Research Article |
Corresponding author: Erik van Nieukerken ( nieukerken@naturalis.nl ) Academic editor: Thomas Simonsen
© 2016 Erik van Nieukerken, Camiel Doorenweerd, Kenji Nishida, Chris Snyers.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
van Nieukerken E, Doorenweerd C, Nishida K, Snyers C (2016) New taxa, including three new genera show uniqueness of Neotropical Nepticulidae (Lepidoptera). ZooKeys 628: 1-63. https://doi.org/10.3897/zookeys.628.9805
|
After finding distinct clades in a molecular phylogeny for Nepticulidae that could not be placed in any known genera and discovering clear apomorphic characters that define these clades, as well as a number of Neotropical species that could be placed in known genera but were undescribed, three new genera and nine new species are here described from the Neotropics: Stigmella gallicola van Nieukerken & Nishida, sp. n. reared from galls on Hampea appendiculata (Malvaceae) in Costa Rica, representing the first example of a gall making Stigmella; S. schinivora van Nieukerken, sp. n. reared from leafmines on Schinus terebinthifolia (Anacardiaceae) in Argentina, Misiones; S. costaricensis van Nieukerken & Nishida, sp. n. and S. intronia van Nieukerken & Nishida, sp. n. each from a single specimen collected the same night in Costa Rica, Parque Nacional Chirripó; S. molinensis van Nieukerken & Snyers, sp. n. reared from leafmines on Salix humboldtiana, Peru, Lima, the first Neotropical species of the Stigmella salicis group sensu stricto; Ozadelpha van Nieukerken, gen. n. with type species O. conostegiae van Nieukerken & Nishida, sp. n., reared from leafmines on Conostegia oerstediana (Melastomataceae) from Costa Rica; Neotrifurcula van Nieukerken, gen. n. with type species N. gielisorum van Nieukerken, sp. n. from Chile; Hesperolyra van Nieukerken, gen. n.. with type species Fomoria diskusi Puplesis & Robinson, 2000; Hesperolyra saopaulensis van Nieukerken, sp. n., reared from an unidentified Myrtaceae, Sao Paulo, Brasil; and Acalyptris janzeni van Nieukerken & Nishida, sp. n. from Costa Rica, Guanacaste. Five new combinations are made: Ozadelpha ovata (Puplesis & Robinson, 2000), comb. n. and Ozadelpha guajavae (Puplesis & Diškus, 2002), comb. n., Hesperolyra diskusi (Puplesis & Robinson, 2000), comb. n., Hesperolyra molybditis (Zeller, 1877), comb. n. and Hesperolyra repanda (Puplesis & Diškus, 2002), comb. n. Three specimens are briefly described, but left unnamed: Ozadelpha specimen EvN4680, Neotrifurcula specimen EvN4504 and Neotrifurcula specimen RH2.
New species, new genus, taxonomy, leafmines, gall, molecular phylogeny
The nature of Latin America is generally known to be both very diverse and peculiar in its composition and Lepidoptera are no exception to this phenomenon. Biogeographically the region is usually known as the Neotropical region, or the Neotropics, although especially botanists often exclude the southern – non tropical – part (
Until fairly recently the nepticulid fauna of the Neotropics was almost completely unstudied, and twelve of the species described between 1877 and 1962 were tentatively placed in the large global genus Stigmella Schrank, 1802, with just a single species in the – at the time – monotypic genus Enteucha Meyrick, 1915 (
Even though almost all Neotropical species up to now have been placed in known nepticulid genera, which have been described from other parts of the world, several species show unique combinations of characters, and their placement is therefore disputable. Only the genera Enteucha Meyrick, 1915 and Manoneura Davis, 1979 were described originally from the Neotropics (respectively British Guyana and southern Florida), but Enteucha is not confined to the Neotropics, and also occurs in the Palearctic and Oriental regions (
While working on the molecular phylogeny of global Nepticulidae, to be published almost simultaneously (
Even though we could still only include a limited set of material compared to the described diversity, the analyses based on DNA sequences yielded several surprises. Three clades with distinctly long branches in the phylogeny could not be placed in any known genus, and other Neotropical species that were studied and could be placed in known genera also appeared to be hitherto unnamed. Taxonomists working on Nepticulidae fortunately have the tradition to be reluctant in describing new genera, preventing the systematic chaos that has been evident in various other groups of Lepidoptera. It is only after careful consideration of the morphological and molecular evidence that we here name these genera as well as several species, of which the common denominators are Neotropics and availability of DNA sequences (minimally a DNA barcode), and often provide additional interesting information on biology.
We describe three new genera and nine new species, including the first nepticulid species from Brazil and Costa Rica and the first known gall-forming Stigmella species.
Material was either collected by one of the authors or loaned from various institutions. The Costa Rica material was collected by Kenji Nishida, and made available as loan from the Museo de Zoología, Escuela de Biología,
Abbreviations for depositories etc.
BIN Barcode Index Number (
BMNH
BOLD Barcode of Life data Systems (http://www.barcodinglife.com/)
MZUCR
Museo de Zoología,
UNALM Museo de Entomología “Klaus Raven Büller”, Universidad Agraria La Molina, Lima, Peru
Collecting and rearing methods varied depending on the collector, but generally followed commonly used methods for the collection of leaf miners.
