Phylogenomic analysis provides diagnostic tools for the identification of Anastrepha fraterculus (Diptera: Tephritidae) species complex.

Insect pests cause tremendous impact to agriculture worldwide. Species identification is crucial for implementing appropriate measures of pest control but can be challenging in closely related species. True fruit flies of the genus Anastrepha Schiner (Diptera: Tephritidae) include some of the most serious agricultural pests in the Americas, with the Anastrepha fraterculus (Wiedemann) complex being one of the most important due to its extreme polyphagy and wide distribution across most of the New World tropics and subtropics. The eight morphotypes described for this complex as well as other closely related species are classified in the fraterculus species group, whose evolutionary relationships are unresolved due to incomplete lineage sorting and introgression. We performed multifaceted phylogenomic approaches using thousands of genes to unravel the evolutionary relationships within the A. fraterculus complex to provide a baseline for molecular diagnosis of these pests. We used a methodology that accommodates variable sources of data (transcriptome, genome, and whole-genome shotgun sequencing) and developed a tool to align and filter orthologs, generating reliable datasets for phylogenetic studies. We inferred 3031 gene trees that displayed high levels of discordance. Nevertheless, the topologies of the inferred coalescent species trees were consistent across methods and datasets, except for one lineage in the A. fraterculus complex. Furthermore, network analysis indicated introgression across lineages in the fraterculus group. We present a robust phylogeny of the group that provides insights into the intricate patterns of evolution of the A. fraterculus complex supporting the hypothesis that this complex is an assemblage of closely related cryptic lineages that have evolved under interspecific gene flow. Despite this complex evolutionary scenario, our subsampling analysis revealed that a set of as few as 80 loci has a similar phylogenetic resolution as the genome-scale dataset, offering a foundation to develop more efficient diagnostic tools in this species group.

Integrative approach reveals the identity of Brazilian specimens previously recognized as Anastrepha dissimilis Stone, 1942 (Diptera: Tephritidae).

Anastrepha dissimilis is currently considered to be widely distributed in Brazil, occurring in 20 of 27 states. However, morphological differences between the holotype (from Plaisance, Haiti) and a paratype (from Pernambuco, Brazil) suggest that the Brazilian specimens are not A. dissimilis, because their aculeus tip is similar to the paratype not to the holotype. Therefore, considering the importance of integrative taxonomy for species delimitation, we used geometric and linear morphometrics and cytochrome c oxidase subunit I sequences integrated with the morphology of the aculeus tip to clarify the identity of populations previously identified as A. dissimilis from multiple Brazilian localities. Morphological data show a uniform pattern among the Brazilian populations, with some variation among specimens from the south and northeast. In addition, the geometric and linear morphometrics suggest considerable geographic variation among these populations, suggesting the existence of at least two morphs. The molecular analysis revealed that specimens from Brazil previously identified as A. dissimilis belong to Anastrepha chiclayae Greene, with a genetic distance ranging from 0.00 to 0.015%. According to our integrative analyses, specimens from Brazil formerly identified as A. dissimilis actually are A. chiclayae. Therefore, this is the first record of A. chiclayae in Brazil, and we also report that A. dissimilis does not occur in Brazil.

Exceptional larval morphology of nine species of the Anastrephamucronota species group (Diptera, Tephritidae).

Anastrepha is the most diverse and economically important genus of Tephritidae in the American tropics and subtropics. The striking morphology of the third instars of Anastrephacaballeroi Norrbom, Anastrephacrebra Stone, Anastrephahaplacantha Norrbom & Korytkowski, Anastrephakorytkowskii Norrbom, Anastrephanolazcoae Norrbom & Korytkowski, and three newly discovered and as yet formally unnamed species (Anastrepha sp. Peru-82, Anastrephasp.nr.protuberans, and Anastrepha sp. Sur-16), and the more typical morphology of Anastrephaaphelocentema Stone, are described using light and scanning electron microscopy. To contribute to a better understanding of the interspecific and intraspecific variation among species in the mucronota species group and facilitate phylogenetic studies, we integrate molecular and morphological techniques to confirm the identity and describe third instars. Larva-adult associations and the identification of described larvae were confirmed using DNA barcodes. We provide diagnostic characters to distinguish larvae among these nine species of the mucronota group and separate them from those of the 29 other Anastrepha species previously described. We introduce the vertical comb-like processes on the oral margin as a novel character, and the unusual character states, including position and shape of the preoral lobe, and dentate or fringed posterior margins of the oral ridges and accessory plates. Our comparative morphology concurs with most previously inferred phylogenetic relationships within the mucronota group.

