caterpillars. These are the only two species of agathidine braconids known to attack butterfly larvae. It is hoped that publication of this information in a lepidopterological journal will stimulate the recording of these parasitoids when they are encountered while studying caterpillars. The Area de Conservacion Guanacaste (ACG) is 88,000 ha of dry forests and associated wetter ecosystems conserved for ecosystem and biodiversity services in northwestem Costa Rica (Janzen, 1988a, b, 1993). As part of its Biodiversity Development, the ACG is conducting a thorough inventory of its biodiversity so as to set up that biodiversity for non-damaging use (e.g., Janzen, 1996a, b). Such inventory encounters undescribed species and simultaneously reveals a sketchy outline of their natural history (e.g., Burns, 1996; Danger-field et al., 1996; Gauld and Janzen, 1994; Sharkey and Janzen, 1995; Woodley and Janzen, 1995). Here we name one of these species, the previously undescribed Bassus brooksi Sharkey n. sp., so that it and its natural history can be included in the greater global taxonomic understanding of braconid parasitoids (e.g., Sharkey, 1985, 1988; Bums and Janzen, 1999), and so that ecological papers can refer to it (e.g., Camargo, 1999). This new species has been commonly collected in two distinct ways. Since the mid-l980's, I. D. Gauld and co-workers have conducted a Malaise trap inventory of the parasitoid wasp fauna of the ACG (e.g., Gauld and Janzen, 1985; Janzen and Gauld, 1997) and have frequently captured specimens of Bassus brooksi. Also, Bassus brooksi larvae frequently emerge from wild-caught skipper caterpillars reared in captivity as part of the ongoing biodiversity inventory (Janzen and Hallwachs, 1998). Here we describe the natural history of the interaction of this distinctive parasite with its hesperiid host caterpillars in a journal about Lepidoptera, with the intent that others will add to this story through their caterpillar rearings.