Genitalia were prepared according to our standard procedures, usually including DNA extraction, and described earlier in detail (
Measurements. Measurements of genitalia were obtained from digital images, using calibrated scaling in the Zeiss AxioVision software, see below, 20× objective for male genitalia and 10× or 20× for females. Capsule length was measured from vinculum to middle of uncus; valva length from tip of posterior process to ventral edge, excluding the sublateral process; phallus length was measured from the sclerotized tube, from tip of ventral process/carina, excluding any protruding vesica parts. Total bursa length includes all of the internal genitalia from cloaca to anterior edge of bursa. Genitalia measurements are usually rounded off to nearest 5µm. Forewing length was measured from tip of fringe to attachment on thorax, usually at magnification of 20×, also preferably on photographs. Antennal segment counts include scape and pedicel; they were counted on photographs or directly using the stereo microscope.
Morphological terms. The terminology used largely follows our earlier treatments and other recent papers on Nepticulidae (
Photographs of mounted moths were made with an AxioCam digital camera attached to a motorized Zeiss SteREO Discovery V12, using the Module Extended Focus, Zeiss AxioVision software, to prepare a picture in full focus from a Z-stack of 10 to 35 individual photos. Leafmines were photographed by EvN with a similar camera on a Zeiss Stemi SV11 stereo-microscope, without extended focus; other illustrated leafmines are from field photographs. Genitalia and wing slides were photographed with a similar camera on a manually operated Zeiss Axioskop H, usually with just a single exposure. Photographs were edited with Adobe Photoshop, avoiding any change to the real object, but backgrounds are cleaned from excess debris and artefacts by using healing brush and clone tools; tone and contrast are adjusted and a little sharpening is used in some photos. Photographs of venation, some mines and some female genitalia were taken in sections, and combined with the photomerge tool in Photoshop.
The photographs in this paper were taken by: Figs
Molecular methods were described in detail in previous studies (
Stigmella Schrank, 1802
Stigmella gallicola van Nieukerken & Nishida, sp. n.
Stigmella prunifoliella group
Stigmella schinivora van Nieukerken, sp. n.
Stigmella costaricensis van Nieukerken & Nishida, sp. n.
Stigmella intronia van Nieukerken & Nishida, sp. n.
Stigmella salicis group
Stigmella molinensis van Nieukerken & Snyers, sp. n.
Ozadelpha van Nieukerken, gen. n.
Ozadelpha conostegiae van Nieukerken & Nishida, sp. n.
Ozadelpha guajavae (Puplesis & Diškus, 2002), comb. n.
Ozadelpha ovata (Puplesis & Robinson, 2000), comb. n.
Ozadelpha specimen EvN4680
Neotrifurcula van Nieukerken, gen. n.
Neotrifurcula gielisorum van Nieukerken, sp. n.
Neotrifurcula specimen EvN4504
Neotrifurcula specimen RH2
Hesperolyra van Nieukerken, gen. n.
Hesperolyra diskusi (Puplesis & Robinson, 2000), comb. n.
Hesperolyra molybditis (Zeller, 1877), comb.n.
Hesperolyra repanda (Puplesis & Diškus, 2002), comb.n.
Hesperolyra saopaulensis van Nieukerken, sp. n.
Hesperolyra species 29122 (
Acalyptris Meyrick, 1921
Acalyptris janzeni van Nieukerken & Nishida, sp. n.
Costa Rica, Puntarenas province, Monteverde, Estación Biológica Monteverde, 10°19'06.9"N, 084°48'29.3"W, 1530 m, 26.x.2001 (adult emergence), col./rear: Kenji Nishida; host plant: Hampea appendiculata (Malvaceae), gall inducer on young leaf veins, pupate outside of the gall, cocoon spun 20.x.2001;
A large rather uniform moth, shining fuscous brown on almost all parts, with an orange frontal tuft and edged eye caps.
Male (Figs
Stigmella species, adult habitus. 1 S. gallicola, male holotype 2 S. gallicola, female Heredia Province, Puerto Viejo de Sarapiquí 3 S. schinivora, male holotype 4 S. schinivora, female
Female (Fig.
Measurements. Male: forewing length 2.5 mm (HT), wingspan: 6.0 mm. Female: forewing length 2.5 mm (n=2), wingspan ca. 5.8 mm.
Male genitalia (Figs
Female genitalia (Figs
Host plant (Figs
Stigmella gallicola, habitat and host plant. 17 General habitat at Alto Masis/Tierras Morenas, a premountain rainforest in Parque Nacional Volcán Tenorio, 13 June 2007 18 same place, 1 June 2007 19, 20 Habitat and host, Hampea appendiculata at same place, 955 m, 14 June 2007 21, 22 Mature galls on primary leaf veins, respectively adaxial and abaxial view, Estación Biológica Monteverde 1530 m, 22 April 2016, arrows pointing at nectar glands.
Gall (Figs
Stigmella gallicola, galls and larvae in Hampea appendiculata, details, all at Estación Biológica Monteverde, 1530 m. 23, 24 Galls on primary leaf veins of young leaf, respectively adaxial and abaxial view, 8 June 2016 25 Old gall found at axillary leaf bud, same date 26 Longitudinal cut of mature gall with late instar larva in situ, 20 October 2001 27 Transversely cut leaf (young gall in middle) showing gall chamber and frass mass line, 24 October 2001 28 Late instar larva in situ, note packed frass in gall chamber, same date 29 Late instar larva, dorsal view, slightly ex situ after gall dissection, 4 April 2015.