New species and host plants of Anastrepha (Diptera: Tephritidae) primarily from Suriname and Par, Brazil.

Seventeen new species of Anastrepha, primarily from Suriname, French Guiana and Par, Brazil, are described and illustrated: A. aithogaster Norrbom from Brazil (Par), French Guiana, and Suriname; A. aliesae Norrbom from Suriname; A. brownsbergiensis Norrbom from Suriname; A. crassaculeus Norrbom Rodriguez Clavijo from Colombia (Magdalena, Norte de Santander) and Suriname; A. curvivenis Norrbom from Brazil (Amazonas), Ecuador (Zamora-Chinchipe), Peru (San Martn), and Suriname; A. fuscoalata Norrbom from Brazil (Par), French Guiana, and Suriname; A. gangadini Norrbom from Suriname; A. juxtalanceola Norrbom from Brazil (Par) and Suriname; A. microstrepha Norrbom from Brazil (Bahia) and Suriname; A. mitaraka Norrbom from French Guiana; A. neptis Norrbom from Brazil (Par), Ecuador (Orellana), Peru (Loreto) and Suriname; A. sobrina Norrbom from Brazil (Par), French Guiana, and Suriname; A. surinamensis Norrbom from Suriname; A. tenebrosa Norrbom from Brazil (Par) and Peru (Loreto); A. triangularis Norrbom from Suriname; A. wachiperi Norrbom from French Guiana and Peru (Cusco); and A. wittiensis Norrbom from Suriname. The following host plant records are reported: A. aithogaster from fruit of Parahancornia fasciculata (Poir.) Benoist (Apocynaceae); A. aliesae from fruit of Passiflora coccinea Aubl. and P. glandulosa Cav. (Passifloraceae); A. crassaculeus from fruit of an undetermined species of Pouteria (Sapotaceae); A. fuscoalata from fruit of Trymatococcus oligandrus (Benoist) Lanj. (Moraceae); A. sobrina from fruit of Eugenia lambertiana DC. (Myrtaceae); and A. wittiensis from fruit of Manilkara bidentata (A. DC.) A. Chev. (Sapotaceae).

New species and host plant records for Neotropical Rhagoletis Loew (Diptera: Tephritidae).

Rhagoletis antioquiensis Rodriguez Norrbom, a new species of fruit fly in the striatella species group of Rhagoletis Loew, is described and illustrated. New distribution records are reported for two other species of the striatella group: R. nicaraguensis Herndez-Ortiz Fras, 2000 is reported from Costa Rica, and R. solanophaga Herndez-Ortiz Fras, 2000 from Bolivia. A lectotype is designated for Urophora scutellaris Macquart, 1851 (= R. macquartii (Loew)). New host plant relationships for three species of Rhagoletis, all reared from fruits, are reported: Acnistus arborescens (L.) Schrtdl. and Witheringia solanacea LHer for R. jamaicensis Foote, 1981 from Colombia; Lycianthes pseudolycioides (Chod. Hassl.) Bitter for R. solanophaga from Bolivia; and Solanum pseudocapsicum L., Solanum tuberosum L. subsp. andigenum (Juz. Burkasov) Hawkes, Solanum interandinum Bitter from Colombia and Solanum americanum Mill. in Peru for undetermined species of the psalida group. The key to species of the striatella group of Herndez-Ortiz Fras (2000) is modified to include R. antioquiensis.

New species and new records of Anastrepha (Diptera: Tephritidae) from Colombia.

Four new species of Anastrepha are described and illustrated: A. arevaloi Rodriguez Norrbom, A. cheslavoi Rodriguez Norrbom, A. coronis Rodriguez Norrbom, and A. pseudacris Rodriguez Norrbom. Additionally, A. buscki Stone, A. concava Greene, A. lutea Stone and A. zacharyi Norrbom are reported from Colombia for the first time, the occurrence of A. spatulata Stone in the country is confirmed and new distribution records of A. flavipennis Grenne and A. littoralis (Blanchard) are reported.

The Bolivian fauna of the genus Anastrepha Schiner (Diptera: Tephritidae).

The general aim of this study is to contribute to and summarize knowledge of the Bolivian fauna of the genus Anastrepha Schiner (Tephritidae) which includes species of both ecological and economic importance. In addition to compiling data from the literature, we report the results of fruit fly sampling using McPhail or multilure traps in the Tropic of Cochabamba region and at the private natural reserve of Potrerillo del Guendá in Santa Cruz de la Sierra, as well as records from various other sites based on specimens in museum collections. Fifty-seven named species and three unnamed species of Anastrepha are recorded from Bolivia. Distribution maps for all of these species are provided. Numerous new department records are reported as well as the first records for Bolivia of A. castanea Norrbom, A. dissimilis Stone, A. elegans Blanchard, A. haywardi Blanchard, A. macrura Hendel, A. montei Lima, A. punctata Hendel, and A. rosilloi Blanchard. Pacouria boliviensis (Markgr.) A. Chev. (Apocynaceae) is reported as a host plant of A. woodleyi Norrbom Korytkowski, and Myrciaria floribunda (H. West ex Willd.) Berg (Myrtaceae) and Pouteria glomerata (Miq.) Radlk. (Sapotaceae) as host plants of A. fraterculus (Wiedemann). This distribution and host information will be useful to monitor and manage species that damage fruit crops in Bolivia.

Host shifting and host sharing in a genus of specialist flies diversifying alongside their sunflower hosts.

Congeneric parasites are unlikely to specialize on the same tissues of the same host species, likely because of strong multifarious selection against niche overlap. Exceptions where >1 congeneric species use the same tissues reveal important insights into ecological factors underlying the origins and maintenance of diversity. Larvae of sunflower maggot flies in the genus Strauzia feed on plants in the family Asteraceae. Although Strauzia tend to be host specialists, some species specialize on the same hosts. To resolve the origins of host sharing among these specialist flies, we used reduced representation genomic sequencing to infer the first multilocus phylogeny of genus Strauzia. Our results show that Helianthus tuberosus and Helianthus grosseserratus each host three different Strauzia species and that the flies co-occurring on a host are not one another's closest relatives. Though this pattern implies that host sharing is most likely the result of host shifts, these may not all be host shifts in the conventional sense of an insect moving onto an entirely new plant. Many hosts of Strauzia belong to a clade of perennial sunflowers that arose 1-2 MYA and are noted for frequent introgression and hybrid speciation events. Our divergence time estimates for all of the Helianthus-associated Strauzia are within this same time window (<1 MYA), suggesting that rapid and recent adaptive introgression and speciation in Helianthus may have instigated the diversification of Strauzia, with some flies converging upon a single plant host after their respective ancestral host plants hybridized to form a new sunflower species.

Descripción de una nueva especie de Anastrepha grupo curvicauda (Diptera: Tephritidae) / Description of a new species of the Anastrepha curvicauda species group (Diptera: Tephritidae)

RESUMEN Las moscas del género Anastrepha, se consideran plagas de gran importancia económica, no solo por el daño directo que causan a la producción de fruta sino, también, por las restricciones de cuarentena impuestas a los países exportadores de fruta. Por lo tanto, preparar descripciones y generar herramientas de identificación taxonómica, que permitan un reconocimiento adecuado y oportuno de las especies de importancia agrícola, es una actividad básica para la sanidad de las plantas en los países productores y exportadores de frutas. En este trabajo, se describe e ilustra Anastrepha ligiae, una nueva especie del grupo Anastrepha curvicauda, capturada con trampas McPhail, en los municipios de Albán y Anolaima (Cundinamarca) y Vélez (Santander), Colombia y criados en la fruta de la papayuela Vasconcellea pubescens A. DC. (Caricaceae). V. pubescens, se registra por primera vez como una planta hospedante para el género Anastrepha.

ABSTRACT Fruit flies of the genus Anastrepha are considered pests of great economic importance because they do not only cause direct damage to the fruit production, but also due to the quarantine restrictions imposed to fruit exporting countries. Therefore, preparing descriptions and generating taxonomic identification tools that allow an appropriate and accurate recognition of the species of agricultural importance are basic plant health activities in the fruit producing and exporting countries. In this work, we describe and illustrate Anastrepha ligiae, a new species of the Anastrepha curvicauda group captured with McPhail traps in the municipalities of Albán and Anolaima (Cundinamarca) and Vélez (Santander), Colombia, and reared from papaya fruit Vasconcellea pubescens A. DC. (Caricaceae). V. pubescens is the first time recorded as a host plant of the genus Anastrepha.

New records of Anastrepha (Diptera: Tephritidae) from Paraguay.