Stigmella gallicola, life history details on Hampea appendiculata, all at Estación Biológica Monteverde 1530 m. 30 Larval exit hole, 20 October 2001 31 Cocoon spun on host plant under rearing conditions, same date 32 Recently exited mature larva, 29 April 2016 33 Reared holotype male resting on host leaf, 26 October 2001.
Egg. Egg position unknown. Probably laid near or at foliar nectaries (nectar glands) abaxially on primary veins of leaf buds or very young leaves.
Larva (Figs
Cocoon (Fig.
Voltinism and habits. Larvae were found inside the galls in March, May and June. Adult emergence has been recorded in January, April, May, June and October.
Parasitoids. Braconidae: Adelinae: Adelius sp., endo-parasitoid, koinobiont of host larva and pupating inside host cocoon. It was reared from the Alto Masis Station site (n=2). Adelinae are specialised parasitoids of Nepticulidae (
Costa Rica: Alajuela, Heredia and Puntarenas provinces. Galls have been recorded mostly from Caribbean/Atlantic slope in lowland Costa Rica. The type locality is near the continental divide on the Pacific slope.
The Holotype (
The genitalia of Stigmella gallicola resemble somewhat those of the S. betulicola group, but in our multi-gene molecular analysis it groups in the clade which contains the S. prunifoliella and S. ultima groups, without a strongly supported position.
There are only few cases of galling in Nepticulidae known, and this species is the first known in the genus Stigmella. Galling has evolved independently several times in Nepticulidae: the North American Ectoedemia populella Busck, 1907 makes a petiole gall (
The specific name is a noun in apposition, derived from the Latin noun galla (= gall) and suffix –cola, “dweller in”.
6♂, 3♀, 5 adults, galls, larvae. Costa Rica: 1♂, Heredia Province, Puerto Viejo de Sarapiquí, Trimbina Biological Reserve, 10°24'59.81"N, 084°7'27.82"W, 161 m, gall inducer on young leaf veins of Hampea appendiculata, adult emergence18.i.2003, Kenji Nishida, Genitalia slide JCK15024; 2♀, Heredia Province, Puerto Viejo de Sarapiquí, OTS La Selva Station, main bridge, 10°25'53.42"N, 084°0'13.22"W, 50 m, gall inducer on young leaf veins of Hampea appendiculata, e.l. 2–5.v.2002, Kenji Nishida, Genitalia slide EvN3739,
[specimen not examined, same BIN]. 1 adult, Costa Rica, Guanacaste, Area de Conservacion Guanacaste, Sector San Cristobal, Estacion San Gerardo, 10.88 -85.389, 575 m, 17–24.ii.2014, DH Janzen, W Hallwachs, Malaise trap GMP#05845, laguna, BIOUG24701-G07 (
Argentina (Misiones), Cataratas del Iguazú. 27.viii.2000, 21J YM5645, E.J. van Nieukerken; leafmines, rather cultivated part of park; Schinus terebinthifolius, e.l. 11–12.ix.2000,
A completely leaden coloured species, with orange head and leaden edged scape, collar also leaden. The genitalia do not resemble any other Neotropical Stigmella species closely.
Male (Fig.
Female (Fig.
Measurements. Male: forewing length 1.7–1.8 mm (n=2), wingspan: 4.0 mm (n=2). Female: forewing length 1.7– 2.0 mm (n=2), wingspan 3.9–4.2 mm (n=2).
Male genitalia (Figs
Female genitalia (Figs
Host plants. Anacardiaceae: Schinus terebinthifolia Raddi, the Brazilian pepper tree.
Leafmines (Figs
Egg. The egg is always deposited on leaf upperside, frequently against a lateral vein.
Larva: green.
Voltinism and habits. Larvae were collected in late August, adults emerged indoors in September, and were caught in malaise traps from late September to late October. We collected the mines on planted trees, together with large numbers of the then still undescribed Leurocephala schinusae Davis & McKay (
Argentina: Misiones.
We barcoded two specimens from our reared series (not the Holotype), that appeared to match with a large number of records from the Malaise traps in Misiones, giving a total of 26 barcodes, all in BINBOLD:ACN0764. One specimen was also sequenced for other genes and will be used in a forthcoming analytical paper. Sequences may be retrieved in BOLD and later also in Genbank under voucher/sample ID
Molecular analysis suggest that S. schinivora is closely related to the North American Anacardiaceae feeding species in the Stigmella prunifoliella group, which suggests a single host-shift from Rosaceae or Rhamnaceae, but in some analyses it also groups with the European S. diniensis (Klimesch, 1975) that feeds on Cistaceae. It is currently the only known Neotropical species of this group of species.
Schinivora: an adjective, derived from the Latin noun Schinus (host genus), stem schin-, and verb voro (=devour).
4♂, 3♀, 4 larvae, leafmines. Argentina: 1♂, 1♀, 3 larvae in alcohol, leafmines, Misiones, Cataratas del Iguazú. 27.viii. 2000, leafmines, rather cultivated part of park; leafmines on Schinus terebinthifolia, e.l. 11–12.ix.2000,
[specimens not examined, same BIN]. 21 adults, in ethanol 96%, Argentina, Misiones, Obera, CIAR, 23.v.–24.x.2013, Pablo Tubaro, Malaise trap: 1, trap GMP#05146, 23–30.v.2013, BIOUG12989-H09; 2 ad., trap GMP#05149, 26–4.vii.2013, BIOUG23823-E01, BIOUG23823-E11; 2 ad., trap GMP#05150, 11–18.vii.2013, BIOUG24501-C12, BIOUG24505-B08; 1 ad., trap GMP#04812, 18–25.vii.2013, BIOUG13317-B04; 4 ad., trap GMP#05151, 25–1.viii.2013, BIOUG24746-D11, BIOUG24746-E04, BIOUG24746-E06, BIOUG24746-H10; 5 ad., trap GMP#04813, 1–8.viii.2013, BIOUG13418-F06, BIOUG13418-G07, BIOUG13418-H11, BIOUG13420-A11, BIOUG13421-G02; 1 ad., trap GMP#04815, 22–29.viii.2013, BIOUG24913-G01; 2 ad., trap GMP#04816, 5–12.ix.2013, BIOUG25104-F01, BIOUG25104-F02; 1 ad., trap GMP#04817, 19–26.ix.2013, BIOUG25154-H01; 2 ad., trap GMP#04819, 17–24.x.2013, BIOUG24948-E10, BIOUG24952-A07 (all in
Costa Rica, San José Province, Parque Nacional Chirripó, Llano Bonito, Refugio, 09°27'16"N, 083°32'41"W, 2492 m, 19–20.ii.2009, Light, Leg: Kenji Nishida. Genitalia slide EvN4037♂,
See under S. intronia.
Male (Fig.
Female. Unknown.
Measurements. Male: forewing length 3.5 mm (n=1), wingspan: ca 7.5 mm.
Male genitalia (Figs
Host plants. unknown.
Voltinism and habits. The moth was collected in February at a light sheet.
Costa Rica: San José Province: Chirripó National Park: Llano Bonito area, a cloud forest surrounded by large oak trees.
Holotype BIN: BOLD:ACG8765. The holotype was also sequenced for three other genes and will be used in a forthcoming analytical paper. Sequences may be retrieved in BOLD and later also in Genbank under voucher/sample ID
This species and the next were collected the same night. They both belong in core Stigmella, in the clade with the S. lapponica, S. rhamnella and S. sanguisorbae groups, but do not belong to any of these groups.
Costaricensis: an adjective, derived from the country name Costa Rica and the suffix –ensis, indicating geographical origin.
Costa Rica, San José Province, Parque Nacional Chirripó, Llano Bonito, Refugio, 09°27'16"N– 083°32'41"W, 2492 m, 19–20.ii.2009, Light, Leg: Kenji Nishida. Genitalia slide EvN4036♂,
Externally S. intronia and S. costaricensis are very similar, but the fascia in intronia seems a bit wider and is placed more anteriorly. Both species resemble somewhat the North American S. slingerlandella (Kearfott, 1908). The species are best separated by the shape and spinosity of the valva and form of uncus and gnathos.
Male (Fig.
Female. Unknown.
Measurements. Male: forewing length 2.8 mm (n=1), wingspan: ca 6.3 mm.
Male genitalia (Figs
Host plants. Unknown.
Voltinism and habits. The moth was collected in February at a light sheet.
Costa Rica: San José Province: Chirripó National Park: Llano Bonito area, a cloud forest surrounded by large oak trees.
Holotype BIN: BOLD:ACG8514. The holotype was also sequenced for other genes and used in the molecular phylogeny (
See under S. costaricensis.
Intronia, a noun in apposition, arbitrarily derived from the word Intron (based on English: intragenic region), because of the presence of several introns in a copy of the gene Elongation Factor 1α (
Peru, Lima, Universidad Agraria la Molina, 240 m, -12.0869, -76.9444, 1.xii.2010, leafmines on Salix humboldtiana, e.l. 16.xii.2010, C. Snyers, genitalia slide EvN4218,
Externally S. molinensis can be confused with other Neotropical Stigmella without fascia, with pale head and collar, such as S. hamata Puplesis & Robinson, 2000, S. montanotropica Puplesis & Diškus, 2002 and S. austroamericana Puplesis & Diškus, 2002, but the male genitalia are characteristic.
Male (Figs
Female (Fig.
Measurements. Male: forewing length ca 1.9–2.0 mm (n=2), wingspan: 4.4–4.5 mm. Female: forewing length 2.3 mm, wingspan 5.1 mm.
Male genitalia (Figs
Female genitalia (Figs
Host plant (Figs
Stigmella molinensis, type locality Lima, Universidad Agraria la Molina, 240 m, host plant and life history. 58 Row of tall Salix humboldtiana trees, 9 December 2013 59 Chris Snyers, collecting leafmines in Salix humboldtiana, 16 December 2013 60–63 Vacated leafmines and leafmine with larva, 18 January 2010 64–65 Male, emerged 18 January 2010.
Leafmines (Figs
Egg. Always deposited on leaf upperside close to the midrib.
Larva yellow.
Voltinism and habits. Larvae were always present between December and late January. Adults usually emerged after two to three weeks after the cocoon was spun, suggesting multiple generations. The species has only been searched for between early December and late January, we have no information on other seasons. Mines were always found at the shady side of the trees.
Peru: Lima.
We barcoded three specimens: the Holotype, the female
This species clearly belongs to the Stigmella salicis group s.str., that is widespread in the Holarctic region and of which all but one species feed on Salicaceae (sensu stricto). However, morphologically it differs by the presence of numerous small cornuti in the phallus, whereas all other species have a reduced number of differently shaped cornuti; the latter thus is a good apomorphy for the Holarctic members of the group. The female shares the characteristic apomorphy: a band of signa around the bursa copulatrix. In our molecular phylogeny (
The specific name, an adjective, is derived from La Molina, Spanish and Latin for Mill, and also the name of the district in Lima and the University where the species was collected.