This work presents new distribution records in Paraguay for 19 fruit fly species of the genus Anastrepha Schiner, including four new records for the country: A. alveatoides Blanchard, A. australis (Blanchard), A. obliqua (Macquart) and A. pastranai Blanchard. Additionally, an updated list of the 26 Anastrepha species present in Paraguay is provided, as well as illustrations (wings, aculeus tips) of the majority of species.

Two new species of Anastrepha (Diptera: Tephritidae) from Ecuador.

Despite the fact that many species are economically important agricultural pests, the fruit fly fauna of Ecuador is still relatively poorly known. To better understand this fauna and the distribution and host plants of the pest species, the Proyecto Nacional de Manejo de las Moscas de la Fruta of AGROCALIDAD has conducted trapping surveys in many parts of the country. Two new species of Anastrepha from Ecuador that were collected by this project are described and illustrated: A. quadripuncta Troya Norrbom and A. tinctobasis Troya Norrbom. A new species group, the morvasi group, is recognized for A. morvasi Zucchi and A. quadripuncta.

Tephritidae flies associated with Chuquiraga avellanedae (Asteraceae) in Patagonia, Argentina.

In Patagonia, knowledge about the interaction among tephritids and the native flora is very scarce. In this study we identified for the first time two tephritid species (Cecidochares sp. and Neosphaeniscus m-nigrum) associated with the capitula of Chuquiraga avellanedae. This is the first record of a host plant for the genus Neosphaeniscus. Cecidochares sp. was more abundant and had a shorter development time than N. m-nigrum. Also, two families of parasitoid wasps (Pteromalidae and Eurytomidae) were registered. Further studies are needed to understand the impact of these tephritids on C. avellanedae fitness and their potential to control its populations.

Revision of the genus Euarestoides Benjamin (Diptera: Tephritidae).

Species of the genus Euarestoides Benjamin (Diptera: Tephritidae) are revised. Euarestoides comprises six species: E. abstersus (Loew) (Bahamas, eastern Canada and USA); E. acutangulus (Thomson) (Canada, Chile, Colombia, Cuba, Dominican Republic, Ecuador, Mexico, Peru, Puerto Rico, Trinidad Tobago, USA, and Venezuela); E. bimaculatus Savaris Norrbom, n. sp. (Peru); E. dreisbachi Foote (Guatemala, Mexico, Peru); E. pereirai Savaris Norrbom, n. sp. (Brazil); and E. rionegrensis Savaris Norrbom, n. sp. (Colombia). A key to the known species of the genus is provided, as well as descriptions, illustrations, distributions, notes on the biology and host plant data for each species.

Comprehensive inventory of true flies (Diptera) at a tropical site.

Estimations of tropical insect diversity generally suffer from lack of known groups or faunas against which extrapolations can be made, and have seriously underestimated the diversity of some taxa. Here we report the intensive inventory of a four-hectare tropical cloud forest in Costa Rica for one year, which yielded 4332 species of Diptera, providing the first verifiable basis for diversity of a major group of insects at a single site in the tropics. In total 73 families were present, all of which were studied to the species level, providing potentially complete coverage of all families of the order likely to be present at the site. Even so, extrapolations based on our data indicate that with further sampling, the actual total for the site could be closer to 8000 species. Efforts to completely sample a site, although resource-intensive and time-consuming, are needed to better ground estimations of world biodiversity based on limited sampling.

Remarkable fly (Diptera) diversity in a patch of Costa Rican cloud forest: Why inventory is a vital science.