Adults and leafmines: Same data as Holotype, Peru, 1♂, 18.i.2010, e.l. 1.ii.2010, damaged specimen (see Figs
Ozadelpha conostegiae van Nieukerken & Nishida, sp. n. by present designation.
Ozadelpha is recognised by the collar with lamellar scales (as in Enteucha and Stigmella), usually two fascia’s on forewing; forewing venation without closed cell, but usually a very long separate CuA. Male genitalia are characterised by separate vinculum and tegumen, large vinculum, bilobed uncus and V-shaped gnathos.
Adult (Figs
Male genitalia. (Figs
Female genitalia (Figs
Larva. Green, where known.
Leafminers on Melastomataceae and Myrtaceae, both belonging to the order Myrtales. See also
Central and South America, known from Costa Rica, Ecuador and Argentina, probably widespread.
We describe here one new species, O. conostegiae, as type species, plus a second one from the same host that we do not name. Both Stigmella ovata Puplesis & Robinson, 2000 and Enteucha guajavae Puplesis & Diškus, 2002 share many characters with the type species: an almost identical venation (albeit without recognisable CuA in the drawing of E. guajavae) and very similar male genitalia. We therefore transfer both to this genus as Ozadelpha ovata (Puplesis & Robinson, 2000), comb. n. and O. guajavae (Puplesis & Diškus, 2002), comb. n. It is possible that some species associated with the Myrtaceae genus Myrceugenia from Patagonia (
Ozadelpha, a noun. The name is based on a combination of the Greek adelphe (αδελφη), meaning sister with the colloquial abbreviation “Oz” as often used for Australia. This to indicate the sister group relationship between Ozadelpha and the Australian clade with Roscidotoga Hoare, 2000, Pectinivalva Scoble, 1983, Menurella Hoare, 2013 and Casanovula Hoare, 2013. The name is to be treated as feminine.
The series reared from Conostegia that KN reared and sent via the Museo de Zoología, Universidad de Costa Rica, to EvN was at first considered to constitute a single species. We were only able to successfully amplify several genes from one large female (
In the morphology Ozadelpha shows similarities with both Stigmella, Enteucha and the Australian genera. The venation resembles Stigmella, but also Pectinivalva, apart for the thickened A in forewing. In our molecular phylogeny Ozadelpha always groups with the Australian genera, either as sister to all of them together, or as sister to Roscidotoga (
Costa Rica, Puntarenas Province, Monteverde, Estación Biológica Monteverde, 10°19'06.9"N, 084°48'29.3"W, 1530 m, 2.iii.2012 collected leafmines, 31.iii.2012 adult emergence, host plant: Conostegia oerstediana (Melastomataceae), photos/leg/rear: Kenji Nishida,
Externally recognised by leaden collar of lamellar scales and forewing with two fasciae, the first indistinctly joined to basal leaden area. Male genitalia unique and can not be confused with other Nepticulidae. However, there are as yet unnamed rather similar species. See also next species.
Male (Figs
Female (Fig.
Measurements. Male: forewing length 1.7–1.9 mm (n=2), wingspan: 3.8–4.3 mm. Female: forewing length 1.7–1.8 mm (n=3), wingspan 3.9–4.0 mm.
Male genitalia (Figs
Female genitalia (Figs
Host plants (Fig.
Ozadelpha conostegiae, habitat, host plant, Conostegia oerstediana and leafmines at Costa Rica, Estación Biológica Monteverde 1530 m. 80 Habitat, a cloud forest (lower mountain wet forest), 8 June 2016 81 Treelets, infested with some leaf-mining larvae 82 Old leaf mines on host leaf, 24 March 2016 83 Old leaf and young mines, 25 April 2016 84, 85 Late instar larva in situ, upper epidermis removed to show larva, calcium oxalate crystals (druses) are not eaten and left behind, 18 May 2016 86 Close-up of leaf mine, upper mine is of late instar larva, note the zigzag frass line, lower mine is of early instar, 7 June 2016 87 Egg scale, dorsal view, attached to upperside of host leaf, 7 June 2016.
Leafmines (Figs
Ozadelpha conostegiae, life history details, Costa Rica, Estación Biológica Monteverde 1530 m. 88 Leaf mines, 2 cocoons and 3 adults on midrib on upperside host plant leaf, 29 March 2012 89 Close up of one adult male 90 Adult female resting on plastic bag under rearing conditions, ventral view, 1 December 2013 91 Pupa, ventral view, cocoon detached and removed, 7 June 2016 92 Cocoon spun on upperside on secondary vein under captive conditions, 7 June 2016 93 Cocoon-spinning mature larva, outer layer cocoon is removed to show larva, 19 June 2016 94 Ozadelpha specimen EvN4680, female, resting on upper side leaf along one of the primary veins, 3 February 2012.
Egg (Fig.
Larva (Figs
Cocoon (Figs
Pupa (Fig.
Voltinism and habits. Old and young leaf mines were seen all year round on treelets found along trails in forest understory at the Estación Biológica Monteverde. The larvae pupate away from the mine and host plant under natural conditions; however, it is unknown where the larvae spin their cocoons except for a single cocoon that was found on the underside of a mined leaf, but this was parasitized by a parasitoid wasp. Late stage to mature mines collected on 2.iii.2016 produced adults on 28–29.iii.2016. Mines collected on 21.v.2016 produced mature pupae (n=2) and mature pupae of a parasitoid wasp inside the moth cocoon (n=2) on 7.vi.2016. Mines collected on 25.v.2016 produced 6 cocoons between 2–6.vi.2016. Two eggs on a single leaf collected on 7.vi.2016 produced very early mines of ca. 1.2 mm and 8.6 mm long on 19.vi.2016. White, circular, pellet-like micro objects were found inside mines surrounding mining larvae (n=2) (Figs
Parasitoid. Eulophidae: Entedoninae: Cornugon diabolos Hansson (
Costa Rica: Alajuela, Guanacaste and Puntarenas Provinces.
We failed to produce a DNA barcode from the DNA extracts, but we obtained sequences for 28S and COII from the holotype,
Ozadelpha conostegiae represents the first published record of a Nepticulidae feeding on Melastomataceae. However, we also collected and reared unnamed Acalyptris species from the genus Melastoma from Australia: Queensland and Indonesia: Borneo. The genus Conostegia comprises 77 species of shrubs and trees in Central America, northern South America and the Carribean (
The epithet conostegiae is a noun in genitive case, derived from the generic name of the host plant Conostegia.
8♂, 6♀. Costa Rica: 1♂, Alajuela Province, Grecia, Reserva Forestal Grecia - Bosque del Niño, 10°.08'32.1"N, 084°.14'49.5"W, 1678 m, leafmines on Conostegia oerstediana, xii.2012 adult emergence, Kenji Nishida (
Externally very similar to O. conostegiae, but markedly larger (forewing length 2.8 mm against 1.8–2.0). Female genitalia recognised by longer anterior apophyses and very short bursa.
(Figs
Measurements. Female: forewing length 2.8 mm (n=1).
Female genitalia (Figs
Host plants. Unknown. The moth was found resting on the upperside of a Conostegia oerstediana (Melastomataceae) leaf along one of the main veins, thus this may well be the host.
Voltinism and habits. The moth was collected on March 2nd, 2012.
Costa Rica: Puntarenas Province.
We leave this species presently unnamed, as we only have a single female and no data on life history. There appear to be more closely related species of Ozadelpha feeding on Conostegia in Costa Rica, and we therefore rather await more material in order to be able to discriminate the various species better.
We barcoded the single specimen, currently the only DNA barcode we obtained from the genus Ozadelpha. This specimen was also sequenced for other genes and used in the molecular phylogeny (
Costa Rica: 1♀, Puntarenas Province, Monteverde, Estación Biológica Monteverde, 1530 m, 10°19'06.9"N, 084°48'29.3"W, 2.iii.2012 resting on upperside of Conostegia oerstediana leaf along the vein, Kenji Nishida & Yuriko Demura; genitalia slide ♀ EvN4680,
Neotrifurcula gielisorum van Nieukerken sp. n. by present designation.
Neotrifurcula can be recognised by the hindwing venation with trifurcate Rs+M, and a very long and separate CuA in forewing, collar with hairscales, in the genitalia male phallus with a long curved flagellum-like appendix; female with reticulate signa and complex vaginal sclerite. Glaucolepis has a similar venation, but usually a velvet patch of special scales on hindwing and three pairs of anal tufts.
Adult (Figs
Male genitalia. (Figs
Female genitalia (Figs
Hostplant and immature stages unknown. Adults collected in Nothofagus forests from November to January.
Chile and Argentina, southern parts at low and medial altitudes.
Next to the type species, we include two unnamed species, for one of which we only have one female that was sequenced, another one only one worn male on loan from Copenhagen (
Neotrifurcula, a noun, a combination of the prefix neo-, new, here derived from Neotropics, and the Latin noun Trifurcula (= a three-pronged fork), another nepticulid genus with a 3-forked Rs+M in the hindwing. The name is to be treated as feminine.
The anterior sclerite of sternum 2 (S2A) has anterior apodemes similar to Bohemannia Stainton, 1859. The venation also has several similarities to Bohemannia, although the latter seems more reduced by the fusing of CuA with Rs+M and the reduction of the closed cell. This supports the possible sistergroup relationship to Bohemannia or Bohemannia + Hesperolyra that we found in our molecular analyses (
Chile, Ñuble, Bio-Bio (VIII), 2 km N Las Trancas, 70 km E Chilan, 1400 m, 36.54S-71.28W, 6.i.2001, C. Gielis & H. W. van der Wolf, sta 53, genitalia slide EvN4503,
One of the largest nepticulids with a wingspan of almost 10 mm. Recognised by very broad cream fascia, male genitalia characteristic by flagellum-like appendix on phallus, but several closely related, but smaller species have very similar genitalia.
Male (Figs
Female. Unknown.
Measurements. Male: forewing length 4.0–4.8 mm (n=3), wingspan: 8.7–10.1 mm.
Male genitalia (Figs
Host plants. Unknown.
Voltinism and habits. The moth was collected from mid-December to mid-January.
Chile: Curoco, Ñuble and Valparaiso. In both localities with dense forest of large Nothofagus trees at middle altitudes (1100–1500 m) (Fig.
Neotrifurcula sp., habitats in Chile 105 (top), Type locality N. gielisorum, Nuble, Bio-Bio, 2 km N Las Trancas, 1400 m, 6 January 2001, edge of Nothofagus forest 106 (bottom) Locality for Neotrifurcula specimen EvN4504 Valparaiso, Parque Nat. La Campana, 450 m, Nothofagus forest near brooklet, photo 8 November 2000, specimen collected here 19 February 2001.