Study of all flies (Diptera) collected for one year from a four-hectare (150 x 266 meter) patch of cloud forest at 1,600 meters above sea level at Zurquí de Moravia, San José Province, Costa Rica (hereafter referred to as Zurquí), revealed an astounding 4,332 species. This amounts to more than half the number of named species of flies for all of Central America. Specimens were collected with two Malaise traps running continuously and with a wide array of supplementary collecting methods for three days of each month. All morphospecies from all 73 families recorded were fully curated by technicians before submission to an international team of 59 taxonomic experts for identification.        Overall, a Malaise trap on the forest edge captured 1,988 species or 51% of all collected dipteran taxa (other than of Phoridae, subsampled only from this and one other Malaise trap). A Malaise trap in the forest sampled 906 species. Of other sampling methods, the combination of four other Malaise traps and an intercept trap, aerial/hand collecting, 10 emergence traps, and four CDC light traps added the greatest number of species to our inventory. This complement of sampling methods was an effective combination for retrieving substantial numbers of species of Diptera. Comparison of select sampling methods (considering 3,487 species of non-phorid Diptera) provided further details regarding how many species were sampled by various methods.        Comparison of species numbers from each of two permanent Malaise traps from Zurquí with those of single Malaise traps at each of Tapantí and Las Alturas, 40 and 180 km distant from Zurquí respectively, suggested significant species turnover. Comparison of the greater number of species collected in all traps from Zurquí did not markedly change the degree of similarity between the three sites, although the actual number of species shared did increase.        Comparisons of the total number of named and unnamed species of Diptera from four hectares at Zurquí is equivalent to 51% of all flies named from Central America, greater than all the named fly fauna of Colombia, equivalent to 14% of named Neotropical species and equal to about 2.7% of all named Diptera worldwide. Clearly the number of species of Diptera in tropical regions has been severely underestimated and the actual number may surpass the number of species of Coleoptera.        Various published extrapolations from limited data to estimate total numbers of species of larger taxonomic categories (e.g., Hexapoda, Arthropoda, Eukaryota, etc.) are highly questionable, and certainly will remain uncertain until we have more exhaustive surveys of all and diverse taxa (like Diptera) from multiple tropical sites.        Morphological characterization of species in inventories provides identifications placed in the context of taxonomy, phylogeny, form, and ecology. DNA barcoding species is a valuable tool to estimate species numbers but used alone fails to provide a broader context for the species identified.

New records of Anastrepha (Diptera: Tephritidae) primarily from Colombia.

New distribution information, primarily from Colombia, is provided for 60 species of Anastrepha, including the first records of 33 species from Colombia: A. acca Norrbom, A. acris Stone, A. amita Zucchi, A. amplidentata Norrbom, A. atrox (Aldrich), A. barbiellinii Lima, A. bezzii Lima, A. canalis Stone, A. cocorae Norrbom Korytkowski, A. compressa Stone, A. cordata Aldrich, A. crebra Stone, A. cryptostrepha Hendel, A. cryptostrephoides Norrbom Korytkowski, A. furcata Lima, A. fuscicauda Norrbom Korytkowski, A. galbina Stone, A. grandicarina Norrbom Korytkowski, A. hamata (Loew), A. katiyari Norrbom, A. macrura Hendel, A. margarita Caraballo, A. minuta Stone, A. nigripalpis Hendel, A. normalis Norrbom, A. pastranai Blanchard, A. pseudanomala Norrbom, A. pulchra Stone, A. similis Greene, A. speciosa Stone, A. urichi Greene, A. willei Korytkowski, and A. zuelaniae Stone. In addition, A. acris is reported from Costa Rica, A. alveata Stone from El Salvador, A. antunesi Lima and A. bahiensis Lima from Bolivia, A. barbiellinii from Trinidad Tobago, A. bezzii, A. canalis, A. coronilli Carrejo González, A. cryptostrepha and A. minuta from Bolivia, and A. willei from Argentina and Bolivia. A list of the 83 species of Anastrepha now known to occur in Colombia is provided, as well as illustrations of at least the wing and aculeus tip or male genitalia for 60 species.

A global checklist of the 932 fruit fly species in the tribe Dacini (Diptera, Tephritidae).

The correct application of the scientific names of species is neither easy nor trivial. Mistakes can lead to the wrong interpretation of research results or, when pest species are involved, inappropriate regulations and limits on trade, and possibly quarantine failures that permit the invasion of new pest species. Names are particularly challenging to manage when groups of organisms encompass a large number of species, when different workers employ different philosophical views, or when species are in a state of taxonomic flux. The fruit fly tribe Dacini is a species-rich taxon within Tephritidae and contains around a fifth of all known species in the family. About 10% of the 932 currently recognized species are pests of commercial fruits and vegetables, precipitating quarantines and trade embargos. Authoritative species lists consist largely of scattered regional treatments and outdated online resources. The checklist presented here is the first global overview of valid species names for the Dacini in almost two decades, and includes new lure records. By publishing this list both in paper and digitally, we aim to provide a resource for those studying fruit flies as well as researchers studying components of their impact on agriculture. The list is largely a consolidation of previous works, but following the results from recent phylogenetic work, we transfer one subgenus and eight species to different genera: members of the Bactrocera subgenus Javadacus Hardy, considered to belong to the Zeugodacus group of subgenera, are transferred to genus Zeugodacus; Bactrocera pseudocucurbitae White, 1999, stat. rev., is transferred back to Bactrocera from Zeugodacus; Zeugodacus arisanicus Shiraki, 1933, stat. rev., is transferred back to Zeugodacus from Bactrocera; and Z. brevipunctatus (David & Hancock, 2017), comb. n.; Z. javanensis (Perkins, 1938), comb. n.; Z. montanus (Hardy, 1983), comb. n.; Z. papuaensis (Malloch, 1939), comb. n.; Z. scutellarius (Bezzi, 1916), comb. n.; Z. semisurstyli (Drew & Romig, 2013), comb. n.; and Z. trilineatus (Hardy, 1955), comb. n. are transferred from Bactrocera to Zeugodacus.