We barcoded two specimens, including the holotype, both in BINBOLD ACG8607. One specimen was also sequenced for other genes and used in the molecular phylogeny (
The specific name gielisorum is a noun in plural genitive, based on the family name Gielis, to honour Cees and Siska Gielis for their efforts not only to collect this species, but to explore and collect Microlepidoptera widely in South America, and to publish in particular about the plume moths, Pterophoridae.
Chile: 1♂, Curico, Maule (VII), 60 km SE Molina, RN Radal Seite Tazas, 1100 m, 18–19.xii.2000, 35.28S-71.00W, C. Gielis & FK Gielis, sta. 45, genitalia + wing slide EvN4703 (
Externally similar to N. gielisorum, but no distinct fascia. Female genitalia characterised by ductus spermathecae with 3.5 convolutions, distinct reticulate signa and omega-shaped sclerotisation in vestibulum.
Male. Unknown.
Female (Fig.
Measurements. Female: forewing length 3.8mm, wingspan 8.2 mm.
Female genitalia (Figs
Host plant. Unknown.
Voltinism and habits. The moth was collected in February.
Chile: Valparaiso. Collected in open shrubby habitat, with shrubby species of Nothofagus (Fig.
We obtained a full barcode of the specimen, with BINBOLD:ACU6693, showing a distance to barcodes of N. gielisorum of ca 12%.
The huge barcode distance to N. gielisorum and the differences in the wing pattern show clearly the species status of this specimen. It could possibly be the female of the next species, but difference in size and number of antennal segments make this rather unlikely.
1♀, Chile: Valparaiso, Parque Nat. La Campana, 6 km E of Olmue, 450 m, 33.00S-71.03W, 19.ii.2001, R.T.A. Schouten & H. W. van der Wolf, station 90, Nothofagus forest near brooklet, genitalia slide EvN4504 (
The moth is markedly smaller than the previous two species, but too worn to see external diagnostics. Male genitalia similar to N. gielisorum, but smaller, valva more triangular and more distinct and cornuti larger.
Male Antenna with 42 segments, forewing length 2.7 mm. Head pale, forewings and thorax brown, very worn, pattern not visible.
Female unknown.
Measurements. Male: forewing length 2.7mm.
Male genitalia (Figs
Host plant. Unknown.
Voltinism and habits. The moth was collected in November, around the conifer Podocarpus salignus D. Don (on the label incorrectly cited as saligna). It is unlikely that this represents the host.
Chile: Valdivia.
The moth had been studied by Robert Hoare for his thesis, and is part of a larger series that is under study with the group of J.R. Stonis (Vilnius) and probably will be described by them.
1♂, Chile, Valdivia, 20 km S Valdivia, Rincon de la Piedra, 15.xi.1981, Nielsen & Karsholt, station 15, caught around Podocarpus saligna, Genitalia and wing slide R. Hoare South America 2 (
Fomoria
molybditis
group
Fomoria diskusi Puplesis & Robinson, 2000: 43, by present designation.
There are no obvious external characters apart from the venation: broad forewing with a straight main vein with 4 branches, CuA absent, rather resembling Acalyptris, but no vestigial closed cell present and wings broader. The most obvious characters are in the male genitalia: bifurcate pseuduncus, deeply divided valva and long extended lyre-shaped transtilla. Female genitalia without reticulate signa, small bursa.
Adult (Figs
Male genitalia. (Figs
Female genitalia (Figs
Pupa (Figs
So far the host plant is only known for one species, H. saopaulensis: Myrtaceae, on which the larva makes normal gallery mines (Figs
Only known from the Neotropics: Belize, Brazil, Colombia and Ecuador.
We recombine here the species previously placed in the Fomoria molybditis group: Hesperolyra diskusi (Puplesis & Robinson, 2000), comb. n., H. molybditis (Zeller, 1877), comb. n., H. repanda (Puplesis & Diškus, 2002), comb. n. and describe one new species, Hesperolyra saopaulensis van Nieukerken sp. n. One unnamed species also belongs here: Hesperolyra species 29122 (
The name Hesperolyra is a combination of Hespero- from Hesperus (the evening star), referring to the occurrence in the western hemisphere, and lyra (lyre), referring to the lyre shaped transtilla, a common character for species where the male is known. The name is to be treated as feminine.
The present genus was recognised first in our molecular analysis by the new species H. saopaulensis, that consistently grouped outside any known genus, often together with Neotrifurcula, but at a large distance. Since we did not have a male of H. saopaulensis, a generic description seemed problematic, until we noticed similarity to the recently obtained DNA barcode of Fomoria diskusi and the unusual venation of both species, quite different from any other Fomoria. Fomoria diskusi was placed with some other species in the Fomoria molybditis group (
Fomoria diskusi Puplesis & Robinson, 2000: 43. Holotype ♂: BELIZE: Cayo Distr., Chiquibul For. Res., Las Cuevas, 3–16.iv.1998, R. Puplesis & S. Hill, Genitalia slide BM28844 (BMNH). [examined]
Hesperolyra diskusi is easily recognisable by its striking pattern: the pale costal streak that turns into an outward oblique fascia at 2/3. In the male genitalia the characteristic long spines separate it from congeners.
Male (Fig.
Female (Fig.
Measurements. Male: forewing length 1.8–2.3 mm (n=2), wingspan 4.0–5.0 mm. Female: forewing length 2.1 mm (n=1), wingspan 4.6 mm.
Male genitalia (Figs
Female genitalia (Figs
Host plant unknown. Collected at light in April, during the dry season in secondary forest (
Only known from Belize (ca.N16.732, W88.988).
We barcoded one paratype (
4♂, 1♀, paratypes: Belize: Cayo District, Chiquibul Forest Reserve, Las Cuevas, 3–16.iv.1998, R. Puplesis & S. Hill, Genitalia slides ♂ EvN4501 (+ wings), BM28845, BM29130, ♀ BM28846, wing slide BM28847, resp.
Brazil, São Paulo, Garulhos, park near airport terminal, 19.viii.2000, EvN no 2000133 [rearing number], leafmines on unidentified Myrtaceae, E.J. van Nieukerken, adult emergence 11–14.ix.2000,
Externally recognised by silver to leaden basal half of forewing and postmedial silvery fascia, with cilia line present and slightly edged scape. Female genitalia simple, without signa.
Male. Unknown.
Female (Figs
Measurements. Female: forewing length 1.9–2.1 mm (2.0 ± 0.1, 14), wingspan 4.0–4.6 mm.
Female genitalia (Figs
Host plants. Myrtaceae: unidentified tree (probably an Eugenia or Myrcia sp.).
Leafmines (Figs
Egg. The egg is deposited on leaf upperside, often on a lateral vein.
Larva. Not described.
Cocoon (Fig.
Voltinism and habits. Larvae found in August, moths emerged in September.
Brazil: São Paulo.
We barcoded both specimens, resulting in BIN: BOLD:ACG9057. The Holotype was also sequenced for other genes and used in the molecular phylogeny (
Even though we have no males, we decided to name this species, of which we know the biology, several genes, and which is characteristic both externally and in female genitalia. Furthermore it is the species that was decisive in erecting the new genus Hesperolyra. Collecting new material should be easy, in the small park forest in front of the entrance of Garulhos airport.
Saopaulensis: an adjective, derived from the province and city name São Paulo and the suffix –ensis, indicating geographical origin.
1♀, leafmines, Brazil, São Paulo, Garulhos, park near airport terminal, 19.viii.2000, EvN no 2000133 [rearing number], leafmines on unidentified Myrtaceae, E.J. van Nieukerken, adult emergence 11–14.ix.2000, genitalia slide Ev4505,
Costa Rica, Guanacaste Province, ACG Santa Rosa Station, 10°50'22.30"N, 085°37'6.43"W, 293 m, 22.vi.2003, light sheet, Kenji Nishida, Genitalia slide EvN3673,
Externally a dull species, not distinguishable from others without obvious colour pattern. The male genitalia are characterised by the bifid pseuduncus, lateral support rods and several large curved carinal spines.
Male (Figs
Female. Unknown.
Measurements. Male: forewing length 1.7 mm, wingspan: 4.2 mm.
Male genitalia (Figs
Host plant. Unknown.
Voltinism and habits. Adults found from late June to early August and again November to mid-March. Collected at light and in malaise traps.
Costa Rica: Guanacaste Province, Área de Conservación Guanacaste. Coordinates type locality: N10.83400, W85.61200. It is a sub-humid to humid tropical dry (deciduous) forest with five to six months of dry season (
We have DNA barcodes of all ten known specimens, the holotype differs 2.3% from the other specimens.
The combination of the venation, absence of transverse bar of transtilla and presence of lateral support rods makes this a typical New World Acalyptris species, probably belonging to the A. scirpi group. The DNA barcode does not place it close to any Acalyptris species of which the barcode is known. Morphologically there are similarities to several species described from Belize: A. bifidus Puplesis & Robinson, 2000 and A. unicornis Puplesis & Robinson, 2000. It is remarkable that the holotype barcode has a 2.3% distance to the Malaise trapped specimens, found in almost the same locality. More material is needed to see whether this is a case of cryptic species, or a unusual high variation in the population.
Janzeni: a noun in genitive case, based on the family name Janzen, to honour Daniel H. Janzen, collector of part of the material, for his long time dedication to study the tropical Lepidoptera fauna of the Guanacaste Conservation area in great detail, both ecologically and taxonomically, and his enthusiastic support of DNA barcoding (eg.
6♂ [DNA barcoded ethanol material]: Costa Rica, Guanacaste, Área de Conservación Guanacaste, Sector Santa Rosa, Bosque San Emilio, Forest, Malaise trap, 300 m, 10.8438, -85.6138, Dan Janzen: 1♂, trap GMP#00624, 31.vii–6.viii.2012, BIOUG05419-H06, genitalia slide JCK8232; 1♂, trap GMP#01813, 6–13.xi.2012, BIOUG09432-B01; 2♂, trap GMP#01815, 20–27.xi.2012, BIOUG09436-B12, BIOUG09436-C06; 1♂, trap GMP#01817, 4–11.xii.2012, BIOUG09441-C02; 1♂, trap GMP#01825, 29.i–5.ii.2013, BIOUG10108-C05 (
[specimens not examined, same BIN] 3 adults, same locality: trap GMP#01824, 22–29.i.2013, BIOUG18276-D11; trap GMP#01826, 5–12.ii.2013, BIOUG18337-A10; trap GMP#01830, 12–19.iii.2013, BIOUG18605-F04 (
We would like to thank the following persons for lending or donating material for study: Alison Brown and Angela Telfer (both
Specimen data for Neotropical Nepticulidae with BOLD data
Data type: Microsoft Excel (xls)
Explanation note: The 87 records are all records of the new taxa described in this paper. Records are listed alphabetically by species, country, stateProvince, AdminDivision2, AdminDivision3, Locality.