Identifying Anastrepha (Diptera; Tephritidae) Species Using DNA Barcodes.

Molecular identification of fruit flies in the genus Anastrepha (Diptera; Tephritidae) is important to support plant pest exclusion, suppression, and outbreak eradication. Morphological methods of identification of this economically important genus are often not sufficient to identify species when detected as immature life stages. DNA barcoding a segment of the mitochondrial cytochrome oxidase I gene has been proposed as a method to identify pests in the genus. The identification process for these fruit flies, however, has not been explained in prior DNA barcode studies. DNA barcode methods assume that available DNA sequence records are biologically meaningful. These records, however, can be limited to the most common species or lack population-level measurements of diversity for pests. In such cases, the available data used as a reference are insufficient for completing an accurate identification. Using 539 DNA sequence records from 74 species of Anastrepha, we demonstrate that our barcoding data can distinguish four plant pests: Anastrepha grandis (Macquart) (Diptera; Tephritidae), Anastrepha ludens (Loew), Anastrepha serpentina (Wiedemann), and Anastrepha striata Schiner. This is based on genetic distances of barcode records for the pests and expert evaluation of species and population representation in the data set. DNA barcoding of the cytochrome oxidase I gene alone cannot reliably diagnose the pests Anastrepha fraterculus (Wiedemann), Anastrepha obliqua (Macquart), and Anastrepha suspensa (Loew).

New records of Rhagoletis (Diptera: Tephritidae) in Colombia, with discussion on the morphological variations of some species.

Rhagoletis jamaicensis Foote, 1981 and an undetermined species of the psalida group, are recorded for the first time in Colombia. The morphological variations of the Colombian specimens of these species and of other species closely related are discussed.

Phylogenetic relationships of the tribe Toxotrypanini (Diptera: Tephritidae) based on molecular characters.

Current hypotheses of relationship among the species of the fruit fly genera Anastrepha and Toxotrypana are tested using sequence data from six DNA regions: the mitochondrial regions 16S, CAD, and COI, and the nuclear regions EF1a, PER, and PGD. DNA sequences were obtained from 146 species of Anastrepha, representing 19 of the 21 species groups as well as five of the six clades of the robusta group, and four species of Toxotrypana in addition to species of Hexachaeta, Pseudophorellia, Alujamyia, and 13 other tephritid genera used as outgroups. The results indicate that Hexachaeta is more closely related to the Molynocoelia group than to Toxotrypana and Anastrepha, and it is removed from the tribe Toxotrypanini. The group Anastrepha+Toxotrypana and the genus Toxotrypana are strongly supported as monophyletic, consistent with previous studies, but Toxotrypana arises within Anastrepha, confirming that Anastrepha as currently defined is paraphyletic. The placement of Toxotrypana within Anastrepha is clearly defined for the first time with high support, as the sister group to the cryptostrepha clade of the robusta group of Anastrepha. Within Anastrepha, the daciformis, dentata, leptozona, raveni, and striata species groups are highly supported clades. The serpentina group is recognized with lower support, and the fraterculus and pseudoparallela groups are supported with minor alterations. The robusta group is resolved as polyphyletic, but four of the six species clades within it are recovered monophyletic (one clade is not represented and another is represented by one species). The punctata and panamensis groups are resolved together in a clade. At least some species of the mucronota group are related, however this group requires further study. The benjamini, grandis, and spatulata groups appear to be polyphyletic. Relationships among the species groups are generally poorly resolved, with the following exceptions: (1) the lineage including Toxotrypana, the cryptostrepha clade, and the tripunctata group; (2) the sister group relationship of the daciformis+dentata groups; (3) a clade comprising the punctata and panamensis groups; and (4) the large clade comprising the pseudoparallela+spatulata+ramosa+grandis+serpentina+striata+fraterculus groups.